Showing papers by "Kettering University published in 1969"

Environmental factors of cancer of the colon and rectum II. Japanese epidemiological data

Abstract: This is an epidemiological study of 107 patients with large bowel cancer and 307 control patients interviewed in Japan. Colon cancer is significantly less common in Japan, but rectal cancer is as common in Japan as it is in the United States. Japanese patients with cancer of the colon have a higher socioeconomic status than rectal cancer patients. Associated with the higher status is a more Western style diet. It is suggested that perhaps dietary fat influences the make-up of the bacterial flora and thus affects the pathogenesis of cancer of the colon. No significant relationship was found to medical and surgical diseases nor to the cholesterol and weight levels.

Treatment of adult acute leukemia with arabinosylcytosine and thioguanine.

Abstract: Forty unselected adults with acute leukemia were treated with CA (2–3 mg/kg) by daily rapid intravenous injection and TG (2.5 mg/kg) orally. Two patients were judged to have had an inadequate trial, having died after 3 and 4 days. Of 38 adequately treated patients, 2 had acute lymphoblastic and 36 acute myeloblastic, monoblastic, or myelomonoblastic leukemia. Twenty-eight patients had had previous chemotherapy (19 with either CA or 6-MP alone) but had failed to respond or had relapsed and were refractory. The 38 adequately treated patients had 1 or more courses of CA and TG of 8–32 days each as limited by individual tolerance. The drugs caused moderate or severe pancytopenia and marrow hypoplasia in all but 3 patients treated. Twenty-one of 38 patients had remission with M-1 marrows lasting an average 6 (1.5–13) months with 6 patients still in remission. Seventeen of 21 patients had H-1 peripheral blood ratings (normal platelet, leukocyte and hemoglobin values) and 4 had H-2 ratings, all because of low hemoglobin. Twenty patients in remission were given maintenance therapy; the dosage varied but is currently TG 2.5 mg/kg 4–5x/wk + CA 2–3 mg/kg 1-2x/wk.

Antigenic structure of cell surfaces, an immunoferritin study of the occurrence and topography of h-2, theta, and tl alloantigens on mouse cells.

Abstract: The representation of mouse alloantigens belonging to three systems, H-2, θ and TL, on the surface of cells from thymus, spleen, lymph nodes, and peritoneal cavity, was studied by electron microscopy with ferritin-labeled antibody. As expected from earlier serological data, TL was confined to thymocytes, θ was found on thymocytes and lymphocytes, and H-2 occurred to some extent on all cell types observed. On reticular cells, lymphocytes, plasma cells, and eosinophils, the majority of the cell surface was occupied by H-2; thymocytes had considerably less H-2, and erythrocytes and peritoneal macrophages least of all. In every instance the representation of antigen was discontinuous, the fraction of the cell surface covered being characteristic both of the antigen and of the type of cell. H-2 and θ provide a striking example of this; H-2 is present in far higher amounts on lymphocytes than on thymocytes, whereas the converse is true of θ. Within areas positive for H-2 or θ, protuberances of the surface membrane were often antigen-negative. A better definition of cell surface structure, gained from studies such as this, is necessary for further inquiry into how the cell surface is assembled, and into selective gene action in relation to cellular differentiation.

Photoreduction of 2,6-Dichlorophenolindophenol by Diphenylcarbazide: A Photosystem 2 Reaction Catalyzed by Tris-Washed Chloroplasts and Subchloroplast Fragments

Abstract: The use of diphenylcarbazide as an electron donor coupled to the photoreduction of 2,6-dichlorophenolindophenol by tris-washed chloroplasts or subchloroplast fragments provides a simple and sensitive assay for photosystem 2 of chloroplasts. By varying the concentration of tris buffer at pH 8.0 during an incubation period it is shown that the destruction of oxygen evolution activity is accompanied by a corresponding emergence of an ability to photooxidize diphenylcarbazide, as evidenced by absorbance changes due to diphenylcarbazide at 300 nm. The temperature-sensitive oxidation of diphenylcarbazide is inhibited by DCMU and by high ionic strengths. This activity appears to measure the primary photochemical reaction of photosystem 2.

Epidemiology of Cancer of the Ovary

Abstract: Etiology and epidemiology are often intertwined. Therefore an introductory discussion is devoted to these subjects.

Epidemiologic investigation of multiple primary cancer of the upper alimentary and respiratory tracts. I. A retrospective study

Abstract: In a retrospective investigation of 104 patients with multiple primary tumors of the upper alimentary and respiratory tracts, the data suggest that patients who smoke heavily before developing cancer are more likely to develop second, primary tumors. Continued smoking after initial diagnosis or radiation therapy for the first primary is associated with an increased frequency of second primary tumors. Discontinuance of smoking and drinking after the first primary provides no assurance against the development of a second primary cancer.

Feline leukemia virus: occurrence of viral antigen in the tissues of cats with lymphosarcoma and other diseases.

Abstract: Felinie leutkemia viruts antigen is demonstrable by immunodiffusion with rabbit precipitating antiserum to purified felinie leukemia virus. The felinie leukemia virus antigen was found in the tissues of 25 of 33 cats with lymphosarcoma and of 5 of 13 cats with infectious peritonitis. Its presence was correlated with the occurrence of felinie leukemia virus demonstrable by electron microscopy. The one clinically normal cat giving a positive test for feline leukemia virus antigen belonged to a household in which two cats had developed lymphosarcoma. With the exception of a dog with lymphosarcoma induced by feline leukemia virus, the antigent was absent from lymphosarcoma and nonlymphomatous tumors of other species (man, dog, cow, goat, or pig).

Relationship between Absorption Spectrum and Molecular Conformations of 11-cis-Retinal

Abstract: THE absorption spectrum of 11-cis-retinal is strongly influenced by the presence of a cis bond between carbon atoms 11 and 12. The cis bond in this position results in two important spatial properties of the molecule: first, steric hindrance between the C13-methyl group and the C10-hydrogen atom; second, a bend in the system of conjugated double bonds. Both the steric hindrance and the bend in the molecule give rise to particular absorption bands. We now report the results of experiments which allow the identification of these absorption bands in the 11-cis-retinal spectrum.

Identification of women at high risk for breast cancer

Abstract: This paper reviews factors which appear to affect a womans risk of developing breast cancer. The low rate in Japan as compared to the West is noted particularly in the post-menopausal period. The stability of mortality in the U.S.A. has been ascribed to an increase in survival balanced by an increase in incidence especially among younger women. In New York City breast cancer morbidity is particularly high among Jewish women. Cancer of the breast is less common among American Negroes. In Asian countries breast cancer is more frequent among upper income groups. It has been suggested that single as well as infertile married women have a somewhat higher risk than fertile married women as much as 2-fold. Pregnancy below 20 years of age may result in a lower risk of the disease than pregnancy after 25. Prolonged nursing particularly over a total of 36 months reduces the risk of breast cancer. Castration of a woman before age of 37 may reduce the risk. A patient with fibrocystic disease of the breast requires special surveilance. Obesity in post-menopausal women is a controversial parameter. Women who have already had cancer of the breast have a 7 per 1000 per year chance of getting cancer in the other breast. Women whose mothers or sisters have developed breast cancer are also a high risk group. Colon cancer may have some association and long-term estrogen therapy is being considered as of possible significance. Laboratory studies of hormone steroids and glucose tolerance have been of value in estimating risk. The postulation that a womans fat intake may be a factor is supported by some animal experiments. Dietary factors especially in terms of lipids may play an important role. With the exception of women with previous breast cancer and those with breast cancer in the immediate family no particular group appears to require more surveillance than the average woman.

Studies on the respiratory system of aerobically (dark) and anaerobically (light) grown Rhodospirillum rubrum.

Abstract: 1. A major part of the respiratory activity of light grown cells of Rhodospirillum rubrum is associated with a system identical with that found in dark grown cells. 2. The specific activity of NADH and succinate dehydrogenase and cytochrome c reductase on a protein basis is the same in the particulate fraction from photosynthetic and aerobic cells. In contrast, the NADH and succinate oxidase is more than four times higher in the aerobic cells. Since the amount of particulate protein per cell is almost four-fold less in the aerobic cells, the total particle bound capacity for NADH and succinate oxidation per cell is approximately the same regardless of whether the cells were grown photosynthetically or aerobically. 3. The effect of various inhibitors of NADH oxidation was compared in the particulate fraction from aerobically and anaerobically grown cells. 4,5,6,7-Tetrachloro-2-trifluoromethylbenzimidazole (TTFB) was found to be a potent inhibitor of NADH and succinate oxidation; one-half maximal inhibition was observed at 0.5 μM. Rotenone and amytal inhibited about 80%. 4. In particles from photosynthetically grown cells respiration was inhibited to a maximum of 45–70% by KCN and TTFB; whereas in aerobically grown cells these inhibitors consistently inhibited maximally by more than 70%. 5. NADH oxidation was inhibited 35–50% by light in chromatophore preparations. Antimycin a relieved the light inhibition. In the presence of antimycin a and m-chlorocarbonyl cyanide phenylhydrazone, light stimulated NADH oxidation and the oxidation was not sensitive to KCN. 6. 2-Hydroxy-3-(ω-cyclohexyloctyl)-1,4-naphthoquinone which had little effect on dark NADH oxidation inhibited the light stimulated oxidation. 7. These results indicate that at least three pathways exist for NADH oxidation in chromatophores: a) KCN sensitive which is characteristic of aerobically grown cells; b) KCN insensitive which appears to be part of the cyclic photochemical pathway; and c) the rotenone insensitive pathway.

α-Naphthylamine and β-Naphthylamine in Cigarette Smoke

Abstract: EPIDEMIOLOGICAL data suggest an association between cigarette smoking and urinary bladder cancer in men1–3. In mice experimental application of cigarette smoke has led only to inflammation and hyperplasia of the bladder mucosa, but not to histologically proven bladder tumours4. Recently, however, tumours were reported in the renal pelvis of mice painted with “refined cigarette tar”5. These observations suggest the need for investigations of bladder carcinogens in cigarette smoke. Two earlier attempts failed to prove the presence of some of the known experimental bladder carcinogens in the group of aromatic amines6,7. Recently, naphthylamines were found to be formed on pyrolysis from amino-acids8. This communication describes the identification and quantitative determination of α and β-naphthylamine in cigarette smoke.

Ultrastructure of human labial salivary glands. II. Intranuclear inclusions in the acinar secretory cells.

Abstract: Human labial salivary glands, obtained by biopsy from 32 subjects, were studied by light and electron microscopy. Intranuclear inclusions, unrelated to nucleoli, were present in many of the acinar nuclei in glands from 16 of the 32 donors. More than one inclusion was sometimes observed within a single nucleus. They measured about 1 μ in diameter, and were stainable in a variety of ways. They were eosinophilic, some were stained by Nile blue sulphate, some were PAS-positive, and all were Feulgen-negative. They were bounded by a single membrane, which never exhibited continuity with the nuclear envelope, and they showed considerable morphological variation. The more complex inclusions consisted of alternating shells of light and dark material with tiny dense granules embedded in the latter. The intranuclear inclusions, which apparently were non-viral in origin, were in some way related to the secretory cycle of the mucous cells, since they were found only in immature cells, and never in cells in which secretory products were abundant.

New visual markers of antibody for electron microscopy.

Abstract: SINGER1 introduced ferritin-labelling of specific antibody as a means of identifying antigens in electron micrographs. This technique has been widely applied to the location of cellular, bacterial and viral antigens2. Recently we have modified the technique by preparing hybrid F(ab′)2 antibody, permitting the development of a multiple-marker system and obviating the chemical coupling of ferritin to antibody (which engenders heterogeneous products and causes considerable loss of antibody activity). In the indirect version of this modified test3, as used for the location of mouse cell surface antigens, alloantibody is applied to the viable cells in suspension, followed by hybrid antibody with one combining site for mouse γG and the other for ferritin, and finally by ferritin. Thus the reaction sequence is surface alloantigen: alloantibody (mouse γ)G : anti-mouse γG/anti-ferritin hybrid antibody : ferritin. Labelling in this way is highly specific and reproducible.

Heterotransplantation of human cell lines from Burkitt's tumors and acute leukemia into newborn rats

Abstract: The heterotransplantability of 22 human cell lines was studied by intravenous inoculation into newborn rats. The cell lines included nine derived from Burkitt (African) lymphoma, one from reticulum cell sarcoma, ten from leukemia, one from infectious mononucleosis and one from a normal lymph node. All were cultivated in vitro as suspension cultures. The usual inoculum for each rat was 50 or 10 or 2 million cells. At the two higher doses four of the Burkitt tumor lines (EB‐2, EB‐3, SL1, B35M), three of the leukemia lines (SK‐L2, SK‐L5, SK‐L9), the infectious mononucleosis line C566, and the cell line from a normal lymph node (SK‐LN1) propagated and caused lethal lesions in approximately half of the recipients. Inocula containing 2 million cells of lines EB‐2 or SL1 grew in a few rats but none of the other cell lines grew at this dose. Brain, eye and kidney were the most common sites of growth. Lung, adrenal, liver, and interscapular fat pad were less frequently involved, and other tissues rarely. The results are discussed in relation to the biologic significance of heterotransplantability and differences in the tissue distribution of implants of these and previously studied human cell lines.

Association of inorganic pyrophosphatase activity with normal calcification of rat costal cartilage in vivo

Abstract: 1. Dialysed extracts of rat costal cartilage were shown to possess an enzyme that hydrolyses inorganic pyrophosphate. 2. Inorganic pyrophosphatase activity assayed in the presence of 2mm substrate was maximal at pH6·8. 3. Mg2+ was essential for activity, which was greatest with 10mm or higher concentrations of Mg2+. 4. Extracts prepared from cartilage taken from suckling rats (<20g.) showed little or no hydrolytic activity, but as rat weight increased inorganic pyrophosphatase activity was detected, increased to a maximum in tissue from animals weighing about 40g., and then rapidly declined. 5. The increase in inorganic pyrophosphatase activity was associated with an increase in the uptake of 45Ca by the cartilage in vivo. 6. Accumulation of calcium, inorganic phosphate and magnesium occurred when inorganic pyrophosphatase activity was at its maximum. 7. Alkaline phosphatase activity, measured in the same extracts used to determine pyrophosphatase activity, was highest in the tissues of the animals weighing <20g., and decreased as inorganic pyrophosphatase activity increased to its maximum. 8. There was no direct relationship between alkaline phosphatase activity and the onset of calcification.

Hemmung maligner Neoplasien des Menschen durch L-Asparaginase

Abstract: 88 Kranke mit malignen Neoplasien wurden mit dem ausE. coli hergestellten Enzym L-Asparaginase behandelt. Das therapeutische Ergebnis konnte bei 60 Kranken ausgewertet werden. Bei 20 von 33 Kranken mit akuter Lymphoblastenleukamie, bei 1 Kranken mit Lymphosarkom in leukamischer Phase und bei 1 von 5 Kranken mit akuter Myeloblasten- oder Myelomonocytenleukamie wurde eine Knochenmarksremission erzielt. Die Remissionsdauer betrug 1–8 Monate. Von 21 Kranken mit malignen Lymphomen, Sarkomen und Carcinomen sprach nur 1 Patient mit malignem Melanom auf die Behandlung mit Asparaginase an. Die Enzympraparation bewirkte in der bisher angewandten Dosis keine Knochenmarkshemmung. Reversible Nebenwirkungen waren gekennzeichnet durch Fieber, Gewichtsverlust, abnorme Leberfunktionsproben, Hypoalbuminamie, Hypofibrinogenamie, Hypolipidamie, Hyperlipidamie und Uberempfindlichkeitsreaktionen gegenuber der Enzympraparation. Die Asparaginabhangigkeit der neoplastischen Zellen lies sich auchin vitro nachweisen. Dieser Test erlaubt jedoch noch keine absolut sichere Vorhersage des Behandlungsergebnisses. Wenn sich wahrend der Behandlung einer primar asparaginase-empfindlichen Leukamie Resistenz entwickelte, waren die Leukamiezellen auchin vitro nicht mehr asparaginabhangig.

Procedures for control of mycoplasma contamination of tissue cultures

Abstract: Mycoplasma contamination of cell and tissue cultures is still a problem in many laboratories despite the availability of improved methods for detection of the contaminants and the availability of better information on the subject. Awareness of the problem has been accentuated recently as shown by the number of requests we have received for testing of cultures and for advice on how to proceed if there is evidence of mycoplasma. Results of reported attempts to trace the source of mycoplasma contamination have been inconcl~sive;’*’~~’~ often the source is not established. Biological materials employed in cell culture work or laboratory technicians performing the work details necessary for cell culture maintenance are suspected, but to this date no foolproof method of prevention of contamination has been proposed. In most cases, mycoplasma contamination occurs suddenly and unexpectedly and may not be detected by either gross or microscopical observation of the cultures. Many mycoplasma effects on mammalian cells in culture reported during the last few years’ so closely resemble effects of viruses that misinterpretation of the causative agent is understandable. However, misinterpretation could have been avoided if methods for demonstration of mycoplasma contamination had been available in the laboratory and if routine testing of cell cultures had been performed. A number of methods for eliminating mycoplasma from contaminated cell cultures have been proposed!, ’ 4,’ ’, ’’,’ 920 Their degree of efficiency has varied for different situations of contamination depending upon the type of cell and mycoplasma. Attempts to eliminate mycoplasma contamination permanently have often failed. Both mammalian cells and mycoplasma vary considerably in their sensitivity to physical exposure and chemical agents. Nevertheless, elimination of the contamination has been efficiently accomplished in many cases under treatment conditions which have not caused apparent serious damage to the mammalian cells. The severity of the contamination problem, however, appears to have grown, not only due to the increasing frequency of contamination accidents in research and diagnostic laboratories, but also to the recognition of additional effects of mycoplasma on mammalian cells. Recently, a most serious consequence of mycoplasma contamination has been reported? When FL human amnion cells were cultured in the presence of mycoplasma of the HT strain for long periods of time, profound changes in the genetic constitution of the cells occurred, resulting in

Cytotoxic Test Automation: A Live-Dead Cell Differential Counter

Abstract: An appropriately modified cell spectrophotometer was used successfully for performing automatic counts of live and dead cells in the cytotoxic test, with trypan blue staining as an indicator of dead cells and light scattering to identify viable cells.

In vitro Interaction of LSD with Purified Calf Thymus DNA

Abstract: THE observation of broken chromosomes in test animals and humans treated with the hallucinogen lysergic acid diethylamide (LSD) has been well documented1–8. Although there has been some speculation about the mode of action of the drug, no specific studies of the mechanism of LSD-induced chromosomal damage have, as yet, been presented. Such studies would be most useful in determining whether LSD interacts directly with the chromosome which becomes broken or whether chromosomal breakage is the result of a series of biochemical events begun by LSD at some site distant from the chromosome. This communication describes circular dichroism experiments which indicate that LSD interacts directly with purified calf thymus DNA, probably by intercalation, causing conformational changes in the DNA. It is suggested that this observed interaction may serve as a model for the action of LSD on the chromosomal material of intact cells.