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Query: UNIPROT:P06889 (Mol)
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To evaluate a short-term epithelial cell assay system to detect respiratory carcinogens, primary cultures of rat tracheal epithelial cells were exposed to a series of 17 compounds and scored for morphologically transformed cell colonies 28 days later. The test compounds included known carcinogens and noncarcinogens in volatile or liquids form. Tracheal epithelial cells were isolated from F344 rats, plated onto collagen-coated dishes, and exposed to the test compounds on day 1 for 24 hours. At day 30 the cultures were fixed, stained, and scored for colonies having a density greater than 1,300 cells/min2. With standardized protocols, such colonies are very infrequent in media and solvent control cultures. Concentration levels for each chemical were chosen over a range from nontoxic to toxic levels. Highly positive compounds in this assay included benzo(a)pyrene, benzo(l)acean-thyrlene, 3-methylcholanthrene, and formaldehyde. Compounds which were negative in this assay included pyrene, benzo(e)pyrene, and 4-nitroquinoline-N-oxide. Examining the concordance of in vitro results with whole animal carcinogenesis studies revealed an accuracy of 88% with one false-positive and one false-negative compound. The results of these studies indicate that the rat tracheal epithelial cell assay may be useful in identifying potential respiratory carcinogens in our environment.
Environ Mol Mutagen 1989
PMID:Evaluation of a rat tracheal epithelial cell culture assay system to identify respiratory carcinogens. 275 28

Using "protein-image" hybridization technique combined with various crosslinking methods, for formaldehyde-prefixed nuclei we have analysed changes induced by activation in the chromatin structure of HSP-70 genes. From the crosslinking data it follows that chromatin of actively transcribed genes undergoes some structural rearrangements resulting in certain weakening of the contacts between DNA and the globular parts of histones so that the histones remain bound to DNA through their N-terminal regions. In addition, there have been found two specific regions with a reduced content of histones: the 5'-promoter of HSP-70 gene and a region distanced by approximately 1 k.b. from the 3'-end of the HSP-70 gene.
Mol Biol (Mosk)
PMID:[Structural changes in chromatin of heat shock protein 70 gene of Drosophila during transcription]. 277 Jul 46

Immunization of DMBA-treated mice by the glycoprotein fraction from mammary glands of mice BALB/c did not decrease the frequencies of induction of mammary tumors. This is in contrast to the results obtained during immunization by the formaldehyde treated preparation of Mouse Mammary Tumor Virus (MMTV). EcoRI and HindIII cleaved DNA from the DMBA-induced mammary tumors did not contain the additional virus specific fragments. In mammary tumors the expression of p27 MMTV was registered in contrast to normal mammary glands and mammary epithelium cultures in which the proteins of MMTV are not expressed even after induction.
Mol Gen Mikrobiol Virusol 1987 Aug
PMID:[Endogenous MMTV in the normal mammary gland and in dimethylanthracene-induced mammary cancer in BALB/c mice]. 282 5

Epidermal "dark cells" (DC) are believed to play a specific role in the so-called promotion phase of experimental skin carcinogenesis. They are recognized by their morphological features both at the light and the electron microscopical level. The possible effects of fixation on the morphology of epidermal cells and hence on the number of DC have not yet been thoroughly studied. In the present light microscopical study we used a semiquantitative method together with simple cell counting to evaluate the influence of fixation on the specific cellular morphology which is traditionally used to determine the number of DC. The use of cacodylate vehicled prefixatives, either formaldehyde or glutaraldehyde, led to a higher incidence of DC, and furthermore both to an increased width of the intercellular spaces (ICS) and a more heavy staining of the keratinocytes than when s-collidine vehicled glutaraldehyde was used. Differences in yield of DC solely due to the prefixative itself (formaldehyde or glutaraldehyde) were not detected. Exposure to TPA or the use of a hyperosmolal prefixative vehicle both yielded higher DC numbers than did controls or conventional prefixative vehicles, respectively. After prefixation with hyperosmolal vehicles, however, TPA treatment did not induce higher DC yield than in a control series. Phenomena usually accompanying exposure to TPA, such as intercellular oedema (widening of the ICS) and cytoplasmic vacuolization, varied in parallel to the number of DC. Hence, there is reason to believe that the induction of epidermal DC is mainly associated with volume reduction of keratinocytes. Such shrinkage may be due to the cytotoxic properties of TPA and degenerative phenomena appearing during tissue processing.
Virchows Arch B Cell Pathol Incl Mol Pathol 1986
PMID:The influence of different fixatives and a tumor promoter, 12-0-tetradecanoyl-phorbol-13-acetate (TPA), on the induction of so-called dark cells in mouse epidermis. A light microscopical study. 287 May 86

Immunohistochemical techniques have been used to localize clotting factor XIII subunit A in human reactive lymphoid follicles. The follicular dendritic reticulum cells (DRCs) were identified by the monoclonal antibodies R4/23 and OKB-7 as well as by their 5'-nucleotidase positivity. Follicular histiocytic reticulum cells (HRCs) were demonstrated by their acid phosphatase and non-specific esterase reactions. Capillaries were selectively visualized by adenosine triphosphatase. The immunohistochemical demonstration of F-XIIIa was preferably carried out in combination with one or two of the above marker techniques, on the same cryostat section. The subunit A of factor XIII is present in follicular DRCs. Their selective immunohistochemical demonstration with antibody against F-XIIIa requires formaldehyde fixation of cryostat sections. Similar fixation, however, is inappropriate for the demonstration of F-XIIIa reactivity of DRCs in paraffin sections. For this purpose, acetic acid-formalin fixation is useful. Follicular HRCs are consistently negative for F-XIIIa, contrary to the F-XIIIa positivity of sinusoidal and interfollicular HRCs. Developmental and functional implications of F-XIIIa reactivity in DRCs and HRCs are suggested.
Virchows Arch B Cell Pathol Incl Mol Pathol 1987
PMID:Selective visualization of human dendritic reticulum cells in reactive lymphoid follicles by the immunohistochemical demonstration of the subunit A of factor XIII (F-XIIIa). 288 67

We undertook an immunohistochemical analysis of human bronchopulmonary epithelial neoplasms and pleural mesotheliomas using a monoclonal antibody which recognizes ras oncogene products (p21ras). The monoclonal antibody, RAP-5, recognizes both unaltered and certain mutated p21ras. Formalin fixed and paraffin embedded tissue samples of 187 lung epithelial tumors and 27 pleural mesotheliomas were investigated; normal and bronchiectatic lungs were similarly studied. Normal lung and pleural tissue did not immunostain except for occasional type II pneumocytes. Reactive type II pneumocytes adjacent to carcinomas and bronchiectasis immunostained consistently. Twenty four/34 (71%) squamous carcinomas immunostained. Only 8/50 (16%) adenocarcinomas immunostained focally and weakly whereas 19/24 (79%) bronchioloalveolar carcinomas immunostained. Eleven/18 (61%) large cell carcinomas immunostained with variable intensity. Eleven/13 (85%) carcinoids, 6/7 (85%) well differentiated neuroendocrine carcinomas, and 18/21 (86%) intermediate cell neuroendocrine carcinomas immunostained while none of 20 small cell neuroendocrine carcinomas immunostained. Only a few mesotheliomas were immunostained focally. Two/14 (14%) epithelial type and 1/9 (11%) biphasic type mesotheliomas immunostained weakly; none of 4 spindle cell mesotheliomas immunostained. We conclude that while at least occasional cases of most types of pulmonary epithelial neoplasms express p21ras, the frequency and intensity of the expression are distinctly greater in certain tumor types such as squamous, bronchioloalveolar, and neuroendocrine neoplasm except for the small cell type. Contrary to these lung epithelial neoplasms, most mesotheliomas did not immunostain for p21ras. Whether the enhanced p21ras expression may point to a different mechanism of transformation or may merely reflect differentiation features remains undetermined.
Virchows Arch B Cell Pathol Incl Mol Pathol 1987
PMID:Immunohistochemical evaluation of ras oncogene expression in pulmonary and pleural neoplasms. 288 32

The molecular nature of formaldehyde (HCHO)-induced mutations was studied in both human lymphoblasts and E. coli. Thirty HPRT- human lymphoblast colonies induced by eight repetitive 150 microM HCHO treatments were characterized by Southern blot analysis. Fourteen of these mutants (47%) had visible deletions of some or all of the X-linked HPRT bands, indicating that HCHO can induce large losses of DNA in human lymphoblasts. In E. coli, DNA alterations induced by HCHO were characterized with use of the xanthine guanine phosphoribosyl transferase (gpt) gene as the genetic target. Exposure of E. coli to 4 mM HCHO for 1 hr induced large insertions (41%), large deletions (18%), and point mutations (41%). Dideoxy DNA sequencing revealed that most of the point mutations were transversions at GC base pairs. In contrast, exposure of E. coli to 40 mM HCHO for 1 hr produced 92% point mutations, 62% of which were transitions at a single AT base pair in the gene. Therefore, HCHO is capable of producing different genetic alterations in E. coli at different concentrations, suggesting fundamental differences in the mutagenic mechanisms operating at the two concentrations used. Naked pSV2gpt plasmid DNA was exposed to 3.3 or 10 mM HCHO and transformed into E. coli. Most of the resulting mutations were frameshifts, again suggesting a different mutagenic mechanism.
Environ Mol Mutagen 1988
PMID:Molecular analysis of formaldehyde-induced mutations in human lymphoblasts and E. coli. 290 Jul 62

An in situ hybridization technique was developed for the strand-specific detection of yellow fever virus (YFV) RNA. An 35S-labeled, transcribed RNA probe was used to detect positive-sense polarity YFV genomic RNA in infected C6/36 (Aedes albopictus) cells, dissected mosquito tissues, and sections of plastic-embedded, YFV-infected Aedes aegypti mosquitoes. Mosquito tissues fixed in buffered Formalin retained morphological integrity. The low concentrations of probe used yielded high specific signal on infected specimens and low background signal on uninfected specimens.
Mol Cell Probes 1988 Dec
PMID:Detection of yellow fever virus nucleic acid in infected mosquitoes by RNA:RNA in situ hybridization. 290 76

Treatment of rats with the cytochrome P-450 suicide substrate, 3,5-diethoxycarbonyl-2,6-dimethyl-4-ethyl-1,4-dihydropyridine (DDEP), produced a 95% inhibition of the in vivo demethylation of either aminopyrine or morphine within 2 hr. One-carbon metabolism of formaldehyde or formate to carbon dioxide was not altered. DDEP also produced a time-dependent decrease in total hepatic microsomal cytochrome P-450 but had no effect on either NADPH-cytochrome c reductase or p-nitrophenol glucuronyl-transferase activities up to 24 hr after administration. A rapid decrease in rat liver microsomal aniline hydroxylation and ethoxyresorufin deethylation was observed in vitro following DDEP administration. Although in vitro testosterone metabolism to 16 alpha-, 16 beta-, and 2 alpha-hydroxy metabolites was depressed profoundly by DDEP in microsomes from untreated and 3-methylcholanthrene-treated animals, 7 alpha-hydroxylation of testosterone was much less affected. Immunochemical quantification of various microsomal cytochrome P-450 protein moieties showed that cytochromes P-450 beta NF-B, P-450UT-A, P-450PCN-E, and P-450PB-C were decreased in hepatic microsomes from DDEP-treated rats. However, the protein moiety of cytochrome P-450UT-H was not diminished and the immunoreactive protein for cytochromes P-450UT-F, P-450PB-B, and P-450ISF-G was only slightly decreased. These results show that DDEP treatment leads to marked decreases in holoprotein and apoproteins of many but not all hepatic microsomal cytochrome P-450 isozymes.
Mol Pharmacol 1986 Jan
PMID:Effect of the suicide substrate 3,5-diethoxycarbonyl-2,6-dimethyl-4-ethyl-1,4-dihydropyridine on the metabolism of xenobiotics and on cytochrome P-450 apoproteins. 308 Jun 74

The common approach is developed for isolation of mutants deficient in key enzymes of ribulose monophosphate pathway for formaldehyde oxidation and assimilation by obligate methylotrophic bacteria. The approach is based on total isolation of temperature sensitive mutants and their biochemical characterization. A number of ts- mutants of obligate methylotroph M. flagellatum KT is isolated following nitrosoguanidine induced mutagenesis. The modified screening method was developed and used for identification of mutants deficient in the key enzymes of ribulose monophosphate pathway. The mutant deficient in glucose-6-phosphate dehydrogenase (zwf) was identified. The NAD-dependent activity of glucose-6-phosphate dehydrogenase was not measurable under nonpermissive temperature while the level of NADP-dependent activity was only four-fold less comparing with wild type strain. It was concluded that growth limitation of zwf mutant of M. flagellatum KT (designated T623) at 42 degrees C results from the absence of NAD-dependent activity of glucose-6-phosphate dehydrogenase.
Mol Gen Mikrobiol Virusol 1987 Jul
PMID:[Temperature-sensitive mutant of the obligate methylotroph Methylobacillus flagellatum KT deficient in glucose-6-phosphate dehydrogenase]. 311 3


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