Showing papers by "Kettering University published in 1987"

Incidence of cancer in 98 patients with common varied immunodeficiency.

Abstract: Ninety-eight patients with common varied immunodeficiency have been observed for periods of 1–13 years. In 1986, 78 were alive, 19 had died, and 1 could not be located. Eleven patients in the group had developed cancer; two patients had had two cancers. Of the total number of neoplastic malignancies, seven were non-Hodgkin's lymphoma, one patient had a Waldenstrom's macroglobulinemia, and nine of the patients who developed cancer were female. Cancer developed in the fifth or sixth decade of life for 10 of the 11 patients. These data show an 8- to 13-fold increase in cancer in general for patients who have this immunodeficiency and a 438-fold increase in lymphoma for females.

Characterization of cellular proteins recognizing the HIV enhancer using a microscale DNA-affinity precipitation assay

Abstract: Numerous nucleic acid sequence motifs have been identified as transcriptional regulatory elements, and proteins involved in the control of gene replication and transcription have been successfully purified using nucleic acid affinity procedures1–5. We have developed an assay that allows direct characterization of cellular proteins binding to the enhancer element of the human immunodeficiency virus (HIV), which is analogous to immunoprecipitation techniques. In extracts of H9 cells, a human CD4+ lymphoblast line clonally selected for its ability to support HIV replication6,7, we find a class of proteins that interact specifically with the HIV enhancer. Reproducible high-resolution, two-dimensional gel electrophoresis has allowed us to resolve these proteins into two major sets. One set is common to H9 cells, two human B-lymphoblast lines, and a phytohaemagglutinin (PHA)-stimulated human CD4+ lymphoblast line (Jurkat). A second set of proteins is found only in H9 cells. Thus, with this assay we have identified HIV enhancer-binding proteins that are constitutive or inducible and that are cell-type specific.

Production of Pili (Fimbriae) by Pseudomonas fluorescens and Correlation with Attachment to Corn Roots.

Abstract: Pseudomonas fluorescens isolates 13525 and 2-79 were grown in Luria broth and low-nutrient medium (LNM). Pililike fibrils were very rarely produced in Luria broth but were abundantly produced in LNM. In LNM the pili were peritrichously distributed and had diameters ranging from 3 to 8 nm. Pili were purified from strain 2-79, and the pilin subunit was found to have a molecular weight of about 34,000. Strain 2-79 produced two colony types on Luria agar, nonmucoidal and mucoidal. Cells in LNM cultures of the nonmucoidal colony type were highly piliated, and cells from the mucoidal type were nearly devoid of pili. The presence of pili on nonmucoidal isolate 2-79 was quantitatively correlated with hydrophobic attachment to polystyrene, hemagglutination, and attachment to corn roots.

Evidence against Ha-ras-1 involvement in sporadic and familial melanoma.

Abstract: It was recently reported that different rare alleles at the Ha-ras-1 locus occurred at a significantly higher combined frequency in cancer patients than in an unaffected population1 In particular, melanoma patients were reported to have a significantly higher frequency of such alleles We have examined the frequency of rare Ha-ras-1 alleles in a large number of cases of sporadic melanoma Our results indicate that the distribution of rare alleles in this population does not differ from that found in normal populations Also, to test the hypothesis that a hereditary predisposition to melanoma could be inherited via an allele at the Ha-ras-1 locus, we examined the transmission of the segment of the short arm of chromosome 11 (11p) carrying the Ha-ras-1 locus in a number of families previously shown to exhibit a hereditary predisposition to melanoma and its precursor lesion, the dysplastic nevus syndrome (DNS)2,3 Our genetic linkage results thus obtained strongly exclude the association of a predisposition to melanoma or the precursor lesion with the inheritance of the Ha-ras-1 locus or the segment of chromosome 11 on which it is located These results imply that hereditary predisposition to melanoma is associated with genes other than the Ha-ras-1 locus, contradicting the original suggestion of Krontiris et al1, made on the basis of either an inadequate sample size or other misleading experimental factors

Interactions of acridine orange with double stranded nucleic acids. Spectral and affinity studies.

Abstract: Spectral properties of acridine orange (AO) alone or in complexes with natural and synthetic nucleic acids of various base composition have been studied in aqueous solutions by absorption and fluorescence spectroscopy. The dimerization constant and absorption spectra of the dye in monomelic and dimeric form were established; dimerization of AO resulted in quenching of its fluorescence. Complexes of the dye with synthetic nucleic acids differed in the degree of enhancement of fluorescence quantum yield, varying between 1.42 to 2.38 fold as compared to AO monomer; these differences, however, were not base-dependent. Affinity of the dye to natural and synthetic polymers was studied and analyzed using McGhee-von Hippel model of polymer-ligand interactions. Because the sterical requirement for intercalative binding assumes interaction of dye monomer, the correction for AO dimerization was made in all calculations. All studied DNAs (natural and synthetic ones, the latter being homopolymer pairs or alte...

Senescence in oat leaves: Changes in translatable mRNAs

Abstract: Changes in translatable mRNA populations during the senescence of oat (Avena sativa L. cv. Victory) leaves were examined by analyzing the in vitro translation products of isolated RNA. Total RNA was isolated from oat leaves of 7-day-old seedlings, and also after these leaves were aged for different lengths of time under various conditions. Polypeptides from in vitro translations were separated by two-dimensional gel electrophoresis to estimate any changes in translatable mRNA populations associated with senescence. Corresponding leaf samples were monitored for loss of chlorophyll as a measure of the extent of senescence. The aging of excised leaves in the light for 4 days resulted in the disappearance or substantial quantitative decrease of a number of mRNA species, while only five new translatable mRNA species were produced. Three of these mRNAs were unique to aging of leaves under light. Two of these mRNA species were also produced during the early stages of senescence in attached leaves of seedlings grown under light. The translatable mRNA populations of leaves aged for 4 days either on intact seedlings or detached and kept in the light in the presence of kinetin were very similar. Aging of excised leaves in the dark on water for 24 h resulted in very extensive changes in translatable mRNA populations. Over thirty polypeptides disappeared or were substantially reduced in quantity, while about an equal number appeared de novo or were substantially increased in quantity. Aging of these leaves for an additional 24 or 48 h resulted in only a few additional changes in translatable mRNAs. The presence of kinetin during aging of excised leaves in the dark inhibited few of the numerous changes in mRNAs that occured during the first 24 h, but did inhibit most of the changes that occured after 48 or 72 h of aging in the dark. When leaves were first aged in the dark and then returned to light, most of the initial changes in translatable mRNAs expression were reversed. Such changes in mRNAs thus appear to be light-regulated and not necessarily associated with senescence.

Application of pyronin Y(G) in cytochemistry of nucleic acids.

Abstract: Chinese hamster ovary (CHO) cells or isolated nuclei were stained with pyronin Y(PY) and analyzed by absorption or fluorescence microscopy, as well as by flow cytometry. Specificity of the staining reaction was assayed by testing sensitivity of the stainable material to RNase or DNase. The colored complexes detected by light absorption in fixed cells stained with PY are nonfluorescent and are most likely the products of condensation of single-stranded (ss) RNA by PY; the poly(rA) and poly(rA,rG) are the most sensitive to condensation. The products of PY interaction with double-stranded (ds) nucleic acids are fluorescent and can be detected in cells by cytofluorometry. PY used alone stains both DNA and RNA, and the staining capabilities of these nucleic acids vary depending upon the PY concentration at equilibrium; at a concentration above 330 microM, the RNA stainability decreases, perhaps due to its denaturation and condensation caused by the dye. In the presence of Hoechst 33342, PY can specifically stain RNA in fixed cells or isolated cell nuclei. Because only complexes of PY with ds RNA are fluorescent, this dye can be used as a probe of RNA conformation, e.g., to monitor denaturation of RNA in situ. The RNA stainability of mitotic cells is about 25% lower than that of cells in G2 phase, which indicates that during mitosis proportionately less cellular RNA is in the ds conformation. The advantages and limitations of the two cytochemical methods for DNA/RNA detection, one based on the use of Hoechst 33342 and PY, and another employing the metachromatic properties of acridine orange, are compared.

Nutrient density: an important and useful tool for laboratory animal studies

Abstract: Nutritional deficiencies or imbalances are suspected contributory factors to several types of human cancers, and perhaps other human diseases related to long-term metabolic derangements. In order to study these more effectively in laboratory animals, it is suggested that the laboratory diets more closely mimic the nutrient density of suspect human diets. Toxicology and carcinogenic data obtained in animal study using diets based on human nutrient density might be more readily applicable in relation to humans.

Interactions of pyronin Y(G) with nucleic acids.

Abstract: Spectral properties of pyronin Y(PY) alone or in complexes with natural and synthetic nucleic acids of various base compositions have been studied in aqueous solution containing 10 or 150 mM NaCl and 5 mM Hepes at pH 7.0. The dimerization constant (KD = 6.27 X 10(3), M-1) and the absorption spectra of the dye in monomeric and dimeric form were established. The complexes of PY with single-stranded (ss) nucleic acids show a hypsochromic shift in absorption, and their fluorescence is quenched by over 90% compared to free dye. In contrast, complexes with double-stranded (ds) RNA or DNA (binding by intercalation) exhibit a bathochromic shift in their absorption (excitation) spectrum, and their fluorescence is correlated with the base composition of the binding site. Namely, guanine quenches fluorescence of PY by up to 90%, whereas A, C, I, T, and U bases exert a rather minor effect on the fluorescence quantum yield of the dye. The intrinsic association constant of the dye to ds RNA (Ki = 6.96 X 10(4), M-1) and to ds DNA (Ki = 1.74 X 10(4), M-1) was measured in 150 mM NaCl; the binding site size was 2-3 base pair for both polymers. Implications of these findings for qualitative and quantitative cytochemistry of nucleic acids are discussed.

Selective up-regulation by interferon-γ of surface molecules of the Ly-6 complex in resting T cells: the Ly-6A/E and TAP antigens are preferentially enhanced

Abstract: Surface molecules encoded by the murine Ly-6 locus can transduce triggering signals in T cells and thus may play important roles in T cell function. Previously, we found that Ly-6 molecules are up-regulated by interferon (IFN)-alpha/beta in resting T cells. Here, we examined the possible influence of IFN-gamma on these molecules. Purified T cells from C57BL/6 (Ly-6.2) and BALB/c (Ly-6.1) mice were incubated in vitro with recombinant murine IFN-gamma and the expression of Ly-6 antigens was measured by flow cytofluorometry. It was found that both Ly-6A/E and T cell-activating protein (TAP) molecules are markedly enhanced while Ly-6C is less affected. Under the same conditions, other T cell surface molecules showed no or marginal changes. The effect of IFN-gamma on Ly-6A/E and TAP expression reached a maximum with as little as 10 U/ml and required only 18-24 h of incubation. Moreover, the enhancement of Ly-6A expression induced by IFN-gamma was stable for at least 5 days. Analysis of T cell subsets further revealed that IFN-gamma-induced augmentation of Ly-6A (C57BL/6 mice) involves both Lyt-2+ and L3T4+ cells while the increase of Ly-6E (BALB/c mice) is more pronounced in Lyt-2+ cells. The functional consequence of these phenotypic alterations was evaluated by studying the mitogenic responses of T cells to antibody-mediated Ly-6 cross-linking in the presence of phorbol myristate acetate. Pretreatment of resting T cells with IFN-gamma dramatically increased the responses to anti-Ly-6A and anti-Ly-6E monoclonal antibodies. IFN-gamma treatment also boosted the stimulation induced by anti-TAP monoclonal antibody when this stimulation was performed under suboptimal conditions. Therefore, IFN-gamma selectively up-regulates the Ly-6A/E and TAP activation pathways in resting T cells. We speculate that this effect may contribute to the immunoregulatory activities of IFN-gamma.

Effect of recombinant tumor necrosis factor on HL-60 cells: cell-cycle specificity and synergism with actinomycin D.

Abstract: The tumor necrosis factor (TNF) exhibits a multitude of activities depending on the type of target cells. We characterized the cytostatic and cytotoxic effects of recombinant TNF, alone and in combination with actinomycin D (AMD), on the human leukemic cell line HL-60. Because HL-60 cells, when triggered to monocytic differentiation by phorbol esters, are known to produce and secrete TNF, their sensitivity to the factor could indicate an autocrine function of TNF in this cell system. Indeed, HL-60 cells were affected by TNF; their doubling time was increased by about 50% and progression through the cell cycle was perturbed. Initially, (up to 8 h) TNF induced a temporary arrest in G2 while later (24–48 h) it delayed progression through the G1 phase. Also, a transient increase in RNA content peaking at 6–8 h was apparent. The cytotoxicity of TNF alone was low. Thus, TNF may be involved in the regulation of the cell cycle of HL-60 cells during early stages of their differentiation. The cytotoxicity of TNF was markedly potentiated in the presence of AMD; the effect was AMD but not TNF concentration-dependent. Whereas at 20 and 50 ng/ml of AMD alone nonviable cells did not exceed 20% during the first 24 h of treatment, their proportion increased to 80 and 90%, respectively, in the presence of TNF. The most sensitive were cells in the S phase of the cell cycle. The observed synergistic effect of TNF and AMD does not appear to be caused by the action of TNF increasing the permeability of the cell membrane to AMD. The results indicate that HL-60 cells, ordinarily resistant to the cytotoxic action of TNF, can be rendered sensitive by treatment with AMD. This implies that a combination of TNF and AMD may be considered in oncology for treatment of tumors otherwise nonresponding to TNF alone.

Synthesis and biological effects of 2'-fluoro-5-ethyl-1-beta-D-arabinofuranosyluracil.

Abstract: 29-Fluoro-5-ethyl-1-beta-D-arabinofuranosyluracil (FEAU) was synthesized, and its biological activities were compared with those of 29-fluoro-5-methyl-1-beta-D-arabinofuranosyluracil (FMAU). Earlier studies indicated that both compounds showed potent anti-herpes simplex virus activity, with a 50% effective dose (ED50) of less than 0.25 microM. In the present study the cell growth inhibitory activity of FEAU (ED50, 200 to 2,060 microM) was found to be about 100-fold less than that of FMAU. With an ED50 ranging from 630 to 3,700 microM, FEAU only weakly inhibited thymidine incorporation into DNA, as compared with FMAU with an ED50 of 9 to 28 microM. Following exposure to [2-14C]FEAU (100 microM), 0.48 pmol/10(6) cells per h was incorporated into the DNA of herpes simplex virus type 1-infected Vero cells, whereas no detectable incorporation was found in uninfected Vero cells or L1210 cells. The Ki of FEAU for thymidine kinase purified from human leukemic cells was greater than 150 microM. For herpes simplex virus type 1- and 2-encoded thymidine kinases, the Kis were 0.6 and 0.74 microM, respectively. Both FEAU and FMAU were relatively nontoxic for mice, with a 50% lethal dose of greater than 800 mg/kg per day (four intraperitoneal doses). However, the lethal dose of FEAU for dogs was 100 mg/kg per day (10 intravenous doses), a dose which is 40- to 80-fold greater than the toxic dose of FMAU. These results suggest that FEAU is a worthy candidate for further development as an antiherpetic agent.

Nodulin gene expression during soybean ( Glycine max ) nodule development

Abstract: In vitro translation products of total RNA isolated from soybean nodules at successive stages of nodule development were analyzed by two-dimensional gel electrophoresis. In that way the occurrence of over 20 mRNAs specifically transcribed from nodulin genes was detected. The nodulin genes could be divided into two classes according to the time of expression during nodule development. Class A comprises at least 4 nodulin mRNAs which are found when a globular meristem is present in the root cortex. These class A nodulin genes have a transient expression. Class B nodulin genes are expressed when the formation of a nodule structure has been completed. Bradyrhizobium japonicum nod + fix-mutants, with large deletions spanning the nif H,DK region, still induced nodules showing normal expression of all nodulin genes, indicating that the nif H,DK region is not involved in the induction of nodulin genes. In nodules induced by Bradyrhizobium japonicum nod + fix-mutant HS124 the bacteria are rarely released from the infection thread and the few infected cells appear to be collapsed. All class A and class B nodulin genes are expressed in HS124 nodules with the exception of 5 class B genes.

Binding of Fe 2+ by Mammalian Ferritin

Abstract: An average of 140 Fe2+ ions bind to mammalian ferritin that has an average core content of 1876 Fe3+ ions per molecule. The Fe2+ ions enter the protein interior and exchange electrons with the core Fe3+ ions. A non-homogeneous model for the iron-containing core of ferritin is proposed.

Aziridinyl putrescine containing compositions and their uses in treating prostate cancer

Abstract: This invention relates to methods of inhibiting the proliferation of prostate cancer cells comprising contacting the cells with an effective amount of a cytotoxic polyamine compound. The invention also relates to methods of inhibiting the proliferation of prostate cancer cells in a subject afflicted with prostate cancer. The invention further concerns therapeutic compositions comprising an effective prostate cancer cell proliferation inhibiting amount of 1-(4-aminobutyl) aziridine and a pharmaceutically acceptable carrier. The invention also concerns two-component therapeutic compositions.

Nucleosides. 146. 1-Methyl-5-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)uracil, the C-nucleoside isostere of the potent antiviral agent 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)thymine (FMAU). Studies directed toward the synthesis of 2'-deoxy-2'-substituted arabino nucleosides. 6.

Abstract: The synthesis of 5-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-1-methyluracil (1, C-FMAU), an isostere of the potent antiviral and antitumor nucleoside 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)thymine (2'-fluoro-5-methyl-ara-U or FMAU), was achieved. Pseudouridine (2) was converted into 4,5'-anhydro-3'-O-acetyl-2'-O-triflylpseudouridine (4), which was treated with tris(dimethylamino)sulfur (1+) difluorotrimethylsilicate (TASF) to give 4,5'-anhydro-5-(3-O-acetyl-2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-1- methyluracil (5b) in 40% yield. Acid hydrolysis of the 4,5'-anhydro linkage of 5b with Dowex 50 (H+) afforded C-FMAU. The inhibitory activity of C-FMAU against HSV-1 and HSV-2 was about 10-fold less than that of FMAU in tissue culture. This compound, however, did not show significant activity in mice inoculated with HSV-1 or HSV-2.

Bacterial leaf symbiosis in dioscorea sansibarensis morphology and ultrastructure of the acuminate leaf glands

Abstract: The symbiosis between a bacterium and the West African rain forest yam Dioscorea sansibarensis is described for the first time at the ultrastructural level. The bacteria are harboured in glands which run the entire length of the pronounced ‘drip-tip’ leaf acumenae of the host plant. Each acumen, which may be up to 12 cm long in very large mature leaves, contains from two to six bacterial glands. The glands are kidney-shaped in cross section and contain numerous multicellular simple trichomes which arise from the epidermis of the gland floor and project into the lumen of the gland. The bacteria are Gram-negative and variously rod, ovoid, and coccoid in shape. The bacterial cells contain mesosomes, polyhydroxybutyrate granules and large electron-dense bodies. Bacteria-free plants grow more slowly and produce one yellowish-green leaf per node in contrast to the vigorous growth habit of infected plants, with two deep green leaves per node. Infected plants exposed to a variety of atmospheres containing acetylene, both in the light and in the dark, failed to reduce acetylene to ethylene, indicating that nitrogen fixation is not a function of this symbiosis.

Compositions and method for treatment of cancer using monoclonal antibody against GD3 ganglioside together with IL-2

Abstract: Cell surface gangliosides are presumed to play a role in cell growth and differentiation. Using monoclonal antibodies directed against GD3, a disialoganglioside expressed predominantly by cells of neuroectodermal origin, we have found that GD3 is expressed by a subpopulation of cells of the immune system including: a) fetal thymocytes in subcortical regions and near vessels, 2) lymph node lymphocytes in interfollicular areas and near vessels, and 3) a small subset of T cells in the peripheral blood. Mouse monoclonal antibodies (two IgGs, one IgM and F(ab')2 fragments) reacting with GD3 were found to stimulate proliferation of T cells derived from peripheral blood. Proliferation induced by binding to GD3 could be augmented by exogenous IL-2, PMA, PHA or Protein A.

Definition of response and remission in children over one year of age with advanced neuroblastoma: proposition for a scoring system.

Abstract: A two-step evaluation system is proposed to analyze the results of therapy for stage IV neuro-blastoma in children over 1 year of age. Since most protocols consist of first-line chemotherapy followed by surgery and often high-dose chemotherapy with bone marrow rescue, “response” should assess the effect of chemotherapy alone and “remission” indicate the status after an attempt to remove the primary tumor. A scoring system is proposed for both steps. The comparability of different studies depends on the use of such common criteria to define patient groups and treatment results.

Human monoclonal antibodies to cell surface antigens of melanoma

Abstract: This invention provides a monoclonal antibody, produced by the hybridoma cell line designated GXM1, which specifically binds to a human class 1tumor antigen. This invention also provides a human monoclonal antibody, produced by a hybridoma cell line designated HJM1, which specifically binds to each of the ganglioside antigens GD2, GD3, GM3 and GD1b. This invention further provides a human monoclonal antibody, produced by a hybridoma cell line designated FCM1, which specifically binds to the ganglioside antigens GM3 and GD1a. Finally, this invention provides a human monoclonal antibody, produced by a hybridoma cell line designated DSM1, which specifically binds to a human class 2 tumor protein antigen.

Monoclonal antibodies to human lung cancers and method

Abstract: A panel of monoclonal antibodies produced from normal human lung fibroblasts and human lung tumors as immunogen is used to diagnose the presence of lung tumors and differentiate between those which are benign and those which are cancerous.

Vaccine for stimulating or enhancing production of antibodies directed against gm2

Abstract: Vaccine for stimulating or enhancing in a subject to whom the vaccine is administered, production of antibodies directed against GM2. The vaccine comprises an amount of purified GM2 effective to stimulate or enhance antibody production in the subject, an effective amount of a microbial adjuvant and a pharmaceutically acceptable carrier. This invention also provides a method for stimulating or enhancing in a subject production of antibodies directed against GM2. The method comprises administering to the subject an effective dose of the vaccine of this invention. The microbial adjuvant may be bacillus Calmette-Guerin or Salmonella minnesota R595. This invention further provides a method for treating cancer in a subject affected with cancer. The method comprises administering to the subject an effective dose of the vaccine of this invention. Finally, this invention provides a method for preventing cancer in a subject affected with cancer. The method comprises administering to the subject an effective dose of the vaccine of this invention.

Job characteristics and satisfaction of pharmacy technicians.

Abstract: Perceptions of job characteristics and job satisfaction of central-area technicians and drug-administration technicians at The Ohio State University Hospitals were analyzed. A questionnaire was administered to 79 pharmacy drug-administration technicians; 44 central-area technicians; 10 pharmacy residents, who served as objective raters (5 in the central area and 5 in the decentral area); 13 central-area pharmacists; and 17 decentral-area pharmacists. Perceived job characteristics were measured with the Job Characteristics Inventory; job satisfaction was measured by the Minnesota Job Satisfaction Questionnaire. The two groups of technicians differed significantly in their perceptions of task identity, task significance, and dealing with others. Significantly greater autonomy in technicians' jobs was perceived to exist by pharmacists and raters in both areas than by technicians. Significantly greater task identity was perceived by the central-area technicians than by their raters, and significantly greater task importance was perceived by both groups of technicians than by their pharmacists and raters. Friendship opportunities were perceived to exist to a significantly greater degree by decentral pharmacists and raters than by the drug-administration technicians. In the institution studied, both central-area and drug-administration technicians tended to be dissatisfied with their jobs. Central-area technicians' satisfaction was influenced most by the technicians' relationships with their supervisors and the feelings of accomplishment they gained. Drug-administration technicians were most satisfied if they believed they had opportunities to use their abilities. Efforts to increase job satisfaction among pharmacy technicians should focus on increasing feedback and task identity.

Alloantigens stimulating graft-v-host disease.

Abstract: This paper describes the Madison experience in pancreas transplantation. With the use of pancreaticocystostomy, overall mortality could be decreased to a level similar to the results reported for kidney transplantation in diabetic patients. Particularly encouraging are the results obtained in combined kidney and pancreas transplantation, with current actuarial graft survival rates for kidneys of 95% and pancreatic grafts of 85%. Overall, the quality of life for patients receiving a pancreatic transplant has been good, and in cases where no postoperative complications were encountered, a tremendous feeling of well-being from a physiological and psychological standpoint was achieved. It is our conclusion that pancreatic transplantation should be pursued as a viable form of therapy in selected patients and centers with large experience in the transplantation of diabetic patients.