Biomedical subjects
Publications and source records attributed to I Chen.
Potent 4-anilido-substituted quinazolines which potently inhibit epidermal growth factor receptor (EGFR) kinase were prepared. Structure-activity relationship studies reveal high sensitivity to substitution at the aniline ring.
The potential estrogenic activities of atrazine and simazine were investigated in vivo using the immature female Sprague-Dawley rat uterus and in vitro using the estrogen-responsive MCF-7 human breast cancer cell line and the estrogen-dependent recombinant yeast strain PL3. Animals that were dosed with 50, 150, or 300 mg/kg of atrazine or simazine alone for 3 consecutive days did not exhibit any significant increases in uterine wet weight while decreases in cytosolic progesterone receptor (PR) binding levels and uterine peroxidase activity were observed. 17 beta-estradiol (E2)-induced increases in uterine wet weight were not significantly affected by cotreatment with either chemical; however, some dose-independent decreases in E2-induced cytosolic PR binding and uterine peroxidase activity were observed. In vitro, atrazine and simazine did not affect basal or E2-induced MCF-7 cell proliferation or the formation of nuclear PR-DNA complexes as determined by gel electrophoretic mobility shift assays. In addition, these chloro-S-triazines did not display agonist activity or antagonize E2-induced luciferase activity in MCF-7 cells transiently transfected with a Gal4-human estrogen receptor chimera (Gal4-HEGO) and a Gal4-regulated luciferase reporter gene (17m5-G-Luc). Moreover, the estrogen-dependent PL3 yeast strain was not capable of growth on minimal media supplemented with atrazine or simazine in place of E2. Collectively, these results indicate that the reported estrogenic and antiestrogenic effects elicited by these chemicals are not mediated by the estrogen receptor.
Treatment of immature 21-day-old female Sprague-Dawley rats with 17 beta-estradiol (E2) (0.5 microgram/rat) caused a significant increase in uterine wet weight, DNA synthesis, progesterone receptor (PR) binding, and peroxidase activity. At doses as high as 40 mg/rat, the bioflavonoid naringenin did not cause a significant increase in any of these E2-induced responses. However, in rats cotreated with E2 (0.5 microgram/rat) plus naringenin (30 mg/rat); there was a significant decrease in E2-induced uterine wet weight, DNA synthesis, PR binding, and peroxidase activity, indicating that naringenin exhibits antiestrogenic activity in the immature rodent uterus. The binding of uterine nuclear extracts to a 32P-labeled estrogen responsive element (ERE) or progesterone responsive element (PRE) was determined using gel electrophoretic band shift assays. Incubation of [32P]ERE with uterine nuclear extracts from rats treated with naringenin or E2 resulted in the formation of estrogen receptor (ER):ERE complexes; a higher mobility complex was prominent in the extracts from E2-treated rats, whereas a lower mobility complex was observed using nuclear extracts from naringenin-treated animals. There was a significant decrease in the intensity of the E2-induced complex using nuclear extracts from rats treated with E2 plus naringenin. In contrast, transformed cytosol from control rats gave an intense ER:ERE complex, whereas the intensity of the band was decreased markedly using transformed uterine cytosol from treated rats. Formation of a PR:PRE complex was also determined using transformed uterine cytosol. Cytosol from E2-treated rats gave an intense retarded band, whereas only weak bands were observed using cytosols from DMSO- (solvent), naringenin-, or naringenin plus E2-treated cells. The results of in vitro studies showed that 1 nM E2 increased (3- to 4-fold) the growth of MCF-7 human breast cancer cells, whereas 1-1000 nM naringenin had no effect on cell proliferation. In cells cotreated with 1 nM E2 plus 1000 nM naringenin, there was a significant decrease in E2-induced cell growth. In MCF-7 cells transiently transfected with a pS2 promoter-regulated luciferase reporter gene, naringenin exhibited weak estrogenic activity. In cells cotreated with 0.1 or 1.0 microM naringenin plus 1 nM E2, naringenin inhibited E2-induced luciferase activity. The results of these studies confirmed that naringenin is a weak estrogen that also exhibits partial antiestrogenic activity in the female rat uterus and MCF-7 human breast cancer cells.
An alpha-amylase gene from Streptomyces sp WL6 was cloned on a 3.1kb DNA fragment, which was completely sequenced. The 3088 nucleotide sequence obtained contains three putative coding regions in the same orientation. The one corresponding to the structural region of the alpha-amylase gene has a deduced amino acid sequence of 459 residues, showing up to 71% identity to other alpha-amylases. An incomplete ORF was identified upstream the alpha-amylase gene, and the deduced product presents some homology to proteins involved in catabolic regulation.
Studies investigating the erosion mechanism of the newly developed poly (fatty acid dimer: sebacic acid) polyanhydride (p:[FAD:SA]) are described. The overall erosion of different monomer compositions of p(FAD:SA) copolymers was examined to determine whether and to what extent copolymer properties affected polymer erosion. Increasing the hydrophobic monomer (FAD) content up to 50 wt% in the copolymer resulted in longer erosion, whereas further increases up to 70 wt% decreased the erosion period. Polymer crystallinity depended on copolymer FAD content. Copolymer degradation was studied by examining anhydride bond hydrolysis using infrared spectroscopy. Much faster hydrolysis was found in p(FAD:SA) 70:30 compared with more crystalline copolymers of higher SA content. Light microscopy indicates the presence of an erosion zone, a distinct area where mass loss occurs. This erosion zone moves from the outside toward the interior of the polymer matrix. It plays an important role in erosion because any water or monomer must diffuse through this eroded layer.
Chronic stress can injure hippocampal neurons as well as alter hippocampal function. The adenylyl cyclase (AC) signal transduction system is an important modulator of neurotransmission in this brain region. This study was conducted to begin to understand the effects of chronic stress on the hippocampal AC system. To assess dependence of type I and type II AC mRNA expression on adrenal integrity, total RNA was prepared from the hippocampus of nonstressed rats 7 days following either sham surgery or adrenalectomy (ADX). Adrenalectomy resulted in a 60% diminution in steady-state expression of type II AC mRNA (p < .005) and a nonsignificant fall in type I AC mRNA expression. Both sham and adrenalectomized rats were then exposed to 1-hour sessions of restraint, twice per day (0800 to 0900 and 1500 to 1600 hours) for 4 days. Following the stress paradigm, RNA was prepared from the hippocampus and type I and type II AC mRNA levels determined by Northern blot. Densitometic analysis showed that in comparison to unstressed sham rats, stressed sham rats had a significant twofold increase in steady-state levels of type I and type II mRNA. Stressed adrenalectomized rats also had increased expression in type II AC mRNA but no significant stress-induced change in expression of type I AC mRNA. To assess dependence of hippocampal membrane G-Proteins on adrenal integrity, hippocampal membranes from nonstressed rats 7 days following either sham surgery or adrenalectomy (ADX) were analyzed by immunoblot.(ABSTRACT TRUNCATED AT 250 WORDS)
The relationship between viral burden, timing of transmission, and clinical progression was investigated in 110 children at risk for vertical human immunodeficiency virus (HIV) infection using quantitative polymerase chain reaction, coculture, and immune complex-dissociated p24 antigen assay. In a cross-sectional study, the mean HIV DNA copy number in 19 symptomatic children was significantly higher than in 31 infected, asymptomatic children (420 +/- 125 vs. 87 +/- 78; P < .0001). In a second group of 8 vertically infected infants followed prospectively from birth, 4 defined as infected in utero showed a more rapid increase in virus load, an accelerated loss of CD4 cells, and early progression to symptomatic disease (3-12 weeks) compared with 4 children with late in utero or intrapartum transmission (10-31 months). These data suggest that a direct relationship exists between HIV replication, the timing of transmission and onset and progression of HIV disease in vertically infected children.
An autopsied patient with Menkes' kinky hair disease, who showed unusually long survival until the age of five years with typical neuropathological changes, was examined for distribution of neuronal depletion in the cerebral cortex, and the cerebellar changes were compared morphologically and immunohistochemically with those found in a younger patient (1 year 8 months old) reported previously. Neuronal loss in the cerebral cortex in the both cases, which was ill-defined and unassociated with gliosis, was preferentially distributed in the fifth and sixth layers, especially of the gyral bottom in almost all lobes in the older case. Therefore, this change was thought to be secondary to local ischemia caused by mechanical distortion at the stage of gyrus formation in addition to abnormal development. Ultrastructurally, a prominent increase of confronting cisternae (CC) complexes was found in the perikaryon and processes of Purkinje cells in both cases, and in the older patient CC complexes were arranged more densely and were transformed into concentric lamellar structures in the swollen dendrites. Immunohistochemically, the stainability of neurofilaments (NF, 200 kDa) in Purkinje cells, with or without somatic sprouts was faint or negative in the older patient compared with the marked or moderate positivity in the younger patient and age-matched controls. Empty baskets were absent and NF-positive axonal terminals and synaptophysin-positive granules on Purkinje cells were markedly decreased in both cases. These changes suggest that Purkinje cells degenerate progressively with time and that basket cells also are simultaneously involved.
Adult male rats fed defined diets containing various fiber supplements or cholestyramine for 4 wk were surgically provided with lymphatic drainage catheters and starved overnight. After duodenal administration of a standard lipid test emulsion, absorption rates and lipoprotein distributions of cholesterol and oleic acid were determined. Prefeeding diets containing cellulose or alfalfa had no significant effect on oleic acid absorption. Diets containing pectin, guar gum, metamucil, mixed fibers (Fibyrax), or cholestyramine caused decreased lymphatic recovery in the initial period; except for the metamucil diet, no decrease was caused in the 24-h recovery, suggesting delayed but not impaired absorption. Fatty acid distribution among lipoproteins and chylomicron size were not altered by diet. All supplements caused a significant reduction in cholesterol absorption during the initial period, and cholesterol absorption remained depressed in animals prefed pectin, guar gum, mixed fibers, metamucil, and cholestyramine.
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Normal human breast lobules, freshly isolated by precision microdissection of tissue stained with methylene blue chloride, were assayed for their ability to induce neovascularization (angiogenesis) in rabbit irises. Histologically, normal lobules from cancerous breast induced angiogenesis twice as often as lobules from noncancerous breasts, suggesting that preneoplastic transformation is diffuse.
Peripheral blood mononuclear cells from a patient with polycythemia vera were cultured in a methylcellulose system employing human serum. Electron microscopy documented the appearance of mixed colonies containing lymphocytes, granulocytes, megakaryocytes, and erythrocytes. In vitro culture characteristics were similar to those seen for other patients with polycythemia ver, ie, colonies grew in the absence of added erythropoietin or other pathway-specific regulators. Plating efficiency was linearly related to the number of cells plated, which supports the concept that each colony arose from a single cell. The appearance of mixed myeloid-lymphoid colonies points to the existence of a primitive stem cell capable of giving rise to multiple hematopoietic cell lines.
Reticuloendotheliosis virus strain T (REV-T) is a highly oncogenic avian retrovirus which causes a rapid neoplastic disease of the lymphoreticular system. We derived six cell lines (1-3, 1-5, 2-10, 2-14, 2-16, and 2-20) from chicken spleen cells infected with REV-T. These cells can produce both the REV-T and its associated reticuloendotheliosis helper virus, REV-A. Histochemical analyses of these cells indicate that, while they are not stained by benzidine, peroxidase, beta-glucuronidase or acid alpha-naphthyl acetate esterase, they contain a high proportion (95%) of cells positive for acid phosphatase. Light and electron microscopic studies of these cells also revealed morphologies of lymphoblasts or activated lymphocytes with irregular nuclei and dispersed chromatin. Immunochemical analyses indicate that essentially all (90 to 100%) of the cells contain the surface marker Ia, but no cytoplasmic immunoglobulin M and immunoglobulin G could be detected by immunofluorescence staining. Results also show that some of these cell lines contain a low level of terminal transferase (0.02 to 0.17 unit/10(9) cells), and a proportion (3 to 35%) of these cells can be stained by an antiserum directed against chicken bursa cells. These results are consistent with the conclusion that the cells transformed by the highly oncogenic REV-T are lymphoid in nature. In addition, at least some of these cell clones may contain features characteristic of activated B-lymphocytes. Analysis of these cell clones indicates that some cell lines contain an adherent and nonadherent population with some differences in morphologies. In addition, electron microscopic examination revealed that, while the non-adherent cells are actively producing type C viruses, type C viruses are either absent or very rare in the adherent cell populations. These results support the conclusion that some of these cell lines are heterogeneous and contain subpopulations of cells with differences in their ability to produce viruses.
Radioiodinated alpha-bungarotoxin (alpha-Bgt) was used to localize alpha-Bgt-acetylcholine receptors in the carotid body of the rat. The gamma spectrometer analyses indicated a high uptake of [125I] alpha-Bgt in carotid bodies incubated in vitro (1.51 fmole per organ). Incorporation of the isotope was effectively blocked by pretreatment of carotid bodies with d-tubocurarine and unlabeled alpha-Bgt, but not by atropine. Light microscopic autoradiography showed a heavy labeling of some parenchymal cells. Electron-microscopic autoradiography revealed that labeling was localized along the interface between parenchymal cells, especially where their cytoplasmic processes engage in complex interdigitations. The silver grain counts on electron-microscopic autoradiographs suggest that labelings are preferentially associated with the plasma membrane of certain Type I cells. It is suggested that these Type I cells in the rat's carotid body probably are provided with nicotinic acetylcholine receptors on their plasma membranes.
Proximal portions of the left coronary arteries were examined microscopically in aging female normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). The age-related intimal alterations in SHR were largely limited to endothelial cells, which demonstrated a proliferation of organelles (most notably Weibel-Palade bodies). In the media, degenerative alterations appeared to be most marked near the medio-adventitial junction. Compared to WKY and increasing with age, the media of the SHR epicardial artery demonstrated an accumulation of extracellular elements that included basement membrane-like material, collagen fibers and debris. Complex carbohydrates, as determined with the silver methenamine reaction, were noted to accumulate in a lamellar fashion. Smooth muscle cells demonstrated an age-related tendency (which was exaggerated in SHR) toward development of invaginations and irregular profiles. These observations indicate that age-related structural alterations in epicardial arteries of SHR are progressive, and they support the concept that structural alterations in SHR coronary arteries may represent accelerated aging phenomena.