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components from tobacco prod-", + "uct samples.", + "B. Cellular and Subcellular Systems", + "Objective: To determine the fundamental mechanisms of", + "specific responses of intact animals to smoke.", + "C. Bioassay", + "Objective: To use those responses of nonhuman biological", + "systems which can be quantitated and are rel-", + "evant to human health problems, for the routine", + "examination of various tobacco product samples.", + "COMMENTS", + "Smoke exposure of intact living animals, using inhalation techniques,", + "will be emphasized in this section of the program, since smoke exposure in", + "humans normally occurs through inhalation. By determining the response of", + "experimental animals in this way, it should be easier to correlate the re-", + "sponses obtained with those of humans exposed to smoke from the same samples.", + "For such comparisons to be most meaningful, the composition of smoke must", + "correspond to the smoke consumed by man, and one must have exact control", + "over the exposure conditions.", + "By working with cellular and subcellular systems, it should be possible", + "to obtain a better understanding of the underlying mechanisms of both short-", + "term and long-term responses noted in the intact animal.", + "Attempts will be made to quantitate the responses, and if this is pos-", + "sible, certain of these may subsequently be utilized for routine bioassay.", + "The extent to which a response- can be changed by product modifications can", + "then be studied on a larger number of samples. This is being done in mouse", + "skin painting bioassay. Primary emphasis will be given to those responses", + "believed to have some relationship to a human medical problem.", + "F/C-- 6029119", + "64", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0ob§", + "0090631" + ] + }, + { + "bbox": [ + [ + 0.293147, + 0.138432, + 0.293752, + 0.009081 + ], + [ + 0.324915, + 0.151376, + 0.238399, + 0.009542 + ], + [ + 0.25292, + 0.176244, + 0.38878, + 0.011531 + ], + [ + 0.127244, + 0.201356, + 0.670834, + 0.011328 + ], + [ + 0.091992, + 0.213945, + 0.698156, + 0.011578 + ], + [ + 0.091928, + 0.226349, + 0.714341, + 0.011172 + ], + [ + 0.092163, + 0.238146, + 0.698431, + 0.011588 + ], + [ + 0.091989, + 0.250151, + 0.357698, + 0.011188 + ], + [ + 0.127648, + 0.274847, + 0.663312, + 0.011297 + ], + [ + 0.091425, + 0.286273, + 0.722956, + 0.010918 + ], + [ + 0.091821, + 0.298386, + 0.70772, + 0.011619 + ], + [ + 0.091418, + 0.311087, + 0.700143, + 0.011664 + ], + [ + 0.091689, + 0.323267, + 0.708279, + 0.010722 + ], + [ + 0.092027, + 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M. Donnelly, Ph.D., Department of Microbiology", + "Almost every speaker in each of the previous sections has stated his need to know the", + "biological activity of a particular tobacco or tobacco component. Consequently, there is", + "little need to introduce this section or to outline its importance to the program. Instead", + "let us look at a film showing the induction of ciliostasis in hamster trachea produced by", + "the water soluble portion of smoke condensate.", + "This motion picture, made in our laboratory, begins by showing ciliary beating in an", + "untreated, or control ring; that is, before the action of the ciliostat. This film was shot", + "at 32 franes/second, and is now being projected at 16 frames/second, so this rapid beating", + "which you see is actually half the speed at which the cilia are beating. The short blank", + "has signaled the addition of the water soluble extract of tobacco smoke condensate. There", + "is now an obvious slowing of the ciliary beating, and shortly we note the complete cessation", + "of ciliary beating. This part of the film has been edited down to about 2 minutes. The", + "actual cessation time was about 20 minutes.", + "For this ciliary monitoring technique a transverse section, or ring, is cut from the", + "trachea of a young hamster and is placed in a balanced salt solution. It is then examined", + "with low power (100 to 150 X) bright field microscopy and the ciliary beating can be readily", + "observed. Tobacco smoke fractions or other ciliostatic agents of known concentration can", + "be presented to the tracheal ring, and the time required to bring about complete ciliostasis", + "in the entire tissue can be measured. Complete ciliostasis serves as a useful endpoint, and", + "the dose-response relations of the ciliostat can be elucidated by measuring ciliostasis time", + "for a range of concentrations. The tracheal ring system can thus be utilized as a bioassay", + "and toxicity screening device, and the effect of product changes on ciliostasis can be", + "determined.", + "Dr. Rees will now present a paper describing his research on chromosomal aberrations-", + "a technique which has the potential for development into a bioassay procedure. This will", + "be followed by a paper by Dr. White describing the use of \"Membrane Formation as a Test", + "System for Biological Activity of Tobacco Smoke Components.'", + "CHROMOSOMAL ABERRATIONS INDUCED BY BENZO[A]PYRENE AND OTHER", + "POLYNUCLEAR AROMATIC HYDROCARBONS: CORRELATION WITH CANCER INDUCTION", + "E.", + "D. Rees, M.D., Department of Medicine and Pharmacology", + "ABSTRACT", + "When given intravenously in a lipid emulsion, ,12-dimethylbenz{a)anthracene (DMBA) and", + ",8,12-trimethylbenz[a)anthracene (TMBA) cause breaks in rat marrow cell chromosomes. The", + "maximum incidence of breaks is found within 6-24 hours; 43% and 55% of metaphase cells have", + "chromosomes with breaks following DMBA and TMBA, respectively. Benzo[a]pyrene (BaP) and", + "benzo[e]pyrene (BeP) have been similarly studied and have incidences of 21% and 11%, re-", + "spectively. The distribution of breaks on various chromosomes and the position of breaks", + "on the chromatids of individual chromosomes have been determined and summarized for each of", + "these compounds. Although damage is inflicted on chromosomes of all sizes and morphology,", + "the damage is nonrandom in that members of the S1 and T1 chromosome pairs are involved to", + "an extent greater than expected on the basis of their size and number. In addition, the", + "average number of breaks per chromosome for each of the individually identifiable chromo-", + "somes was calculated. Sharp differences between the four compounds were revealed for the", + "S1 and T1 chromosomes; TMBA was more damaging to the S1 chromosome by 1.25, 1.6 and 10 times,", + "respectively, than was DMBA, BaP, and BeP. These rankings (TMBA > DMBA BaP BeP) are", + "in", + "accord with our data for the carcinogenicity for these compounds. Thus, evidence is accu-", + "mulating which suggests that the ability of a polynuclear aromatic hydrocarbon to cause", + "cancer is related to its ability to inflict damage on certain chromosomes.", + "65", + "RC- 6029120", + "LG", + "0090632", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.174964, + 0.143655, + 0.682101, + 0.01332 + ], + [ + 0.135241, + 0.157606, + 0.748713, + 0.013365 + ], + [ + 0.135436, + 0.168814, + 0.705817, + 0.012219 + ], + [ + 0.135713, + 0.182937, + 0.735486, + 0.013384 + ], + [ + 0.135404, + 0.19633, + 0.750351, + 0.013379 + ], + [ + 0.135443, + 0.208212, + 0.743378, + 0.012607 + ], + [ + 0.135249, + 0.220622, + 0.690899, + 0.01257 + ], + [ + 0.135527, + 0.233189, + 0.73019, + 0.011506 + ], + [ + 0.135172, + 0.245869, + 0.714847, + 0.01167 + ], + [ + 0.135845, + 0.258611, + 0.728986, + 0.011813 + ], + [ + 0.13568, + 0.271094, + 0.722149, + 0.012698 + ], + [ + 0.135064, + 0.283678, + 0.722956, + 0.012626 + ], 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The mouse skin pro-", + "cedure has been described by Van Duuren (1) as follows: \"These tests, in order to be", + "ingful, must be carried out for at least one year, or preferably, for the lifetime of the", + "test group. It is frequently overlooked that tobacco smoke condensate is a weak carcinogen", + "and this fact adds to the problems of bioassay. High doses, 25 mg or more per application", + "applied three times weekly on mouse skin, are required, and even then do not result", + "tumors until after one year.\" Since over 1200 compounds have been identified in tobacco", + "smoke, such a testing system is almost prohibitively demanding of material, labor, and", + "facilities. Obviously, more rapid and convenient methods are needed. The objectives of", + "the present project are: a) to test the possibility of using chromosomal aberrations as", + "means of assessing the carcinogenicity of tobacco smoke components, and b) to study the", + "effect of carcinogens on chromosomes with the hope that insight might be obtained as to the", + "mechanism of polynuclear aromatic hydrocarbon carcinogenesis. In regard to (b), a major", + "problem at present in devising a method of testing compounds for carcinogenicity is that", + "we do not know with any certainty the subcellular and/or molecular details of the mecha-", + "nism(s) of chemical carcinogenesis. Presumably an accurate and appropriate carcinegen", + "assay could be readily devised if we had this knowledge.", + "RESEARCH METHODOLOGY", + "Chromosome Studies. The polynuclear aromatic hydrocarbons used in this study are", + "listed in Table I. Preparation of the lipid emulsion is based upon the procedure of Schurr", + "(2) ingredients are as follows: dextrose 0.15% w/w, Pluronic F-68 flakes 0.3% w/w, lecithin", + "(Upjohn) 1.2% w/w, sesame oil 15.0% w/w, water 78.85% w/w, and polynuclear aromatic hydro-", + "carbon (when added) 0.5% w/w (which provides 5 mg per ml of final emulsion) Emulsification", + "is carried out by high-speed (40,000 rpm) blending. The emulsion is administered by tail", + "vein injection to young rats of the Long-Evans and Sprague-Dawley strains in a dose of 5 mg/", + "100 g body weight. Chromosomes are prepared from the marrow cells of tibias and femurs of", + "colchicine-treated rats by a hypotonic citrate procedure.", + "Leukemia and Cancer Induction. Male and female rats of the Long-Evans and Sprague-", + "Dawley strain received four intravenous injections of one of the ten different polynuclear", + "aromatic hydrocarbons tested. For most of the compounds, three different groups of 10-15", + "rats each have been, or are being, followed for 6-8 months for appearance of leukemia,", + "mammary carcinomas, and other neoplasms.", + "RESEARCH RESULTS", + "Early changes in marrow cell chromosomes following intravenous DMBA. Following DMBA", + "injections, the most striking and distinctive observation is the presence of chromatid and", + "to a lesser extent isochromatid breaks. The appearance time of these lesions is relatively", + "rapid with 45% and 43% of metaphase cells having chromosomes with breaks at 6 and 24 hours,", + "respectively. Breaks are observed only rarely (less than 1%) in untreated rats. Control", + "rats receiving intravenous emulsion with all of the ingredients except DMBA have less than", + "a 1% incidence of breaks. In contrast to breaks it is not unusual to find gaps (narrow,", + "sharp, achromatic bands) on a chromosome of some untreated rats. Breaks are visually dis-", + "tinguished from gaps by displacement (greater separation or malalignment) of two pieces", + "chromatid and by unevenness of the edge on each side of the break.", + "In Fig. 1 the percent of cells having at least one chromosome with a break or gap is", + "plotted (separately for gaps and for breaks) versus time after injection.", + "dose, 2-3 days are required for the incidence of breaks to decay to one-half the initial", + "24 hour value. By the tenth day the incidence approaches the level of controls. Subse-", + "quent injections on day 10 and on day 20 again increase the incidence, but after the third", + "injection many animals show a persistent elevation of breaks. This is of considerable in-", + "terest, since at least three and preferably four to six injections are needed to índuce", + "66", + "Source: :https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "INTRODUCTION", + "leu", + "rat", + "DMI", + "att", + "mean-", + "in", + "a", + ":", + "b", + "cl", + "i:", + "t", + "F", + "d", + "c", + "t", + "E", + "s", + "1", + "P", + "1", + "1", + ".", + "of", + "first", + "the", + "After", + "RO--- 6029121", + "0090633", + "LG" + ] + }, + { + "bbox": [ + [ + 0.107153, + 0.14363, + 0.674177, + 0.015743 + ], + [ + 0.107418, + 0.155829, + 0.682175, + 0.014425 + ], + [ + 0.107015, + 0.16882, + 0.690019, + 0.012926 + ], + [ + 0.10702, + 0.18169, + 0.177007, + 0.014332 + ], + [ + 0.429976, + 0.205984, + 0.383195, + 0.009915 + ], + [ + 0.392257, + 0.218086, + 0.420748, + 0.010719 + ], + [ + 0.184541, + 0.222924, + 0.011807, + 0.006736 + ], + [ + 0.280891, + 0.223367, + 0.050902, + 0.007361 + ], + [ + 0.156953, + 0.223945, + 0.012123, + 0.00684 + ], + [ + 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A single injection does not lead to leukemia except in very", + "rate instances (3). . The incidence of chromosomal aberrations is dependent on size of the", + "DMBA dose as indicated in a dose-response curve obtained by Mr. Denis Becker in our labor-", + "atory (also, cf. ref. 4)", + "Incidence of aberrations induced by compounds re-", + "lated to DMBA. Nine other polynuclear aromatic hydro-", + "3rd", + "(o CONTROLS", + "2nd", + "ist", + "carbons, varying from strongly carcinogenic to non-", + ".", + "AFTER Ist INJ.", + "-", + "carcinogenic, and prepared in the same type of emul-", + "AFTER 2nd INJ.", + "sion and given in the same dose as DMBA, were injected", + "AFTER 3rd INJ.", + "in order to assess the relation between carcinogenic", + "potency and the ability to produce breaks and gaps", + "(Table I). Carcinogenic potency in the rat was esti-", + "mated from our own experience and from published work", + "of others on mammary cancer induction and sarcoma", + "production in other tissues following intragastric or", + "subcutaneous injection of the compounds. The incidence", + "of breaks does correlate roughly with oncogenic power.", + "The incidence of gaps also increased, but there is", + "I", + "*", + "great variability in gap incidence, and gaps commonly", + "20", + "40", + "10", + "50", + "70", + "30", + "60", + "DAYS AFTER FIRST DMBA INJECTION", + "occur even in untreated rats.", + "Effect of TMBA, DMBA, BaP, and BeP on individual", + "chromosomes and position of aberrations on chromatids.", + "Fig. 1", + "Of the ten compounds, TMBA, DMBA, BaP, and BeP have", + "been studied in greatest detail. We recorded the location of breaks and gaps on individual", + "chromosomes, as well as the position of the lesion on the chromatid. The chromosome number-", + "ing system is presented in Fig. 2 and follows the convention of Ijio and Levan (5). Karyo-", + "types of the Sprague-Dawley and Long-Evans rats have been described by Rees, et al (6)", + "For each compound data on all chromosomes with breaks and with gaps were assembled and the", + "distribution among the chromosomes or groups of chromosomes (i.e. I and M groups) was cal-", + "culated (Table II). Members of the S1 and T1 chromosome pairs had more breaks and gaps", + "than expected from probability considerations based on their size and number. Possibly a", + "more revealing statistic is obtained by dividing the number of breaks observed on chromo-", + "somes of a given type by the total number of the chromosomes studied (Fig. 3). This calcu-", + "lation not only sharply delineates the damaging ability of each of the compounds, but also", + "points out clearly the preferential effect of TMBA and DMBA on the S1 and T1 chromosomes.", + "The", + "ranking is clearly TMBA DMBA BaP BeP. In this regard it should be noted that", + "a", + "karyologic abnormality (i.e. trisomy and/or aneuploidy) involving T1 is found in almost half", + "of the DMBA-induced leukemic rats studied by Sugiyami et al (4,7) and by Rees et al (8,9).", + "Though most of the breaks and gaps are of the chromatid type, there is a significant number", + "TABLE I", + "Incidence 24 hours after i.v.", + "Carcinogen", + "% cells/gaps", + "% cell/breaks", + "Potency (rat)", + "Cells", + "Rats", + "TMBA", + "67%", + "-", + "7", + "55%", + "130", + "++++", + "DMBA", + "52%", + "43%", + "15", + "266", + "++++", + "33%", + "28%", + "79", + "3", + "+++", + "46%", + "enz(a)anthracene", + "22%", + "63", + "3", + "+", + "BaP", + "47%", + "21%", + "146", + "8", + "++", + "49%", + "19%", + "aminochrysene", + "65", + "4", + "++", + "benzolghi)fluoranthene", + "25%", + "48", + "10%", + "2", + "o", + "BeP", + "39%", + "6", + "11%", + "92", + "+", + "Chrysene", + "37%", + "8%", + "84", + "4", + "+", + 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"30.1", + "13.3", + "7.8", + "3.6", + "5.4", + "0.6", + "36.7", + "2.4", + "BaP (155)", + "32.2", + "21.9", + "1.9", + "5.2", + "1.9", + "0.6", + "34.3", + "1.9", + "BeP (80)", + "18.8", + "18.8", + "7.5", + "7.5", + "3.8", + "1.3", + "33.7", + "8.8", + "of the isochromatid type (TableIII) The position of breaks", + "Holtzmon", + "and gaps on chromatids of the individual chromosomes is pre-", + "R", + "R", + "sented in Fig. 4, the number of breaks or gaps observed in", + "each chromatid quarter is listed and the distribution (in", + "S1", + "11", + "S2", + "percent) is plotted. The M chromosomes are not presented,", + "0", + "p", + "a", + "n", + "P", + "since the centromere is in a median or near median region and", + "T CROUP", + "longer or shorter arms cannot be dependably distinguished.", + "Of necessity, the T chromosomes (other than T1) are lumped", + "n", + "A", + "A", + "together making interpretation difficult within this group.", + "S3", + "S,", + "55", + "Y", + "The first (next to centromere) and fourth quarters of the S1", + "=", + "=", + "y.", + "chromatids seem most vulnerable to breaks (and gaps) by TMBA,", + "M CROUP", + "DMBA, and BaP. (BeP did not elicit a sufficient number of", + "I", + "breaks for statistical analysis). In the case of T1, breaks", + "(but not gaps) were elicited most frequently by TMBA in the", + "Long-Evons", + "first and third quarters and by DMBA in the first.", + "in 10 All", + "o\"", + "S1", + "T1", + "Leukemia and mammary cancer inducing ability of the", + "various polynuclear aromatic hydrocarbons. Dose, sex, strain,", + "CADDRANO", + "age, and route of administration are important in carcinogen-", + "T", + "GROUP", + "esis by these compounds. Rats of both the Long-Evans and", + ",", + "2", + ".", + "Sprague-Dawley strain develop leukemia following intravenous", + "doses of DMBA and TMBA. The incidence of leukemia is very", + "53", + "S", + "S5", + "Y", + "much lower when the compounds are given intragastrically.", + "The Sprague-Dawley female is quite susceptible to mammary", + "M CROUP", + "cancer induction, but males are relatively resistant as shown", + "Fig. 2", + "by Sydnor, et al (10). Even females of the Long-Evans strain", + "are resistant to DMBA given intragastrically, but as shown by", + "et al (11) this resistance can .be overcome by giving DMBA intravenously. The results most", + "germane to this paper, however, are summarized in Fig. 5. (The incidence of mammary cancers", + "is for Sprague-Dawley females; the incidence of leukemia is for both males and females of", + "the Long-Evans strain) The data presented indicate that the compounds which are most", + "potent in damaging chromosomes are also the most potent cancer producers. However, for", + "both mammary cancers and leukemia (especially) it seems necessary for a compound to exceed", + "a threshold level of chromosomal aberrations before a substantial incidence of cancer", + "occurs. It is not known whether these findings are a peculiarity of polynuclear aromatic", + "hydrocarbons; other types of compounds have not yet been tested.", + "RO- 6029123", + "68", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgchoob§", + "0090635" + ] + }, + { + "bbox": [ + [ + 0.468256, + 0.124505, + 0.008911, + 0.00456 + ], + [ + 0.58018, + 0.125596, + 0.040341, + 0.004758 + ], + [ + 0.459152, + 0.129378, + 0.007668, + 0.006416 + ], + [ + 0.468419, + 0.132838, + 0.009307, + 0.005426 + ], + [ + 0.470617, + 0.143005, + 0.007528, + 0.004876 + ], + [ + 0.130738, + 0.146142, + 0.009113, + 0.006083 + ], + [ + 0.736977, + 0.156789, + 0.004452, + 0.006399 + ], + [ + 0.696268, + 0.157232, + 0.008449, + 0.009103 + ], + [ + 0.65571, + 0.157435, + 0.005646, + 0.006318 + ], + [ + 0.260147, + 0.165831, + 0.012541, + 0.006173 + ], + [ + 0.128642, + 0.165926, + 0.010096, + 0.006737 + ], + [ + 0.733802, + 0.187365, + 0.015966, + 0.004381 + ], + [ + 0.129184, + 0.188005, + 0.009082, + 0.005613 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Instead of DMBA or one of the", + "are:", + "other compounds, the neutral aromatic frac-", + "isogaps", + "isobreaks", + "tion of tobacco smoke condensate was used", + "28.2%", + "32.7%", + "TMBA", + "to prepare emulsions containing 5, 25, or", + "24.6%", + "27.1%", + "DMBA", + "50 mg of condensate per ml of emulsion. Con-", + "23.4%", + "21.1%", + "BaP", + "densate in a concentration of 5 mg/ml dis-", + "18.8%", + "7.1%", + "BeP", + "solved readily in sesame oil and emulsified", + "well, and when given at a dose of 5 mg per", + "100 8 rat weight, the rats showed no adverse", + "effects. The 25 mg dose, however, immedi-", + "ately made the animal inactive and this", + "TMBA", + "DMBA", + "lasted for about 40 minutes. The 50 mg dose", + ".", + "also led to immediate inactivity and the", + "animal died hours later (autopsy was not re-", + "3-MC", + "vealing though pulmonary embolism was sus-", + "20", + "pected). The 50 mg/ml emulsion was not", + "stable; the 25 mg/ml preparation was barely", + "sn", + "not", + "so. At both the 5 mg and 25 mg levels, chro-", + "forr", + "mosome preparations at 24 hours indicated a", + "DEBA", + "decreased mitotic index and the chromosomes", + "INDUCTION 11. 1. MUECTION)", + "CANCER", + "%", + "50", + "did not spread as satisfactorily as they", + "TMBA", + "DMSA", + "40", + "should have. With the 25 mg dose, the chro-", + "mosomes appeared \"battered.\" At 96 hours,", + "30", + "3-MC", + "too few metaphase cells were seen for chro-", + "mosome study; the paucity of mitotic cells", + "BaP", + "20", + "at this time again indicates an inhibitory", + "effect of the condensate on cell division.", + "CWR", + "% INDUCTION 11. Y. 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It sees", + "most probable, as has been suggested by Nichols and Levan (12), that more subtle chromosomal", + "lesions (which may include point mutations) occur as well as the breaks. To cause", + "cancer,", + "a subtle lesion undoubtedly would have to afflict an appropriate chromosome in the right PO-", + "sition in a manner analogous to that proposed in the target theory of radiation injury.", + "Since our data indicate that breaks are unduly frequent on S1 and T1 chromosomes (possibly", + "due to a greater \"affinity\" of these chromosomes for TMBA and DMBA, or a greater intrinsic", + "vulnerability of these chromosomes to damage by these compounds), subtle lesions presumably", + "would also be more common on these chromosomes. However, this concept requires that damage", + "must not only be inflicted in the proper place(s) on the proper chromosome (s), but that the", + "cell must not be rendered inviable or nonproliferative by additional damage elsewhere. These", + "are difficult requirements; even though the S1 and T1 chromosomes are preferentially involved,", + "they are not selectively involved. In fact, our findings indicate rather widespread damage", + "to other chromosomes, and the extent of nonchromosomal damage to the cell is not known. Also,", + "the \"leukemicized\" cell must not succumb to host defenses, if leukemia is to occur. Such", + "considerations suggest that the proper combination of cell damage and favorable environment", + "must be a rare event; a view which is in accord with the observation that*, despite many cells", + "in the marrow and a high incidence of damage to these cells following DMBA or TMBA, only one", + "abnormal stemline of cells is generally present in the leukemic host. In all, there are", + "ample experimental and theoretical reasons for exploring further the hypothesis that chromo-", + "somal damage is involved in chemical carcinogenesis--a least insofar as the polynuclear", + "aromatic hydrocarbons are concerned.", + "CONCLUSIONS AND RECOMMENDATIONS", + "It is still necessary to analyse in more detail the types of damage that the polynu-", + "clear aromatic hydrocarbons inflict on chromosomes and to test more extensively the hypoth-", + "esis that the chromosomal lesions are associated, or at least reflect in some manner the", + "carcinogenic process. This is required in order to establish the validity or invalidity of", + "using the incidence of chromosomal breaks (or other lesions) as an index of carcinogenicity.", + "If valid, such a test should be much faster and less demanding of test material and facil-", + "ities than the standard mouse skin bioassay which is customarily used in assessing the car-", + "cinogenicity of tobacco smoke fractions and components. As a fringe benefit, chromosome", + "work would also be more exciting than skin painting.", + "Data from experiments of the following types would be valuable in understanding chemi-", + "cal carcinogenesis and in developing a bioassay based on chromosome damage: a) The effect", + "of other_ types of compounds found in tobacco smoke (e.g. nitrosamines, alkaloids, etc.)", + "need to be tested by the intravenous emulsion procedure for both cancer-producing and chro-", + "mosome-damaging effects, since it is important to ascertain whether the observations above", + "pertain to compounds other than the polynuclear aromatic hydrocarbons. b) The localization", + "of tritiated polynuclear aromatic hydrocarbons on chromosomes can be carried out by auto-", + "radiography and the distribution of tritium could then be correlated with the distribution", + "of breaks and gaps. c) By impairing host defenses with immunosuppressives, a more sensitive", + "test for carcinogenicity might be achieved.", + "ACKNOWLEDGEMENTS", + "The excellent assistance of Mrs. Amy Shuck, Mr. John Lowry and Mrs. Helen Lipscomb is", + "gratefully acknowledged. Emulsions of TMBA and DMBA were generously provided by Professor", + "Charles Huggins, Ben May Laboratory for Cancer Research, University of Chicago. Emulsions", + "of DMBA were kindly provided by Dr. Paul E. Schurr of the Upjohn Company. Initial work", + "for this study was sponsored by an AMA-ERF grant.", + "RC.", + "70", + "LG 0090637", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.413598, + 0.137706, + 0.077451, + 0.007602 + ], + [ + 0.118133, + 0.159501, + 0.57279, + 0.014835 + ], + [ + 0.117613, + 0.179732, + 0.011469, + 0.00883 + ], + [ + 0.143358, + 0.174216, + 0.648808, + 0.014752 + ], + [ + 0.14283, + 0.186894, + 0.207384, + 0.012514 + ], + [ + 0.118057, + 0.19809, + 0.699459, + 0.014423 + ], + [ + 0.14283, + 0.210045, + 0.547771, + 0.012976 + ], + [ + 0.11804, + 0.226744, + 0.011717, + 0.00869 + ], + [ + 0.142593, + 0.222198, + 0.667581, + 0.013549 + ], + [ + 0.142897, + 0.234158, + 0.650391, + 0.012302 + ], + [ + 0.118368, + 0.250539, + 0.010909, + 0.008932 + ], + [ + 0.143323, + 0.249087, + 0.033248, + 0.010662 + ], + [ + 0.184511, + 0.246218, + 0.625998, + 0.011104 + ], + [ + 0.142832, + 0.257815, + 0.261468, + 0.01233 + ], + [ + 0.118265, + 0.270197, + 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In the S. aureus", + "system, study of the potentiation of BaP toxicity by smoke components can now be analyzed in", + "detail.", + "INTRODUCTION", + "Vertebrate cells generate energy in a complex membrane-organized electron transport-", + "respiratory system. Neoplastic cells and some bacteria generate energy in a loosely organ-", + "ized, inefficient glycolytic system. One of the constant features of the neoplastic trans-", + "formation is the progressive loss of the respiratory system and the increasing dependence on", + "the glycolytic enzymes for energy. Agents which effect the formation of the membrane-bound", + "electron transport system could select cells with increasing independence from the respiratory", + "system (neoplastic cells), and could also so interfere with the normal cellular process as to", + "change the adsorption properties of the epithelium and damage the normally efficient pulmo-", + "hary which clearance mechanisms to allow a greater exposure to noxious agents. A microbial system", + "grows glycolytically but in which the membrane-bound electron transport system can be", + "71", + "RO- 6029126", + "LG", + "0090638", + "Source: ttps://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.14633, + 0.123455, + 0.750618, + 0.011431 + ], + [ + 0.146406, + 0.134773, + 0.744761, + 0.013124 + ], + [ + 0.146567, + 0.147668, + 0.750595, + 0.014114 + ], + [ + 0.147015, + 0.161616, + 0.743631, + 0.013153 + ], + [ + 0.146813, + 0.173653, + 0.736369, + 0.014066 + ], + [ + 0.147033, + 0.187302, + 0.737614, + 0.012249 + ], + [ + 0.147024, + 0.201303, + 0.168947, + 0.01066 + ], + [ + 0.440471, + 0.224825, + 0.164856, + 0.008718 + ], + [ + 0.196258, + 0.248863, + 0.695567, + 0.011736 + ], + [ + 0.146426, + 0.261555, + 0.739591, + 0.012776 + ], + [ + 0.146071, + 0.274227, + 0.74633, + 0.012393 + ], + [ + 0.147151, + 0.286341, + 0.732246, + 0.011565 + ], + [ + 0.146347, + 0.299486, + 0.726185, + 0.011427 + ], + [ + 0.147314, + 0.311826, 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In addition great numbers of mutants can be examined to study rare", + "genetic events if selection systems can be developed. These mutants can be used to construct", + "genetic maps of various lesions by efficient means. Such a microbial system has been devel-", + "oped. The response of Staphylococcus aureus to smoke components has been shown to parallel", + "the responses in mammalian tissue. This gives validity to its use in the bioassay of smoke", + "and smoke components.", + "RESEARCH METHODOLOGY", + "S. aureus cannot grow in nutrient broth unless oxygen and a functional electron trans-", + "port system are present (1). This observation formed the basis of a selective technique in", + "which mutants unable to form an electron transport system form minute colonies. If the agar", + "surface is seeded heavily and the inhibitors of the electron transport system are added to", + "wells cut in the surface, inhibition of growth can be readily detected (Fig. 1). Mutants", + "can be analyzed genetically by use of transducing phage 53 (1). The electron transport sys-", + "tem can be induced by adding air to anaerobically growing cultures (2). Cytochromes, complex", + "lipids and carotenoids have beei. characterized and methods for their accurate assay developed", + "(3,4). The metabolism of membrane components was measured by 3rowing the culture in a medium", + "containing suitable labeled compounds and then examining the radioactivity in components dur-", + "ing further growth in nonradioactive medium (5). This pulse-chase type of experiment has", + "great sensitivity to retabolic interactions.", + "Fig. 1. Inhibition of growth of S. aureus in nutrient agar by", + "0.01M crystal violet (top), benzo (e) pyrene (left) and benzo (a) pyrene", + "(right). The components were added in ethanol solution.", + "the growth of S. aureus under conditions in which the noncarcinogenic isomer BeP had no", + "effect (Fig. 2). BaP had no effect on glycolytic growth in this system. The fact is that", + "the BaP effect parallels the changes accompanying the formation of the electron transport", + "system. The electron transport system can be induced by adding air to anaerobically grow-", + "ing cells (Fig. 3). In the course of this induction there was extensive change in the", + "metabolism of the membrane lipids (2) and a striking synthesis of the rubixanthin carote-", + "noids (Fig. 4). Inhibition of the synthesis of vitamin K2 or polar carotenoids by 50 to", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "RESEARCH RESULTS", + "Using selective media as a bioassay system", + "in which growth is dependent on the function of", + "the membrane-bound electron transport system,", + "the following smoke components were shown to", + "have no effect on growth at 10mM: pyrocatechol,", + "nicotine, harmine, anabasine, 2,6-lutidine,", + "phenol, toluene safrole, dihydrosafrole, es-", + "culetin, p-dihydroxybenzene, benzene or aged", + "water soluble smoke condensate. Inhibition was", + "shown by benzo[a]pyrene (BaP), benzo[e]pyrene", + "(BeP), dibenz[a,i]acridine, alpha-naphthylamine", + "and beta-naphthylamine. If the inhibition was", + "dependent on the function of the electron trans-", + "port system, then the addition of glucose to", + "the medium should reverse the inhibition. Only", + "BaP showed a decreased zone of inhibition when", + "glucose was present indicating that BaP might", + "be somehow involved with the electron transport", + "system. The specific phospholipid inhibitor", + "4 '-4\" Bis ,5,6,-tetrahydro-2-pyrimidinyl)-", + "terephthalanilide (TPA) has the same inhibitory", + "properties as BaP. Very fresh smoke extracts", + "also had marked inhibitory activity. Stable", + "genetic changes resulting in defective ability", + "to form a functioning electron transport system", + "have been selected after growth with BaP.", + "BaP at a smoke carcinogen, inhibited", + "72", + "0090639", + "LG", + "RO- 6029127" + ] + }, + { + "bbox": [ + [ + 0.448086, + 0.136593, + 0.373195, + 0.015163 + ], + [ + 0.136525, + 0.158452, + 0.012781, + 0.007089 + ], + [ + 0.21643, + 0.155954, + 0.006859, + 0.005579 + ], + [ + 0.257652, + 0.156066, + 0.01413, + 0.0068 + ], + [ + 0.448648, + 0.15027, + 0.371531, + 0.014487 + ], + [ + 0.448519, + 0.163225, + 0.355987, + 0.014428 + ], + [ + 0.449042, + 0.174805, + 0.370944, + 0.016483 + ], + [ + 0.449325, + 0.18842, + 0.379633, + 0.016183 + ], + [ + 0.136461, + 0.205711, + 0.011891, + 0.006431 + ], + [ + 0.257416, + 0.203332, + 0.01348, + 0.006581 + ], + [ + 0.449485, + 0.200074, + 0.380178, + 0.016265 + ], + [ + 0.4499, + 0.212122, + 0.363557, + 0.015634 + ], + [ + 0.449384, + 0.224781, + 0.380218, + 0.014359 + ], + [ + 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synthesis. The major por-", + "tion of the rubixanthins in S. aureus were syn-", + "05", + "05", + "thesized from -carotene by aerobic hydroxyla-", + "tion. This hydroxylation is inhibited by the", + "specific inhibitors of mammalian mixed function", + "oxidase systems (8). A P450 pigment which is", + "characteristic of mammalian mixed function oxi-", + "dase systems has been detected in S. aureus (8).", + "This pigment is induced by aeration (Fig. 3).", + "Synthesis of the P450 microsomal mixed function", + "oxidase system in mammalian liver is induced by", + "or", + "or", + "BaP (9) and BaP toxicity in mice is markedly", + "elevated in the presence of mixed function oxi-", + "SNUTRIENT BROTH", + "50mM GLUCOSE", + "NUTRIENT BROTH", + "dase inhibitors (10). BaP at reasonable concen-", + "trations is toxic to S. aureus only under condi-", + "005", + "2", + "3 4", + "oos", + "I", + "o", + "2", + "4", + "6", + "tions when the P450 system is active.", + "HOURS", + "HOURS", + "CONTROL", + "DISCUSSION", + "1,2 BENZPYRENE", + "0.01mM 3,4 BENZPYRENE", + "The demonstration that BaP, a known tobacco", + "Fig. 2. Growth rate of S. aureus measured as the absor-", + "smoke carcinogen, is highly inhibitory to S.", + "bance at 750 mu in 13 mm round test tubes in nutrient", + "aureus under conditions when the noncarcinogenic", + "broth (left) and autrient broth plus 50 mM glucose", + "(right) in the presence of benzo(e]pyrene or benzo(a)-", + "smoke isomer BeP is not, represents a striking", + "pyrene, The carcinogens were added in ethanol such", + "observation. In addition BaP greatly increases", + "that the final ethanol concentration was less than 1%.", + "A similar amount of ethanol was added to the control.", + "the frequency with which stable genetic changes", + "resulting in mutants unable to form a membrane-", + "bound electron transport system appear. BaP is", + "04", + "inhibitory in S. aureus only when the recently", + "discovered (8) mixed function oxidase system is", + "active. Since BaP induces this system in mamma-", + "02", + "lian cells (9), further work could lead to the", + "details of its interaction with the membrane.", + "Of", + "This should be an ideal system to see if BaP is", + "0081", + "modified during its biological activity.", + "-", + "o", + "I", + "2", + ".", + "Piperonyl butoxide (PB) is a very specific in-", + "10", + "e", + "hibitor of the mixed function oxidase in S.", + "o", + "80", + "aureus (8), and it is widely used in agriculture", + "e", + "PROTOHEME", + "60", + "as a potentiator of insecticides. As much as", + "10 ppm have been found in fruit and cereals at", + "40", + "harvest (10). PB increases the mortality after", + "2", + "20", + ".", + "exposure to BaP by 34% in weanling mice (10).", + "-1", + "o", + "I", + "2", + "An aromatic ether like PB, myristicin has been", + "detected in tobacco smoke (11) and could very", + "D", + "14", + "o", + "a", + "I", + "likely potentiate the effect of BaP on pulmonary", + "450", + "tissue. Carbon monoxide (co) is a very effec-", + "%", + "tive inhibitor of both the mammalian and S.", + "to", + "o", + ",", + "aureus P450 system. co could further potentiate", + "DO", + "the toxic effects of BaP. A biological system", + "os", + "has been developed in which these potentiating", + "effects can be readily tested. The applica-", + "bility of the conclusions from work with S.", + "0.2", + "aureus to the tobacco and health program is", + "-", + "o", + "I", + "2", + "based on the parallelism between the effects on", + "MOURS", + "both systems and makes further work an exciting", + "Figure 3. Cytochrome levels during a switch", + "possibility.", + "from anaerobic to serobic growth. The time", + "of switch is indicated by the arrow. Cyto-", + "RO- 6029128", + "chromes were measured by difference spectra", + "after reduction in the presence of L-lactate,", + "73", + "LG", + "0090640", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.557648, + 0.140918, + 0.249591, + 0.009364 + ], + [ + 0.224481, + 0.152854, + 0.006008, + 0.007959 + ], + [ + 0.224053, + 0.165001, + 0.007051, + 0.007595 + ], + [ + 0.493229, + 0.167014, + 0.385193, + 0.010089 + ], + [ + 0.305508, + 0.174439, + 0.008278, + 0.004436 + ], + [ + 0.248246, + 0.179884, + 0.004801, + 0.003813 + ], + [ + 0.35828, + 0.17865, + 0.004423, + 0.004687 + ], + [ + 0.305119, + 0.182016, + 0.040685, + 0.004886 + ], + [ + 0.373302, + 0.183002, + 0.004853, + 0.007148 + ], + [ + 0.452814, + 0.179004, + 0.431576, + 0.011733 + ], + [ + 0.249059, + 0.190528, + 0.004857, + 0.005248 + ], + [ + 0.453254, + 0.192151, + 0.430845, + 0.009975 + ], + [ + 0.248156, + 0.205521, + 0.004435, + 0.00417 + ], + [ + 0.357253, + 0.204669, + 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The S. aureus system offers", + ",", + "-", + "the opportunity to examine the action of BaP especially", + "os", + "7", + "o", + "*,", + "a", + "$", + "KOURS", + "with the potentiating agents co and PB or fresh smoke", + "PRYTOENE", + "itself to find how this carcinogen could induce genetic", + "4", + "change. The changes induced in this organism are strik-", + "ingly like the changes in the neoplastic transformation.", + "2", + "s", + "Since this assay is relatively easy, economical, and", + "RUBIXANTHINS", + "rapid, modifications of the cigarettes or filters can be", + "3", + "C-CAROTENE", + "examined in attempts to reduce the potentiation.", + "$", + "REFERENCES", + "4", + "PHYTOFLUENOL", + "1. Tien, W. C. and White, D. c. Linear sequential ar-", + "2", + "rangement genes for the biosynthetic pathway of", + "is", + "protoheme in Staphlococcus aureus. Proc. Nat. Acad.", + "Sci. U. S. 61:1392-8, 1969.", + "6", + "2.", + "Frerman, F. E. and White, D. C. Membrane lipid", + "4", + "changes during formation of a functional electron", + "transport system in Staphlococcus aureus. J.", + "2", + "Bacteriol. 94:1868-74, 1967.", + "\"1", + "o", + "of", + "*2", + "3. White, D. C. and Frerman, F. E. Extraction, charac-", + "MOURS", + "terization and cellular localization of the lipids of", + "Figure 4. Carotenoid levels during a", + "Staphylococcus aureus. J. Bacteriol. 94:1854-67, 1967.", + "svitch from anaerobic to aerobic growth.", + "4.", + "Hammond, R. K. and White, D. C. Carotenoid formation", + "The svitch from anaerobic growth is in-", + "dicated by an arrow on the abscissa.", + "by Staphylococcus aureus. J. Bacteriol. 101: in press,", + "The carotencids were purified, assayed", + "1970.", + "as described (4). Growth is illustrated", + "in the upper part of the figure.", + "5. White, D. c. and Tucker, A. N. Phospholipid metabolism", + "during bacterial growth. J. Lipid Res. 10:220-3, 1969.", + "6. Yesair, D. W. ; Rodgers, W. I. 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Science 151:96-7, 1966.", + "DISCUSSION HIGHLIGHTS", + "SMITH: Dr. Rees, how many animals do you need to develop data shown on your graphs?", + "REES: Our usual procedure is to score 20 cells in each animal, and, depending upon the kind", + "RO- 6029129", + ":", + "74", + "LG", + "0090641", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.103063, + 0.127923, + 0.687613, + 0.015933 + ], + [ + 0.103176, + 0.144993, + 0.052542, + 0.011261 + ], + [ + 0.171194, + 0.142996, + 0.023447, + 0.009257 + ], + [ + 0.202458, + 0.139304, + 0.620514, + 0.013798 + ], + [ + 0.103226, + 0.152027, + 0.704245, + 0.015301 + ], + [ + 0.103463, + 0.165532, + 0.703639, + 0.014696 + ], + [ + 0.102867, + 0.18146, + 0.067364, + 0.011067 + ], + [ + 0.102856, + 0.203458, + 0.712954, + 0.012734 + ], + [ + 0.10271, + 0.215007, + 0.511692, + 0.013665 + ], + [ + 0.102002, + 0.240409, + 0.705593, + 0.012693 + ], + [ + 0.101648, + 0.254041, + 0.099777, + 0.010906 + ], + [ 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about four", + "acimals.", + "The", + "decay curve, which involved studying animals over a time period beyond injec-", + "tion, required about ten animals and the scoring of about 200 cells. When merely scoring", + "for the presence of breaks, four or five rats are enough, and two or three rats a day can", + "be scored.", + "COLLINS: You mentioned that you have to give three-six doses a day. Can you give more in", + "one dose or would it be too toxic? Is there a cumulative effect?", + "REES: It would be toxic in higher doses. You can lose quite a few animals with DMBA and", + "TMBA as it is.", + "COLLINS: But if you distribute the same dose over say 15 applications, would this be simi-", + "lar to the same dose given in six applications? Or does it act like small doses when ap-", + "plied this way?", + "REES: I don't know that. I do know that when giving intragastrically it is more effective", + "to give several small amounts than one large one. But in these experiments the application", + "has been intravenous.", + "COLLINS: Are you ready to make a guess about low dosages and cumulative effect as it may", + "apply to tobacco smoke", + "even though the doses you are using are much larger than would", + "be obtained even from quite a few cigarettes?", + "REES: Yes. Tobacco smoke is weakly carcinogenic, and as Van Duuren points out, this is a", + "difficulty in bioassay. It takes a large number of animals over long periods of time. If", + "you lower the dose, two things happen as far as mammary cancer induction is concerned: fewer", + "animals get cancer; and it takes a lot longer for those animals to get cancer. I think all", + "of the environmental carcinogens that we're dealing with are low doses over long periods of", + "time, and it must also involve individual susceptability. This makes for real problems in", + "understanding the cause of human cancer.", + "DONNELLY: In regard to the graphs which you presented, have you given an interpretation to", + "these chromosome break data?", + "REES: Most breaks probably are either lethal to the cell, or else the cell cannot go on to", + "divide again. In the case of the marrow, these cells leave the marrow and never return.", + "The break itself is probably lethal and is not the actual cause of the leukemia. If the", + "break occurs in the right part of the chromosome at the right time, then you may have cancer.", + "There are a lot of independent factors in carcinogenesis, and a big element of fate.", + "ALEEM: Could you give us more information about how you use the emulsion?", + "REES: We mix the lipid emulsion with 5 milligrams of each one of the compounds per milli-", + "liter of the emulsion, and we inject 5 milligrams per 100 grams of rat body weight. We give", + "the first injections when the rats are about 23-24 days old, because the tail veins are nice", + "at this age.", + "SMITH: Is this a practical bioassay for smoke condensate? Since there is not much BaP", + "in", + "the condensate, how much condensate do you have to inject into a rat to get some response", + "to the carcinogen?", + "REES: We've started injecting smoke condensate, but we haven't done enough to make me want", + "to go on record. If this system of ours proves to be a valid assay, then we will be able", + "to ask the quantitative question.", + "DONNELLY: Dr. Rees, is the DNA synthesis sequence along the chromosomes of the rat well", + "understood? The point to my question stems from the fact that you notice breaks six hours", + "after the DMBA administration. This suggests that the breaks you see probably occurred", + "within the last three hours of DNA synthesis. You might be able to thereby determine", + "RO- 6029130", + "75", + "LG", + "0090642", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.170416, + 0.13928, + 0.600574, + 0.012176 + ], + [ + 0.171213, + 0.16467, + 0.278853, + 0.010505 + ], + [ + 0.171655, + 0.190377, + 0.720173, + 0.011228 + ], + [ + 0.171188, + 0.203466, + 0.217111, + 0.011253 + ], + [ + 0.172397, + 0.227893, + 0.697554, + 0.012137 + ], + [ + 0.172196, + 0.240259, + 0.72009, + 0.01202 + ], + [ + 0.172582, + 0.252382, + 0.720665, + 0.01136 + ], + [ + 0.171939, + 0.264514, + 0.690091, + 0.011933 + ], + [ + 0.171255, + 0.276614, + 0.722077, + 0.011227 + ], + [ + 0.172322, + 0.288974, + 0.687273, + 0.011483 + ], + [ + 0.172584, + 0.313949, + 0.719393, + 0.011028 + ], + [ + 0.173079, + 0.326266, + 0.577474, + 0.011523 + ], + [ + 0.173089, + 0.351209, + 0.743678, + 0.01108 + ], + [ + 0.173373, + 0.363801, + 0.705994, + 0.011896 + ], + [ + 0.17321, + 0.375804, + 0.713604, + 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associated with leu-", + "kemia cells; is that right?", + "REES: Yes, except that the S1 and T1 chromosomes do seem to be the most susceptible to", + "damage. They are the ones which are abnormal with the greatest frequency, sometimes 40 to", + "50% of the time. But I don't think the breaks cause the leukemia. The most promising ex-", + "periment is to use tritiated DMBA and see if it can be located on the chromosomes, and", + "whether it is located on the chromosomes at the site of the breaks and gaps. The same can", + "be done for BaP. These are quite feasible experiments, but we have not done them yet.", + "SHEEN: How does your rat bone marrow system compare with human tissue? When you add BaP,", + "do you get aberrations at the same frequency as one would in HeLa cells?", + "REES: HeLa cells have abnormal chromosomes to begin with. In tissue culture things are quite", + "different, and the problem of delivery is important. Since BaP and all related compounds", + "are not soluble in water; you have to have a liquid emulsion. The nice thing about using", + "the whole animal is that the animal does a lot of work for you in the way of delivering the", + "material to the vulnerable site, and that's a big help. A lot of things happen to the chro-", + "mosomes by just putting cells in culture. In the case of X-rays I don't think there is too", + "much difference between the species as far as the effect on the chromosomes is concerned.", + "ALEEM: Dr. White, does BaP have an effect on the cytochrome oxidase system?", + "WHITE: Not much effect, but it cannot be excluded, because it is in the particulate phase.", + "DONNELLY: Do you mean you cannot use the volatile phase effectively?", + "WHITE: I'm waiting for the new smoking machine.", + "DONNELLY: Dr. Yoneyama, do you see any high incidence of abnormality associated with spe-", + "cific hydrocarbons?", + "YONEYAMA: We have noticed BaP carcinoma and also benign tumors in surviving animals.", + "ALEEM: What is your thought about the inhibitor in the very fresh smoke? Is it nitric", + "oxide?", + "WHITE: No, it is more potent than that. It is more potent than anything we've ever looked", + "at.", + "ALEEM: But you have been unable to identify it for certain?", + "WHITE: It is very labile.", + "GRUNWALD: Dr. Donnelly, with your cillostasis system, couldn't you be cutting out the oxi-", + "dative phosphorylation system which must be involved in cilia movement? Are you not in", + "effect testing the same thing that Dr. White is testing?", + "DONNELLY: I don't know. No one is quite sure what controls ciliary beating. There are", + "many things in the water soluble condensate which would be expected to kill cells, and", + "death would certainly turn off cilia movement. Many of these agents bring about loss of", + "cells from tissue. When cilia stop beating in the trachea, materials pile up in the lungs,", + "and major histological changes occur in the respiratory tissues.", + "GRUNWALD: Dr. White, how important do you really think oxidative phosphorylation is in the", + "overall problem of cancer. Might this be the starting point for chromosomal breakage within", + "the precancerous cell?", + "76", + "RO-- 6029131", + "LG", + "0090643", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.128133, + 0.143913, + 0.704711, + 0.013682 + ], + [ + 0.129899, + 0.157535, + 0.695226, + 0.012734 + ], + [ + 0.129284, + 0.169649, + 0.72661, + 0.014379 + ], + [ + 0.129214, + 0.182726, + 0.695402, + 0.012526 + ], + [ + 0.13072, + 0.195639, + 0.678217, + 0.011377 + ], + [ + 0.130817, + 0.208037, + 0.692352, + 0.011436 + ], + [ + 0.130452, + 0.232817, + 0.511808, + 0.011215 + ], + [ + 0.130186, + 0.257102, + 0.719053, + 0.011458 + ], + [ + 0.13108, + 0.269627, + 0.462745, + 0.012017 + ], + [ + 0.130413, + 0.293886, + 0.528127, + 0.011817 + ], + [ + 0.130463, + 0.318681, + 0.703543, + 0.011426 + ], + [ + 0.131138, + 0.331252, + 0.669507, + 0.011504 + ], + [ + 0.13111, + 0.355483, + 0.718692, + 0.010738 + ], + [ + 0.131367, + 0.368081, + 0.703438, + 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Of course, if there is no oxidative phos-", + "phorylation, then the cell dies, and there is no chance for chromosomal repair. Many agents", + "which are inhibitors of the electron transport system are also carcinogens. But many of", + "the effects could also be indirect, such as inhibition of defense mechanisms. Ciliary", + "clearance of the lungs, for example, is dependent upon the ciliary requirements for ATP.", + "ISSIDORIES: Dr. Donnelly, which are the main ciliostatic agents?", + "DONNELLY: Most of the agents which have been examined are those in the vapor phase, and of", + "these phenols and acrolein are the most highly ciliostatic.", + "ISSIDORIES: Is the phenomenon of ciliostasis a reversible process?", + "DONNELLY: Reversibility would also be time dependent. This is a very different question", + "since we are measuring ciliostasis on a population basis and not on a per cell basis.", + "SHEEN: Since cilia movement is inhibited by specific compounds, some of which inhibit oxi-", + "dative phosphorylation, is your system better than one which used microorganisms or lower", + "animals such as the paramecium?", + "DONNELLY: These systems have been used in determining and screening ciliostatic compounds.", + "Cilia occur throughout the plant and animal kingdoms, except in angiosperms, I believe.", + "Cilia formation can occur in almost any cell type; the only apparent requirement is the", + "presence of a centriole, because the centriole becomes the basal body which gives rise to", + "the cilium. All cilia which move probably have the same 9 + 2 pattern, and there is a close", + "relationship to mitotic events, the mitotic spindle, and the whole array of microtubules.", + "Cilia are thus part of a very basic and broad biological phenomenon. The mode of action of", + "the ciliostats is not known, but one would certainly presume that oxidative phosphorylation", + "is involved. But what you're really asking is whether the tracheal system is in fact the", + "preferred system when there are so many other systems available. The tracheal system offers", + "several advantages. For example, if you want to develop dose-response information, you", + "recognize that there is also some temperature dependence. If one is using a paramecium or", + "algal or clam gill system, mammalian body temperature is probably not optimal, so one more", + "variable must be added in trying to correlate the response with the human condition. With", + "the trachea, we can study ciliostasis, and, in the same piece of tissue study the histology", + "of the respiratory epithelium. This tissue is after all the target tissue with which this", + "whole smoking question is concerned. Invertebrate systems require an even greater stretch", + "of the correlations found in mouse back painting. Here at least we are in a mammalian sys-", + "tem and in the target tissue as well. I think this is ample justification, even though", + "we", + "may never produce cancer with our present arrangement.", + "GRUNWALD: Maybe Dr. White would like to give a justification of his system. Do you think", + "it's valid to make comparisons between your system and mammalian systems?", + "WHITE: I think one must show that it is. We can't make a tumor with Staph that's for sure.", + "But Dr. Donnelly can't make genetic observations at the level of 1 X 10-9 which I can do.", + "REES: 1 was wondering, Dr. Donnelly, if you have ever taken rats like those of Dr. Sydnor,", + "treated with the aqueous extracts for long periods of time, and then made tracheal rings", + "to see what the cilia function was.", + "DONNELLY: One of the problems is that her rats are older, which means the tracheal diameter", + "is much greater. We use a very unusual method of histological preparation. For light", + "microscopy studies we prepare the tissues as one does for electron microscopy. We are", + "limited in the diameter of tracheal tissues that we can cut. By the time the rat is 30 dpys", + "old, his tracheal diameter is too big for us to handle. The size of the section would ex-", + "ceed the size of the glass knife. You just can't cut thin sections that way and partial", + "sections make orientation difficult. You need to examine the ring in toto because there is", + "so much damage when the trachea is used under the best of conditions. We are using hamsters", + "RO- 6029132", + "77", + "LG", + "0090644", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.17419, + 0.130823, + 0.730295, + 0.012201 + ], + [ + 0.174627, + 0.144413, + 0.528419, + 0.012312 + ], + [ + 0.174788, + 0.169713, + 0.657194, + 0.011604 + ], + [ + 0.174553, + 0.196177, + 0.668437, + 0.010499 + ], + [ + 0.851136, + 0.200095, + 0.02398, + 0.007054 + ], + [ + 0.882343, + 0.1988, + 0.036633, + 0.011618 + ], + [ + 0.174874, + 0.208245, + 0.755816, + 0.014644 + ], + [ + 0.174537, + 0.221003, + 0.750898, + 0.012761 + ], + [ + 0.1741, + 0.233719, + 0.736694, + 0.01259 + ], + [ + 0.174129, + 0.246077, + 0.73805, + 0.012387 + ], + [ + 0.174626, + 0.258309, + 0.744881, + 0.01282 + ], + [ + 0.173985, + 0.271028, + 0.731091, + 0.012636 + ], + [ + 0.174541, + 0.28376, + 0.738367, + 0.013641 + ], + [ + 0.173841, + 0.296256, + 0.746268, + 0.011774 + ], + [ + 0.174989, + 0.308626, + 0.731186, + 0.012585 + ], + [ + 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"analyzed because many of ours have been exposed to smoke condensate for six months or longer.", + "The only preliminary things that we can talk about here are what we do see in the whole ani-", + "mal. We are using whole animals and give intranasal applications into the bronchial tree.", + "The one fantastic change in the cell turnover has been with tobacco smoke where the normal", + "range in a high spontaneous tumor animal (producing a tumor at approximately 600 days) will", + "now go on until approximately 2000 days. In other words, there is a cell population that", + "doesn't synthesize cells. They are not necrotic; they'r not inflamed; they're not losing", + "weight. These animals are healthy. We will correlate, of course, the anatomic differences", + "and we have a built in control in their swallowing. We have studied the esophagus at the", + "same time. The genetically-determined cell turnover in the mouse can be altered", + "chemically", + "by tobacco smoke and turnover time is totally different from that for inzo[a]pyrene as well", + "as from acrolein, which we have used also. I can't give you the exact figures. We have", + "given massive doses concentrating on cancer. We have produced lung tumors at an age when", + "there should be no tumors at all, and we have actually produced lung tumors in our animals", + "that normally never get lung tumors. There are a number of things occurring in the whole", + "animal which are very pertinent to applying cigarette smoke to this model, and a biologic", + "assay which allows us to test the level quanitatively is very important.", + "YONEYAMA: From the standpoint of pathology I feel that mouse back painting efforts and", + "lung cancer are very different things. We have been observing tumors and cancers in the", + "skin as well as in the lung from the very small ones to very large ones; and the process of", + "developing a cancer in the skin and in the lung are very different. Even though we can pro-", + "duce the skin cancer by the condensate of the smoke, I don't think condensate can produce", + "lung cancer. We have to have some kind of system with which we can simulate human cancer", + "development in the lung.", + "DONNELLY : Another plea for inhalation studies.", + "GRIFFITH: Before considering inhalation studies, let me give an analysis which will indi-", + "cate that the members of this panel may be much closer than they perhaps realize to answer-", + "some of the critical questions pertaining to the relationship between tobacco and health.", + "Dr. Donnelly is perhaps closest to having a viable bioassay system which can measure", + "differences in the biological activity of different products. This test system is concerned", + "with the gas phase components of smoke, and she only needs the new smoking machine to put", + "her test into routine operation.", + "Dr. White is also close to having a bioassay system in that he has been able to demon-", + "strate", + "an effect of fresh smoke on the growth of his bacteria. With the new smoking machine", + "he should be able to develop this into a test which will allow him to test products routinely", + "and to use the test system to. identify the compound(s) responsible for the effect on cyto-", + "chrome oxidase. This test system will be most concerned with the gas phase components of", + "smoke.", + "The chromosome breakage work of Dr. Rees is potentially useful as a bioassay in which", + "the number of chromosome breaks could be used as a measure of the biological activity of", + "different products. His test would probably be correlatable with cancer initiation and", + "hopefully would be able to predict the level of mouse skin painting activity. He should", + "be able to correlate his work with some of that carried on by Dr. Sydnor.", + "Dr. Sydnor, unfortunately, could not be here this afternoon. Her work has a potential", + "for replacing the mouse skin painting technique, and she is near the point where her tech-", + "niques could be used to measure quantitative differences in biological activity.", + "Dr. Roeckel is obtaining interesting results in her studies. of cell turnover rates, but", + "the chances of a routine bioassay are a bit more remote unless some way can be found to", + "mechanize her measurement procedures--they are simply too time consuming at the present", + "time.", + "The program attempts to develop new bioassay procedures have been successful and we can", + "78", + "RC-- 6029133", + "LG", + "0090645", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.102312, + 0.126651, + 0.728805, + 0.013141 + ], + [ + 0.101944, + 0.139472, + 0.745081, + 0.013911 + ], + [ + 0.102592, + 0.153527, + 0.40013, + 0.011089 + ], + [ + 0.140687, + 0.165352, + 0.706951, + 0.011574 + ], + [ + 0.103385, + 0.177752, + 0.712372, + 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The results which", + "will be obtained should provide guidelines for those doing agricultural research, and should", + "be useful to others in the total research program.", + "In the discussions within the technical committee during the development of the program", + "of research, the need for inhalation research was recognized. We will soon have the new", + "smoking machine and will be able to manipulate smoke in new ways. Assuming that we have", + "this capability, what work do we need to do in inhalation research?", + "REES: There are a lot of physiological functions and a lot of components in smoke; I think", + "we try too hard to get one component to do everything. We probably do make distinctions, as", + "we mentioned earlier. Exactly what is it that we want to study? The transport of smoke", + "through the villi into the blood is probably of interest, but Dr. Mandelstam and I have been", + "mentioning it for some time. Everything may not be inhalation; some things may be coughed", + "up in the sputum, swallowed and delivered into the intestinal tract.", + "GRIFFITH: Let us consider a specific example where inhalation might be useful. What would", + "happen if Dr. Sydnor would inject BaP into the backs of rats and then instead of giving them", + "the water soluble fraction of smoke condensate subject the animals to smoke inhalation?", + "WHITE: To use inhalation in my system, one would have to figure out a way to stop biologi-", + "cal systems instantly. For example, one could immerse mice in liquid nitrogen, look at", + "their electron transport systems, and find out how these have been affected by smoke. This", + "approach would not be very comparable to the human situation.", + "GRIFFITH: To what extent might we expect animal response to smoke inhalation to correspond", + "to the response of humans to smoke?", + "WHITE: It is like painting condensate on mouse skins. You have to put smoke into the ani-", + "mal to develop example, lung tumors in something less than 50 years.", + "GRIFFITH: Can one demonstrate cardiovascular responses? Dr. Aleem, for example, may know", + "that Dr. Gordon has been able to obtain differences in the cardiovascular response of rats", + "receiving smoke from the alkaloid series of samples. Nicotine toxicity is a factor which", + "must be considered in inhalation research.", + "ALEEM: We have had some difficulties in our smoke inhalation studies. We want to isolate", + "the mitochondria as quickly as possible after the animal has been given a brief exposure", + "to smoke. However, we tound that the anesthetic which we were using was also inhibitory to", + "the ATP electron transport system, and that the mitochondria were no longer making ATP. So", + "now we have to look for a new anesthetic. We have looked at electron transport in heart", + "mitochondria and also in yeast mitochondria. Smoke does inhibit the respiratory process,", + "and it does have an effect on the ATP system. We found that nicotine is not inhibitory in", + "the respiratory processes. It may have some effect on the cilia movement in Dr. Donnelly'e", + "system, but nicotine may in some cases enhance respiration. This might be due to an un-", + "coupling effect, that ATP is hydrolyzed to enhance respiration, but we haven't proven this.", + "DONNELLY: Nicotine inhibits ciliary movement at high concentrations, but at the levels in", + "which it is found in commercial cigarettes, nicotine is not ciliostatic.", + "ALEEM: I want to emphasize that our experiments are very preliminary.", + "DONNELLY : I think one thing we can say is that we are ready for samples.", + "REES: I think Dr. Griffith is more concerned about the acute effects of inhalation than", + "might be warranted. It seems to me that the acute effects must be fairly pleasant and not", + "very harmful, and the chronic effects of smoking are more important.", + "GRIFFITH: I have not stated a position concerning the merits of acute vs chronic exposure.", + "Since you have raised the question though, I would like to state the belief that one should", + "start inhalation research using acute exposures and then grade into chronic exposure. 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Inhalation research is important in that", + "one is working with an intact animal, and if one uses smoke composition equivalent to human", + "exposure, one has the best chance of correlating animal response with human response.", + "Source: tps://www.industrydocuments.ucsf.edu/docs/jgchGO", + "80", + "RC- 6029135", + "0090647" + ] + }, + { + "bbox": [ + [ + 0.473775, + 0.455153, + 0.0498, + 0.008762 + ], + [ + 0.360612, + 0.479687, + 0.274639, + 0.00891 + ], + [ + 0.716121, + 0.763605, + 0.120552, + 0.015143 + ], + [ + 0.734905, + 0.970195, + 0.02311, + 0.009549 + ], + [ + 0.79241, + 0.969808, + 0.080098, + 0.009343 + ], + [ + 0.229112, + 0.981099, + 0.540797, + 0.016318 + ] + ], + "poly": [ + [ + { + "X": 0.473775, + "Y": 0.455153 + }, + { + "X": 0.523576, + "Y": 0.455153 + }, + { + "X": 0.523576, + "Y": 0.463915 + }, + { + "X": 0.473775, + "Y": 0.463915 + } + ], + [ + { + "X": 0.360612, + "Y": 0.479687 + }, + { + "X": 0.635252, + "Y": 0.479687 + }, + { + "X": 0.635252, + "Y": 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Smoking Behavior", + "Objective: To quantitate human smoking behavior and de-", + "termine the factors which affect this response.", + "B. Short-tern Effects", + "Objective: To measure physiological, biochemical, pulmo-", + "nary, etc., responses to smoke from tobacco", + "product samples.", + "C. Long-term Effects", + "Objective: To correlate long-term responses to smoke", + "(measured by epidemiological techniques) with", + "smoke behavior and short-term responses", + "COMMENTS", + "Simple observation of smokers establishes the fact that individuals differ", + "widely in their smoking behavior and characteristics--sor people inhale while", + "some do not, some people take frequent puffs while others puff infrequently,", + "etc. These variations in smoking behavior result in variations in the compo-", + "sition of the smoke consumed by different individuals and also have marked", + "effects on their total daily smoke consumption. For meaningful research in-", + "volving human response, it is considered essential to quantitatively define", + "the smoking characteristics and behavior of individuals and populations and", + "to have an understanding of the factors which affect smoking behavior. The", + "basic techniques necessary for this characterization are available within in-", + "dustry but must be perfected further. Meaningful research in this area cannot", + "be conducted until the procedures are made available to the scientific", + "community.", + "When it is possible to quantitatively characterize smoking behavior of", + "individuals, it should then be possible to give these individuals sample cig-", + "arettes with known variabilities, and look for alterations in such short-term", + "responses as pulmonary functions, blood chemistry, metabolism, and to corre-", + "late the results obtained with those from the nonhuman biological systems.", + "By concentrating on subjects deliberately chosen for wide variations in smok-", + "ing behavior, the number of experimental subjects can be minimized if one has", + "knowledge of the proportion of such smokers in the total population.", + "Longer term effects of the response to smoke will require an epidemio-", + "logical approach in which emphasis is given to the smoking behavior of indi-", + "viduals and populations and multi-variate analysis. A thorough study, in", + "quantitative terms, of the smoking behavior of subjects who have medical", + "problems should help establish the nature and magnitude of the relationship", + "between smoking and any disease. Such studies should provide information", + "which will be valuable in establishing the basis for fundamental prospective", + "studies, in correlating nonhuman and human responses, and in determining and", + "predicting the factors important in the development of reduced detrimental", + "biological activity.", + "RO - 6029137", + "82", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgchodd", + "0090649" + ] + }, + { + "bbox": [ + [ + 0.397445, + 0.118006, + 0.166588, + 0.007839 + ], + [ + 0.265488, + 0.14152, + 0.422334, + 0.012638 + ], + [ + 0.151322, + 0.167145, + 0.676188, + 0.012756 + ], + [ + 0.11391, + 0.180191, + 0.740333, + 0.013712 + ], + [ + 0.114682, + 0.193595, + 0.761977, + 0.01266 + ], + [ + 0.114808, + 0.205351, + 0.746254, + 0.012797 + ], + [ + 0.15217, + 0.231106, + 0.702846, + 0.011277 + ], + [ + 0.114026, + 0.243687, + 0.717467, + 0.012879 + ], + [ + 0.114221, + 0.255866, + 0.741467, + 0.012721 + ], + [ + 0.114454, + 0.268967, + 0.748878, + 0.013858 + ], + [ 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W. Chick, M.D., Department of Community Medicine", + "The session this morning is concerned with the human response to tobacco products.", + "Previous sections in this conference have pointed out that tobaccos do differ. Strains of", + "tobacco vary from each other in their chemical composition and in their final smoke products.", + "Even the same strain may vary from crop year to crop year depending upon growth conditions.", + "It is evident that individuals may vary in their smoking habits. With two such major", + "variables, individual smoking habits, and tobacco variances, the study for associations", + "between smoking and health becomes quite complicated. This is the challenge to this morn-", + "ing's session--to point out what is known from scientific observation, removed of emotional", + "overlays, and to discuss possible avenues for future study.", + "Dr. Griffith will begin the session with an overview of the associations between smok-", + "ing and health, highlighting imperative as yet unanswered questions concerning these", + "associations.", + "HUMAN RESPONSE TO SMOKE", + "R. B. Griffith, Ph.D., Director, Tobacco and Health Research Institute", + "ABSTRACT", + "Procedures for determining in quantitative terms the smoking behavior of an individual or", + "population are described. Alterations in smoking parameters within the range of human ex-", + "periences are shown to have effects on smoke yields and composition which are large enough", + "to have biological significance. These procedures and the procedure for determining total", + "daily smoke intake described by Wakeham (17) are considered in relationship to epidemiolog-", + "ical studies and to the potential for determining human response to smoke under experimental", + "conditions. The capability of characterizing human smoking behavior, combined with the", + "ability to manipulate and control smoke composition provides the experimental conditions", + "necessary for a determination of the nature and magnitude of the tobacco and health question.", + "INTRODUCTION", + "One of the simplest and yet complete presentations concerning tobacco and health, en-", + "titled \"Chart Book on Smoking, Tobacco and Health\" (1), was published by the Public Health", + "Service in 1969. The data given summarizes rather concisely the magnitude and ramifications", + "of the tobacco industry. The epidemiological evidence for the apparent relationship between", + "smoking and health must certainly be viewed seriously when one considers that about 42% of", + "the adult population or some 51,300,000 people are presumably at risk because they smoke.", + "Few will question the fact that almost all of the epidemiological studies indicate a", + "relationship between smoking and health, but rather sharp differences of opinion exist con-", + "cerning the validity and interpretation of the epidemiological evidence.", + "Emotion", + "is", + "often", + "more evident than objective judgments. This is not too hard to understand when one con-", + "siders the magnitude of the financial interests involved and the daily confrontations with", + "tragedy encountered by the physician in his contact with patients. It is sometimes diffi-", + "cult for even the trained scientist to remember that he has been taught to avoid wishful", + "thinking. The normal tendency is to jump to conclusions on the basis of inadequate informa-", + "tion.", + "It is not the purpose of this presentation to evaluate the evidence or present the ar-", + "guments pro or con for a relationship between smoking and health. Such evaluations can be", + "found in Smoking and Health, Report of the Advisory Committee to the Surgeon General of the", + "Public Health Service (2), or the more recently issued Report of the Standing Committee on", + "83", + "RC-- 6029138", + "LG", + "0090650", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.159211, + 0.124846, + 0.736405, + 0.025216 + ], + [ + 0.160403, + 0.135356, + 0.72078, + 0.026528 + ], + [ + 0.160958, + 0.150226, + 0.7208, + 0.024526 + ], + [ + 0.16089, + 0.162701, + 0.720406, + 0.026325 + ], + [ + 0.160362, + 0.174091, + 0.728823, + 0.02563 + ], + [ + 0.161348, + 0.188774, + 0.656666, + 0.022334 + ], + [ + 0.340301, + 0.214492, + 0.376516, + 0.016401 + ], + [ + 0.201605, + 0.237386, + 0.702125, + 0.026431 + ], + [ + 0.160534, + 0.25001, + 0.71474, + 0.025434 + ], + [ + 0.160803, + 0.262779, + 0.743359, + 0.026034 + ], + [ + 0.159705, + 0.275724, + 0.74497, + 0.023177 + ], + [ + 0.161643, + 0.287688, + 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Instead, let us focus our", + "attention on some relatively new and thus far little used techniques which may be of help", + "in obtaining a more precise understanding of the nature and magnitude of the relationship", + "between smoking and health. We will then consider how these might be used in the research", + "program of tobacco and health and some of the implications of the data presented.", + "THE CHARACTERIZATION OF HUMAN SMOKING BEHAVIOR", + "The need to know as much as possible about the smoking behavior of individuals and pop-", + "ulations has been recognized and a number of studies have been reported (4-15). Average", + "values given in some of these studies are presented in Table I. Data obtained in laboratory", + "measurements usually represent small populations and the measurements are made under labora-", + "tory conditions--ar artificial environment for most smokers. In view of the fact that the", + "studies reported were made in Germany, France, and the United States, the agreement in most", + "values is remarkable. Lack of agreement for puff interval values is probably the result of", + "the experimental situation, and this is the reason also for the difference between labora-", + "tory measurements and field observations for this parameter.", + "Field observations are made on smokers who do not know that they are being observed,", + "and one would expect that these results would be more accurate for some values than would", + "laboratory measurements. Puff interval can be rather readily determined by observation, but", + "puff duration would be much more difficult. Puff volume cannot be determined directly by", + "field observations and must be calculated by making assumptions. This probably accounts", + "for the low puff volume values in two reports (11 & 12). In a later study made in England", + "by Adams (14) puff volumes were found to be in line with the others given for laboratory", + "measurements.", + "In all studies the very large range in values for individuals becomes apparent. Some", + "concept of the range in some parameters for individuals is given in Table II. Values for", + "the peak height and the latter two sets of values are based on pressure-time curves obtained", + "during a puff. Techniques for measuring the smoking characteristics of individuals have been", + "constantly improved within the industry and have reached a more advanced stage than is indi-", + "cated by the references cited. It is important to note that the differences between indi-", + "viduals in the way they smoke is large for all parameters measured and one can question, \"Is", + "the difference in the way people smoke important in connection with the tobacco and health", + "question?\"", + "TABLE I", + "REPORTED VALUES FOR SMOKING PARAMETERS", + "SMOKING PARAMETERS AND", + "Smoking", + "Butt", + "Number", + "Puff", + "Puff", + "Puff", + "SMOKE COMPOSITION", + "Time", + "Length", + "of", + "Interval", + "Duration", + "Volume", + "(sec.)", + "(mm)", + "Puffs", + "(sec.)", + "(sec.)", + "(cc)", + "The variations in the smok-", + "Laboratory Measurements", + "ing parameters of individuals", + "Pfyl (4)", + "42.5", + "was considered in establishing", + "-", + "the standard conditions for the", + "Schur and Rickards (5)", + "1.58", + "42", + "routine determination of tar", + "Cuzin (6)", + "19.0", + "1.7", + "34.4", + "and nicotine (18) and an attempt", + "1.91", + "was made to adopt the average", + "Keith (7)", + "31.4", + "11.4", + "29.6", + "43.7", + "values for the population. The", + "Griffith (15)", + "383", + "36.5", + "10.4", + "46.8", + "1.96", + "-", + "values of one puff per minute", + "Field Observations", + "of 2 seconds duration are prob-", + "ably very close to the popula-", + "Hilding (8)", + "495", + "36.2", + "9.9", + "-", + "2.0", + "-", + "tion average. The puff volume", + "Waltz (9)", + "528", + "24", + "9.2", + "75.9", + "-", + "38.5", + "of 35 ml adopted as a standard", + "is probably low, and the true", + "TMSC-Todd (10)", + "635", + "24", + "13.2", + "-", + "-", + "-", + "value is probably nearer 42 ml", + "Cigarette Components Ltd. (11)", + "per puff. The effect on tar", + "-", + "17.4", + "-", + "64", + "-", + "25", + "and nicotine yields of various", + "TMSC-Bentley & Burgan (12)", + "smoking parameters have been", + "-", + "-", + "-", + "57", + "-", + "25", + "studied by some (19). The", + "Shepherd (13)", + "-", + "-", + "-", + "67", + "-", + "-", + "84", + "RC- 6029139", + "LG", + "0090651", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.501161, + 0.113889, + 0.317198, + 0.015786 + ], + [ + 0.276055, + 0.129097, + 0.046321, + 0.007363 + ], + [ + 0.501339, + 0.124804, + 0.349284, + 0.016071 + ], + [ + 0.188218, + 0.146191, + 0.208922, + 0.010762 + ], + [ + 0.503095, + 0.137808, + 0.340674, + 0.017324 + ], + [ + 0.194824, + 0.15517, + 0.201613, + 0.010834 + ], + [ + 0.502974, + 0.151037, + 0.348857, + 0.017805 + ], + [ + 0.502417, + 0.163288, + 0.333017, + 0.015899 + ], + [ + 0.275807, + 0.182594, + 0.041484, + 0.00741 + ], + [ + 0.356895, + 0.183135, + 0.040411, + 0.00748 + ], + [ + 0.430588, + 0.182117, + 0.041993, + 0.007653 + ], + [ + 0.502314, + 0.17635, + 0.332958, + 0.016468 + ], + [ + 0.281596, + 0.191243, + 0.02993, + 0.007225 + ], + [ + 0.362437, + 0.191385, + 0.029111, + 0.006546 + ], + [ + 0.43619, + 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The", + "Minimum", + "Average", + "Maximum", + "effects are large and it is evident that", + "Value", + "Value", + "Value", + "Total Smoking Time (min.)", + "2.9", + "6.6", + "10.3", + "alterations in these parameters, all within", + "the range of human experience, do affect", + "Length Smoked (mm)", + "22", + "49", + "60", + "smoke yields.", + "Number of Puffs", + "4", + "9.8", + "23", + "Interval Between Puffs (min.)", + "The effect of alterations in smoking", + "parameters on smoke composition are virtu-", + "Average/cig.", + "0.20", + "0.74", + "2.06", + "ally nonexistent in the literature, but one", + "Puff Duration (seconds)", + "0.91", + "1.96", + "3.41", + "can logically expect that such information", + "Maximum Peak Ht.", + "has been obtained. One can examine this", + "8.0", + "30", + "54.4", + "point by calculating the pgs of phenol or", + ": of Duration to Maximum", + "15.9", + "34", + "65.4", + "cyanide per mg of nicotine. Such data are", + "% of Maximum at 3/6 Duration", + "16.8", + "44", + "83.1", + "presented in Table IV. Once again it is", + "obvious that smoking parameters within the", + "range of human experience have significant", + "effects on smoke composition. The indi-", + "vidual who would take a 15 ml puff of 3 seconds duration would consume about twice as much", + "phenol and 1/2 as much cyanide per milligram of nicotine consumed as the individual who", + "took a 35 ml puff of 1 second duration. These differences are large enough to have biolog-", + "ical significance, but here again data are virtually nonexistent in the literature. There", + "is the probability that a report will be issued in the not too distant future which will", + "show an effect of puff volume variations on the specific activity of tar in mouse skin", + "tests--a low volume puff is more active than a high volume puff.", + "TABLE III", + "Further work (by Heideman, 16) deter-", + "\"Free Smoking\" Regime", + "mined that puff frequency, puff profile,", + "Puff", + "draft around the cigarette, moisture content", + "Duration", + "Puff Volume (ml)", + "of the tobacco, and in fact every variable", + "(sec.)", + "15", + "25", + "35", + "Avg.", + "examined has measurable effects on smoke", + "Mg. Nicotine", + "yields and smoke composition. The need to", + "1", + "2.09", + "2.93", + "3.29", + "2.77", + "have a high degree of control on smoking", + "2", + "2.56", + "3.36", + "3.83", + "3.25", + "3", + "2.64", + "3.82", + "4.18", + "3.55", + "TABLE IV", + "Average", + "2.43", + "3.37", + "3.77", + "RATIO OF TOTAL PHENOLS AND TOTAL CYANIDE TO NICOTINE", + "pg. Total Phenole", + "1", + "144", + "198", + "217", + "186", + "\"Free\" Smoking Regime", + "2", + "218", + "288", + "250", + "252", + "Puff", + "3", + "276", + "330", + "398", + "335", + "Duration", + "Puff Volume (ml)", + "Average", + "213", + "272", + "288", + "(sec.)", + "15", + "25", + "35", + "Avg.", + "ug- Total Cyanides", + "ug. Total Phenols/mg. 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if one is to obtain meaningful smoke data or conduct meaningful inhala-", + "tion studies.", + "QUANTITY OF SMOKE CONSUMED", + "In most epidemiological studies, smoke exposure is related to the number of cigarettes", + "consumed per day. The range in values is recognized as large and one can plot a distribution", + "curve (1). By classifying people on the basis of the number of cigarettes smoked per day,", + "one can calculate the effect of this variable on the relationship with a particular disease.", + "The crudeness of this approach should be obvious in view of the variability of individuals", + "and the effects which such variability can have on smoke composition.", + "During his recent visit to the University of Kentucky, Wakeham (17) briefly described", + "a", + "technique for determing total daily exposure of an individual and presented a slide showing", + "daily tar intake in a rather large population. The procedure was based on nicotine determi-", + "nations in cigarette butts and on calculation of the tar which passed through the filter,", + "using knowledge of the filtration properties of the cigarette butt. For this technique to", + "be valid one would have to establish that the known variables in the smoking parameters of", + "human beings have no effect on the results or that one could correct for any such discrep-", + "ancies. It is reasonable to assume that this procedure is under control, and, if so, this", + "is one of the most powerful and potentially useful techniques developed to date.", + "In his talk he gave an example involving the use of this technique. Sixty smokers were", + "given a cigarette with a normal tar delivery of about 20 mg/cigarette, and their total daily", + "intake was determined. One half of the group was then given a cigarette with a tar delivery", + "of", + "15 mg/cigarette, and the other half was given a 25 mg/cigarette. The change in tar de-", + "livery on an individual cigarette basis did not alter total daily consumption of particulate", + "matter or nicotine. However, because an acetate filter does not remove some smoke components", + "such as carbon monoxide or nitrogen oxides selectively, those smokers given the 15 mg/ciga-", + "rette received significantly more of these gas components in a day than did those smoking the", + "25 mg cigarette. The implications of this finding are a bit frightening. The big emphasis", + "on reducing tar and nicotine content of cigarettes must assume that the smoker will not in-", + "crease the number of cigarettes per day for a reduction in the tar delivery of a cigarette", + "to have any effect on any health risk which might be associated with tar. If total daily tar", + "intake remains the same and one receivés a much larger quantity of the gas phase components.", + "one has not reduced the risk from tar and may have increased significantly the risk from any", + "health problem which may be associated with gas phase components. In their zeal to do good", + "have the proponents of reduced tar and nicotine cigarettes jumped to a conclusion on the", + "basis of inadequate information, and created an emphysema-chronic bronchitis problem of in-", + "creasing concern? Is the present concern with air pollution another factor which must be", + "considered? Has the lung cancer problem been helped?", + "SMOKING BEHAVIOR AND EPIDEMIOLOGY", + "Thus far we have described techniques for characterizing the smoking behavior of an", + "individual or population in quantitative terms, have given data that show significant altera-", + "tions in smoke composition with changing smoke parameters within the range of human experi-", + "ence, and discussed the quantitative procedure for determining daily smoke intake described", + "by", + "Dr. Wakeham (17). Let us now consider the possible use of these techniques in epidemio-", + "logical studies related to the smoking and health question. This part of the presentation", + "will raise, deliberately, more questions than it will provide answers for, and it is hoped", + "that the experts in community medicine will give appropriate comments to clarify the points", + "made.", + "Going back to data from the first reference (1) we note that the total adult popula-", + "tion of the United States in 1965 can be divided into the following groups:", + "RC- 6029141", + "86", + "LG 0090653", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.369814, + 0.121543, + 0.025496, + 0.007562 + ], + [ + 0.526227, + 0.120787, + 0.042445, + 0.007881 + ], + [ + 0.693157, + 0.120396, + 0.042866, + 0.008251 + ], + [ + 0.252658, + 0.146474, + 0.043266, + 0.009567 + ], + [ + 0.337974, + 0.146306, + 0.082848, + 0.01096 + ], + [ + 0.502455, + 0.145609, + 0.084389, + 0.011226 + ], + [ + 0.663658, + 0.145562, + 0.088607, + 0.010947 + ], + [ + 0.187502, + 0.171635, + 0.100311, + 0.009629 + ], + [ + 0.337275, 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"Women", + "Total", + "Total", + "57,585,000", + "64,501,000", + "122,086,000", + "Never Smoked", + "17,274,500 (30%)", + "37,410,580 (58%)", + "54,685,000", + "Former Smokers", + "10,941,150 (19%)", + "5,160,080 (8%)", + "16,101,230", + "Present Smokers", + "29,369,350 (51%)", + "21,930,340 (34%)", + "51,299,690", + "Except for the chance distribution of a single chromosome and a modification of the numbers", + "involved because of war and other external forces, the groups are the result of the freedom", + "of", + "choice--to smoke, to quit smoking, or to never smoke. Chance distribution results in es-", + "sentially equal numbers--freedom of choice results in as much as a seven-fold difference in", + "the size of the sub-groupings involved. Are these groups the same in all characteristics", + "except those shown? What are the conditions which must be established in order to make valid", + "comparisons between the groups? Is the susceptibility factor for lung cancer, emphysema or", + "any other disease present in the same proportion in each group? Can this be established?", + "Why did the former smokers quit smoking? Is the former smoker more concerned about health", + "naturally and therefore more likely to have a different life pattern than the smoker? Have", + "the groups reacted the same to the shift from an active rural environment to the inactive,", + "pressure prone environment of present society? Must one consider factors other than smoking", + "in evaluating the relationship of smoking to lung cancer or any other disease?", + "In the epidemiological studies thus far made, clarification has been sought in estab-", + "lishing the relationship between smoking and disease by considering the number of cigarettes", + "smoked, place of residence, number of years smoked, age when smoking started, etc. An evalua-", + "tion of the variability of individuals suggests that the correlation between number of ciga-", + "rettes smoked and total daily tar intake may be unacceptable, and an answer to some of the", + "above questions may suggest that important factors may not have been considered. Will the", + "more quantitative techniques permit more meaningful groupings and lead to a better under-", + "standing of the nature and magnitude of the relationship between smoking and health? Can the", + "techniques be used in determining by epidemiological means the long-term effects of major", + "product modifications? Do other techniques need to be developed for more meaningful studies?", + "Will improved groupings permit the use of smaller populations?", + "The question of equal susceptibility of the individuals in any groupings will probably", + "be one of the more difficult things to establish. Can the quantitative characterization of", + "smoking behavior of individuals with medical problems, with more attention given to a multi-", + "factorial analysis approach, lead to the definition of the factors which need to be con-", + "sidered in prospective epidemiological studies?", + "SMOKING BEHAVIOR AND THE EXPERIMENTAL SITUATION", + "The establishment of quantitative terms for smoking behavior of individuals permits", + "us to consider further refinements in the experimental approaches which can be made in to-", + "bacco and health research. Smoking machine technology has progressed to the point where the", + "smoking behavior of any individual can be duplicated exactly and the actual quantity of many", + "smoke components taken in by the individual can be determined. Furthermore, it will soon be", + "possible to present an experimental animal or a sub-cellular bioassay system to smoke of the", + "exact composition of that taken in by any individual.", + "The potential for product modifications by alterations in the tobaccos used and/or the", + "way in which the product is manufactured has been pointed out by others in this conference.", + "These developments lead to a consideration of the possibilities of determining human response", + "to smoke in an experimental situation.", + "Dr. Benner and Mr. Atkinson described the manufacture and characteristics of the sam-", + "ples now available in the University sample series. 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It is now possible to give these samples to an individual or", + "individuals and determine if they smoke the samples in the same or different ways and the", + "total quantity of smoke taken in by the individual in any given time. This leads to tne", + "question of what, if any, measurements could be made to determine the response of the intact", + "human system to the smoke from this or any other sample system. Can one measure alterations", + "in:", + "1. pulmonary functions", + "2. physiological responses", + "3. cardiovascular responses", + "4. psychological responses", + "Can one correlate the human response to the response of a nonhuman system? If the answer is", + "yes, we are more than halfway to our goal for we can then proceed with confidence that we can", + "develop any product modification required to minimize or eliminate any undesirable component", + "identified.", + "REFERENCES", + "1. 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To date an etiologic basis has not been dem-", + "onstrated. There was general acceptance of the observation that the more a person smokes,", + "and the longer the period he smokes, the greater his risk of developing bronchitis, emphy-", + "sema, lung cancer, or cardiovascular problems. It was further recognized that the current", + "health problems in smoking adults may be associated with their smoking experiences 10 to 15", + "years previously. There was some discussion of the differences in rates of these associated", + "health problems in different countries of the world and that they may be related to the use", + "of different tobacco products. There was also a discussion on the possible interrelation-", + "ships between smoking and moving into an area with significant air pollution. Certainly", + "the possibility of viral etiology or mediation cannot be excluded at this time. Similarly", + "the role of genetic determination is unsolved; one can theorize a predisposition which is", + "augmented by smoking and reversed by quitting smoking.", + "A great deal of discussion centered around the way in which a person smokes, the fre-", + "quency with which he smokes, the duration over which he smokes, and the variation in the", + "composition of what he smokes. Comments from the audience broadened the perspective.", + "There was general agreement that the measurement of an individual's smoking habits by", + "\"packs per day, etc.' is relatively meaningless. Individuals have notable differences in", + "their smoking patterns and habits. There was agreement that a more proper definition of", + "smoking patterns of individuals can be obtained from their puff profiles. How deeply do", + "they draw on the cigarette? How long do they draw? To what degree do they inhale back into", + "the lungs? Does an individual's smoking habits change over a period of time, as may be in-", + "dicated by the observation that older individuals may tend to switch to mentholated ciga-", + "rettes?", + "In considering data presented previously in the program, the panel discussed at length", + "the implications of variations. and composition of cigarettes. Are the associated health", + "problems related to specific factors in tobacco or tobacco smoke? Retrospective search for", + "such a factor or factors would be complicated by the fact that different tobacco strains", + "have been used over the years in the manufacture of tobacco products; the chemical composi-", + "tion of the same strain of tobacco may vary year to year, that the proportion of different", + "tobaccos used in cigarettes may well have changed over the years; and finally that a variety", + "of different additives have been used in the manufacture of tobacco products. If a factor", + "or factors could be identified with individually associated health problems, or collectively,", + "it would be possible for the tobacco industry to rid their product of that factor or factors.", + "For example, it has been shown that the addition of filters to cigarettes will remove 90%", + "of the phenols, and that the gas phase can be significantly modified by the use of charcoal", + "filters.", + "The panel recognized that emotionalism has hampered the collection of objective data", + "on the association between certain health conditions and smoking. Further, most of the", + "studies which have been done have been retrospective and subject to the limitations of any", + "retrospective technique, namely, the possible bias of the investigator, the limitations of", + "patient recall, the limitations of existing records on individuals, etc. A number of", + "studies are based on death certificate diagnosis. The limitations here are obvious when", + "one considers that death certificates may be signed by nonmedical personnel in some states,", + "by a physician (medical examiner or other) who has not previously seen the patient, and", + "frequently is not verified by autopsy studies. Clinical studies do not always offer stan-", + "dardized criteria for diagnosis of the associated health problems. All too often the smoker", + "who does not develop one of the associated health problems is not studied as intensively as", + "the smoker who does develop these associated problems. Cohort studies have not been fully", + "utilized.", + "Experimental laboratory studies are open to question in many instances. There is the", + "age-old question of how similar is the experimental rat, rabbit, etc., to the human and to", + "89", + "RC- 6029144", + "LG", + "0090656", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.099152, + 0.124746, + 0.742616, + 0.011811 + ], + [ + 0.1003, + 0.136622, + 0.719253, + 0.011951 + ], + [ + 0.099201, + 0.150027, + 0.72051, + 0.011882 + ], + [ + 0.100769, + 0.162447, + 0.703982, + 0.01173 + ], + [ + 0.811754, + 0.163438, + 0.014858, + 0.00915 + ], + [ + 0.101727, + 0.174404, + 0.725645, + 0.013764 + ], + [ + 0.101038, + 0.188147, + 0.630271, + 0.012695 + ], + [ + 0.140713, + 0.212696, + 0.671096, + 0.012435 + ], + [ + 0.100798, + 0.225953, + 0.719025, + 0.011469 + ], + [ + 0.100362, + 0.23812, + 0.719676, + 0.011126 + ], + [ + 0.101016, + 0.24996, + 0.726835, + 0.012244 + ], + [ + 0.100862, + 0.262594, + 0.712224, + 0.011762 + ], + [ + 0.101003, + 0.275115, + 0.74095, + 0.012094 + ], + [ + 0.100678, + 0.28756, + 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Is the mechanism of pro-", + "duction of tumors on the mouse back by painting with tars or concentrates similar to that", + "which happens in the bronchial mucosa of the human smoker? There was unanimous agreement", + "that the development of an appropriate animal model which could be used under a variety", + "of", + "conditions simulating different smoking patterns and utilizing tobacco products of varying", + "composition would greatly potentiate the study of smoking and health problems.", + "A human prospective study would be highly desirable. Such a study should encompass", + "paired individuals whose only variable, insofar as possible, would be whether they smoked", + "or did not smoke: There should be regular appraisal of their smoking profiles and habits", + "as well as analysis of the tobacco product smoked. Standardized criteria for the recogni-", + "tion of possible associated health conditions should be maintained. Such a study over a", + "long period of years could provide needed insight. The use of standard reference cigarettes", + "and experimental cigarettes which contain graded amounts of nicotine, phenols, tars, etc.,", + "from tobaccos that are genetically controlled or from filters or other product modifications", + "would provide more precise identification of an individual factor or multiple factors in to-", + "bacco potentially related to health problems.", + "RC- 6029145", + "90", + "LG 0090657", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.464158, + 0.438962, + 0.058271, + 0.008986 + ], + [ + 0.374435, + 0.464209, + 0.245159, + 0.008692 + ], + [ + 0.754335, + 0.827816, + 0.119554, + 0.012177 + ], + [ + 0.750519, + 0.967475, + 0.024071, + 0.009597 + ], + [ + 0.808332, + 0.965972, + 0.081982, + 0.010509 + ], + [ + 0.226084, + 0.981018, + 0.550723, + 0.016399 + ] + ], + "poly": [ + [ + { + "X": 0.464158, + "Y": 0.438962 + }, + { + "X": 0.522429, + "Y": 0.438962 + }, + { + "X": 0.522429, + "Y": 0.447948 + }, + 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Biological", + "B. Chemical", + "C. Physical", + "COMMENTS", + "In the previous sections of the program products are manufactured,", + "characterized, and specific biological responses to these products are de-", + "termined. Presumably, product modifications will alter biological response", + "and one should be able to develop ways to minimize any adverse biological", + "responses noted.", + "Alternatively, one can consider, for example, the basic nature of a", + "particular disease in relationship to the nature of tobacco products, and", + "develop information of importance. Thus, studies of mouse skin activity", + "have indicated that smoke condensate exhibits weak carcinogenic properties,", + "and chemical studies have shown that smoke condensate contains known chem-", + "ical carcinogens. Fundamental studies involving the metabolism, rates of", + "absorption, and relationship to viruses of chemical carcinogens and smoke", + "condensate could be just as important as the response-oriented program.", + "Both types of studies are required for a balanced program of research.", + "RC- 6029147", + "92", + "LG 0090659", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.4308, + 0.127451, + 0.163888, + 0.009553 + ], + [ + 0.316783, + 0.152307, + 0.39157, + 0.013653 + ], + [ + 0.185477, + 0.177324, + 0.668455, + 0.015159 + ], + [ + 0.146934, + 0.189913, + 0.715533, + 0.015973 + ], + [ + 0.147768, + 0.201413, + 0.715168, + 0.017087 + ], + [ + 0.147956, + 0.214789, + 0.41236, + 0.015252 + ], + [ + 0.185433, + 0.238835, + 0.701993, + 0.016788 + ], + [ + 0.149337, + 0.251155, + 0.706776, + 0.016771 + ], + [ + 0.149206, + 0.263828, + 0.705797, + 0.017725 + ], + [ + 0.148763, + 0.27628, + 0.692334, + 0.01578 + ], + [ + 0.148385, + 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[ + "INTRODUCTORY REMARKS", + "J. W. Flesher, Ph.D., Department of Pharmacology", + "The traditional role of academic research is to investigate fundamental questions.", + "These studies have yielded basic information, sometimes of great theoretical importance,", + "but are generally regarded by the individual scientist as having little practical value,", + "since usually no pretence of usefulness is implied.", + "Recently, universities have been asked to undertake numerous studies of a more practi-", + "cal nature, perhaps due to the urgency of many problems. As more and more work of this", + "type is undertaken, it seems clear that less and less basic research will be conducted.", + "Disinterested science is increasingly difficult to justify despite the fact that such", + "studies are the basis of future investigations in applied research. It is to be hoped that", + "fundamental studies, the primary aim of which is to understand natural phenomena, will be", + "an expanding part of our program in spite of the present climate of reseach which demands", + "immediate relevance.", + "In our discussions so far we have seen a great deal of variation and variability, and", + "we really don't know exactly where to begin our attack on the problem. I'm certain that", + "fundamental studies will provide the necessary information which will lay the foundation", + "for some meaningful applied research within this program.", + "MECHANISM OF INTESTINAL ABSORPTION OF BENZO[a]PYRENE", + "P. Mandelstam, M.D., Ph.D., and E. D. Rees, M.D., Department of Medicine", + "ABSTRACT", + "Little is known about the mechanism of absorption of polynuclear aromatic hydrocarbon car-", + "cinogens through the gastrointestinal tract. We have studied the absorption of several of", + "these compounds, benzo[a]pyrene (BaP) in most detail, in the rat by in vitro and in vivo", + "techniques. With the in vitro everted sac technique, the uptake of BaP by sac tissue is", + "not appreciably influenced by inhibition of respiration and/or glycolysis. Accumulation", + "in intestinal sac tissue increases exponentially with increased incubation medium concentra-", + "tion. We postulate that initially unilayer (Langmuir) adsorption occurs on the mucosal", + "surface of the intestine and then multilayer (Polanyi) adsorption follows. The passage of", + "polynuclear aromatic hydrocarbons from sac tissue into the fluid within the sac occurs in", + "direct proportion to sac tissue concentration, a finding consistent with passive diffusion.", + "In vivo experiments show that an exponential relationship exists between the intragastric", + "dose of compound administered in sesame oil and accumulation of compound in the adipose", + "tissue and manmary glands. Adsorption of BaP on string was found to increase logarithmi-", + "cally with an arithmetic increase in medium concentration.", + "INTRODUCTION", + "Although it is well established that the polycyclic aromatic hydrocarbon carcinogens", + "are absorbed via the gastrointestinal tract, little is known of the mechanism of absorption", + "of these highly hydrophobic and lipophilic compounds. The study reported here concerns", + "primarily the mechanism of absorption of BaP, a potent carcinogen which is found in tobacco", + "smoke.", + "RESEARCH METHODOLOGY", + "Both in vitro and in vivo studies were performed.", + "The in vitro studies were carried out for the most part with a modified Wilson-Wiseman", + "everted small intestinal sac technique. In this preparation (Fig. 1), the gut is everted,", + "so that the rapidly metabolizing mucosa is exposed directly to the incubation medium through", + "93", + "RC- 6029148", + ":-", + "LG", + "0090660", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.467691, + 0.129101, + 0.361991, + 0.009767 + ], + [ + 0.467722, + 0.140506, + 0.397412, + 0.016181 + ], + [ + 0.469013, + 0.153624, + 0.38908, + 0.012052 + ], + [ + 0.468273, + 0.166934, + 0.360979, + 0.009731 + ], + [ + 0.468155, + 0.177845, + 0.395346, + 0.014561 + ], + [ + 0.4679, + 0.190613, + 0.368942, + 0.011653 + ], + [ + 0.468372, + 0.20341, + 0.400643, + 0.011835 + ], + [ + 0.468824, + 0.214601, + 0.375682, + 0.012041 + ], + [ + 0.468737, + 0.227939, + 0.396216, + 0.010835 + ], + [ + 0.469043, + 0.239594, + 0.381779, + 0.01191 + ], + [ + 0.468922, + 0.251588, + 0.373766, + 0.010442 + ], + [ + 0.468396, + 0.263852, + 0.402637, + 0.011226 + ], + [ + 0.468585, + 0.275943, + 0.395819, 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Tissue culture", + "medium (Eagles Minimal Essential Medium) containing", + "20% hypo gamma calf served as the basic incubation", + "medium. The electrolyte concentration of this", + "medium corresponded to that of extracellular fluid,", + "but, in addition, vitamins and amino acids were", + "present. The BaP was added in the following manner.", + "One part of Tween 20, one part lecithin, and ten", + "parts sesame oil containing the BaP were emulsified", + "at 40,000 rpm with a VirTis blender. One part of", + "this emulsion was utilized as incubation medium.", + "The standard incubation was carried out in a Dubnoff", + "metabolic shaker, rate 100/minute, for 1 hour at 37", + "degrees C, in a 95% oxygen-5% carbon dioxide atmos-", + "phere.", + "For the in vivo studies the requisite amount", + "of BaP was administered intragastrically in 1 ml of", + "sesame oil to rats, some of which had previously", + "undergone thoracic duct cannulation.", + "Fig. 1", + "At the termination of both the in vitro and in vivo experiments, the appropriate sam-", + "ples were homogenized in sulfuric acid, the polycyclic hydrocarbon extracted into chloro-", + "form, and concentration determined spectrofluorometrically. Excitation wave length was", + "300 nm and fluorescence was measured at 425 nm.", + "RESEARCH RESULTS", + "Figure 2 illustrates recovery experiments in which BaP, dissolved in 0.1 ml chloroform,", + "was applied directly to rat small intestine and allowed to soak for 30 minutes. There was", + "six fold range in the quantity of BaP utilized. With the exception of one experiment, in", + "a", + "which recovery was 75%, almost the total amount of applied BaP was recovered.", + "Figure 3 illustrates the most striking finding in this study, i.e., the mode of accumu-", + "lation of BaP in everted sac tissue. On the abscissa is plotted the outside medium concen-", + "tration and on the ordinate, BaP concentration in sac tissue (points clustered about upper", + "regression line) and in medium inside the sacs (points clustered about lower regression", + "line) At outside medium concentrations of 200 micromoles/1 and above, as the medium", + "100", + "50", + "BaP ADDED", + "10 5000 UG", + ". 833 UG", + "fested", + "ber", + "2", + "6", + "4", + "SAC SIZE (CMS)", + "Eig. 2", + "Fig. 3", + "94", + "LG 0090661", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "-", + "I", + "2500", + "RO- 6029149" + ] + }, + { + "bbox": [ + [ + 0.144193, + 0.12332, + 0.724094, + 0.010952 + ], + [ + 0.143447, + 0.134866, + 0.742185, + 0.013529 + ], + [ + 0.144284, + 0.148983, + 0.740226, + 0.012763 + ], + [ + 0.144038, + 0.161626, + 0.756387, + 0.012404 + ], + [ + 0.144504, + 0.173633, + 0.442882, + 0.012273 + ], + [ + 0.185317, + 0.199087, + 0.716098, + 0.011483 + ], + [ + 0.144163, + 0.211531, 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The parallel relationship between the regression lines denotes that the quantity", + "of BaP transported through the sac tissue into the medium inside the sacs is directly pro-", + "portional to the sac tissue accumulation. This finding is consistent with passive diffusion", + "of carcinogen from sac tissue to fluid inside the sac.", + "The age of the animals, and whether the animals were fasted or fed, made little differ-", + "ence. The accumulation of BaP in everted sac tissue exceeded by 1 1/2-2 fold the accumula-", + "tion in sacs prepared without eversion. Anaerobiosis (95% nitrogen-5% carbon dioxide atmos-", + "phere) did not influence BaP accumulation in tissue, nor did 0.1 M iodoacetate, which in-", + "hibits glycolysis by reacting with the sulfhydryl groups of phosphoglyceraldehyde dehydro-", + "genase. Adsorption on the mucosal surface of sacs was found to be, at least in part, re-", + "versible. In a typical experiment, carried out with an incubation medium concentration of", + "570 micromole/1 BaP, incubation of 2 sacs was carried out for one hour. One sac was then", + "prepared for analysis. The other sac was incubated for an additional hour in medium identi-", + "cal except for the absence of BaP. This additional incubation diminished the tissue content", + "of BaP by 26%.", + "Figure 4 demonstrates the time course of BaP uptake in sac tissue. It is noteworthy", + "that in the first half minute there is very rapid uptake, and that at higher incubation", + "medium concentrations equilibrium is not complete in one hour.", + "Figure 5 illustrates incubation experiments in which the accumulation of BaP on string", + "was studied. As in small intestinal sac tissue, with arithmetic increase in medium concen-", + "tration, there is logarithmic increase in accumulation on string.", + "$00", + "190 220 BC/ML", + "600", + "130-170 UG/ML", + ".....", + "400", + "-1000", + "200", + "110 130 DC/m", + "30", + "60", + "90", + "120", + "150", + "180", + "300", + "SAC INCUBATION TIME CHINUTES)", + "SN", + "OUTSIDE", + "Fig. 4", + "Fig. 5", + "Figure 6 delineates BaP accumulation, on string as a function of incubation time.", + "Note the similarity of the curves to those time studies with small intestinal sacs as shown", + "in Figure 4.", + "Figure 7 demonstrates a typical study of the time course of appearance in thoracic", + "duct lymph of intragastrically administered BaP. Note that the greater part of the carcin-", + "ogen which is to appear in the lymph has appeared by the sixth hour.", + "Figure 8 delineates experiments in which varying doses of BaP were given to rats sacri-", + "ficed 18 hours thereafter, in which tissue concentrations were measured in breast and in", + "adipose tissue. Note that with arithmetic increases in dose of hydrocarbon there is loga-", + "rithmic increase in accumulation in these tissues.", + "95", + "RC- 6029150", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgchoot§", + "0090662" + ] + }, + { + "bbox": [ + [ + 0.167512, + 0.747837, + 0.729936, + 0.011877 + ], + [ + 0.208704, + 0.810627, + 0.696739, + 0.013068 + ], + [ + 0.169802, + 0.824211, + 0.751434, + 0.011348 + ], + [ + 0.168889, + 0.836815, + 0.751553, + 0.013292 + ], + [ + 0.16843, + 0.849565, + 0.736959, + 0.013028 + ], + [ + 0.169903, + 0.861659, + 0.727224, + 0.012287 + ], + [ + 0.17017, + 0.87415, + 0.750806, + 0.012201 + ], + [ + 0.224572, + 0.981309, + 0.553034, + 0.016348 + ], + [ + 0.202521, + 0.14102, + 0.014103, + 0.008237 + ], + [ + 0.236577, + 0.148918, + 0.057381, + 0.009595 + ], + [ + 0.624287, + 0.149472, + 0.012006, + 0.008071 + ], + [ + 0.430041, + 0.152354, + 0.050481, + 0.00872 + ], + [ + 0.729479, + 0.162114, + 0.075566, + 0.008938 + ], + [ + 0.237434, + 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sac tissue accumulation and concentration of BaP in the incubation", + "medium. First, the exponential relationship implies that as the concentration of polycyclic", + "hydrocarbon to which the intestinal mucosa is exposed is increased, the rate of absorption", + "accelerates. For example, if a certain increase in concentration of BaP results in a 10-", + "fold increase in absorption, then a second linear increase of the same amount would increase", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "150", + "White string", + "70", + "280 UC/ML", + "BENZO(a)PYRENE", + "7 cm, 28 30 mg", + "to MC. I.C.", + "60", + "50", + "e", + "100", + "200 UG/ML", + "40", + "o", + "30", + "e", + "110 UG/ML", + "50", + "e", + "20", + "€", + "9", + "70 UG/ML", + ".", + "10", + "o", + "150", + "60", + "120", + "2", + "30", + "90", + "4", + "6", + "8", + "10", + "12", + "NOUR INTERVALS", + "STRING INCUBATION TIME (MINUTES)", + "Fig. 6", + "Fig. 7", + "DISCUSSION", + "at \" \"OURS:", + "In many respects, the results of our experiments", + "ADIPOSE .", + "100", + "are similar to those reported by others for cholesterol", + "BREAST .", + "and fatty acid uptake by intestinal tissues. Uptake", + "of these compounds is also not blocked by anaerobiasis", + "nor by inhibitors of glycolysis. Unlike our results", + "with polyaromatic hydrocarbon carcinogens, however, the", + "10", + "tissue concentration of cholesterol and fatty acid did", + "not increase logarithmically with arithmetic increase", + "cur", + "in medium concentration.", + "Our exponential accumulation can be accounted for", + "by multilayer adsorption. The first step in the in-", + "testinal absorption of BaP can be considered as mole-", + "cules of BaP adsorbing onto surface sites on the intes-", + "tinal mucosa according to the Langmuir isotherm. Sat-", + "uration of the sites can be considered to represent", + "completion of a unilayer. Increases in medium concen-", + "15", + "tration above 200 micromoles/1 leads to multilayer", + "BOSE NC/100 6", + "adsorption according to Polanyi potential theory.", + "Fig. 8", + "The second step in the process of intestinal", + "absorption of BaP, i.e., from the sac tissue into the", + "simple diffusion.", + "CONCLUSIONS AND RECOMMENDATIONS", + "96", + "0090663", + "LG", + "RL- 6029151" + ] + }, + { + "bbox": [ + [ + 0.146154, + 0.121212, + 0.682067, + 0.012181 + ], + [ + 0.147036, + 0.133172, + 0.721138, + 0.01173 + ], + [ + 0.146828, + 0.145935, + 0.729777, + 0.010994 + ], + [ + 0.147484, + 0.15825, + 0.712274, + 0.010931 + ], + [ + 0.147869, + 0.170476, + 0.711769, + 0.012691 + ], + [ + 0.147238, + 0.18366, + 0.697209, + 0.012038 + ], + [ + 0.147883, + 0.196278, + 0.711755, + 0.012113 + ], + [ + 0.147791, + 0.20769, + 0.696849, + 0.011345 + ], + [ + 0.147932, + 0.220444, + 0.614325, + 0.011587 + ], + [ + 0.290988, + 0.257982, + 0.450474, + 0.009507 + ], + [ + 0.380413, + 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This stresses, for reasons not previously", + "recognized, the importance of maintaining low concentrations of such substances in the en-", + "vironment to which man and his tissues are exposed. Second, the ability to absorb BaP from", + "the extracellular fluid (including blood) to the cells in at least some of the tissues of", + "the body (breast and fat) also has an exponential relationship; the rate of absorption of", + "polycyclic hydrocarbons from the intestine is therefore very important. If the rate of", + "absorption is rapid, then the level of the extracellular fluid would become high and with", + "arithmetic increase in extracellular fluid concentration, the amount deposited in those", + "tissues exhibiting the exponential relationship would increase exponentially.", + "INVESTIGATIONS ON THE EFFECT OF TOBACCO SMOKE COMPONENTS", + "ON PLANT CELLS IN TISSUE CULTURE", + "P. S. Sabharwal, Ph.D., and P. R. Bhalla, Ph.D., Department of Botany", + "ABSTRACT", + "Studies were conducted to determine the effects of tobacco smoke components on the callus", + "tissue from tobacco pith maintained in tissue culture. Modified Murashige's medium con-", + "taining LAA (2 ppm) kinetin (2 pom) and sucrose (2%) served as the control. Different", + "concentrations (ranging from 10-4M to 10-10M) of the following smoke components were uti-", + "lized: acrolein, benzo[alpyrene, benzole]pyrene, phenol and pyrene. About 350-500 mg of the", + "tobacco pith callus was used as inoculum. The cultures were observed for five weeks for", + "effects on growth caused by the presence of tobacco smoke components in the medium. Benzo-", + "[a}pyrene (10-6M to 10-9M) caused callusing of the tissue. Other tobacco smoke components", + "have proved ineffective on this system. These observations clearly demonstrate that plant", + "tissue cultures can be used for rapid and sensitive bioassay of benzo[a]pyrene.", + "INTRODUCTION", + "Since plant and animal cells are often physiologically and biochemically similar, it", + "is possible to correlate research findings in one with similar responses in the other. So", + "far, animals have been used almost exclusively to study the effect of tobacco smoke compo-", + "nents. Since experimentation with plants is easier than with animals, investigations were", + "conducted to study the effect of tobacco smoke components on plant cells grown in vitro.", + "An attempt was made to ascertain the feasibility of the use of plant tissues as rapid bio-", + "assay materials for tobacco smoke components. A rapid bioassay of tobacco smoke or its com-", + "ponents should aid in expanding utilization of tobacco.", + "RESEARCH METHODOLOGY", + "Colorless and green calluses of Nicotiana tabacum were used to study the effect of", + "tobacco smoke components. The callus was produced from pith explants raised on modified", + "Murashige's medium containing the following (in mg/litre) : NH4NO2 1650, KNO3 1900, H3BO4", + "170, KH2PO4 170, KI 0.83, Na2Mo04 2H20 0.25, CoC12*7H2 0.025, MgS04*H2O 22.3, MnSO4*H2O", + "22.3, 2ns04.7H2 8.6, CuS04-5H20 0.25, EDTA 22.4, Fe2(504)3 10.67, CaC12*2H20 440.0,", + "Thiamine HC1 0.1, Nicotinic acid 0.5, Pyridoxine HC1 0.5, Glycine 2.0, NAA 0.2, Kinetin 0.2,", + "Sucrose 20,000 and Agar 8,000.", + "About 350-500 mg of the tobacco pith callus was used as the inoculum. Modified", + "Murashige's medium containing 2 ppm of IAA, 2 ppm of kinetin, 2% sucrose and 0.8% agar", + "served as the control. Different concentrations of tobacco smoke components ranging from", + "10-4M to 10-10M were incorporated into the medium. The pH of the medium was adjusted to", + "5.8. Equal amounts of medium were dispensed into each of the culture flasks or culture", + "tubes. Sterilization was achieved by autoclaving at 18 1b psi for 20 minutes. For each", + "concentration at least two sets of 24 cultures were maintained at 25° + 1°C under continuous", + "cool-white fluorescent light with ca 250 footcandles intensity. 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The callus retained its original color during this period.", + "It is apparent from Table I that the addition of acrolein, benzo[e]pyrene, phenol, or", + "pyrene to the control medium did not induce growth in the colorless tobacco callus. How-", + "ever, when benzo[a]pyrene was incorporated into the medium, proliferation was observed two", + "weeks after inoculation. The callus continued to grow for another 3 weeks after which the", + "growth ceased. The tissue ultimately turned brown. At the very low concentration (10-8M)", + "of benzo (a)pyrene the fresh weight of the callus increased from 350-500 mg to 5500-6000 mg", + "within 5 weeks. Lower (10-10M) or higher (10-4M) concentrations of this smoke component", + "were less effective.", + "The green callus showed no growth when the tobacco smoke components (Table I) were", + "added to the control medium.", + "TABLE 1", + "Tobacco Smoke", + "Concentrations", + "Effective", + "Growth Response", + "Growth Response", + "Components", + "(Molar)", + "Concentrations", + "of", + "of", + "(Holar)", + "Colorless Callus", + "Green Callus", + "Acrolein", + "10-4 to 10-10", + "-", + "0*", + "0*", + "Benzo(a)pyrene", + "\"", + "10-6 to 10-9", + "+", + "0*", + "Benzo(e)pyrene", + "\"", + "-", + "0*", + "0*", + "Phencl", + "\"", + "-", + "0*", + "0*", + "Pyrene", + "\"", + "-", + "0*", + "0*", + "*The Callus did not grow and turned brown.", + "+The callus started growing two weeks after inoculation.", + "DISCUSSION", + "The importance of nonhuman bioassay systems for routine examination of various tobacco", + "products has long been felt. Since 1953 various investigators have succeeded in producing", + "cancer in animals with tobacco products and provided proof that some tobacco smoke components", + "are tumorigenic to a variety of experimental animals (1).", + "Wynder et al (2) summarized experiments which utilized the mouse skin as a test sys-", + "tem for carcinogenic activity while Graham et al (3) presented similar data for rabbit", + "epidermis. According to these investigations, when animals were used as bioassay materials,", + "it took several months before the effects of tobacco smoke components were detected. In", + "the present investigation, a new approach was utilized to study the effects of tobacco smoke", + "components on living systems. Since the plants are inexpensive, immobile and can be easily", + "grown in the controlled conditions, they should prove excellent experimental materials as", + "compared to the animals.", + "Colorless and green tobacco callus tissues raised in vitro were used to study the", + "effect of some tobacco smoke components. Acrolein, benzo[e]pyrene, phenol, and pyrene", + "proved toxic at the concentrations of 10-4M to 10-10M. However, benzo[a]pyrene induced", + "marked growth of colorless callus at the concentration of 10-8M within five weeks. The", + "present observations demonstrate that plant tissue cultures can be used for a rapid and", + "sensitive bioassay of benzo[a]pyrene, a tobacco smoke component. We are currently working", + "on other smoke components to find out if this system is specific to benzo [a]pyrene or can", + "be used for other smoke components. The results will be reported subsequently.", + "RO- 6029153", + "98", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgchod6s", + "0090665" + ] + }, + { + "bbox": [ + [ + 0.456608, + 0.128603, + 0.080675, + 0.007918 + ], + [ + 0.141143, + 0.15159, + 0.709849, + 0.012939 + ], + [ + 0.171646, + 0.165576, + 0.085918, + 0.010657 + ], + [ + 0.141641, + 0.174923, + 0.73317, + 0.013037 + ], + [ + 0.171612, + 0.188916, + 0.381953, + 0.012518 + ], + [ + 0.141642, + 0.200716, + 0.717849, + 0.012554 + ], + [ + 0.173603, + 0.212639, + 0.667966, + 0.011177 + ], + [ + 0.172975, + 0.22532, + 0.230275, + 0.011005 + ], + [ + 0.33828, + 0.263233, + 0.329795, + 0.009574 + ], + [ + 0.338807, + 0.276002, + 0.328698, + 0.009187 + ], + [ + 0.306847, + 0.300586, + 0.385834, + 0.011293 + ], + [ + 0.466404, + 0.32591, + 0.065277, + 0.008425 + ], + [ + 0.140553, + 0.349223, + 0.70305, + 0.011727 + ], + [ + 0.140908, + 0.361596, + 0.710431, + 0.012225 + ], + [ + 0.141467, + 0.377157, + 0.007884, + 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Wynder, E. L. and Hoffmann, D., Ed. '5, Tobacco and Tobacco Smoke. Academic Press, New", + "York, 1967.", + "2. Wynder, E. L.; Graham, E. A.; and Croninger, A. B. Experimental production of carcinoma", + "with cigarette tar. Cancer Res. 13:855-64, 1953.", + "3. Graham, S.; Crouch, S. Levin, M. L.; and Bock, F. G. Variation in amounts of tobacco", + "tar retrieved from selected models of smoking behavior simulated by smoking machine.", + "Cancer Res. 23:1025-30, 1963.", + "THE POSSIBLE ROLE OF REACTIVE METABOLITES", + "OF POLYCYCLIC HYDROCARBONS IN ONCOGENESIS", + "J. W. Flesher, Ph.D., Department of Pharmacology", + "ABSTRACT", + "Previous experiments have established that -hydroxy-dimethylbenz[a]anthracene is formed", + "from dimethylbenz{a] anthracene (DMBA) by rat liver homogenates. Its formation represents", + "a", + "toxification reaction, as it is clearly more toxic than DMBA. The compound is also", + "a", + "potent carcinogen. Studies of its distribution showed that far less of the hydroxy deriva-", + "tive than DMBA reaches the mammary gland and other fatty tissue. This was in accordance", + "with the finding of its lower carcinogenicity for mammary tissue. A hypothesis that the", + "ultimate carcinogen is a highly reactive carbonium ion was tested. In conformity with this", + "hypothesis it has been found that tumors are rapidly formed following repeated subcuteneous", + "injection of compounds which would be expected to be precursors of carbonium ions. Each", + "compound was administered in 1 mg doses (in sesame oil) by subcutaneous injection on alter-", + "nate days for 20 doses. -7-Iodomethyl-12-methylbenz{a)anthracene, 7-bromomethyl-12-methyl-", + "benz[a]anthracene,7-benzoyloxy-methyl-12-methylbenz{a)anthracenewere the most active and", + "rapidly induced sarcomata at the site of injection. 7-Hydroxymethyl-12-methylbenz[a]anthra-", + "cene, 7-formyl-12-methylbenz[a)anthracene, and 7-methoxymethyl-12-methylbenz[a]anthracene", + "were less active; 4-hydroxy-7,12-DMBA and 4-methoxy-7,12-DMBA were inactive under these", + "experimental conditions. Only DMBA and its 7-hydroxymethyl metabolite induced breast cancer", + "in addition to subcutaneous sarcomata. The high degree of carcinogenicity of the most", + "active compounds, considered together with their chemical structure, suggest that conversion", + "to more reactive intermediates must occur in vivo.", + "The discovery that a metabolite of DMBA is a potent general carcinogen and the studies re-", + "ported here suggest a new approach in the search for polycyclic aromatic compounds which can", + "transform a normal cell into a cancer cell.", + "INTRODUCTION", + "A fundamental assumption in pharmacology is that in order for drugs to exert their", + "effects, interactions with cellular constituents must occur. The presumption is that dur-", + "ing chemical oncogenesis, the flow of genetic information coded in the base sequence of", + "DNA is somehow modified, thereby transforming the normal cell into a cancer cell. It is", + "the primary purpose of the present paper to focus attention on the initial steps in this", + "chain of events.", + "The simplest imaginable mechanism one can propose would suggest the reversible inter-", + "action of the parent hydrocarbon with some critical cellular component. The complex so", + "formed would alter the function of the cell sufficiently to transform it into a cancer cell.", + "Alternatively, one might suppose that the parent hydrocarbon is metabolized in vivo to a", + "form capable of reacting with cellular constituents. Polycyclic aromatic hydrocarbons have", + "long been regarded as direct acting carcinogens because microgram quantities are capable of", + "evoking cancers at the site of application. Ten years ago Huggins and his co-workers (1)", + "found that under optimal conditions the incidence of mammary cancer after a single 20 mg", + "meal of DMBA was 100%. Such data suggest that mammary cancer might be the result of a", + "RC- 6029154", + "99", + "i", + "LG 0090666", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.14674, + 0.135341, + 0.74665, + 0.016105 + ], + [ + 0.147977, + 0.150283, + 0.405436, + 0.013032 + ], + [ + 0.190104, + 0.174854, + 0.726839, + 0.014936 + ], + [ + 0.14796, + 0.188098, + 0.754318, + 0.013956 + ], + [ + 0.148366, + 0.200513, + 0.755007, + 0.014062 + ], + [ + 0.148457, + 0.216732, + 0.081805, + 0.01193 + ], + [ + 0.190983, + 0.238388, + 0.689599, + 0.013168 + ], + [ + 0.14915, + 0.250888, + 0.754219, + 0.013338 + ], + [ + 0.148724, + 0.262828, + 0.768146, + 0.014739 + ], + [ + 0.148646, + 0.276306, + 0.746403, + 0.012681 + ], + [ + 0.148582, + 0.289027, + 0.740247, + 0.012904 + ], + [ + 0.150443, + 0.30167, + 0.708533, + 0.014057 + ], + [ + 0.86604, + 0.301358, + 0.015716, + 0.009549 + ], + [ + 0.149807, + 0.314726, + 0.673196, + 0.012041 + ], + [ + 0.191204, + 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If the existence of such a metabolic product is a possibility,", + "it must be isolated, identified, and synthesized.", + "Several years ago I decided to investigate the distribution and metabolism of polycyclic", + "aromatic hydrocarbons in the rat. This species possessed a number of attractive advantages", + "for the experimental analysis of the relation between metabolism of the hydrocarbon and on-", + "cogenesis.", + "In the first place, the data accumulated by Huggins demonstrated that under optimal", + "conditions every female rat of the Sprague-Dawley strain would have manmary cancer in a re-", + "markably short time. Secondly, the experiments of Sydnor et al (2) demonstrated that various", + "rat strains differ in their susceptibility to cancer induction by polycyclic hydrocarbons.", + "Here we have the first suggestion that a metabolite could play the role of initiating the", + "complicated series of events leading to cancer formation. The intriguing implication", + "of", + "these observations made it mandatory that they be extended in several directions.", + "The existence of metabolites formed in vivo in the tissues of rats has been demonstrated", + "by several groups of investigators. In a study of the distribution and metabolism of DMBA", + "in the female Sprague-Dawley rat I found evidence for the formation of metabolites (3). In", + "the absence of further information it was possible to interpret these results in terms of", + "the \"direct-action\" hypothesis with the formation of metabolites representing a detoxifica-", + "tion process. It was clearly of importance to provide more definite evidence for the exis-", + "tence of a metabolite capable of inducing cancer. The unequivocal proof of the existence", + "of such a metabolite would immediately generate a host of new experimental possibilities.", + "As a convenient guide to the following discussion, we may set down the questions which stim-", + "ulated us to the performance of the experiments we will describe.", + "1. Is the hydrocarbon bound to cellular components by a covalent bond?", + "2. Is it possible to isolate, identify, and synthesize a carcinogenic", + "metabolite of the hydrocarbon?", + "3. Are some metabolites biologically active whereas others are not?", + "4.", + "Can one find evidence of other biological activity, predicted by the", + "assumption that a metabolite which is more closely related to the ulti-", + "mate carcinogen than the parent hydrocarbon would be expected to have", + "other biological activities?", + "5. What is the nature of the ultimate carcinogen?", + "6. Can one synthesize model compounds capable of generating the ultimate", + "carcinogen directly?", + "It will be noted that posing each of these questions, if answered in the affirmative,", + "suggests", + "the next one. I will now describe the experiments which led us to our conclusions.", + "THE BINDING OF HYDROCARBONS TO CELLULAR COMPONENTS", + "I undertook to obtain information pertinent to the first two questions raised.", + "Twenty-four hours after the oral administration of DMBA to rats, the tissue was homogenized", + "in 70% acetone and the homogenate centrifuged. The acetone soluble fraction was evaporated", + "under reduced pressure and the remaining aqueous mixture extracted with ether. The results", + "are illustrated in Figure 1.* The binding to the acetone insoluble fraction suggests the", + "formation of a covalent bond with cellular components. Although these results were en-", + "couraging, I hoped to establish the extent of binding to DNA, RNA and protein. Twenty-four", + "hours after the oral administration of DMBA mixed nucleic acids were isolated by the phenol", + "procedure of Kirby (4). Radioactivity was determined by oxygen-flask combustion to furnish", + "tritiated water which was counted in a liquid scintillation spectrometer (5). The data", + "so", + "obtained should detect differences in the degree of binding to these cellular components.", + "Care was exercised in dissolving and reprecipitating the nucleic acids. It was found that", + "no change in specific activity occurred, indicating the firm nature of the binding The", + "fact that these preparations resist extraction with organic solvents suggest the formation", + "of a covalent bond. Most significantly, it should be noted that practically no radioactivity", + "*Both ether and water-soluble forms. are present which demonstrates the formation of", + "metabolites.", + "100", + "RC- 6029155", + "/", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgchodos", + "0090667" + ] + }, + { + "bbox": [ + [ + 0.231649, + 0.168542, + 0.061451, + 0.007912 + ], + [ + 0.232035, + 0.18619, + 0.011616, + 0.005616 + ], + [ + 0.232651, + 0.216475, + 0.011799, + 0.00535 + ], + [ + 0.233658, + 0.24771, + 0.010958, + 0.005573 + ], + [ + 0.297077, + 0.269555, + 0.013633, + 0.004933 + ], + [ + 0.23132, + 0.278131, + 0.011855, + 0.005589 + ], + [ + 0.231298, + 0.309768, + 0.010912, + 0.005588 + ], + [ + 0.361255, + 0.324478, + 0.014686, + 0.006894 + ], + [ + 0.2319, + 0.339893, + 0.011191, + 0.005218 + ], + [ + 0.295933, + 0.352682, + 0.015383, + 0.006223 + ], + [ + 0.234851, + 0.371176, + 0.008306, + 0.005316 + ], + [ + 0.234141, + 0.401593, + 0.009265, + 0.005224 + ], + [ + 0.236235, 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AFTER ORAL", + "ADMINISTRATION OF 3H-DMBA (20 mg, 1120 pc)", + "Molecules of hydrocarbon bound to each", + "molecule of nucleic acid or protein", + "Nucleic acids", + "1st PPt.", + "2nd ppt.", + "Protein", + "to have oncogenic properties (9).", + "by", + "12-methylDMBA as a metabolite of DMBA. Many years ago Fieser (8) suggested that metabolism", + "of polycyclic hydrocarbons was probably necessary for carcinogenesis. This line of reason-", + "ing led Boyland and Sims to compare synthetic 7-hydroxy-DMBA prepared by Professor J. W.", + "Cook with the metabolites obtained when DMBA is incubated with rat liver homogenates.", + "7-Hydroxy-DMBA was identifiéd as a metabolite, and in addition, with Huggins, it was shown", + "We immediately recognized that the 7-iodomethyl-12-methyl compound, previously described", + "Fieser as an intermediate in the synthesis of DMBA, would provide a convenient interme-", + "diate for the introduction of the hydroxyl group. We have here then methods for synthesiz-", + "ing this interesting metabolite and some related compounds.", + "Flesher, Soedigdo, and Kelley (10) employed these methods to examine for the carcinc-", + "genicity of compounds which were identified as metabolites in rat-liver hemogenates (Table", + "II) . Compounds were dissolved in sesame oil, and a single oral dose was administered to", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "was bound when comparable amounts of", + "radioactivity were added to the tissue", + "prior to processing. It is clear that", + "if a covalent bond is involved, a meta-", + "bolic transformation must have occurred.", + "IDENTIFICATION AND SYNTHESIS", + "OF METABOLITES OF DMBA", + "The procedures employed in the in-", + "vestigations just summarized led to the", + "separation of tissue metabolites of DMBA", + "and further suggested that reactive me-", + "tabolites of DMBA were generated leading", + "to the formation of covalent bonds with", + "cellular components. It seemed unlikely", + "that a highly reactive form could be", + "isolated, but the metabolites separated", + "on paper and thin-layer chromatograms", + "appeared to be reasonably stable. The", + "SAT", + "central issue of the significance and", + "meaning of these metabolites is whether", + "or not they are carcinogenic. To answer", + "H2O", + "this question would require the isolation", + "and synthesis of these metabolites. A", + "direct attack on this question by com-", + "plete structure determinations of metab-", + "olites isolated from tissues was and is", + "LIVEA", + "not technically feasible. However, the", + "FAT", + "finding of Dickens of a product in the", + "urine of rats dosed with DMBA which", + "could be converted to 4-methoxy-DMBA by", + "methylation with dimethyl sulfate sug-", + "gested the possibility that 4-hydroxy-", + "DMBA might be one of the tissue metab-", + "olites (6).", + "We therefore undertook the synthesis", + "of 4-hydroxy-DMBA for comparison with me-", + "tabolites isolated from tissues and for", + "determination of pharmacological proper-", + "0.034", + "ties. The scheme employed is outlined", + "0.040", + "in Figure 2. At the time this synthesis", + "0.00008", + "(3)", + "was completed, Boyland and Sims (7) re-", + "ported the isolation of 7-hydroxymethy1-", + "101", + "RC- 6029156", + "0090668", + "LG" + ] + }, + { + "bbox": [ + [ + 0.668942, + 0.115737, + 0.241324, + 0.011366 + ], + [ + 0.669025, + 0.128077, + 0.24199, + 0.011202 + ], + [ + 0.669075, + 0.141229, + 0.206146, + 0.010782 + ], + [ + 0.669501, + 0.154189, + 0.2285, + 0.011143 + ], + [ + 0.669466, + 0.166328, + 0.247712, + 0.011909 + ], + [ + 0.669453, + 0.179833, + 0.236637, + 0.011018 + ], + [ + 0.669674, + 0.192433, + 0.203316, + 0.008623 + ], + [ + 0.44475, + 0.204318, + 0.055848, + 0.011713 + ], + [ + 0.709993, + 0.21735, + 0.195944, + 0.009636 + ], + [ + 0.670117, + 0.229734, + 0.229658, + 0.010808 + ], + [ + 0.669706, + 0.241841, + 0.254887, + 0.011032 + ], + [ + 0.669039, + 0.254942, + 0.259834, + 0.012239 + ], + [ + 0.670537, + 0.267928, + 0.242409, + 0.010497 + ], + [ + 0.315769, + 0.275918, + 0.019836, + 0.007975 + ], + [ + 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"age. The only metabolite with", + "carcinogenic activity was", + "7-hydroxy-DMBA. The methoxy", + "and acetoxy compounds which are", + "converted into 7-hydroxy-DMBA", + "in vivo also were active.", + "sairon", + "The data obtained in the", + "course of this investigation", + "showed that a metabolite of DMBA", + "has the ability to induce marmary", + "cancers. Further, this is the", + "OMa", + "OK", + "OH", + "first instance of a hydrocarbon", + "metabolite with oncogenic prop-", + "erties. This fact suggests that", + "DMBA must be converted to a me-", + "tabolite to transform a normal", + "cell into a cancer cell. How-", + "OMk", + "OM", + "OH", + "ever, it will also be noted that", + "the tumor incidence was lower", + "CHI", + "and the latent period longer for", + "7-hydroxy than for DMBA. It was", + "concluded, therefore, that the", + "most likely reason for this", + "Figure 2. Scheme for the synthesis of", + "lower activity is its lower lipid", + " 4-hydroxy-7,12-dimethylbenz[a)anthracene - -", + "solubility.", + "EVIDENCE OF OTHER BIOLOGICAL", + "ACTIVITY OF THE METABOLITE", + "The fact that evidence", + "could be provided for a carcino-", + "CHAON", + "genic metabolite of DMBA lends", + "+", + "obvious support to the hypoth-", + "esis that the normal process of", + "CH.ON", + "CHEON", + "transforming a normal cell into", + "a tumor cell involves a mechanism", + "whereby metabolism activates the", + "parent hydrocarbon. If the gen-", + "8", + "eration of a reactive interme-", + "diate is involved, it should be", + "possible to find evidence of", + "CHAI", + "other biological activity.", + "An opportunity to test the", + "validity of this prediction came", + "CH2OME", + "with the discovery of biological", + "CH2OH", + "CH2OAc", + "effects of DMBA on the hemo -", + "poietic system of the rat by", + "Cawein and Sydnor (11). These", + "investigators found that DMBA", + "induced maturation arrest of de-", + "Figure 3. Scheme for the synthesis of 7-hydroxymethyl-12-", + "methylbenz[a]anthracene and related compounds. The", + "veloping cells in bone marrow.", + "top", + "portion of the figure shows the conversion of DMBA to the", + "The peripheral blood showed", + "monohydroxymethyl and dihydroxymethyl compounds with lead", + "lymphocytopenia, granulocyto-", + "tetraacetate according to Cook.", + "phenia, and thrombocytopenia.", + "In a comparative study, Sydnor", + "and Flesher (12) examined the", + "effects of DMBA, 7-hydroxy-DMBA,", + "102", + "RC- 6029157", + "LG", + "0090669", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.320836, + 0.132809, + 0.045523, + 0.006607 + ], + [ + 0.596098, + 0.125246, + 0.268922, + 0.012941 + ], + [ + 0.592813, + 0.138035, + 0.247109, + 0.012389 + ], + [ + 0.18167, + 0.145455, + 0.329504, + 0.012821 + ], + [ + 0.593771, + 0.150931, + 0.264617, + 0.01067 + ], + [ + 0.593646, + 0.163367, + 0.272048, + 0.013079 + ], + [ + 0.377422, + 0.175231, + 0.067624, + 0.007792 + ], + [ + 0.592568, + 0.175928, + 0.241406, + 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Both DMBA and 7-", + "tumors/no. of", + "Appearance of", + "Benz(a)anthracene", + "Dose,mg", + "animals", + "tumors, days", + "hydroxy-DMBA induced anemia as", + "Range", + "Mean", + "shown by hematocrit determination,", + "7-Hydroxymethyl-12-methyl", + "20", + "6/8", + "83-150", + "120", + "but the 7-hydroxymethyl compound", + "is by far the most active (Figure", + "7-Methoxymethyl-12-methyl-", + "20", + "1/10", + "110", + "-", + "4). There was little influence", + "7-Acetoxymethyl-12-methy", + "20", + "2/10", + "110-114", + "112", + "on the hematocrit when either", + "1,12-dihydroxy-DMBA or 4-methoxy-", + "7,12-Dihydroxymethyl-", + "10", + "0/10", + "(180 days of", + "observation)", + "DMBA were fed and the values found", + "are indistinguishable from controls", + "4-Hydroxy-7,12-dimethyl-", + "20", + "0/10", + "(180 days of", + "observation)", + "given sesame oil alone. 7-Hydroxy-", + "DMBA produced profound thrombocy-", + "7.12-Epidoxy-7.12-dimethyl-", + "20", + "0/10", + "(180 days of", + "observation)", + "topenia which was most severe nine", + "days after treatment (Figure 5).", + "7,12-Dimethyl-", + "20", + "6/6", + "64-160", + "87", + "DMBA also induced a severe depres-", + "sion, but the decrease was only", + "The hydrocarbon, dissolved in 1 ml of sesame oil was administered by", + "to about 50% of the normal value", + "gastric instillation to female Sprague-Dawley rats, age 50 days. The ani-", + "mals vere examined for appearance of palpable tumors tvice weekly. (3)", + "whereas the values obtained for", + "groups given 4-methoxy-DMBA or", + ",,12-dihydroxymethyl derivative were not different from the control values.", + "Both DMBA and 7-hydroxy-DMB. induce a comparable degree of granulocytopenia on days", + "five and nine, but by day 14 pronounced granulocytosis had developed in rats fed 7-hydroxy-", + "DMBA whereas rats fed DMBA showed no such behavior (Figure 6). A similar type of response", + "has been observed by Elson (13) with alkylating agents such as chlorambucil. Growth rate", + "(Figure 7) was not influenced by 4-methoxy-DMBA or 7,12-dihydroxy-DMBA. DMBA induced a tem-", + "porary inhibition of growth rate, whereas 7-hydroxy-DMBA caused a significant and prolonged", + "inhibition. These observations strongly support the conclusion that the biological activi-", + "ties of DMBA are due to its metab-", + "olite, 7-hydroxy-DMBA.", + "In experiments which employ", + "-HYOROXY-DMBA", + "tritium-labeled DMBA and 7-hydroxy-", + "DMBA", + "SESAME OIL", + "DMBA, the existence of differing", + "50", + "tissue levels of hydrocarbon would", + "be detected. If the decrease in", + "45", + "lipid solubility is reflected in", + "the tissue level, then clearly", + "40", + "7-hydroxy-DMBA should be lower", + "than DMBA. At 24 hours the values", + "35", + "(Table III) for peripheral blood,", + "adrenal, spleen, thymus, kidney", + "30", + "and bone marrow were nearly iden-", + "tical for the two compounds. In", + "25", + "liver the concentration was lower,", + "but in lung, peri-renal and mammary", + "20", + "fat it was considerably higher for", + "DMBA-3P than for 7-hydroxy-DMBA-", + "o", + "5", + "9", + "14", + "19", + "24", + "3H (Figure 8). At 72 hours radio-", + "DAY", + "activity in all tissues had de-", + "clined appreciably, but the rate", + "of decline appears to be faster", + "Figure 4. Hematocrit on blood from tail vein of Sprague-", + "for DMBA-3H. The relative levels", + "Dawley female rats following administration of a single", + "of radioactivity are all consistent", + "20-mg feeding of benz [alanthracene derivative dissolved", + "with what would be predicted from", + "in sesame oil by gastric tube.", + "lower lipid solubility of the", + "metabolite.", + "RC- 6029158", + "103", + "LG 0090670", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.168942, + 0.376331, + 0.354161, + 0.011003 + ], + [ + 0.167957, + 0.389535, + 0.324182, + 0.011415 + ], + [ + 0.58317, + 0.400818, + 0.282552, + 0.011179 + ], + [ + 0.16877, + 0.401878, + 0.35605, + 0.012022 + ], + [ + 0.585043, + 0.41349, + 0.31407, + 0.01097 + ], + [ + 0.169594, + 0.415136, + 0.355282, + 0.010914 + ], + [ + 0.583988, + 0.426053, + 0.314662, + 0.011127 + ], + [ + 0.583803, + 0.439114, + 0.288158, + 0.009621 + ], + [ + 0.628045, + 0.478724, + 0.280995, + 0.009582 + ], + [ + 0.584784, + 0.491228, + 0.324643, + 0.011065 + ], + 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Serial platelet count in Sprague-", + "Dawley female rats given a single 20-mg", + "Figure 6. Granulocytes in blood of", + "feeding of the benz [a] anthracene derivative", + "Sprague-Dawley rats following a single", + "in sesame oil by gastric tube on day 0 (age", + "20-mg feeding of the benz[alanthracene", + "derivative dissolved in sesame oil.", + "If it is assumed that the level of", + "hydrocarbon in the tissue is a critical", + "factor in the induction of cancer, then", + "the relationship between the dose and the", + "incidence of cancer may be determined.", + "This would presume the tissue level of", + "7 -MYDROXY- DMBA", + "I90", + "170", + "I5O", + "130", + "DAY", + "Figure 7. Mean body weight of Sprague-Dawley", + "female rats given a 20-mg feeding of the", + "benz(alanthracene derivative in sesame oil.", + "The relationship between a single feeding of a 5, 10 or 20 mg. dose of DMBA or a 20 mg. dose", + "of 7-hydroxy-DMBA and the concentration of radioactivity in mammary fat removed 24 hours", + "after feeding the compound is shown in Figure 9.", + "After 5 mg. DMBA-3H the concentration was equivalent to 3.2 g, after 10 mg. to 6.4 1g", + "Source: ttps://www.industrydocuments.ucsf.edu/docs/jgchodo9", + "DAY", + "50 days).", + "230", + "7,12-DIHYDROXY-DMBA", + "hydrocarbon is one of the critical deter-", + "DMBA", + "210", + "minants in the induction of cancer. To", + "test for this possibility the level of", + "DMBA and 7-hydroxy-uMBA in tissue was de-", + "termined for comparison with the dose", + "administered.", + "In agreement with the earlier obser-", + "vations of Huggins et al (1) the incidence", + "of breast cancer following a single feed-", + "ing of DMBA was dose-related as shown in", + "Table IV. Four of 15 (27%) developed", + "FC- 6029159", + "0090671", + "s", + "OMBA", + "x10", + "XID", + "DMSA", + "SESANE OIL", + "5", + "10.", + ".", + "8", + "3", + "6", + "4", + "2", + "NO", + "2", + "-", + "T", + "7.12-DIHYDROXY-DMBA", + "DMBA", + "7-HYOROXY-OMBA", + "o", + "24", + "20", + "12", + "16", + "8", + "o", + "4", + "19", + "24", + "5", + "9", + "0", + "(4", + "DAY", + "&", + "breast cancer after 5 mg., 11 of 15 (72%)", + "o", + "4", + "8", + "12", + "16", + "20", + "24", + "after 10 mg., and 15 of 15 (100%) after", + "20 mg. DMBA. 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(24.0-43.4)", + "3.7 (3.6-3.8)", + "6.1 (5.0-6.9)", + "25.9 (18.7-39.4)", + "Liver", + "served after feeding 5 mg.", + "7.6 (6.1-9.2)", + "2.2 (1.8-2.4)", + "2.2 (2.2-2.3)", + "13.5 (10.8-18.6)", + "Lung", + "DMBA-3H. These values corre-", + "late well with the observed", + "34.4 (28.6-38.2)", + "4.5 (4.4-4.6)", + "6.4 (4.8-7.5)", + "33.1 (26.6-43.3)", + "Kidney", + "concentrations of unchanged", + "2.4 (1.8-3.0)", + "2.3 (1.7-3.3)", + "0.6 (0.4-0.7)", + "0.4 (0.4-0.5)", + "Thymus", + "hydrocarbon in mammary fat", + "3.2 (2.7-3.8)", + "1.2 (1.0-1.3)", + "2.5 (2.0-3.5)", + "0.7 (0.4-0.9)", + "Spleen", + "and clearly indicate that the", + "concentration of hydrocarbon", + "13.7 (11.3-15.7)", + "11.2 (8.8-13.9)", + "Adrenals", + "3.2 (2.7-4.1)", + "3.1 (2.2-3.7)", + "is an important determinant", + "3.1 (1.8-4.3)", + "0.8 (0.7-0.8)", + "4.2 (2.0-5.4)", + "32.4 (25.0-36.9)", + "Fat", + "in the genesis of experimen-", + "tally-induced breast cancer.", + "Peripheral Blood", + "2.6 (2.3-3.0)", + "17.7 (10.5-28.5)", + "17.1 (9.1-36.0)", + "3.2 (2.4-3.7)", + "These observations provide", + "2.5 (1.9-2.8)", + "1.8 (1.7-2.0)", + "4.9 (3.6-6.3)", + "4.4 (3.1-6.5)", + "Cells", + "the most acceptable explana-", + "12.8 (6.9-22.2)", + "Plasma", + "12.7 (6.0-29.5)", + "0.7 (0.5-0.9)", + "0.8 (0.6-1.0)", + "tion for the lower incidence", + "Bone Marrow", + "6.7 (3.5-9.9)", + "7.0 (6.5-7.4)", + "of breast cancer in rats of", + "this strain fed 7-hydroxy-", + "The figures in parentheses represent the range.", + "DMBA. We may conclude, there-", + "fore, that a metabolite exists with potent general carcinogenic propeties. Its formation", + "represents a toxification process as it is clearly more active than DMBA in certain biolog-", + "ical tests. The existence of this biological activity constitutes another link in the chain", + "of evidence supporting the hypothesis that a reactive metabolite is the ultimate carcino-", + "genic agent.", + "SELECTIVE SYNTHESIS OF COMPOUNDS RELATED TO DMBA CAPABLE OF GENERATING CARBONIUM IONS", + "We now turn to the last two questions posed in the introductory paragraphs, namely", + "the nature of the ultimate carcinogen and the selective synthesis of compounds capable of", + "generating the ultimate carcinogen.", + "RADIOACTIVITY (ug/g)-24 HRS.", + "MAMMARY GLAND)-24 HRS.", + "35", + "AFTER ORAL ADMINISTRATION OF CARCINOGEN-¹", + "FAT", + "I", + "T-HYOROXYMETHYL-2-METHTL", + "30", + "7,12-DIMETHYL", + "25", + "20", + "MAMMARY", + "PLASMA", + "15", + "to", + "5", + "Figure 8. Radioactivity levels in mammary", + "gland, fat, and plasma 24 hrs. after 20", + "mg of DMBA-3H or 7-hydroxy-DMBA-3a.", + "105", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "IS", + "10", + "5", + "o", + "S MG", + "10 MG", + "20MG", + "20 NG", + "DMBA", + "OMBA", + "DMBA", + "7-OM", + "RO - 6029160", + "Figure 9", + "0090672", + "LG", + "and after 20 mg. to 15.5 H8", + "per gram wet weight of tissue.", + "Over the range studied, the", + "tissue concentration of", + "radioactivity bears a linear", + "relationship to dose. 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+ "10 mg", + "0", + "3/15", + "11/15", + "81", + "23-170", + "4/15", + "which is postulated to occur", + "15", + "DMBA", + "5 mg", + "0", + "1/15", + "4/15", + "85", + "33-172", + "11/15", + "in metabolism of DMBA.", + "20", + "7-0H-DMBA", + "9", + "4/11", + "5/11", + "126", + "78-185", + "0.11³", + "20 mg", + "Essentially what we are", + "demanding is a carcinogen", + "1Loss due to toxicity of the compound.", + "which permits the formation", + "2Autopsy was performed 30 days after the appearance of first palpable tumor or 200 days", + "of the carbonium ion, with-", + "after feeding the carcinogen.", + "Three rats had lung cancer, one had reticulum-cell leukemia, and three had renal cancer.", + "out the intervening process", + "of metabolism. If this", + "could be achieved, then these compounds would have tremendous advantages in the analysis of", + "the complicated series of events leading to cancer. The proposed mechanism would then sug-", + "gest that the first step in carcinogenesis by DMBA is hydroxylation of the 7-methyl group.", + "The second step is the formation of a derivative which would be expected to be a good leaving", + "group, thereby generating a highly reactive carbonium ion.", + "Several facts emerged which encouraged us to look more carefully into compounds of this", + "type. 7-Bromomethyl-12-methyl-benz[a]anthracene was recently synthesized by Pataki et al", + "(15), and shown to be a carcinogen on injection in the rat. Although the iodomethyl compound", + "was prepared by Fieser more than", + "TABLE V", + "thirty years ago, its biological", + "TIME OF APPEARANCE OF FIRST TUMORS IN SPRAGUE DAWLEY FEMALE RATS GIVEN 1 MG DOSE OF", + "properties have apparently not", + "EACH COMPOUND IN 0.1 ML SESAME OIL BY SUBCUTANEOUS INJECTION ON ALTERNATE DAYS FOR", + "been reported. We had already", + "20 DAYS BEGINNING AT 50 DAYS OF AGE", + "prepared this compound. More", + "importantly though, the compounds", + "Benz(a)anthracene", + "Mammary Tumors", + "Sarcomata", + "derivative", + "tumor an./tan. mean range", + "tumor", + "/#an.", + "we wished to synthesize could", + "an", + "mean", + "range", + "all be prepared from 7-hydroxy-", + "7,12-dimethyl", + "7/10*", + "76", + "56-97", + "10/10", + "97", + "94-102", + "methyl-12-methyl-benz[a]anthra-", + "7-hydroxymethy)", + "cene, and we had the method of", + "4/5", + "**", + "12-methyl", + "83", + "57-97", + "4/5", + "103", + "92-111", + "choice for preparing this", + "1-formyl-12-methyl", + "4/5", + "90", + "77-99", + "4/5", + "105", + "101-113", + "compound.", + "7-benzcyloxymethyl", + "I therefore instituted an", + "12-methyl", + "0/5", + "--", + "5/5", + "87", + "83-92", + "investigation designed to see", + "7-1odomethyl-12-", + "methyl", + "0/5", + "5/5", + "55", + "50-60", + "whether the above expectations", + "were realizable.", + "7-bromothyl-12", + "methyl", + "0/5", + "--", + "3/5", + "90", + "87-91", + "After these syntheses were", + "4-hydroxy-7,12", + "complete, Dr. K. Sydnor joined", + "dimethyl", + "0/5", + "0/5", + "--", + "--", + "me in a preliminary examination", + "4-methoxy-7,12-", + "of their carcinogenic properties.", + "dimethyl", + "0/5", + "--", + "0/5", + "--", + "This is shown in Table V. It", + "7-formyl", + "0/5", + "--", + "--", + "0/5", + "--", + "--", + "will be noted that in each case", + "7-formyl-12-methyl", + "the compound proved to be a", + "0/5", + "--", + "5/5", + "69", + "60-80", + "potent carcinogen. What is even", + "7,12-dibromome chyl", + "0/5", + "--", + "--", + "5/5", + "110", + "54-203", + "more remarkable is the short in-", + "Sesame 011", + "0/5", + "--", + "--", + "0/5", + "--", + "duction time required to induce", + "the formation of a palpable", + "Two rats had ear duct tumor,", + "tumor.", + "*One rat had ear duct tumor.", + "RC- 6029161", + "106", + "LG 0090673", + "Source: :https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.193402, + 0.132969, + 0.66292, + 0.017773 + ], + [ + 0.154453, + 0.146019, + 0.718156, + 0.01831 + ], + [ + 0.154633, + 0.1585, + 0.599584, + 0.019178 + ], + [ + 0.470389, + 0.185796, + 0.087108, + 0.010393 + ], + [ + 0.194652, + 0.206732, + 0.655678, + 0.018758 + ], + [ + 0.155226, + 0.219837, + 0.726259, + 0.017421 + ], + [ + 0.156339, + 0.231905, + 0.715128, + 0.018144 + ], + [ + 0.156015, + 0.244118, + 0.701977, + 0.017515 + ], + [ + 0.156611, + 0.256272, + 0.72312, + 0.019019 + ], + [ + 0.156418, + 0.268451, 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type would be potent carcinogens. It is of obvious interest to continue this investi-", + "gation and see whether other biological properties of DMBA are also present.", + "CONCLUSIONS", + "The present paper records our efforts to gain an understanding of the nature of the", + "ultimate carcinogen which presumably influences the flow of genetic information. The direc-", + "tion taken by the series of investigations described was greatly influenced by two factors.", + "One was the assumption that an intermediary metabolite between DMBA and the ultimate car-", + "cinogen must exist. The second assumes that this intermediary metabolite could be isolated,", + "identified, and synthesized. Before the intermediate 7-hydroxy-DMBA could be accepted as", + "a point of departure for a more extensive search of the ultimate carcinogen, its carcino-", + "genicity had to be established beyond doubt. This was accomplished by oral and subcutane-", + "ous administration. It was further shown that the lower carcinogenicity following oral", + "administration was due to lower concentrations in tissues. Having established 7-hydroxy~", + "DMBA as a general carcinogen and devised a method for its synthesis, it was possible to go", + "to the next stage of investigation and inquire into the nature of the ultinate carcinogen.", + "This was done in terms of selective synthesis of compounds related to DMBA.", + "It was possible to show that tumors were indeed rapidly formed when these agents were", + "administered subcutaneously. The expectation that such compounds were potent carcinogens", + "was fully realized.", + "It would appear from the results summarized here that the search for a carcinogenic", + "metabolite of DMBA has been successful. In addition compounds have been synthesized which", + "appear to be the direct precursors of the ultimate carcinogen of DMBA.", + "REFERENCES", + "1. Huggins, C. B.; Grand, L. C.; and Brillantes, F. P. 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Dipple, A. ; Lawley, P. D.; and Brookes, P. Theory of tumor initiation by chemical car-", + "cinogens: dependence of activity on structure of ultimate carcinogen. European J.", + "Cancer 4:493-502, 1968.", + "15. Pataki, J.i Wlos, R. ; Young-Ja Cho. Adrenocorticolytic derivatives of benz(a)anthra-", + "cene. J. Med. Chem. 11:1083, 1968.", + "DISCUSSION HIGHLIGHTS", + "SYDNOR: Dr. Mandelstam, I would like to know about the concentration of BaP in other", + "tissues. Is the increase in concentration the same as that observed in breast tissue, or", + "are there differences?", + "MANDELSTAM: Our data are clear-cut only with breast and adipose tissue, to which we're con-", + "vinced this exponential relationship applies. We're not at all sure of our data with some", + "other tissues. We cannot draw any firm conclusions from available data.", + "SMITH: What happens if you use BeP? Does it absorb in the same way?", + "MANDELSTAM: Yes. We had hoped in the beginning that this might possible be a bioassay", + "and that we might be able to distinguish the carcinogenic and noncarcinogenic hydrocarbons", + "this way. That hasn't worked out, and this phenomenon has generally prevailed across the", + "whole range of polycyclic aromatic hydrocarbons. We've screened perhaps a dozen of these", + "compounds, and have done more detailed work with three or four of them. Most of our work,", + "however, has been with BaP.", + "FLESHER: Do you find the transfer across the gut wall to be logarithmic?", + "MANDELSTAM: No, that is not logarithmic; it seems to be a passive diffusion process from", + "what actually gets into the tissue.", + "SYDNOR: You start with a known concentration of BaP and by analyzing both inside and out-", + "side of the sac, are you able to account for it all or do you lose something in the analysis?", + "MANDELSTAM: If we take into account the medium concentration, wash off the compound that", + "adheres to the wall, measure our sac, measure our internal medium, measure even the ends", + "of the string that tie these sacs, we can account for about 97 + 4% of what we put into the", + "system.", + "SYDNOR: That's very good recovery. The reason I ask is that with liver we don't get 100%", + "recovery.", + "ALEEM: Is this an enzymatic phenomenon?", + "MANDELSTAM: No, I don't think so; generally we were able to show the same thing on string.", + "ALEEM: Then how is this process regulated? From the kinetics it would look like an enzy-", + "matic reaction.", + "MANDELSTAM: We can speak of our actual analytical results with conviction; however, we have", + "no sound theory as yet to account for -it.", + "ANDERSEN: What is the anatomy of these sacs; can they be likened to a semi-permeable mem-", + "brane, or are they more complicated than that?", + "MANDELSTAM: Yes, I think that they can be considered as semi-permeable membrane. They are", + "membranes that under other circumstances will demonstrate active transport.", + "SYDNOR: Dr. Sabharwal, do you have n owth in the control?", + "SABHARWAL: No appreciable growth has been detected.", + "RQ- 6025163", + "108", + "LG 0090675", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.165199, + 0.135484, + 0.405024, + 0.013581 + ], + [ + 0.165645, + 0.160395, + 0.69065, + 0.016045 + ], + [ + 0.165712, + 0.175471, + 0.125762, + 0.009861 + ], + [ + 0.165832, + 0.198754, + 0.554542, + 0.014586 + ], + [ + 0.16519, + 0.222648, + 0.546496, + 0.013954 + ], + [ + 0.165897, + 0.247378, + 0.690409, + 0.015259 + ], + [ + 0.165275, + 0.260233, + 0.213637, + 0.012137 + ], + [ + 0.166195, + 0.284236, + 0.705106, + 0.014761 + ], + [ + 0.165059, + 0.293912, + 0.688707, + 0.017491 + ], + [ + 0.165707, + 0.321215, + 0.380978, + 0.013431 + ], + [ + 0.164771, + 0.34586, + 0.458742, + 0.013154 + ], + [ + 0.165286, + 0.370678, + 0.397044, + 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I can say that benzo [a]pyrene is a potential cell", + "division factor.", + "FLESHER: Are there any other compounds that produce the same response?", + "SABHARWAL: z[a]anthracene-7, 12-dione also induces similar growth.", + "SYDNOR: When you add BaP to the medium, does it stay in the same concentration for the", + "duration of the experiment?", + "SABHARWAL: The concentration of BaP might change with the age of the culture; we have not", + "measured this. We have tried seven concentrations of BaP ranging from 10-4M to 10-10M.", + "SYDNOR: Is BaP being metabolized by the tissue?", + "SABHARWAL: That is what we are going to investigate next.", + "SYDNOR: When does the tissue start active growth?", + "SABHARWAL: After about two weeks.", + "EHMANN: Have you kept any of your BaP cultures in dark?", + "SABHARWAL: We are planning to do this.", + "SHEEN: How does the BaP-treated tissue compare with the stock cultures?", + "SABHARWAL: I do not see any differences. We are trying to determine if the BaP-treated", + "tissue has become auxinotrophic. If it has become cancerous, it should actively grow with-", + "out BaP.", + "AHMAD: Have you tried to grow fresh callus on a medium containing extract of the tissue", + "grown on BaP medium.", + "SABHARWAL: I have some cultures going now, but we do not have the results yet.", + "SHEEN: Do you know the effect of BaP on chromosomes of these cells?", + "SABHARWAL: We have not made observations on the chromosomes.", + "AHMAD: Dr. Flesher, I don't understand how you visualize a carbonium ion as being respon-", + "sible for carcinogenic activity.", + "FLESHER: In the scheme we visualize, the first step would be hydroxylation of the seven", + "position. The second step is the formation of a derivative which would be expected to be a", + "good leaving group, thus generating a carbonium ion. This reacts with some crucial cellular", + "constituent and is covalently bonded. The binding is of 8 low level, but sufficient to", + "transform a cell.", + "JEWELL: I notice that the blood elements are depressed. What about the lymphocytes; are", + "they also depressed?", + "FLESHER: Very much so. It is possible that the carcinogens suppress immunity, but whether", + "or not that provokes cancer is not clear.", + "ALEEM: Have you considered free radical formation?", + "FLESHER: People have talked about free radicals off and on for years; I haven't ruled out", + "109", + "RC- 6029164", + "LG", + "0090676", + "Source: :https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.223759, + 0.120401, + 0.709386, + 0.013765 + ], + [ + 0.223129, + 0.131647, + 0.734103, + 0.013718 + ], + [ + 0.223945, + 0.145398, + 0.676615, + 0.013673 + ], + [ + 0.224096, + 0.170295, + 0.735588, + 0.012423 + ], + [ + 0.225089, + 0.184011, + 0.414618, + 0.01418 + ], + [ + 0.224525, + 0.207482, + 0.629041, + 0.012905 + ], + [ + 0.224421, + 0.233531, + 0.72607, + 0.012493 + ], + [ + 0.224996, + 0.24628, + 0.723642, + 0.012141 + ], + [ + 0.225556, + 0.259448, + 0.422848, + 0.011535 + ], + [ + 0.224695, + 0.283585, + 0.597221, + 0.012825 + ], + [ + 0.224628, + 0.30836, + 0.734103, 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The", + "next stage of investigation will be to show how well these compounds bind to DNA, where th", + "attack, and what are the cellular mechanisms leading to this whole chain of events.", + "AHMAD: I was wondering why the methyl groups on other positions do not show as strong car.", + "cinogenic activity as the positions studied by you.", + "JEWELL: Have you attempted to locate the site of attachment within the cell?", + "FLESHER: We isolated nucleic acids and protein by the phenol method of Kirby and found a", + "low level of binding; nevertheless, it seemed likely that with highly reactive compounds,", + "reactions would occur with many cellular components.", + "JEWELL: Do you think the ultimate action has to be in the DNA molecules?", + "FLESHER: That's my presumption. Somehow it appears there has to be a modification of the", + "flow of genetic information. I don't think we know all the factors that regulate DNA, RNA,", + "and protein synthesis.", + "JEWELL: The virologists are ahead of the chemical oncologists because they'v demonstrated", + "that the DNA virus is able to insert itself into the DNA of the host cell. This very nicel:", + "explains the oncogenic effect of the virus. There is a lot of speculation as to whether the", + "so-called latent viruses are involved in chemical carcinogenesis. It is, of course, an un-", + "resolved question.", + "SYDNOR: I have a question for Dr. Roeckel; you have two cell lines--one with rapid turnover", + "and another that is quite long. Are there any differences in the two cell lines with re-", + "gard to their response to chemical carcinogens?", + "ROECKEL: First of all, I think you have to define the way carcinogens act. Dr. Flesher has", + "shown that with a single dose of carcinogen the animals develop cancer after a mean of 120", + "days. Yet he also shows that the distribution within 24 hours is to the manmary gland.", + "Could it be postulated that the cells exist for that length of time before becoming cancer-", + "ous? What mechanism does one postulate as being responsible for this tumor? Can it", + "be", + "said that the cells are being changed `genetically? What is the nature of this change?", + "It certainly could be argued that if it takes an average of 120 days, it couldn't be", + "a direct effect of the carcinogen. A possible explanation would be that when you give the", + "carcinogen you have a high cellular concentration; then something happens to the genetic", + "mechanism within the cell. You might postulate that it unmasks the virus which is present", + "initially. You have the malignant potential--a you say, 1% of your rats get spontaneous", + "mammary cancer.", + "I think that the cell turnover studies in which we've been involved have been very in-", + "formative. We did a few studies to find out if there is a difference in the regeneration", + "of bronchial epithelium of genetically defined mice. Turnover is the expression of a cell", + "population defined by mitosis or DNA synthesis. The genetic difference in our animals", + "exists only in the lung.an not in the esophagus. The esophagus has the same replication", + "time in both strains. I think it is very suggestive that we have one strain that is abso-", + "lutely different in the bronchial epithelial turnover from the other strain not spontane-", + "ously growing lung tumors.", + "If the same genetic difference were found in the esophagus, then I would say that per-", + "haps the turnover rate has nothing to do with the development of cancer. Since there is a", + "definite difference only in the lung, and the animals have spontaneous lung tumors, it is", + "appealing evidence that there is something different in the bronchial epithelium of these", + "strains. How does this happen? Does it have something to do with a congenital virus? It", + "would be informative to show that one strain is resistant to the virus whereas the other", + "is not. As to the mechanism of action of chemical enhancement, we could postulate, for", + "example, that in one strain the carcinogen kills the virus. Maybe that's", + "why", + "they", + "don't", + "get liver cancer. Perhaps there is something in the cell that interferes with the virus,", + "as in the mammary glands, and by giving a carcinogen the inhibition is removed.", + "Dr. Flesher gave a number of substances that gave cancer at the site of injection;", + "other substances didn't do it; and others do it in both sites. It is interesting, but it", + "110", + "RC- 6029165", + "LG 0090677", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.146886, + 0.124533, + 0.525872, + 0.015902 + ], + [ + 0.146975, + 0.148188, + 0.734468, + 0.016668 + ], + [ + 0.147704, + 0.162999, + 0.199474, + 0.014165 + ], + [ + 0.147016, + 0.186624, + 0.72657, + 0.016149 + ], + [ + 0.147796, + 0.198048, + 0.710105, + 0.018237 + ], + [ + 0.147772, + 0.210506, + 0.540221, + 0.016504 + ], + [ + 0.147265, + 0.23523, + 0.687029, + 0.016063 + ], + [ + 0.148243, + 0.248198, + 0.717098, + 0.016999 + ], + [ + 0.148344, + 0.260528, + 0.716978, + 0.01735 + ], + [ + 0.148593, + 0.274939, + 0.110689, + 0.011957 + ], + [ + 0.147781, + 0.296843, + 0.726463, + 0.017604 + ], + [ + 0.148123, + 0.309421, + 0.725666, + 0.017794 + ], + [ + 0.147419, + 0.321979, + 0.726221, + 0.017144 + ], + [ + 0.147287, + 0.333957, + 0.703884, + 0.017767 + ], + [ + 0.147808, + 0.346147, + 0.702256, + 0.016677 + ], + [ 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high doses. Why must man", + "smoke so long before getting cancer? Are we looking at the wrong thing by looking at the", + "fast carcinogen. Obviously, these compounds are not acting rapidly.", + "FLESHER: What we must ascertain first is: after a cell or a small number of cells are", + "transformed, how long does it take to manifest itself as a tumor? Suppose we knew immedi-", + "ately when the carcinogenic event occurred; how long would it be before a cancer was actu-", + "ally detected?", + "ROECKEL: The thing that stunned me was that before the animals get cancer (if you're look-", + "ing at synthesis as well as mitosis of bronchial epithelium) it takes 600 days to renew the", + "epithelial covering of the bronchus in cancer-susceptible animals. In the animal that does", + "not get cancer it takes only about 200 days. You either shed the bad cells or kill them", + "off; I don't know which is correct. One analogy that has disturbed me is people who say", + "they haven't chronic lung disease when we know that they do. The cell turnover in the mice", + "with chronic lung disease may go down to 12 days. In man the norm has not been established,", + "but we can demonstrate mitotic figures on biopsies; and if mitotic figures are increased,", + "you would assume that there is an increase in cell turnover. The interesting thing is that", + "the people with chronic lung disease can start cancer which may be histologically indis-", + "tinguishable from smoking cancer. It's practically identical.", + "SABHARWAL: If you give a dose of BaP today, how long does it take for the animals to show", + "tumors?", + "SYDNOR: If you give a single dose of 400 ug of BaP, 15 of 18 rats develop cancer within one", + "year; if you give 3 P8 on alternate days for 30 doses, 18 of 18 develop cancer. The mean", + "tumor induction time is longer for the single dose although the total dose for repeated in-", + "jections was only 90 micrograms.", + "SABHARWAL: Can you detect any BaP at that stage?", + "SYDNOR: I doubt very seriously if you would. If BaP is excreted as rapidly as 3-methyl-", + "cholanthrene or DMBA, you would not. Dr. Flesher may be able to add something to this.", + "FLESHER: The concentration of radioactive hydrocarbons is measurable in the tissue up to", + "one week or ten days. Then after that you find a couple of CPM. It's not zero, but for", + "all practical purposes it is.", + "SABHARWAL: In other words, maybe you already have the formation of so-called proximate", + "carcinogens in that tissue and keep working slowly.", + "FLESHER: I don't know if it keeps working slowly. A carbonium ion is highly reactive", + "just like an x-ray. You can give a single dose of x-ray, then wait for tumors to develop.", + "Obviously, if you don't give any more x-rays, the x-rays must have initiated the chain of", + "events.", + "SYDNOR: Does the vulnerability of the cell influence the response? If this were true, it", + "would explain, in part, some of the discrepancies observed. If the cells were in right", + "phase when the carcinogen was around, then you would get a change. One can imagine that", + "some cells are too damaged to replicate; others would not be in the right phase in the", + "mitotic cycle and wouldn't be damaged at all. Others may be so slightly damaged that it", + "would take many generations of cells before a subline would become autonomous and grow", + "rapidly.", + "MANDELSTAM: Dr. Sydnor, do you examine your cells to see if there are any abnormal chro-", + "mosomes--splitting, aberrations, or abnormal unknown chromosomes?", + "RC- 6029166", + "111", + "LG 0090678", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.170067, + 0.115564, + 0.363796, + 0.00954 + ], + [ + 0.170078, + 0.140135, + 0.713396, + 0.01243 + ], + [ + 0.169721, + 0.152698, + 0.721705, + 0.011672 + ], + [ + 0.169945, + 0.164689, + 0.741963, + 0.01298 + ], + [ + 0.169294, + 0.177891, + 0.737303, + 0.014868 + ], + [ + 0.170554, + 0.191217, + 0.735702, + 0.011999 + ], + [ + 0.170341, + 0.203349, + 0.751023, + 0.014148 + ], + [ + 0.169138, + 0.216154, + 0.722227, + 0.012591 + ], + [ + 0.171453, + 0.229744, + 0.162025, + 0.010458 + ], + [ + 0.169933, + 0.25431, + 0.765872, + 0.012956 + 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I wonder if you could", + "further explain the long latent period by saying that the initial alkylation of the DNA", + "doesn't cause an immediate mutation, but might cause a change in only one base of the DNA.", + "Because of the degeneracy of the genetic code, this change in one base would not have any", + "effect in the next replication process. In later replication processes this change could", + "either revert to the original base sequence and no tumor would result or the initial change", + "would lead to further changes during subsequent replication, and eventually result in a", + "tumor-causing event.", + "FLESHER: There is a lot of thinking along these lines and something like this may be reason-", + "ably close to the actual mechanism. The latent period varies, but again it is related to", + "the dose, and we know it must also be related to metabolism and cellular mechanism. 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B. Griffith, Ph.D., Director, Tobacco and Health Research Institute", + "The conference has been noteworthy for the quality of the papers presented and the way", + "in which the panels and the audience participated in the very worthwhile discussions. More", + "questions were asked than were answered-a good measure of the quality of the discussions.", + "Certainly everyone should have a better appreciation for the complexity of the tobacco and", + "health question and we will hope that new ideas for further research have been generated.", + "Variability was one of the words most frequently used in the conference. We learned", + "of the variability of tobacco and the ways in which the chemical and physical properties of", + "tobacco can be modified by genetics, cultural conditions, weather and curing conditions.", + "This product can be made even more variable, or perhaps less so, by the way tobacco is pro-", + "cessed and the way it is manufactured into products for sale to consumers. The consumers", + "smoke in different manners, thereby creating further fluctuation in smoke yields and compo-", + "sition which must be considered in connection with our search for a true understanding of", + "the nature and magnitude of the smoking and health question.", + "Recognition of at least some of the variables leads logically to a consideration of the", + "ways in which they can be controlled and used in the experimental situation. The University", + "of Kentucky Reference Cigarette and experimental samples, based on nicotine variabilities", + "arising from genetic alterations in tobacco, have been produced and other samples are planned.", + "The chemical and physical properties of the cigarettes and the smoke can be determined and", + "this information can be passed on to the researchers interested in determining biological", + "response. Their work can be made even more meaningful because of the new smoking machine", + "which permits variation and control of smoking parameters in a manner which will insure the", + "use of smoke corresponding in yield and composition with those analagous to the human situa-", + "tion. More information is needed, however, concerning the way in which people smoke and the", + "quantities of smoke consumed to provide the basis for more definitive work with animal", + "models.", + "The creation and characterization of samples has little meaning until the biological", + "significance of the variables can be assessed. As a result of fundamental work conducted", + "at the University during the past five years, we have now reached the point where routine", + "bioassays relating to gas phase components of smoke, carcinogenic activity of smoke and the", + "response of animals to inhaled smoke can be established. These procedures will be used as", + "guidelines for those doing agricultural research, and one can consider other product modifi-", + "cations designed to minimize any responses considered to be detrimental. We can now begin", + "to apply, in a routine way, the techniques developed in the fundamental research program.", + "Recognizing that the emphasis may shift from the fundamental to applied research,", + "Dr. Flesher reminds us that we must not forget fundamental research and expressed the opin-", + "ion that the level of this type of research should be increased. The value of fundamental", + "research is certainly indicated by the progress made in developing bioassay procedures, and", + "one must maintain a balance between basic and applied research. This is especially important", + "when so little is known about carcinogenesis and other fundamental problems in this terribly", + "complex area.", + "A part of Dr. Flesher's concern is undoubtedly related to the traditional role of", + "fundamental research by individual scientists in an academic atmosphere and the problems", + "created in this environment by a multi-disciplinary mission-oriented program which is ex-", + "pected to yield results immediately. The problems, however, should be minimal if one rec-", + "ognizes the need to continue to conduct the fundamental research at the department and", + "college level. The Tobacco and Health Research Institute should provide a service to basic", + "research efforts, coordinate the fundamental efforts, and conduct the applied research nec-", + "essary for a balanced program.", + "RC- 6029158", + "113", + "LG 0090680", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.218418, + 0.9796, + 0.673289, + 0.017967 + ], + [ + 0.289085, + 0.267896, + 0.034883, + 0.008225 + ], + [ + 0.223209, + 0.294381, + 0.184987, + 0.011619 + ], + [ + 0.224817, + 0.320483, + 0.065535, + 0.009447 + ], + [ + 0.223252, + 0.332914, + 0.119163, + 0.009341 + ], + [ + 0.223644, + 0.345073, + 0.117788, + 0.009828 + ], + [ + 0.224695, + 0.357688, + 0.116878, + 0.009573 + ], + [ + 0.224227, + 0.370809, + 0.117726, + 0.009131 + ], + [ + 0.224908, + 0.38374, + 0.092257, + 0.009105 + ], + [ + 0.225147, + 0.396052, + 0.100205, + 0.009264 + ], + [ + 0.225225, + 0.408738, + 0.100587, + 0.009129 + ], + [ + 0.225516, + 0.421756, + 0.108387, + 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Ahmad", + "M. I. H. Aleem", + "R. A. Andersen", + "E. S. Anderson", + "W. o. Atkinson", + "A. Benenson", + "J. F. Benner", + "P. R. Bhalla", + "C. E. Bortner", + "E. V. Brown", + "H. R. Burton", + "L. P. Bush", + "J. Calvert", + "E. W. Chick", + "L. W. Cochran", + "G. B. Collins", + "D. L. Davis", + "S. Diachun", + "G. M. Donnelly", + "W. D. Ehmann", + "J. W. Flesher", + "c. Gairola", + "J. G. Green", + "R. B. Griffith", + "c. Grunwald", + "J. L. Hamilton", + "A. J. Hiatt", + "C. H. Issidories", + "W. Jewell", + "M. J. Kasperbauer", + "P. Legg", + "H. 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ABSTRACTS OF CONTRACTED WORK", + "AREA I", + "EFFECTS ON TOTAL SOLUBLE TOBACCO LEAF PHENOLS OF NEAR ULTRAVIOLET LIGHT AND TEMPERATURE,", + "by R. A. Andersen, Ph.D., and M. J. Kasperbauer, Ph.D., Department of Agronomy [ARS Adjunct", + "Personnel)", + "Phenolic levels in field-grown leaves were higher than in greenhouse-growr tobacco leaves.", + "Near ultraviolet photocontrol was speculated as involved because sunlight includes wave-", + "lengths down to 290 nm while glass completely filters out wavelengths shorter than 315 nm.", + "To test this, two tobaccos, F. C. 402 and Ky. 151, were grown in controlled environment", + "chambers equipped with cool-white fluorescent lamps and filters that removed shorter wave-", + "lengths of light. Light intensity was about 2000 foot candles and the unfiltered lamps", + "(controls) emitted some light of 290-320 nm. Filters were placed about five feet above the", + "treated plants. Leaves were sampled 10, 20, and 30 days after treatment was started, and", + "results are averaged over all the sampling dates and stalk positions. F. C. 402 leaves at", + "75°F that did not receive wavelengths shorter than either 315 nm or 400 nm had 60-62 per-", + "cent of the total phenolic content of controls. Ky. 151 leaves grown at either 75°F or", + "90°F in the absence of wavelengths of light shorter than 400 nm had 62-64 percent the total", + "phenolic content of controls. Ky. 151 leaves grown at 90°F had 75 percent the levels of", + "total phenols as from plants at 75°F in the case of either control or light-filtered", + "plants. The sums of chlorogenic acids (major tobacco phenols) had the same response pat-", + "tern to light wavelengths and temperature as did total phenols.", + "TOBACCO AND HEALTH RELATED RESEARCH ON THE BIOSYNTHESIS, ACCUMULATION AND TRANSLOCATION OF", + "SECONDARY AMINES AND RELATED ALKALOIDS, by L. P. Bush, Ph.D., Department of Agronomy", + "[Contract No. 12-14-100-9523(34)", + "A gas chromatographic procedure has been developed for the separation of the principal", + "tobacco alkaloids. Columns used in this procedure were packed with acid-washed and di-", + "methyldichlorosilane-treated 60-80 mesh Chromosorb W coated with 10% DC550 (a methylphenyl", + "silicone oil). Relative retention times for nicotine, nornicotine, mysomine, anabasine,", + "and anatabine were 1.00, 1.55, 1.87, 2.03, and 2.50, respectively. Resolution among the", + "fractions was at least 95% except between nornicotine and myosmine. Using this procedure,", + "nicotine, nornicotine and anabasine were detected in air-dry mature seed of N. tabacum", + "var. Ky. 16. During germination the alkaloid content of the seed decreased until radicle", + "protrusion and then increased very rapidly. Seed germinated for 144 hr. at 16° contained", + "equal amounts of nicotine, nornicotine, anabasine, and anatabine. Seed germinated for 144", + "hr. at 32° contained the highest percent nicotine but contained less total nicotine than", + "other temperatures except 16°. Greatest accumulation of alkaloids occurred in seed germi-", + "nated at 27°. Comparison of alkaloid content of seedling at 144 hr. showed that 16° and", + "27° germination temperatures differed from all others, but 21° and 32° did not differ from", + "each other.", + "Dimethylamine, methylethylamine, diethylamine, and methylpropylamine were the principal", + "aliphatic secondary amines identified in burley tobacco. In general dimethylamine", + "accounted for 60%, diethylamine and methylpropylamine 15% each, and methylethylamine 5% of", + "the total amines measured. The amines were positively correlated with total nitrogen and", + "total volatile nitrogenous bases and negatively correlated with nitrate nitrogen content", + "of the tissue. Leaf lamina and stalk had the highest amine content. Leaf midribs had the", + "lowest and roots and seed were intermediate-in amine content. Volatile amine content of", + "leaf tissue also increased with higher stalk position. As determined by sample prepara-", + "tion and acid-base extraction procedures, the amines were not present as free forms or", + "their salts, but probably as a complex and liberated during distillation. In general,", + "wild species of Nicotiana had lower accumulations of volatile amines than commercial", + "tobaccos.", + "GENETIC AND BIOCHEMICAL STUDIES OF TOBACCO STEROLS, by D. L. Davis, Ph.D., Department of", + "Agronomy (Contract No. 12-14-100-9521(34)", + "Phytosterol levels in air-cured burley tobacco, Nicotiana tabacum L., ranged from 1.40 to", + "2.10 mg/8 of dry weight. The pattern of distribution of sterols in the tobacco plant was", + "characterized by a linear increase from the base to the apex. Within a leaf, the sterol", + "RC- 6029171", + "117", + "LG 0090683", + "Source: tps://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.138647, + 0.108815, + 0.744433, + 0.016561 + ], + [ + 0.13829, + 0.119372, + 0.724801, + 0.017193 + ], + [ + 0.142145, + 0.132161, + 0.714173, + 0.018485 + ], + [ + 0.140176, + 0.145161, + 0.730666, + 0.01705 + ], + [ + 0.139629, + 0.158183, + 0.738717, + 0.015961 + ], + [ + 0.139313, + 0.170616, + 0.731747, + 0.016489 + ], + [ + 0.139744, + 0.184433, + 0.731631, + 0.016448 + ], + [ + 0.139475, + 0.196102, + 0.724508, + 0.016598 + ], + [ + 0.138409, + 0.208407, + 0.663511, + 0.015581 + ], + [ + 0.809014, + 0.21526, + 0.062842, + 0.009873 + ], + [ + 0.139285, + 0.221576, + 0.739744, + 0.016953 + ], + [ + 0.139194, + 0.234281, + 0.733051, + 0.015633 + ], + [ + 0.139152, + 0.248395, + 0.138343, + 0.011603 + ], + [ + 0.138938, + 0.272413, + 0.748593, + 0.015259 + ], + [ + 0.139089, + 0.284973, + 0.739036, + 0.016793 + ], + [ + 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section to the leaf tip.", + "The major component sterols are cholesterol (8-11%), campesterol (22-26%), 8-sitosterol", + "(16-27%), and stigmasterol (35-47%). Cholesterol and campesterol varied much less than", + "the other two sterols. Stigmasterol was found to increase and P -sitosterol to decrease", + "in the more mature tissues. This represents a double band change in the side chain of the", + "sterol molecule. There were greater differences among component sterols within a leaf at", + "the base of the plant than one at the apex. The component sterol pattern was not greatly", + "influenced by changes in total sterol content. Some degree of genetic control for total", + "sterol levels was observed in burley tobacco. The progeny was influenced by the", + "presence", + "of a high, low or intermediate parental line. There were only slight differences in rela-", + "tive amounts of component sterols in the progeny of a cross involving parental lines with", + "similar patterns.", + "TOBACCO AND HEALTH-RELATED RESEARCH ON THE BIOSYNTHESIS AND METABOLISM OF RUTIN AND CERTAIN", + "STEROLS, by C. Grunwald, Ph.D., and c. Keller, Ph.D., Department of Agronomy [Contract No.", + "12-14-100-8945(34)", + "Two types of tobacco, Coker 139, a flue-cured variety and Burley 21, an air-cured variety,", + "were field-cultured under high and low levels of nitrate nitrogen fertilization and sampled", + "throughout a growing season. Chemical analyses of polyphenols and phytosterols were per-", + "formed on fresh leaves and also on primed leaves which had been air- or oven-dried. A1-", + "though slight repression was observed in content of rutin and chlorogenic acid in both", + "varieties treated with high nitrogen levels, the differences between nitrogen treatments", + "were not significant. However, Coker 139 when sampled fresh or subsequent to air-drying", + "was significantly higher in both compounds than Burley 21 similarly treated. Coker 139 and", + "Burley 21 contained about equal quantities of sterol at time of transplanting. Nitrogen", + "fertilization had no effect on the sterol content of Burley 21, but increased the sterol", + "content in Coker 139. However, nitrogen fertilization influenced the individual sterols in", + "Burley 21 and to a lesser extent in Coker 139.", + "Sterols and phenols were determined in two samples of flue-cured tobacco (Nicotiana tabacum", + "L.) which differed only in that one sample was heavily infested with the molds Aspergillus", + "niger, Aspergillus flavus, and Penicillium. Total sterol content was higher in the moldy", + "tobacco sample; total phenol content was higher in the nonmoldy sample. Differences in", + "amounts of chlorogenic acid, rutin, scopolin, caffeic acid, and minor sterol constituents", + "indicated that cured tobacco subjected to conditions favoring mold development may undergo", + "alteration in a number of its chemical constituents.", + "The enzymes involved in the biosynthesis of rutin and other polyphenols of tobacco were", + "investigated. The presence of the enzymes tyrosine ammonia-lyase (TAL) and phenylalanine", + "ammonia-lyase (PAL), key intermediates in the biosynthesis of polyphenols has been dem-", + "onstrated in tobacco. Specific activity of PAL was at least 10 times greater than TAL in", + "tissue tested. An assay method for PAL which does not require ether extraction wast devised.", + "Rutin was not induced under white light, far-red, red, yellow and blue light. Chlorogenic", + "acid, which was present at the onset of the experiment, was found under all light treat-", + "ments, and plants under blue light retained more chlorogenic acid than those under red,", + "far-red, and yellow. Far-red light caused the greatest loss of chlorogenic acid. The", + "quality of light did not markedly influence the sterol content in Burley 21, but did in-", + "fluence the sterol content in Iso 2.", + "An accurate and precise gas chromatographic procedure for the quantitative determination", + "of cholesterol, campesterol, stigmasterol, and -sitosterol was described. The phytos-", + "terols are separated as free sterols on a glass column packed with 5% OV-101 on Anakrom", + "ABS, 80/90 mesh. The quantification is carried out by adding cholestane as the internal", + "standard and measuring peak ratios of peak areas. This procedure is easily used on a rou-", + "tine basis with biological samples, and has been applied to the quantitative analysis of", + "tobacco sterols.", + "The condensing effect of sterols on phospholipids apparently depends upon several different", + "actions. It appears that many different sterols are capable of forming the closed membrane", + "configuration, and their effectiveness depends upon their molecular configuration and their", + "charge distribution. If the sterols in the plant actually play a role in controlling the", + "permeability of the membrane, then the particular sterol or combination of sterols that", + "118", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch000S", + "]", + "&", + "t", + "T", + "t", + "I", + "L", + "a", + "o", + "a", + "I", + "R", + "C", + "Ti", + "to", + "f:", + "en", + "ar", + "11", + "Wi", + "t:", + "Ky", + "CC", + "RO- 6029172", + "0090684", + "LG" + ] + }, + { + "bbox": [ + [ + 0.101972, + 0.10591, + 0.726831, + 0.011365 + ], + [ + 0.101137, + 0.117757, + 0.727035, + 0.010824 + ], + [ + 0.101305, + 0.128877, + 0.759605, + 0.012633 + ], + [ + 0.102149, + 0.14414, + 0.306417, + 0.012503 + ], + [ + 0.102016, + 0.167725, + 0.756834, + 0.013735 + ], + [ + 0.102465, + 0.181882, + 0.5217, + 0.012427 + ], + [ + 0.101254, + 0.200326, + 0.719671, + 0.010663 + ], + [ + 0.101945, + 0.212666, + 0.759219, + 0.012157 + ], + [ + 0.101935, + 0.225821, + 0.718464, + 0.011619 + ], + [ + 0.101934, + 0.238133, + 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The general impermeability of biological", + "membranes can be attributed in part to the stabilizing influences of compounds like choles-", + "terol, B -sitosterol or stigmasterol.", + "TOBACCO AND HEALTH RELATED RESEARCH STUDYING THE STEROL BALANCE OF TOBACCO, by c. Grunwald,", + "Ph.D., Department of Agronomy [Contract No. 12-14-100-9559(34)1", + "The level of free sterols in Nicotiana tabacum L. var. Burley 21 was found to increase", + "during seed germination commencing at the fourth day of germination. Increase in the chlo-", + "rophyll content, under comparable conditions, was found to start approximately 2.5-3.5", + "days after germination. Of the four major sterols (cholesterol, campesterol. stigmasterol,", + "3-sitosterol), stigmasterol was found to increase most rapidly and became the predominant", + "free sterol at the longest germination period studied. The esterified sterol fraction", + "showed little change during germination; however, relatively large fluctuations in the", + "sterol ester fraction were observed from sample to sample. There appears to be an increase", + "in the ratio of (free sterol)/(base sterols) in the case of stigmasterol and campesterol.", + "No significant shift in the ratio was found in the case of cholesterol.", + "INVESTIGATIONS TO DETERMINE THE FATE OF POLYPHENOLS IN TOBACCO AT HARVEST AND DURING THE", + "PROCESS OF AIR CURING, by R. E. Hampton, Ph.D., Department of Plant Pathology [Contract", + "No. 12-14-100-9540(51)]", + "The objective of this study is to investigate the mechanism of virus-induced increase in", + "polyphenol oxidase activity. If the enzyme could be purified, the plant could be provided", + "with c14-amino acids during the period of rapid increase in enzyme. The specific radio-", + "activity of the enzyme protein would then indicate the extent to which new enzyme synthe-", + "sis is involved in the increased activity.", + "Preliminary investigations suggest that the technique of density labeling is applicable to", + "the problem. The use of growth regulators such as Kinetin, IAA, and 2,4-D for stimulating", + "enzyme activity, while possible to some extent under laboratory conditions, does not seem", + "practical under field conditions. The one exception to this is chlorogenic acid, which", + "stimulates enzyme activity presumably without significantly increasing the phenol level.", + "However, the rarity of this compound might render it impractical.", + "Procedures for achieving a fairly high degree of purification of the enzyme have been de-", + "veloped and several properties of the enzyme have been determined. However, the yield of", + "active enzyme is quite low, and the use of radioisotopes to \"tag\" the enzyme would therefore", + "be of limited value. Much more promising in this respect is density labeling using N15 and", + "measurement of the density increase in the enzyme by cesium chloride density gradient cen-", + "trifugation. As reported here, this technique works well for the system under study.", + "Tobacco polyphenol oxidase, like the enzyme from other sources, has a low oxygen affinity.", + "Unlike the enzyme from other sources, it has a rather narrow substrate range. Chlorogenic", + "acid, isochlorogenic acid, caffeic acid, catechol, and L-3,4-dihydroxyphenylalanine in", + "order of decreasing affinity are oxidized. Tyrosine and o-courmaric acid, both monophenols,", + "are not hydroxylated. It may be concluded that the enzyme is strictly an oxidase.", + "INVESTIGATIONS ON THE AMMONIA-LYASE ENZYMES DESIGNED TO PROVIDE BASIC INFORMATION ON THE", + "REGULATION OF POLYPHENOLIC COMPOUNDS IN TOBACCO PLANTS, by C. Keller, Ph.D., and", + "C. Grunwald, Ph.D., Department of Agronomy [Contract No. 12-14-100-8945(34)", + "The enzyme responsible for the first committed step in phenol and polyphenol synthesis in", + "tobacco, L-phenylalanine ammonia-lyase, was purified approximately 200-fold from lamina of", + "field-grown Burley 21 plants. The most highly purified preparation was at least 70% homog-", + "enous as defined by disc electrophoresis. The optimal pH for the reaction was between 8.0", + "and 8.6. At pH 7.4, the Km for L-phenylalanine was 1.6 x 10-4 M and at pH 8.55 was 2.2 x", + "10-4 M.", + "When plants were analyzed subsequent to appearance of disease symptoms, the specific ac-", + "tivity of phenylalanine ammonia-lyase was lower in leaves and stems of greenhouse-cultured", + "Ky 16 which had been inoculated with purified preparations of tobacco mosaic virus than", + "in", + "comparable uninoculated tissue.", + "RC- 6025173", + "119", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009 0090685" + ] + }, + { + "bbox": [ + [ + 0.176378, + 0.112389, + 0.750999, + 0.012893 + ], + [ + 0.178536, + 0.123762, + 0.189273, + 0.011091 + ], + [ + 0.177729, + 0.1424, + 0.737607, + 0.011874 + ], + [ + 0.178243, + 0.155435, + 0.736642, + 0.011959 + ], + [ + 0.178214, + 0.167101, + 0.75799, + 0.014791 + ], + [ + 0.177829, + 0.180688, + 0.737204, + 0.012846 + ], + [ + 0.177602, + 0.193065, + 0.742933, + 0.011353 + ], + [ + 0.177703, + 0.204742, + 0.758195, + 0.013415 + ], + [ + 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Legg, Ph.D., Department of Agronomy,", + "[ARS Adjunct Personnel]", + "Two low alkaloid lines that do not convert nicotine to nornicotine have been derived from", + "crosses of Cuban cigar strains and burley varieties. By backcrossing low alkaloid plants", + "in E2 populations to Burley 21 and Kentucky 16, burley lines comparable to the two varieties", + "except for alkaloid content have been obtained. Parental, F1- F2, B1, and B2 generations", + "derived from crossing the low alkaloid lines with the two burley varieties were evaluated.", + "These experiments indicated that the low lines differed from the varieties at two important,", + "nonlinked loci. The two loci had different dosage effects and gene action was primarily", + "additive with additive x additive epistisis being of some importance. The low alkaloid", + "burley 21 line has been released to geneticists and other scientists for research purposes.", + "POLYPHENOLS AND PEROXIDASES IN THE GENUS NICOTIANA, by S. J. Sheen, Ph.D., Department of", + "Agronomy [Contract No. 12-14-100-8941(34)1", + "Leaf peroxidase of Nicotiana species, cultivars, autopolyploids, interspecific hybrids, and", + "amphidiploids have been compared by polyacrylamide gel block electrophoresis. At least 19", + "peroxidase bands, four cathodic and 15 anodic, were detected in the species. They varied", + "from two bands in N. arentsii to 12 bands in N. tabacum. The cultivars of the latter", + "species failed to reveal any intraspecific variation. Zymograms from autopolyploids and", + "amphidiploids appeared to be identical to that of diploid parents, suggesting that peroxi-", + "dase banding patterns are independent of ploidy levels. An additive manner of parental", + "peroxidase bands without hybrid enzyme formation in interspecific hybrids and the failure", + "of dissociating peroxidases into subunits lead to a hypothesis that peroxidases in Nicotiana", + "may be controlled by multiple dominant genes and/or codominant alleles in chromosomes of", + "different genomes.", + "Eight wild Nicotiana species, Kostoff's amphdiploid, and 19 cultivars, have been compared", + "for total polyphenol content and peroxidase activity in the leaf extracts by a small plant", + "technique. Greater variations for these chemical constituents occurred in the species than", + "in cultivars. N. tomentosiformis was highest in polyphenol content, while N. sylvestris", + "showed strongest peroxidase activity. The diversity of germ plasm among members of the", + "genus Nicotiana may be useful in the breeding program for altering the level of polyphenols", + "and their oxidative products in commercial tobaccos. Peroxidase banding patterns may be", + "used as genetic markers to facilitate such breeding programs.", + "BREEDING OF POLYPHENOL QUANTITY AND LOCALIZATION OF PEROXIDASE ISOZYMES IN TOBACCO LEAVES,", + "by S. J. Sheen, Ph.D., Department of Agronomy [Contract No. 12-14-100-9561 (34)]", + "A population of the fourth backcross progenies derived from crosses between six tobacco", + "introductions and Burley 21, the latter being used as the recurrent parent, was analyzed", + "for total polyphenol content in green leaves at flowering stage. A range of 2.02 to 23.0", + "mg chlorogenic acid equivalent/gm dry weight was obtained as compared to a 7.45 to 14.70", + "mg/gm variation in Burley 21 which was grown in the same field experimental plot. Burley", + "plants either high or low in polyphenol content were selfed and again backcrossed to", + "Burley 21. Hopefully, breeding lines having agronomical characteristics of Burley 21, but", + "varying in polyphenol level would be derived from this backcross program.", + "Peroxidase isozymes in the leaf laminae, epidermal layers of the vein, endodermis with", + "enclosed vascular tissues, and trichomes of Burley 21, N. sylvestris, N. tomentosiformis,", + "and N. otophora were investigated by, polyacrylamide gel block electrophoresis. Differences", + "in zymogram and enzymatic activity exist among the tissues studied, suggesting that peroxi-", + "dase isozymes have tissue specificity. The major peroxidase isozymes in leaf laminae may", + "bear direct impact on the metabolism of polyphenols because polyphenols are known to be", + "synthesized in this tissue.", + "RC- 6029174", + "120", + "Source: :https://www.industrydocuments.ucsf.edu/docs/jgchO069", + "0090686" + ] + }, + { + "bbox": [ + [ + 0.118155, + 0.11233, + 0.73654, + 0.010894 + ], + [ + 0.118835, + 0.123088, + 0.741545, + 0.010575 + ], + [ + 0.120939, + 0.135351, + 0.261797, + 0.012124 + ], + [ + 0.11848, + 0.155418, + 0.719853, + 0.011704 + ], + [ + 0.119359, + 0.166561, + 0.719325, + 0.012227 + ], + [ + 0.846952, + 0.169805, + 0.017109, + 0.007353 + ], + [ + 0.119439, + 0.179153, + 0.703175, + 0.012598 + ], + [ + 0.11953, + 0.192611, + 0.752, + 0.011207 + ], + [ + 0.118765, + 0.204118, + 0.745021, + 0.012131 + ], + [ + 0.11847, + 0.21691, + 0.729659, + 0.011951 + ], + [ + 0.119593, + 0.229822, + 0.719858, + 0.011377 + ], + [ + 0.118636, + 0.242045, + 0.745125, + 0.011655 + ], + [ + 0.120378, + 0.254408, + 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L. Sims, Ph.D., and W. o. Atkinson, M.S., Department of Agronomy,", + "[Contract No. 12-14-100-8940(34)", + "Burley 21 tobacco was grown in field tests at four locations and on four different soil", + "types to study the influence of soil properties, N fertilization, and growth regulators", + "on", + "the development and chemical composition of Burley tobacco. Dry matter synthesis was", + "slower in high N than low N treated plots on all soils until the topping stage of plant de-", + "velopment; thereafter this trend was reversed and synthesis was greatest in plants treated", + "with high rates of N. Concentration of Mo in the soil solution was inversely related to", + "level of N fertilizer applied to Maury soil. Concentration of Mo in tobacco leaves was", + "directly related to dry matter and protein contents, but inversely related to nitrate con-", + "centration. Nitrate and total N concentration (%) was, respectively, 0.3 and 0.8 percent-", + "age points lower in cured leaf of tobacco grown on Tilsit soil than in leaf produced on the", + "Maury soil. Using growth regulators for sucker control generally resulted in lower concen-", + "trations of nitrate N in cured tobacco than hand suckering.", + "EFFECT OF NITROGEN NUTRITION AND TOBACCO TYPE ON LEVELS OF TOTAL PARTICULATE MATTER AND", + "BENZO[A]PYRENE IN CIGARETTE SMOKE, by J. L. Sims, Ph.D., and W. 0. Atkinson, M.S., Depart-", + "ment of Agronomy [Contract No. 12-14-100-9560(34))", + "Burley 21 tobacco was grown at Lexington, Kentucky, in a replicated field test using vary-", + "ing levels of N fertilization and conventional production practices. Similarly, flue-cured", + "tobacco was produced in North Carolina in two replicated field tests using varying levels", + "of N fertilization. When mature, half of the burley tobacco was quick dried and the remain-", + "ing burley and all the flue-cured was barn cured in a manner conventional for each type.", + "Growth of the burley was normal during much of the growing season, but growth was slow", + "during one period because of poor distribution of rainfall. Leaf yields of burley tobacco,", + "respectively, for the 50-, 100-, 200-, and 400-1b/acre N fertilizer levels were 1761, 1931,", + "1953, and 2207 lbs/acre. Per acre yields of flue-cured tobacco were respectively, 2237,", + "2643, 2903, and 2930 lbs in Experiment 1 and 1771, 2132, 2691, and 2597 lbs in Experiment 2", + "for N fertilizer rates of 0, 30, 45, and 60 lbs N per acre. Laboratory analyses of leaves", + "and smoke will contribute further to the attainment of the major objectives of the project.", + "RC- 6029175", + "121", + "LG", + "0090687", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.499424, + 0.107415, + 0.058863, + 0.010534 + ], + [ + 0.166348, + 0.144176, + 0.72303, + 0.015955 + ], + [ + 0.166507, + 0.155555, + 0.717212, + 0.018462 + ], + [ + 0.168515, + 0.170589, + 0.103182, + 0.011312 + ], + [ + 0.165814, + 0.189737, + 0.025072, + 0.009742 + ], + [ + 0.199929, + 0.189997, + 0.073452, + 0.011242 + ], + [ + 0.280791, + 0.187543, + 0.626488, + 0.016264 + ], + [ + 0.165982, + 0.198391, + 0.701175, + 0.017862 + ], + [ + 0.166654, + 0.211399, + 0.715529, + 0.016096 + ], + [ + 0.167147, + 0.22416, + 0.698279, + 0.017083 + ], + [ + 0.166403, + 0.236419, + 0.701397, + 0.016363 + ], + [ + 0.165705, + 0.249062, + 0.725766, + 0.015788 + ], + [ + 0.16643, + 0.261267, + 0.733188, + 0.017114 + ], + [ + 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Atkinson,", + "M.S., J. Calvert, M.S., and c. Haynes, B.S., Department of Agronomy [Contract No. 12-14-", + "100-9528(73)]", + "The", + "cigarette", + "manufacturing laboratory has achieved a high degree of proficiency during the", + "period covered by Contract IV. It now has the capability of starting with whole cured", + "tobacco leaves and proceeding by a sequence of operations, including stemming, blending,", + "conditioning and cutting, to finished cigarettes manufactured on a Hauni-baby machine.", + "After making, the cigarettes can be sorted into selected weight groups on an automatic", + "balance and the average pressure drop determined for each group. In the smoke condensate", + "laboratory cigarettes are conditioned at 75°F and 60% RH prior to smoking. Cigarettes are", + "smoked to a 23 or 30 mm butt length, using Mason Mk 111 smoking machine standardized to", + "take one 35 ml puff per cigarette per minute. Tobacco smoke condensate is collected in", + "glass traps which are immersed in a dry ice acetone slurry (approx. -80°C). The traps", + "presently used are of two types, 1) plain bottom, partially filled with Raschig rings, or", + "2) modified bottom with a small well into which the glass insert protrudes, nearly touch-", + "ing the bottom, as described by Elmenhorst. Condensate is recovered from flasks contain-", + "ing the Raschig rings by using an organic solvent (acetone) to dissolve the condensate.", + "The solution is transferred to a rotoevaporator and solvent removed in a 35°C water bath.", + "Acetone is added and removed a total of four times to azeotrope with water and produce whole", + "dry condensate which can be used directly or diluted. Condensate is removed from the", + "Elmenhorst type trap by breaking up the \"cake\" formed at the bottom of the flask with", + "a", + "cold spatula and pouring the still frozen condensate into an appropriate container. The", + "flask may be rinsed out with a few ml of acetone and the wash solution added to the por-", + "tion removed initially. Keeping the condensate cold during the removal process increases", + "its solubility and solutions containing 50-60% of condensate can be prepared. Water and", + "acetone contents are determined by gas chromatography and subsequent dilutions made on the", + "basis of dry condensate yield. The water soluble fraction of smoke condensate can be ob-", + "tained by appropriate washing and evaporation and acid extract by extraction with dilute", + "HC1 and ether.", + "DEVELOPMENT AND EVALUATION OF CIGARETTE MODIFIERS WHICH ALTER SMOKE COMPOSITION, by", + "J. F. Benner, Ph.D., and H. R. Burton, Ph.D., Department of Agronomy [Contract No.", + "12-14-100-10279( (73)]", + "Investigations of the effects of various additives on the thermal degradation of tobacco", + "and the composition of the resulting smoke are being conducted. The major objectives of", + "this study are to obtain fundamental information on the thermal degradation of tobacco and", + "on the occurrence and formation of health-related components of cigarette smoke. The", + "ultimate goal is to develop a smoking product which delivers reduced yields of health-", + "related components in the smoke. Recent work has shown that, compared to cigarettes made", + "from untreated tobacco, the levels of phenol varies from 70% for cigarettes made from", + "tobacco treated with ,3-epithiopropyl methyl ether to 206% for cigarettes treated with", + "sodium molybdate. o-Cresol varies from 59% for sodium thiocyanate to 272% for sodium", + "molybdate and enzo[alpyrene varies from 48% for sodium stannate and sodium thiocyanate", + "to 191% for sodium pyrovanadate. The per cigarette yields of nicotine varies from 0.49 mg", + "for sodium vanadate to 2.37 mg for sodium aluminate.", + "A scheme for classifying the effect of an additive on the pyrolysis of tobacco has been", + "developed. This classification is based on the relative weight losses of treated and un-", + "treated tobacco as shown by thermogravimetric analysis. By using this system the levels of", + "certain health-related smoke components from treated tobaccos, such as benzo[a]pyrene and", + "phenol, can often be predicted from the thermogravimetric data.", + "Temperature-yield profiles have been used to study the general mode of action of borate", + "salts on the pyrolysis of tobacco and certain tobacco constituents. This work has shown", + "that one of the major effects of borate is through the formation of complexes with carbo-", + "hydrate and lignin present in the tobacco.", + "RC- 6029176", + "122", + "LG", + "0090688", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.13517, + 0.120153, + 0.741413, + 0.010743 + ], + [ + 0.136809, + 0.131158, + 0.728711, + 0.012758 + ], + [ + 0.136652, + 0.143789, + 0.715525, + 0.01281 + ], + [ + 0.136586, + 0.158088, + 0.714199, + 0.011316 + ], + [ + 0.137733, + 0.16981, + 0.696004, + 0.011595 + ], + [ + 0.13673, + 0.195374, + 0.723812, + 0.010879 + ], + [ + 0.136602, + 0.206686, + 0.395357, + 0.010939 + ], + [ + 0.136067, + 0.224814, + 0.708359, + 0.01372 + ], + [ + 0.137569, + 0.238027, + 0.739616, + 0.011013 + ], + [ + 0.136485, + 0.250027, + 0.699661, + 0.011854 + ], + [ + 0.137309, + 0.262977, + 0.715312, + 0.01139 + ], + [ + 0.137009, + 0.274931, + 0.723197, + 0.011747 + ], + [ + 0.137235, + 0.287685, + 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series of twelve experimental tobaccos has been investigated.", + "The original intent of this study was to ascertain whether thermal analytical data could", + "be used to predict the levels of certain health-related smoke components from \"agronomi-", + "cally feasible\" tobaccos. To date this approach has proven to be a qualified success.", + "RECOVERY OF AFLATOXINS FROM ARTIFICIALLY INOCULATED TOBACCO, by C. Y. Yang, Department of", + "Plant Pathology [Contract No. 12-14-100-9562(51)]", + "Aflatoxin producing strains of Aspergillus parasiticus and Aspergillus flavus (fc3) are", + "the major components of tobacco foliar microflora. Morphologically, these strains resemble", + "Aspergillus flavus strain NRRL 2999; however, their aflatoxin producing capacities are", + "greater. By coupling solvent extraction and chromatographic fractionation procedures, a", + "rapid and simplified separation method was developed for obtaining purified aflatoxins in", + "large quantities. Using a sucrose-yeast extract solution as the culture medium, 1.89 mg", + "of crude extract was obtained per g of substrate. The crude extract consists of 43.7%", + "aflatoxins in the ratio: G1:B1:G2:B2::100:24.6:17:14.1. The major aflatoxin components", + "were characterized by TLC,UV absorption, fluorescence, IR spectra and melting points and", + "their properties were identical with known standards. Duckling bioassay of B1, G1, G2", + "were positive. B2 could not be tested. The unusually high ratio of G1 to B1 provides a", + "source for further study of the physiological significance of G1 on many biological", + "systems. Ninety percent of the Aspergillus flavus isolates from the tobacco foliar flora-", + "were found to possess aflatoxin producing potentials. The recovery of aflatoxins from", + "artificially infested, environmentally-grown tobacco at various curing stages was confirmed", + "experimentally.", + "RC- 6029177", + "123", + "LG 0090689", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.107192, + 0.148577, + 0.696252, + 0.016797 + ], + [ + 0.107568, + 0.160602, + 0.671075, + 0.016984 + ], + [ + 0.10685, + 0.174128, + 0.45561, + 0.015253 + ], + [ + 0.107106, + 0.189137, + 0.73523, + 0.017222 + ], + [ + 0.108545, + 0.200956, + 0.725817, + 0.018576 + ], + [ + 0.108286, + 0.213574, + 0.695964, + 0.021988 + ], + [ + 0.108779, + 0.226606, + 0.726498, + 0.018222 + ], + [ + 0.108131, + 0.238224, + 0.720171, + 0.018958 + ], + [ + 0.108344, + 0.252673, + 0.562317, + 0.01572 + ], + [ + 0.108499, + 0.276763, + 0.736097, + 0.018089 + ], + [ + 0.109776, + 0.288957, + 0.726431, + 0.019982 + ], + [ + 0.110486, + 0.304825, + 0.260268, + 0.012812 + ], + [ + 0.108023, + 0.318487, + 0.682816, + 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Brown, Ph.D., and I. Ahmad, Ph.D.,", + "Department of Chemistry [Contract No. 12-14-100-9477(73)", + "We have subjected the high boiling basic fraction of cigarette smoke tar to column chroma-", + "tography using alumina and florisil followed by thin layer chromatography and gas chroma-", + "tography. A number of compounds have been tentatively identified with the help of Rf", + "values and mass spectrometry. These compounds are: nicotine, r r -dipyridyl, nicotina-", + "mide, anabasine, B-nicotyrene, a,a-dipyridyl, myosamine and 2,2 '-dipyridyliamine. Mass", + "spectra interpretations of several of these are discussed in reports.", + "INVESTIGATIONS ON DETECTION, IDENTIFICATION AND QUANTITATIVE DETERMINATION OF NITROSAMINES", + "IN CIGARETTE SMOKE, by E. V. Brown, Ph.D., and K. W. Yang, Ph.D., Department of Chemistry", + "(Contract No. 12-14-100-9518(73)", + "Previous work by ourselves and others has shown that if nitrosamines are present in", + "cigarette smoke, they are there in very small amounts. We have made progress in investi-", + "gating two methods of identification known for their sensitivity: fluorophotometry and", + "tracer techniques. In 1964 Neurath reported that reduction of nitrosamines followed by", + "formation of the colorful hydrazones of the resulting hydrazines with 5-nitrosalicylaldehyde", + "enabled one to identify individual nitrosamines in a complex mixture. Fluorometry is a", + "much more sensitive analytical tool than colorimetry for measuring concentrations of a", + "substance. Anthracene and phenanthrene are known for their strong fluorescence and their", + "aldehydes are also fluorescent. Nitrosamines have been reduced by our improved reduction", + "method and converted to the hydrazones using anthracene and phenanthrene aldehydes. All", + "hydrazones show strong fluorescence at 350-450 um. These are separated and identified by", + "TLC. This method is presently being applied to cigarette smoke tar. As -concerns the", + "second approach, we are still studying synthetic methods for the incorporation of c14", + "(radioactive carbon) into the nitrosamines.", + "INVESTIGATIONS ON THE IDENTIFICATION AND QUANTITATIVE ESTIMATION OF N-HETEROCYCLIC HYDRO-", + "CARBONS IN TOBACCO SMOKE, by E. V. Brown, Ph.D.; I. Ahmad, Ph.D. and A. J. Poole, Ph.D.", + "Department of Chemistry [Contract No. 12-14-100-10273(73))", + "In our work with the basic (HC1 soluble) fraction of tobacco smoke tar, we have been in-", + "vestigating the possibility of improving our separations by the use of column materials", + "other than those we have previously used. In this connection cellulose columns have been", + "found to be very useful. Materials separated on alumina columns and on cellulose columns", + "were subjected to thin layer chromatography with some success in further dividing these", + "complex mixtures. Finally, we have subjected materials obtained by the above methods to", + "gas chromatography using more thermally stable stationary phases, such as 15% Apiezon W on", + "Anakrom (50/60 AB), 10% Versamide on Chromosorb W and 10% apiezon L on Chromosorb W coated", + "with 5% KOH. The various fractions obtained in this way have been subjected to mass spec-", + "trometry and we are presently studying these data with the hope of obtaining interpretations.", + "Our study of the neutral fraction of tobacco smoke tar is just beginning. We have separated", + "the neutral fraction (insoluble in acid and base) into further fractions based on solvent", + "partition between methanol-water and cyclohexane followed by extraction of the cyclohexane", + "soluble material by nitromethane. These fractions are to be subjected to column chroma-", + "tography using alumina and ion exchange columns.", + "OPERATION OF THE UNIVERSITY OF KENTUCKY TOBACCO AND HEALTH ANALYTICAL SERVICE LABORATORY,", + "by J. G. Green, Ph.D., and J. Crutchfield, B.S., Department of Agronomy [Contract No.", + "12-14-100-9530(73)]", + "During the preceding year (1969), the Tobacco and Health Analytical Service Laboratory was", + "relocated to a renovated, permanent facility. During the year, a total of approximately", + "5,500 individual analyses (excluding internal replication and routine moisture analysis)", + "were performed involving 19 procedures. Not all procedures were available at any given", + "time, and 70 percent of the workload involved only five procedures. A survey of expected", + "utilization indicates that the workload has stabilized at the current level of activity.", + "124", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009", + "AREA III", + "I:", + "e:", + "C1", + "pi", + "Il", + "PT", + "Ch", + "Se", + "dr", + "ar", + "de", + "ic", + "hy", + "an", + "to", + "cr", + "ca", + "cy", + "at", + "A", + "th", + "IN", + "OF", + "De", + "Me", + "wh:", + "etl", + "fra", + "by", + "cal", + "and", + "150", + "dic", + "cy", + "(15", + "die", + "ter", + "The", + "als", + "TRA", + "THE", + "Ehm", + "Abu", + "hav", + "fil", + "fil", + "smo", + "par", + "col", + "Non", + "Ref", + "be", + "was", + "mat", + "log", + "res", + "RC- 6029178", + "LG 0090690" + ] + }, + { + "bbox": [ + [ + 0.116634, + 0.118356, + 0.701388, + 0.018232 + ], + [ + 0.116105, + 0.129912, + 0.726531, + 0.018955 + ], + [ + 0.116118, + 0.144635, + 0.71738, + 0.017828 + ], + [ + 0.116885, + 0.157323, + 0.59381, + 0.018408 + ], + [ + 0.11676, + 0.182645, + 0.701889, + 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Most such projects are in late planning and pro-", + "curement stages. As laboratory resource and scheduling improves, it is anticipated that", + "project orientation in methods development will be an expanding function.", + "INVESTIGATIONS ON THE EFFECT OF STRUCTURAL VARIATIONS ON THE COMPOSITION OF AMINO ACID", + "PYROLYZATES, by J. M. Patterson, Ph.D., and W. T. Smith, Jr., Ph.D., Department of", + "Chemistry [Contract No. 12-14-100-9476(73)]", + "Serine, proline, glutamic acid, valine, alanine, a-aminobutyric acid, phenylalanine anhy-", + "dride (3,6-dibenzy1-2,5-diketopiperazine) and phenethylamine have been pyrolyzed at 650°", + "and 850°. The amounts of neutrals, acids, bases and phenols in the pyrolyzates have been", + "determined. Most of the individual components of the neutral and basic fractions have been", + "identified and their relative amounts determined. Gases formed during pyrolysis, including", + "hydrogen cyanide and carbon monoxide, as well as carbon dioxide, methane, ethane, ethylene", + "and ammonia, have been identified and their amounts determined. Variations in the struc-", + "tures of amino acids affect the chemical composition of the pyrolyzates. Thus, an in-", + "crease in chain length results in an increase in carbon monoxide and polynuclear hydro-", + "carbons. The presence of additional functional groups results in an increase in hydrogen", + "cyanide, heterocyclics and polar compounds (largely nitriles and phenols). Proline, an", + "atypical amino acid, gives relatively large amounts of hydrogen cyanide and heterocyclics.", + "A preliminary determination of benzo[alpyrene in the pyrolyzates of aline indicate that", + "this compound is formed at both temperatures.", + "INVESTIGATIONS ON THE EFFECT OF STRUCTURAL VARIATIONS ON THE COMPOSITION OF THE PYROLYZATES", + "OF SULFUR CONTAINING AMINO ACIDS, by J. M. Patterson, Ph.D., and W. I. Smith, Jr., Ph.D.", + "Department of Chemistry [Contract No. 12-14-100-9575(73)]", + "Methionine and cystine were pyrolyzed in a nitrogen atmosphere at 850°. Salt mixtures", + "which were obtained on pyrolysis were examined by infrared and mass spectroscopy The", + "ether soluble material was separated into neutral, basic, and acidic fractions. The neutral", + "fraction from methionine was separated further by solvent partitioning. Gases were analyzed", + "by infrared spectroscopy. Pyrolysis gases obtained from methionine were carbon disulfide,", + "carbon oxysulfide, carbon dioxide, carbon monoxide, methane, ethylene, acetylene, ammonia,", + "and hydrogen cyanide. The solids obtained on pyrolysis were decomposed on heating to about", + "150°. Gases obtained in this way from methionine (infrared and mass spectra) were carbon", + "dioxide, carbon disulfide, carbon oxysulfide, carbon monoxide, ammonia, and hydrogen", + "cyanide. A similar solid was obtained from the cystine pyrolysis. Thermal decomposition", + "(150°) of the solid produced the following gases: carbon dioxide, carbon oxysulfide, carbon", + "disulfide, ammonia and hydrogen cyanide. These present results are of considerable in-", + "terest because some of the materials produced are usually described as toxic or poisonous.", + "These include carbon monoxide, carbon disulfide and hydrogen cyanide. These materials", + "also probably contribute to the flavor of tobacco smoke.", + "TRANSFERENCE STUDIES OF TRACE ELEMENTS FROM CIGARETTE TOBACCO INTO SMOKE CONDENSATE, AND", + "THEIR DETERMINATION BY NEUTRON ACTIVATION ANALYSIS, by'R. A. Nadkarni, Ph.D., and W. D.", + "Ehmann, Ph.D., Department of Chemistry", + "Abundances of trace elements - La, As, Br, Cr, Se, Ag, Cs, Sc, Zn, Fe, Co, Fe, and Sb -", + "have been determined in tobacco, ashes, and smoke condensate obtained from filter and non-", + "filter varieties of a commercial brand of cigarette. In case of all the elements the", + "filter cigarette was found to prevent the transfer of trace elements from cigarette into", + "smoke condensate more effectively than the nonfilter cigarette. The smoke condensates pre-", + "pared by two different methods - collecting on a Cambridge filter, and collecting in a", + "cold glass-trap - were found to have different concentrations of Fe, Zn, Hg, and Cr.", + "Nondestructive neutron activation analysis was also applied to analyse the Kentucky", + "Reference Cigarette for twelve trace elements. The tobacco from this cigarette appears to", + "be fairly homogenous except for the elements Br and Fe. Hydrated antimony pentoxide (HAP)", + "was used to remove large quantities of 24Na and 42K from the irradiated biological", + "materials. La, As, and Br were also determined in the reference cigarette tobacco and bio-", + "logical standard kale after radiochemical separation of 24Na and 42K on HAP. The latter", + "results are in very good agreement with those obtained by the nondestructive method.", + "RC- 6029179", + "125", + "LG", + "0090691", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.145663, + 0.110282, + 0.736378, + 0.011891 + ], + [ + 0.260319, + 0.121189, + 0.574447, + 0.011626 + ], + [ + 0.145167, + 0.134242, + 0.360811, + 0.013926 + ], + [ + 0.145367, + 0.155066, + 0.735641, + 0.011915 + ], + [ + 0.146825, + 0.167016, + 0.757057, + 0.013786 + ], + [ + 0.146092, + 0.180594, + 0.74264, + 0.013012 + ], + [ + 0.145496, + 0.193621, + 0.736744, + 0.011818 + ], + [ + 0.145719, + 0.205832, + 0.719461, + 0.012105 + ], + [ + 0.146405, + 0.218334, + 0.743999, + 0.011839 + ], + [ + 0.146453, + 0.230949, + 0.702525, + 0.011602 + ], + [ + 0.147045, + 0.243643, + 0.727571, + 0.012226 + ], + [ + 0.152226, + 0.256557, + 0.729993, + 0.012033 + ], + [ + 0.147124, + 0.269195, + 0.72813, + 0.011227 + ], + [ + 0.146379, + 0.283227, + 0.055076, + 0.010606 + ], + [ + 0.802236, + 0.771577, + 0.124432, + 0.013989 + ], + [ + 0.504309, + 0.891914, + 0.024351, + 0.009618 + ], + [ + 0.813736, + 0.97243, + 0.081615, + 0.010917 + ], + [ + 0.219571, + 0.98157, + 0.067782, + 0.012738 + ] + ], + "poly": [ + [ + { + "X": 0.145663, + "Y": 0.110282 + }, + { + "X": 0.882041, + "Y": 0.110282 + }, + { + "X": 0.882041, + "Y": 0.122173 + }, + { + "X": 0.145663, + "Y": 0.122173 + } + ], + [ + { + "X": 0.260319, + "Y": 0.121189 + }, + { + "X": 0.834765, + "Y": 0.121189 + }, + { + "X": 0.834765, + "Y": 0.132814 + }, + { + "X": 0.260319, + "Y": 0.132814 + } + ], + [ + { + "X": 0.145167, + "Y": 0.134242 + }, + { + "X": 0.505977, + "Y": 0.134242 + }, + { + "X": 0.505977, + "Y": 0.148168 + }, + { + "X": 0.145167, + "Y": 0.148168 + } + ], + [ + { + "X": 0.145367, + "Y": 0.155066 + }, + { + "X": 0.881008, + "Y": 0.155066 + }, + { + "X": 0.881008, + "Y": 0.166981 + }, + { + "X": 0.145367, + "Y": 0.166981 + } + ], + [ + { + "X": 0.146825, + "Y": 0.167016 + }, + { + "X": 0.903882, + "Y": 0.167016 + }, + { + "X": 0.903882, + "Y": 0.180801 + }, + { + "X": 0.146825, + "Y": 0.180801 + } + ], + [ + { + "X": 0.146092, + "Y": 0.180594 + }, + { + "X": 0.888731, + "Y": 0.180594 + }, + { + "X": 0.888731, + "Y": 0.193606 + }, + { + "X": 0.146092, + "Y": 0.193606 + } + ], + [ + { + "X": 0.145496, + "Y": 0.193621 + }, + { + "X": 0.88224, + "Y": 0.193621 + }, + { + "X": 0.88224, + "Y": 0.20544 + }, + { + "X": 0.145496, + "Y": 0.20544 + } + ], + [ + { + "X": 0.145719, + "Y": 0.205832 + }, + { + "X": 0.86518, + "Y": 0.205832 + }, + { + "X": 0.86518, + "Y": 0.217936 + }, + { + "X": 0.145719, + "Y": 0.217936 + } + ], + [ + { + "X": 0.146405, + "Y": 0.218334 + }, + { + "X": 0.890404, + "Y": 0.218334 + }, + { + "X": 0.890404, + "Y": 0.230173 + }, + { + "X": 0.146405, + "Y": 0.230173 + } + ], + [ + { + "X": 0.146453, + "Y": 0.230949 + }, + { + "X": 0.848979, + "Y": 0.230949 + }, + { + "X": 0.848979, + "Y": 0.242551 + }, + { + "X": 0.146453, + "Y": 0.242551 + } + ], + [ + { + "X": 0.147045, + "Y": 0.243643 + }, + { + "X": 0.874616, + "Y": 0.243643 + }, + { + "X": 0.874616, + "Y": 0.255869 + }, + { + "X": 0.147045, + "Y": 0.255869 + } + ], + [ + { + "X": 0.152226, + "Y": 0.256557 + }, + { + "X": 0.882219, + "Y": 0.256557 + }, + { + "X": 0.882219, + "Y": 0.26859 + }, + { + "X": 0.152226, + "Y": 0.26859 + } + ], + [ + { + "X": 0.147124, + "Y": 0.269195 + }, + { + "X": 0.875254, + "Y": 0.269195 + }, + { + "X": 0.875254, + "Y": 0.280423 + }, + { + "X": 0.147124, + "Y": 0.280423 + } + ], + [ + { + "X": 0.146379, + "Y": 0.283227 + }, + { + "X": 0.201455, + "Y": 0.283227 + }, + { + "X": 0.201455, + "Y": 0.293834 + }, + { + "X": 0.146379, + "Y": 0.293834 + } + ], + [ + { + "X": 0.802236, + "Y": 0.771577 + }, + { + "X": 0.926668, + "Y": 0.771577 + }, + { + "X": 0.926668, + "Y": 0.785566 + }, + { + "X": 0.802236, + "Y": 0.785566 + } + ], + [ + { + "X": 0.504309, + "Y": 0.891914 + }, + { + "X": 0.528661, + "Y": 0.891914 + }, + { + "X": 0.528661, + "Y": 0.901532 + }, + { + "X": 0.504309, + "Y": 0.901532 + } + ], + [ + { + "X": 0.813736, + "Y": 0.97243 + }, + { + "X": 0.895352, + "Y": 0.97243 + }, + { + "X": 0.895352, + "Y": 0.983347 + }, + { + "X": 0.813736, + "Y": 0.983347 + } + ], + [ + { + "X": 0.219571, + "Y": 0.98157 + }, + { + "X": 0.287353, + "Y": 0.98157 + }, + { + "X": 0.287353, + "Y": 0.994308 + }, + { + "X": 0.219571, + "Y": 0.994308 + } + ] + ], + "score": [ + 0.9965, + 0.9896, + 0.8836, + 0.9926, + 0.9887, + 0.9953, + 0.9902, + 0.9907, + 0.9949, + 0.9514, + 0.9537, + 0.9986, + 0.9987, + 0.9803, + 0.8493, + 0.9977, + 0.9824, + 0.9834 + ], + "text": [ + "INVESTIGATIONS ON THE COMPOSITION OF THE PYROLYZATES OF MALEIC HYDRAZIDE AND N,N-DIMETHYL", + "by W. T. Smith, Jr., Ph.D., and J. M. Patterson, Ph.D., Department of", + "Chemistry [Contract No. 12-14-100-10271(73))", + "The sucker control agents maleic hydrazide and N,N-dimethyldodecylamine were pyrolyzed at", + "850° in a nitrogen atmosphere. Gases from maleic hydrazide included carbon monoxide, hydro-", + "gen cyanide, ammonia and carbon dioxide. The condensable pyrolyzate consisted of aromatic", + "nitriles such as benzonitrile, tolunitriles, cinnamonitrile, naphthonitriles, nitrogenous", + "bases, phenols and aromatic hydrocarbons such as naphthalene, phenanthrene, anthracene,", + "fluoranthene and pyrene. Both the pyrolyzates from maleic hydrazide and N,N-dimethyldode-", + "lamine contained benzo[alpyrene. The formation of carbon monoxide, hydrogen cyanide,", + "ammonia, phenols from maleic hydrazide and benzo[a]pyrene from maleic hydrazide and N,N-", + "limethyldodecylamine has a direct bearing on the problem of tobacco and health because of", + "the extensive use of these substances for the prevention of sucker production in tobacco", + "plants.", + "RC- 6029180", + "126", + "0090692", + "Source:" + ] + }, + { + "bbox": [ + [ + 0.481301, + 0.136657, + 0.058075, + 0.009965 + ], + [ + 0.150965, + 0.175133, + 0.681023, + 0.01364 + ], + [ + 0.150052, + 0.188592, + 0.69562, + 0.01164 + ], + [ + 0.152392, + 0.200541, + 0.157401, + 0.010126 + ], + [ + 0.150095, + 0.217744, + 0.721196, + 0.011852 + ], + [ + 0.150619, + 0.231064, + 0.704944, + 0.010575 + ], + [ + 0.150732, + 0.243327, + 0.721435, + 0.011065 + ], + [ + 0.150202, + 0.255126, + 0.696794, + 0.012671 + ], + [ + 0.152572, + 0.268465, + 0.686258, + 0.011898 + ], + [ + 0.151135, + 0.280484, + 0.727981, + 0.011233 + ], + [ + 0.149772, + 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"PILOT STUDIES ON THE EFFECT OF TOBACCO SMOKE CONSTITUENTS ON RESPIRATORY ENZYMES, by", + "M. I. H. Aleem, Ph.D., and c. Gairola, Ph.D., Department of Microbiology [Contract No.", + "12-14-100-10280(73)]", + "Yeast (Saccharomyces carlbergensis) was grown aerobically and the effect of tobacco smoke", + "condensates on cell growth and on cellular enzymes was studied. Rat-heart mitochondria", + "and bacterial membrane fractions were included for comparative work. A 75% inhibition in", + "yeast growth occurred in the presence of 2 mg/ml of added acetone-suspended condensate", + "(ASC). A 93% inhibition of the mitochondrial succinoxidase activity was caused by as", + "little as 0.7 mg/ml of ASC. Both the NADH and succinate oxidase activities were inhibited", + "by 80 to 100% upon a one-minute exposure of the mitochondrial preparations to smoke. A1-", + "though the denicotinized condensates inhibited growth of yeast and the respiratory enzymes", + "from yeast and rat-heart mitochondria, nicotine virtually unaffected the respiratory pro-", + "cess, and, in fact, caused a stimulation in respiration. These findings may be directly", + "applicable to the energy metabolism of man as affected by tobacco smoke and/or its con-", + "stituents, and can also be applied for a rapid bioassay for the potency of commercial", + "cigarettes.", + "INVESTIGATIONS ON THE EFFECT OF TOBACCO SMOKE CONSTITUENTS ON RODENT TRACHEAL RINGS GROWN", + "IN TISSUE CULTURE, by Grace M. Donnelly, Ph.D., Department of Microbiology, and Robert", + "Penman, M.D., M.R.C.P. Department of Medicine [Contract Nos. 12-14-100-9480(7 and 12-", + "14-100-10274(73)]", + "Several methods of exposing rodent tracheal rings in vitro to tobacco products have been", + "developed, and these have proven capable of producing measured responses which are time/", + "concentration related. These methods include assessment of histological conditions based", + "on both immediate response and recovery capabilities; measures of ciliostasis induced by", + "aqueous extract and vapor phase; and ciliostasis profiles derived from vapor phase cilio-", + "stasis times. Our method of measuring ciliostasis, whereby we use complete ciliostasis", + "around the entire ring, provides a simple, clean, and nonsubjective endpoint. When com-", + "bined with correlative histological studies, ciliostasis need not be merely an empirical", + "bioassay method, but one which can provide information relative to the biological action of", + "the test material. The ciliostasis methods described in this report could be made immedi-", + "ately applicable as bioassay and/or screening operations useful in the identification of", + "less harmful tobacco products. The choice of endpoint and method of quantification which", + "we have described should increase the overall usefulness of ciliostasis techniques in", + "general.", + "133", + "EFFECTS OF TOBACCO SMOKE INHALATION ON Xe PULMONARY WASHOUT TIME AND ON CARDIAC PARAM-", + "ETERS OF RATS, by H. A. Gordon, M.D., and B. F. Schwartz, B.S., Department of Pharmacology,", + "[Contract No. 12-14-100-10277(73)", + "The purpose of this work was to study the effects of acute tobacco smoke exposure on the", + "pulmonary membrane and on cardiac function of experimental animals. Using an apparatus", + "which simulated conditions of human smoking, anesthesized rats inhaled tobacco smoke of", + "graded nicotine content over a period of 30 min. via a tracheotomic cannula connected to a", + "respirator pump. The pneumogram, electrocardiogram, and the emergence and washout time of", + "intravenously administered 133xe doses were recorded. The observations indicated little", + "or no difference in the emergence or washout time of the label before, during, and after", + "the exposure to various types of smoke. In cardiac function, bradycardia and anomalies of", + "the P waves were observed which in severity appeared concordant to the concentration of", + "nicotine in the inhaled smoke. Previous atropinization of the rats largely prevented the", + "development of such cardiac effects. These results indicate that on first impact of smoke", + "inhalation, cardiac anomalies mediated via the parasympathetic system may be present in", + "the animal host, when the respiratory membrane of the lungs still shows unimpaired function.", + "RO- 6029181", + "127", + "LG 0090693", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.153816, + 0.121426, + 0.728732, + 0.010554 + ], + [ + 0.153433, + 0.133554, + 0.690023, + 0.014696 + ], + [ + 0.153345, + 0.147301, + 0.54931, + 0.012567 + ], + [ + 0.154118, + 0.16478, + 0.754427, + 0.013904 + ], + [ + 0.154003, + 0.177995, + 0.761108, + 0.013792 + ], + [ + 0.154683, + 0.192346, + 0.754006, + 0.010805 + ], + [ + 0.155234, + 0.203837, + 0.745337, + 0.012087 + ], + [ + 0.154712, + 0.216233, + 0.746309, + 0.013068 + ], + [ + 0.154565, + 0.22849, + 0.746896, + 0.011884 + ], + [ + 0.15534, + 0.241742, + 0.746401, + 0.011482 + ], + [ + 0.154581, + 0.253939, + 0.747017, + 0.012984 + ], + [ + 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G. Green, Ph.D., Department of Agronomy, and I. Yoneyama, M.D., Department of", + "Pathology [Contract Nos. 12-14-100-9483(73 and 12-14-100-9574(73)", + "During the past year (1969), renovation of the laboratory facility and procurement of newly", + "authorized equipment was completed. Two dose related acute toxicity studies were completed,", + "and four related long-term tumorigenicity experiments were initiated. The results from one", + "toxicity study indicate that aged condensates (greater than 24 hours old) from the Univer-", + "sity of Kentucky Reference Cigarette (Lot 1R1) will be suitable for mouse skin bioassay at", + "dose levels to 450 mg/wk for whole condensates and to 500 mg/wk for acid extracted conden-", + "sates following suitable acclimatization procedures. The facility has attained a standing", + "population of approximately 2,700 animals. An additional capacity equal to ten percent of", + "the standing population is utilized to house isolated animals which require more space (7", + "to 8 times) than nonisolated populations. Procedures now employed for animal care, treat-", + "ment, and record keeping appear to be adequate.", + "INVESTIGATIONS ON THE INFLUENCE OF CARCINOGENS, IRRITANTS AND CIGARETTE SMOKE ON RESPIRATORY", + "EPITHELIAL CELL TURNOVER RATES IN MICE, by Irene Roeckel, Dr.M.Sc., Department of Pathology", + "[Contract No. 12-14-100-9520(73)", + "Cell turnover in inbred mice is quite distinct in mice with \"high spontaneous\" tumor in-", + "cidence and \"low spontaneous\" tumor incidence. The tumor prone animals have a slower cell", + "turnover than the animal not developing tumors. The intranasal introduction of known car-", + "cinogens, irritants, and tobacco smoke condensate was carried out on these animals and the", + "cell turnover rate determined to permit comparison with untreated animals. It was neces-", + "sary to cover a wide range of applications over as much as 5 months to observe short- and", + "long-term effects. Although at this stage the final characterization of know carcinogens,", + "irritants and tobacco smoke condensate is not complete, some treated animals show a very", + "marked change from their normal cell replication pattern. These changes range from decrease", + "to increase in cell turnover time. The final evaluation and characterization is now in", + "progress.", + "INVESTIGATIONS ON THE EFFECT OF TOBACCO SMOKE COMPONENTS ON PLANT CELLS IN TISSUE CULTURE,", + "by P. s. Sabharwal, Ph.D., and P. R. Bhalla, Ph.D., Department of Botany [Contract No.", + "12-14-100-10278(73)", + "Studies were conducted to determine the effects of tobacco smoke components on the callus", + "tissue from tobacco pith maintained in tissue culture. Modified Murashige's medium con-", + "taining IAA (2 ppm), kinetin (2 ppm), and sucrose (2%) served as the control. Different con-", + "centrations (ranging from 10-4M to 10-10M) of the following smoke components were utilized:", + "acrolein, benzo{a]pyrene, benzo[e]pyrene, pyrene, and phenol. About 350-500 mg of the to-", + "bacco pith callus was used as inoculum. The cultures were observed for 5 weeks", + "for", + "effects", + "on growth caused by the presence of tobacco smoke components in the medium. Benzo[a]pyrene", + "(10-6M--10-9M) caused extensive callusing of the tissue. Other tobacco smoke components", + "have proved ineffective on this system. These observations clearly demonstrate that plant", + "tissue cultures can be used for rapid and sensitive bioassay of benzo[a]pyrene.", + "INVESTIGATIONS ON THE EFFECT OF AQUEOUS EXTRACTS OF CIGARETTE SMOKE ON TUMORIGENESIS IN", + "RATS, by K. L. Sydnor, M.D., Department of Medicine (Contract No. 12-14-100-9527 (73)]", + "The aqueous extract of cigarette smokercondensate from a commercial brand of cigarettes", + "was administered in 5% sugar solution as drinking fluid to Sprague-Dawley strain female", + "rats from 30 days of age until autopsy. (This extract [AECSC] was prepared by the Smoke", + "Laboratory at this University.) Concentrations of AECSC greater than 1 mg/ml 5% sugar", + "solution induced marked loss of body weight as a consequence of dehydration and decreased", + "food intake; a concentration of 1 mg/ml inhibited normal weight gain which could not be", + "attributed to decreased food intake. A concentration of 0.125 mg/ml permitted near optimal", + "weight gain; the average daily dose of AECSC per rat was estimated to be 3.8 mg. Adapta-", + "tion to the foregoing fluid regimen was complete within two weeks and did not change appre-", + "ciably for two months.", + "Four groups of Sprague-Dawley strain female rats, age 30 days, were given 0.125 mg AECSC/ml", + "5% sugar solution and four groups were given a comparable volume of 5% sugar solution alone", + "128", + "RC- 6029182", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch000", + "0090694" + ] + }, + { + "bbox": [ + [ + 0.156048, + 0.127957, + 0.009993, + 0.005507 + ], + [ + 0.17196, + 0.120293, + 0.713246, + 0.016256 + ], + [ + 0.156113, + 0.132055, + 0.715344, + 0.017017 + ], + [ + 0.155446, + 0.146365, + 0.731412, + 0.015669 + ], + [ + 0.15654, + 0.158247, + 0.706206, + 0.016311 + ], + [ + 0.155942, + 0.170351, + 0.723418, + 0.015919 + ], + [ + 0.155619, + 0.184, + 0.691531, + 0.015875 + ], + [ + 0.156244, + 0.19786, + 0.233008, + 0.011694 + ], + [ + 0.156364, + 0.213432, + 0.715011, + 0.016046 + ], + [ + 0.156317, + 0.226501, + 0.542391, + 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Each group consisted of twenty animals and was injected subcutaneously with 400,", + ":00, or 100 micrograms benzo[alpyrene (BaP) in 0.1 ml sesame oil on alternate days for 30", + "loses. An additional group was similarly injected with sesame oil. All rats injected with", + "ia? developed fibrosarcomata at the site(s) of injection. Mean tumor induction time for", + "the three groups drinking AECSC was 80 to 83 days, and for the three groups drinking sugar", + ";olution, 95 to 98 days. No tumors were observed in the two control groups sacrificed", + "after five months observation.", + "The foregoing observations indicated that the doses of BaP were too high for use for bio-", + "issay purposes and that AECSC might accelerate tumor induction time.", + "ENVESTIGATIONS ON THE TUMORIGENIC ACTIVITY OF AQUEOUS EXTRACT OF CIGARETTE SMOKE CONDENSATE,", + "cy K. L. Sydnor, M.D., Department of Medicine [Contract No. 12-14-100-9484(73)]", + "more precisely define the dose-response range of benzo[a]pyrene (BaP) using the develop-", + "nent of cancer at the injection site as a parameter for \"response,\" and to delineate more", + ":learly the influence of the aqueous extract of cigarette smoke condensate (AECSC) on", + ":morigenesis, five groups of 20 Sprague-Dawley female rats, age 30 days, were given AECSC", + ".25 mg/ml of 5% sugar solution as drinking solution from 30 days of age until autopsy.", + "four of the groups were injected with 50, 25, 12.5, or 3 ug BaP dissolved in 0.1 ml sesame", + ":il by the subcutaneous route on alternate days for 30 doses. The fifth group was injected", + "ith sesame oil. Five additional groups of 20 rats were given a comparable volume of 5%", + "sugar solution as drinking fluid, of which four were similarly injected with BaP and the", + "lifth with sesame oil. Tumor-bearing rats were sacrificed 33 + 3 days following detection", + ":f the first palpable tumor; control rats injected with sesame oil were observed 375 + 10", + "lays. All rats given AECSC to drink and injected with BaP developed fibrosarcomata at", + "the", + "site of injection. Mean tumor induction time was 97 + 4.7, 104 + 3.2, 127 + 6.5, and", + ":35 + 15 days for the groups given the 50, 25, 12.5, and 3-microgram doses respectively.", + "in the groups given sugar solution, rats injected with 50, 25, or 12.5 ug BaP developed", + "sarcomata and mean tumor induction time was similar to comparable groups drinking AECSC.", + "in the group injected with 3-pg dose only 9 of 19 rats had developed tumors by 365 days.", + "These observations indicate that the continuous ingestion of \"nontoxic\" quantities of AECSC", + "lecelerates chemical carcinogenesis by mechanism(s yet undetermined. It is presumed that", + "IECSC contains a co-carcinogen.", + "in the other hand, the continuous ingestion of AECSC did not affect the carcinogenic effect", + ":f BaP when 100 mg of the compound were administered as a single or as divided (20 mg q wk)", + "i 5) doses by gastric tube at 50 days of age. Approximately 20-25% of the animals devel-", + "oped mammary cancer. The overall incidence of breast tumors (fibroadenoma and adenocar-", + "cinoma) and other cancers was slightly but not significantly less in the groups fed AECSC.", + "Freliminary data, using tritium-labelled BaP as substrate, indicates that livers from rats", + "riven AECSC in 5% sugar solution metabolize the carcinogen more rapidly than livers from", + "ists given sugar solution alone. Liver menadione reductase activity was likewise lower in", + "the livers from control animals.", + "INVESTIGATIONS ON THE DETERMINATION OF THE EFFECTS OF COMBINATIONS OF CIGARETTE SMOKE", + "AND NONONCOGENIC VIRUSES ON THE DEVELOPMENT OF LUNG TUMORS IN RODENTS, by D. L. Weiss,", + "L.D., Department of Pathology [Contract Nos. 12-14-100-9482(73 and 12-14-100-10276(73)", + "&", + "variety of studies of the application of viral agents, classical chemical carcinogens,", + "and tobacco smoke condensate, separately and together, have been pursued in order to de-", + "velop information pertinent to the construction of alternate means of bioassay for onco-", + "genetic potential.", + "Direct application of tobacco smoke condensate to the shaved mouse back has been studied", + "to attempt to identify the mechanism of tumor formation and the parameters which will first", + "identify the presence of malignancy. To this time, loss of cell adhesion and the constant", + "presence of microbodies interpreted as viral particles have been the major observations in", + "the skin. Frank invasive malignant tumors have not been developed in the mouse-back", + "studies, leaving this area to be investigated when they become available.", + "RC:- 6029183", + "129", + "LG", + "0090695", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.130306, + 0.119456, + 0.733467, + 0.009959 + ], + [ + 0.130387, + 0.130789, + 0.732814, + 0.013162 + ], + [ + 0.130883, + 0.145502, + 0.732281, + 0.011402 + ], + [ + 0.131392, + 0.158361, + 0.146869, + 0.010949 + ], + [ + 0.130721, + 0.1751, + 0.732319, + 0.011997 + ], + [ + 0.131121, + 0.188822, + 0.749572, + 0.011149 + ], + [ + 0.13179, + 0.201014, + 0.756403, + 0.011495 + ], + [ + 0.130763, + 0.213319, + 0.749706, + 0.011826 + ], + [ + 0.131303, + 0.225168, + 0.76257, + 0.012816 + ], + [ + 0.13017, + 0.243768, + 0.734349, + 0.012345 + ], + [ + 0.130318, + 0.257474, + 0.733794, + 0.012179 + ], + [ + 0.130704, + 0.270177, + 0.749983, + 0.011393 + ], + [ + 0.131018, + 0.282447, + 0.630892, 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It must be left open to question whether these changes are a measure of on-", + "cogenic potential.", + "The same material consistently produced microthrombi in the small vessels in the area of", + "inoculation. This is made particularly evident when the vascular bed necessary to support", + "a transplanted bit of pulmonary tissue becomes incompetent because of such microthrombosis.", + "The significance of this biological action of tobacco smoke condensate is discussed in re-", + "ports in relation to chronic lung disease, vascular disease and anaerobic tissue metabolism.", + "Multiple studies of the utility of lung transplantation as a procedure for adaptation to", + "oncogenic bioassay have proven inconclusive. A large amount of basic data regarding the", + "reactions, survival, and interpretation of transplantation with and without potential car-", + "cinogens, and with and without host immunosuppression have been accumulated.", + "The young Syrian hamster has not proven to be a useful bioassay animal for respiratory sys-", + "tem oncogenesis in the single exposure system used. Young mice have proven far more useful", + "for this purpose.", + "Transplantation of cell suspensions from the respiratory tree have been successful and pro-", + "vide an isolated cell matrix in which oncogenicity evaluations might be performed.", + ";", + "RO- 6029184", + "130", + "LG", + "0090696", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.441656, + 0.111554, + 0.059342, + 0.008339 + ], + [ + 0.113657, + 0.146036, + 0.671997, + 0.017513 + ], + [ + 0.113581, + 0.157665, + 0.622066, + 0.016443 + ], + [ + 0.114776, + 0.172231, + 0.253909, + 0.016491 + ], + [ + 0.113849, + 0.189712, + 0.744122, + 0.017696 + ], + [ + 0.119766, + 0.200983, + 0.741042, + 0.019928 + ], + [ + 0.113794, + 0.213429, + 0.731178, + 0.019337 + ], + [ + 0.11434, + 0.226625, + 0.714063, + 0.018018 + ], + [ + 0.114462, + 0.238671, + 0.739502, + 0.019275 + ], + [ + 0.112883, + 0.251322, + 0.724133, + 0.018046 + ], + [ + 0.113996, + 0.264804, + 0.730349, + 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W. Flesher, Ph.D., Department of Pharmacology", + "[Contract No. 12-14-100-9516(73)", + "Tritiun-labelled benzo[alpyrene (BaP) was prepared by the method of acid-catalyzed exchange.", + "rapid and convenient radioassay for the metabolism of the hydrocarbon by rat liver homog-", + "mates has been developed. The radioassay products have been tentatively identified from", + "their mobility on TLC as 3-hydroxybenzo[a)pyrene, --hydroxybenzo[a]pyrene, 3,6-quinone", + "and 1,6-quinone. In addition a number of other products are formed. Conversion of BaP to", + "water-soluble and protein bound forms is also followed by radioactivity measurements and", + "an increase in metabolism of the hydrocarbon results in a corresponding increase in these", + "forms. Although the behavior of BaP appears to parallel that of other carcinogenic hydro-", + ":arbons, much additional study will be required to gain an understanding of its mechanism", + "of action.", + "ELLULAR MECHANISMS OF TOBACCO PRODUCTS CARCINOGENESIS, by William R. Jewell, M.D., Depart-", + "nent of Surgery [Contract Nos. 12-14-100-9481(73) and 12-14-100-10275(73J]", + "This project was initiated to attempt to identify underlying host factors in methylcholan-", + "threne (MC) and tobacco products carcinogenesis. The identification of a human red blood", + "hemadsorption factor unique to MC sarcoma (Sa) suggested a virus or virus-like etiology", + ": line of identity between MC and tobacco products-induced tumors was demonstrated when the", + "latter was found to possess identical hemadsorption properties. A factor present in MCSa", + ":apable of co-carcinogenic or perhaps enhancement of tumor formation was subsequently", + "identified. This factor(s) was shown to be present in two ultracentrifuge factors, i.e.", + "iraction I containing nuclei cell debris and cell membrane (700 x 8 for 10 min.) and in", + "fraction III containing the microsomes (100,000 x 8 for 30 min.) Attempts to induce a", + "talignant transformation in mouse embryo tissue culture (METC) cells with a cell free homog-", + "mate of MCSa plus MC yielded cytologic changes suggestive of neoplastic change, i.e., in-", + ":reased granularity, accelerated growth and a peculiar process of the cell membrane. How-", + "ever, when the METC cells were innoculated into weanling mice no tumor developed. Further", + "evidence of a viral or virus-like factor in MC carcinogenesis was found when an antigen", + "mique to MCSa was identified using Ouchterloney in gel diffusion techniques. Recent", + "studies demonstrating suppression of MCSa formation in mice preinfected with mouse hepati-", + "tis virus, a known producer of viral interferons further substantiates this view.", + "!ECHANISM OF INTESTINAL ABSORPTION OF BENZO[A]PYRENE, by P. Mandelstam, M.D., Ph.D. and", + "E. D. Rees, M.D., Department of Medicine [Contracti No. 12-14-100-9525 (73)]", + "Though the polynuclear aromatic hydrocarbons are readily absorbed through the intestinal", + "tract, little is known about the mechanism of their absorption. Using the everted intesti-", + "nal sac technique, we have studied the absorption of several of these compounds, including", + "enzo[a]pyrene which is representative of these compounds and which has been studied in", + "the most detail. Uptake of these compounds by sac tissue resembles the uptake of fatty", + "acid and cholesterol in that it is not *appreciably influenced by inhibition of respiration", + "and/or glycolysis. However, the polynuclear aromatic hydrocarbons are unique in that accu-", + "zulation in intestinal sac tissue increases exponentially with increase in the incubation", + "sedium concentration. To account for this unexpected finding, we postulate that first", + "uiliayer (Langmuir) adsorption occurs on the mucosal surface of the intestine and then", + "multilayer (Polanyi) adsorption follows. The latter would explain the exponential rela-", + "tionship and may reflect the extreme hydrophobicity and the planar structure of these com-", + "pounds. The passage of compounds from sac tissue into the fluid within the sac occurs in", + "direct proportion to sac tissue concentration, a finding consistent with passive diffusion.", + "In vivo experiments show that an exponential relationship exists between the intragastric", + "dose given in sesame oil and accumulation in the adipose tissue and mammary glands of rats.", + "In addition, we have shown that the feature of exponential adsorption is not limited to", + "living tissue, for adsorption isotherms and time curves of other material incubated in the", + "same media have the same forms as those obtained for the intestinal sacs. These data are", + "consistent with the concept that benzo [alpyrene and related compounds enter other tissues", + "by the same non-specific mechanism observed in intestinal absorption.", + "131", + "RO- 6029185", + "L.G.", + "0090697", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.153053, + 0.118671, + 0.726274, + 0.010402 + ], + [ + 0.153048, + 0.130602, + 0.772935, + 0.012919 + ], + [ + 0.155859, + 0.144819, + 0.267911, + 0.013004 + ], + [ + 0.153451, + 0.161922, + 0.724945, + 0.012996 + ], + [ + 0.153958, + 0.175513, + 0.758759, + 0.012508 + ], + [ + 0.154458, + 0.188961, + 0.750192, + 0.012923 + ], + [ + 0.154235, + 0.203266, + 0.025558, + 0.009346 + ], + [ + 0.186667, + 0.202952, + 0.017163, + 0.008968 + ], + [ + 0.211932, + 0.201019, + 0.674735, + 0.012283 + ], + [ + 0.157514, + 0.21281, + 0.748189, + 0.013727 + ], + [ + 0.154436, + 0.228791, + 0.032877, + 0.008627 + ], + [ + 0.195441, + 0.227744, + 0.083307, + 0.009082 + ], + 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AROMATIC HYDRO-", + "CARBONS: CORRELATION WITH CANCER INDUCTION, by E. Douglas Rees, M.D., Department of Medicine,", + "[Contract No. 12-14-100-9524 (73)1", + "Both ,,12-dimethylbenz[a) anthracene (DMBA) and7,8,12-trimethylbenz[a]anthracene (TMBA)", + "cause breaks in rat marrow cell chromosomes when these compounds are given intravenously in", + "a lipid emulsion. The maximum incidence of breaks is found within 6-24 hours with 43% and", + "55%", + "of", + "metaphase cells having chromosomes with breaks after DMBA and TMBA, respectively.", + "Benzo[a]pyrene (BaP) and Benzo[e]pyrene (BeP) have been similarly studied and have respec-", + "tive", + "incidences", + "of 21% and 11%. The distribution of breaks on the various chromosomes and", + "the position of breaks on individual damaged chromosomes have been determined and summarized", + "for each of thse compounds. Although damage is inflicted on chromosomes of all sizes and", + "morphology, the damage is nonrandom in that members of the #1 and #2 chromosome pairs are", + "damaged to an extent greater than expected on the basis of their size and number. In addi-", + "tion, the total number of breaks per chromosome for the individually identifiable chromo-", + "somes was calculated. Distinct differences between the four compounds were revealed; e.g.", + "TMBA was more damaging to the #1 chromosome by 1.25, 2.6, and 10 times, respectively, than", + "was DMBA, BaP, and BeP. These rankings (TMBA DMBA > BaP BeP) are in accord with our", + "data for the carcinogenicity of these compounds. These findings provide further evidence", + "that the ability of a polynuclear aromatic hydrocarbon to cause cancer may be related to", + "its ability to inflict extensive damage on certain chromosomes.", + "INVESTIGATIONS ON THE EFFECT OF TOBACCO SMOKE ON THE FORMATION OF A MICROBIAL MEMBRANE-", + "BOUND ELECTRON TRANSPORT SYSTEM, by D. C. White, M.D., Ph.D., Department of Biochemistry.", + "[Contract No. 12-14-100-9517(73)", + "Using the S. aureus system developed in Contract II, it has been shown that the smoke car-", + "cinogen benzolalpyrene markedly inhibited aerobic growth without affecting anaerobic growth", + "at 10-PM. The noncarcinogenic isomer was not inhibitory in either condition.", + "Adding air to anaerobic cultures induces the formation of a membrane-bound electron trans-", + "port system. The most striking change in the lipid membrane in this period is the rapid", + "synthesis of the rubixanthin carotenoids. From studies with 14c it appears that most of", + "the rubixanthin is formed by the hydroxylation of the cyclic S-carotene in the presence of", + "oxygen. A minor pathway via phytofluenol and its cyclization that does not require oxygen", + "was also detected. The oxygen requirement suggested a mixed function oxidase might be in-", + "volved. Specific inhibitors of the mammalian mixed function oxidases which hydroxylate", + "drugs, carcinogens like benzo[a]pyrene, or steroids markedly inhibited the aerobic synthe-", + "sis of rubixanthin in S. aureus. Piperonyl butoxide appeared to specifically depress the", + "hydroxylation of S-carotene to rubixanthin without other effects. Other inhibitors showed", + "this as the major effect. The mixed function oxidases in liver microsomes contain a hemo-", + "protein P450. A very similar carbon monoxide binding pigment has been shown to be induced", + "in S. aureus on the initiation of aerobic growth. Since benzo[a]pyrene in mammalian cells", + "is hydroxylated by mixed function oxidases, perhaps the high biological activity of this", + "carcinogen in aerobic cultures results from the hydroxylated carcinogen. This will be", + "tested directly in this system and the biological activity of the carcinogen and its de-", + "rivatives will be examined closely.", + "RC- 6029186", + "1", + "132", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgchob&", + "0090698" + ] + }, + { + "bbox": [ + [ + 0.222738, + 0.11654, + 0.581069, + 0.010419 + ], + [ + 0.492047, + 0.14024, + 0.050494, + 0.009702 + ], + [ + 0.158799, + 0.165482, + 0.735142, + 0.012449 + ], + [ + 0.177307, + 0.177579, + 0.445059, + 0.012251 + ], + [ + 0.159221, + 0.192741, + 0.062549, + 0.009815 + ], + [ + 0.231531, + 0.190922, + 0.645827, + 0.010835 + ], + [ + 0.177409, + 0.20236, + 0.478468, + 0.011307 + ], + [ + 0.160208, + 0.214797, + 0.696866, + 0.013211 + ], + [ + 0.176383, + 0.228105, + 0.729932, + 0.011639 + ], + [ + 0.159525, + 0.240769, + 0.71899, + 0.011737 + ], + [ + 0.176444, + 0.252891, + 0.723409, + 0.011758 + ], + [ + 0.177576, + 0.265311, + 0.363329, + 0.011688 + ], + [ + 0.159531, + 0.277546, + 0.718629, + 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PUBLICATIONS BY TOBACCO AND HEALTH RESEARCH INVESTIGATORS", + "AREA I", + "Andersen, R. A. and Todd, J. R. Estimation of total tobacco plant phenols by their bonding", + "to polyvinylpyrrolidone. Tobacco Sci. XII:107-11, 1968.", + "Andersen,", + "R. A. and Sowers, J. A. Optimum conditions for bonding of plant phenols to in-", + "soluble polyvinylpyrrolidone. Phytochem. 7,2:293-301, 1968.", + "Andersen, R. A.; Lowe, R.; and Vaughn, I. A. Plant phenols and polyphenoloxidase in N.", + "Tabacum during greenhouse growth, field growth and air-curing. Phytochem. 8:2139-47, 1969.", + "Atkinson, W. 0.; Ragland, J. L.; Sims, 3. L.; and Bloomfield, B. J. The influence of ir-", + "rigation on the responses of burley tobacco to nitrogen-fertilization (Abstract). Assoc.", + "of Southern Agric. Workers, Proc.65:64, 1968.", + "Atkinson, W. 0.; Ragland, J. L.; Sims, J. L.; and Bloomfield, B. J. Nitrogen composition", + "of burley tobacco. I. The influence of irrigation on the responses of burley tobacco to", + "nitrogen fertilization. Tobacco Sci. XIII:123-6, 1969.", + "Bush, L. P. and Sims, J. L. Accumulation of volatile aliphatic secondary amines in", + "Nicotiana", + "spp. (Abstract) XI International Botanical Congress, September 2-4, 1969.", + "Bush, L. P. Cation requirement for succinic thiokinase of Nicotiana tabacum (Abstract)", + "Plant Physiol. 43 (Supp.):51, Aug. 20-23, 1968.", + "Bush, L. P. Influence of certain cations on activity of succiny1 CoA synthetase from to-", + "bacco. Plant Physiol. 44:347-50, 1969.", + "Bush, L. P. Succinyl CoA synthetase of tobacco. Physiol. Plantarum. 22:1097-104, 1969.", + "bush, L. P. Succinyl thiokinase of tobacco (Abstract) Agron. Abstr. Nov. 5-10:25, 1967.", + "Cheng, A. L. S.; Chaplin, J. F.; and Iso, I. c. Sterol variation in flue-cured tobacco vari-", + "eties. Tobacco Sci. XII:33-4, 1968.", + "Collins, G. B. and Legg, P. D. Cytogenetics of a burley tobacco breeding line derived from", + "a cross with N. Longiflora cau. Can. J. Genet. 11:382-8, 1969.", + "Grunwald, C. The effect of ribonuclease on the binding of calcium to mitochondria. Planta", + "83:314-22, 1968", + "Grunwald, c. Effect of sterols on the permeability of alcohol-treated red beet tissue.", + "Plant Physiol. 43, 4:484-8, 1968.", + "Grunwald, C. The effects of inhibitors on the efflux of 8-cyanin and aerobic respirn in", + "red beet tissue. Planta 90:1-11, 1970.", + "Keller, c. J.; Bush, L. P.: and Grunwald, c. Changes in content of sterols, alkaloids, and", + "phenols in flue-cured tobacco during conditions favoring infestation of molds. J. Agr.", + "Food Chem. 17:331-4, 1969.", + "Keller, c. J.; Bush, L. P.; and Grunwald, c. The effect of microorganisms on the quantita-", + "tive and qualitative content of alkaloids, sterols and polyphenols in tobacco leaves.", + "(Abstract) Plant Physiol. 43 (Supp.) Aug. 20-23, 1968.", + "Keller, c. J.; Bush, L. P.; and Grunwald, c. Effects of fungus infestation on some chemical", + "constituents of tobacco leaves during post-harvest processing. (Abstract) Agron. Abstr.", + "Nov. 10-18:35, 1968.", + "Keller, c. J. and Kasperbauer, M. J. Effect of storage method on content of chlorogenic", + "acid and rutin in tobacco. J. Agr. Food Chem. 17:327-30, 1969.", + "Legg, P. D.; Chaplin, J. F.; and Collins, G. B. Inheritance of percent total alkaloids in", + "N. tabacum L.--populations derived from crosses of low alkaloid lines with burley and", + "flue-cured varieties. J. Heredity 60:213-17, 1969.", + "Sheen, S. J. Comparative quantities of polyphenols and oxidases in Nicotiana species.", + "Tobacco Sci. XIV:16-9, 1970.", + "Sheen, s. J. The distribution of polyphenols, chlorogenic acid oxidase and peroxidase in", + "different plant parts of tobacco, N. tabacum L. Phytochem. 8:1839-47, 1969.", + "Sheen, S. J.; Calvert, J.; Rebagay, G. R. Effects of nitrogen fertilization of polyphenol", + "content and oxidase activities in tobacco seedlings. Crop Sci. 9:547-50, 1969.", + "Sheen, S. J. and Calvert, J. Polyphenol content activities and isozymes of polyphenol oxi-", + "dase and peroxidase during air-curing in the tobacco types. Plant Physiol. 44,2:199-204,", + "1969.", + "Sheen, S. J. and Calvert, J. Quantitative variation in polyphenol content in the green and", + "air-cured leaves of tobacco (N. Tabacum L.) Tobacco Sci. XII:10-2, 1969.", + "Sheen, S. J. and Calvert, J. Studies on polyphenol content, isozymes and activities of", + "133", + "RC- 6029187", + "LG 0090699", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0009" + ] + }, + { + "bbox": [ + [ + 0.191898, + 0.107976, + 0.744451, + 0.011744 + ], + [ + 0.193643, + 0.120426, + 0.113826, + 0.010205 + ], + [ + 0.168684, + 0.131296, + 0.762709, + 0.014186 + ], + [ + 0.193865, + 0.145621, + 0.694293, + 0.011251 + ], + [ + 0.169201, + 0.15791, + 0.695878, + 0.011979 + ], + [ + 0.195707, + 0.169795, + 0.360736, + 0.013168 + ], + [ + 0.167934, + 0.183711, + 0.763808, + 0.011715 + ], + [ + 0.193072, + 0.195156, + 0.297934, + 0.011686 + ], + [ + 0.516055, + 0.221204, + 0.059662, + 0.00901 + ], + [ + 0.168206, + 0.245871, + 0.72115, + 0.011913 + ], + [ + 0.192094, + 0.258899, + 0.289586, + 0.010721 + ], + [ + 0.508135, + 0.283889, + 0.067819, + 0.009204 + ], + [ + 0.168572, + 0.309029, + 0.739742, + 0.011961 + ], + [ + 0.193029, + 0.321748, + 0.456752, + 0.010993 + ], + [ + 0.169806, + 0.334896, 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M. Inhibition of protein synthesis and cellular damage associated with cilio-", + "stasis in rat tracheal tissue (Abstract). J. Cell. Biol. 43,2 pt. 2:31a, 1969.", + "AREA VI", + "Courant, P. The effect of smoking on the anti-lactobacillus system in saliva. Odont. Rev.", + "18:241-61, 1967.", + "Flesher, J. W. Distribution of radioactivity in the tissues of rats after oral administra-", + "tion of 7,12-dimethylbenz{a]anthracene-3H. Biochem. Pharmacol. 16:1821-31, 1967.", + "Flesher, J. W. and Sydnor, K. L. Distribution of tritiated 7,12-dimethylbenz[a]anthracene", + "(1) and 7-hydroxymethyl-12-methylbenz[a]anthracene (II) in rat. (Abstract) Fed. Proc.", + "27,2:1771, 1968.", + "Frerman, F. E. and White, D. C. Membrane lipid changes during formation of a functional", + "electron transport system in Staphylococcus aureus. J. Bact. 94:1868-74, 1967.", + "Hammond, R. K. and White, D. C. Formation of vitamin K2 isoprenologues by Staphylococcus", + "aureus. J. Bacteriol. 100,2:573-8, 1969.", + "Hammond, R. K. and White, D. C. Separation of vitamin K2 isoprenologues by reversed-phase", + "thin-layer chromatography. J. Chromatog. 45:446-52, 1969.", + "Humphrey, L. J. and Milgron, F. Hemadsorbfactor of malignant tumors. Surg. Forum 16:106-8,", + "1965.", + "Humphrey, L. J.; Milgrom, F.; Tonder, o.; and Witebsky, E. Hemadsorption by murine tumors", + "Int. Arch. Allerg. 30:474, 1966.", + "Humphrey, L. J.; Milgrom, F. Tonder, 0.; and Witebsky, E. Hemadsorption by murine tumors.", + "(Abstract) Fed. Proc. 26:702, 1967.", + "Humphrey, L. J. Immunologic studies on the hemadsorbing factor of malignant tumors. Surg.", + "Forum 18:94-6, 1967.", + "Humphrey, L. J. Studies on passage of the hemadsorbing factor of murine tumors to normal", + "cell cultures. Cancer Res. 27,1:2126-8, 1967.", + "Humphrey, L. J. Studies on the relationship of hemadsorption by normal murine spleen and", + "murine tumors. Cancer Res. 27,1:2123-5, 1967.", + "LaBach, J. P. and White, D. c. Identification of ceramidephosphorylethanolamine and", + "teramidephosphorylglycero] in the lipids of an anaerobic bacterium. J. Lipid Res. 10,", + "Sept. 1969.", + "Lester, R. L. and White, D. c. Quantitative gas-liquid chromatographic analysis of", + "ethanolamine monomethyl ethanolamine, and dimethyl ethanalamine from lipids. J. Lipid", + "Res. 8,6:565-76, 1967.", + "Rees, E. D.; Mandelstam, P.; Linville, W.; and Lipscomb, H. Absorption of benzo{alpyrene", + "by everted sacs of rat small intestine. (Abstract). Fed. Proc. 27:635, 1968.", + "Rees, E. D. Chromosome changes in marrow cells following intravenous administration of a", + "chemical carcinogen. (Abstract) J. Lab. Clin. Med. 74:999, 1969.", + "Rees, E. D. Chromosomal aberrations in rat marrow cells and leukemic cells following intra-", + "venous polycyclic hydrocarbons. (Abstract) J. Clin. Invest. 48:A67, 1969.", + "Rees, E. D. and Mandelstam, P. In vitro and in vivo studies of intestinal absorption of", + "aromatic polycyclic hydrocarbon carcinogens. (Abstract) Fed. Proc. 28,2:749, 1969.", + "Rizza, V.; Sinclair, P. R.; White, D. C.; and Courant, P. R. Electron transport system of", + "the protoheme-requiring anaerobe Bacteroides nelaninogenicus. J. Bact. 96:665-71, 1968.", + "Rizza, V.; Tucker, A. N. ; and White, D. C. Lipids of Bacteroides melaninogenicus. J.", + "Bacteriol. 101,1:84-91, 1970.", + "Short, S. A.; White, D. C.; and Aleem, M. I. H. Phospholipid metabolism in Ferrobacillus", + "ferroxidans. J. Bacteriol. 96:142, 1969.", + "Sinclair, P. R. and White, D. c. The effect of nitrate, fumarate and oxygen on the forma-", + "tion of the membrane-bound electron transport system of Haemophilus. J. Bacteriol.", + "01,2:365-72, 1970.", + "Sydnor, K. L. and Flesher, J. W. Comparative biologic activities of 7,12-dimethylbenz[a]", + "anthracene; 7-hydroxymethyl-12-methylbenz{a)anthracene; 7,12-dihydroxymethylbenz[a]-", + "anthracene and methyoxy-7.12-dimethylbenz[a]anthracene in Sprague-Dawley female rat.", + "Cancer Res. 29:403-8, 1969.", + "Tien, W. C. and White, D. C. Linear sequential arrangement of genes for the biosynthetic", + "pathway of protoheme in Staphylococcus aureus. Proc. Nat. Acad. Sci. 61:1392-98, 1969.", + "White, D. c. and Tucker, A. N. 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Lipids 1,1:56-62, 1970.", + "White, D. C.; Tucker, A. N.; and Sweeley, C. C. Characterization of the isobranched sphin-", + "ganines from the ceramide phosphates of Bacteroides melaninogenicus. Biochim. Biophys.", + "Acta. 187:527-32, 1969.", + "White, D. C. Effect of glucose on the formation of the membrane-bound electron transport", + "system in Haemophilus parainfluenzae. J. Bacteriol. 93:567-73, 1967.", + "White, D. C. and Frerman, F. E. Extraction, characterization and cellular localization", + "of", + "the lipids of Staphylococcus aureus. J. Bact. 94:1854-67, 1967.", + "White,", + "D. C. and Frerman, F. E. Fatty acid composition of the complex lipids of Staphy-", + "lococcus aureus during the formation of the membrane-bound electron transport system.", + "J. Bacteriol. 95:2198-209, 1968.", + "White, D. C. Lipid composition of the electron transport membrane of Haemophilus parainflu-", + "enzae. J. Bacteriol. 96:1159-70, 1968.", + "White, D. C. and Tucker, A. N. Phospholipid metabolism during bacterial growth. J. Lipid", + "Res. 10:220-33, 1969.", + "White, D. C. and Tucker, A. N. Phospholipid metabolism during changes in the proportions of", + "membrane-bound respiratory pigments in Haemophilus parainfluenzae. J. Bacteriol. 97:199-", + "209, 1969.", + "Short, S. A. White, D. C.; and Aleem, M. I. H. Phospholipid metabolism in Ferrobacillus", + "ferrooxidans. J. Bacteriol. 96:142-50, 1969.", + "RC- 6029190", + "136", + "Source: https://www.industrydocuments.ucsf.edu/docs/jgch0bG9", + "0090702" + ] + } + ] +} \ No newline at end of file