Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0432222 (SEM)
 47,337 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The peroxidase-labeled antibody method was employed for immunohistochemical localization of intracellular and extracellular antigens with SEM. With this method, the antigenic sites may be localized with high sensitivity on tissue sections mounted on carbon-coated glass slides by detecting 0s04-DAB complexes using the backscatter mode. The sections may be fresh frozen fixed on slide, frozen sections of fixed tissue, or sections of immunostained tissue embedded in epon. The method introduces another utilization of SEM and the immunoenzyme histochemistry.
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PMID:Intracellular localization of antigens with backscatter mode of SEM using peroxidase-labeled antibodies. 703 73

Characterizing lymphocyte subsets in bronchoalveolar lavage fluid (BALF) by flow cytometry (FC) proper gating of the lymphocyte subpopulation being analyzed is crucial. In order to test lymphocyte gate quality for the first time we used a DNA-dye to evaluate plasmamembrane integrity and thus to mark off fluorescent but not DNA-containing particles (eg, debris). A comparative prospective study between this newly developed FC technique and a standard peroxidase anti-peroxidase (PAP) method was performed. Samples of BALF from 50 patients with various pulmonary diseases were examined. After determination of the total cell yield, a differential cell count was performed. Subsequently, the immunophenotype of pan T lymphocyte CD3-, T-helper lymphocyte CD4-, and T-suppressor lymphocyte CD8-positive lymphocyte subsets was assessed with FC as well as with the PAP method. Both methods showed excellent correlation (CD3: r = 0.81; CD4: r = 0.97; CD8: r = 0.96; p < 0.05, respectively). Comparing the mean +/- SEM, FC tends to overestimate CD3+ cells (90.6 +/- 1.0% vs 85.8 +/- 1.3%). For CD4 (45.0 +/- 3.4% vs 44.4 +/- 3.4%) and CD8 (48.1 +/- 3.5% vs 46.7 +/- 3.5%), there was good agreement. In a clinical setting, the reliability of both methods was equivalent, and FC using a DNA-dye to test lymphocyte gate quality offered a rapid and reliable determination of lymphocyte subsets in BAL.
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PMID:Immunophenotyping of lymphocytes in bronchoalveolar lavage fluid. A new flow cytometric method vs standard immunoperoxidase technique. 763 85

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a newly discovered neuropeptide that is present in high amounts in hypothalamic neuroendocrine neurons and potently stimulates the accumulation of cAMP within cells of the anterior pituitary. We have employed several specific antisera recognizing different parts of the PACAP precursor to elucidate the distribution of PACAP-like immunoreactivities in the hypothalamic components of the hypothalamo-pituitary-adrenocortical axis in sections obtained from normal and colchicine-treated rats. Using immunohistochemistry with avidin-biotin-coupled peroxidase as a reporter system, high numbers of PACAP-immunoreactive perikarya were found in colchicine-pretreated rats in many of the parvicellular subdivisions of the hypothalamic paraventricular nucleus (PVN). A few cells were also found in the magnocellular subdivisions of the nucleus, and a similar small population of cells was observed in the dorsolateral aspect of the supraoptic nucleus. Using indirect immunofluorescence, the relation between CRF- and PACAP-containing neurons in the various parvicellular subnuclei of the PVN was studied, and a high degree of colocalization was demonstrated in the neurons of the medial parvicellular part of PVN. To further study the functional implications of PACAP in the hypothalamo-pituitary-adrenocortical axis, we examined the expression of PACAP messenger RNA (mRNA) in the PVN in response to five different stimulatory paradigms that previously have been shown to stimulate CRF mRNA expression in the medial parvicellular part of the PVN. The stimulatory challenges of adrenalectomy, restraint stress, ip injection of hypertonic saline, ether stress, and intracerebroventricular injection of colchicine induced significant elevations of CRF mRNA expression in the medial parvicellular part of the PVN. In contrast, the expression of PACAP mRNA, which is hardly detectable within the medial parvicellular part of the PVN, was induced only by colchicine treatment (from undetectable levels to 177 +/- 21 dpm/g; mean +/- SEM), whereas PACAP mRNA remained undetectable in this region of the PVN after exposure to any of the other stimulatory paradigms. The onset of colchicine-induced PACAP mRNA expression in the PVN was rapid (3 h), and PACAP mRNA levels remained elevated throughout the 48-h observation period. Considering the different topography and connections of the parvicellular subnuclei of the PVN, the current observations suggest that PACAP present in parvicellular neurons of the PVN may act not only as a neuroendocrine transmitter/modulator in the hypothalamo-pituitary-adrenocortical axis, but also as transmitter mediating neurotransmission conveyed from the PVN to preganglionic neurons of the autonomic system.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Pituitary adenylate cyclase-activating peptide gene expression in corticotropin-releasing factor-containing parvicellular neurons of the rat hypothalamic paraventricular nucleus is induced by colchicine, but not by adrenalectomy, acute osmotic, ether, or restraint stress. 764 20

On their surface, renal tubular cells present intercellular adhesion molecule-1 (ICAM-1) during acute renal allograft rejection. We propose that the extent of ICAM-1 expression by renal tubular cells can be estimated from urine immunocytology. To test this hypothesis, we obtained 52 samples of urine from 31 renal transplant recipients with either acute tubular necrosis, rejection or stable renal function. Cytocentrifuged aliquots of urinary sediment were incubated with monoclonal antibodies to ICAM-1 in an avidin-biotin-peroxidase technique. To corroborate our findings, biopsy specimens were obtained for conventional and immunohistology one hour following vascular anastomosis and during rejection episodes. The proportion of renal tubular cells that expressed ICAM-1 was low in patients with acute tubular necrosis (23.8 +/- 3.6%) and high in patients with rejection (53.1 +/- 4.4% [SEM]) (P < .001). In 11 patients who recovered from rejection, the proportion of ICAM-1-positive renal tubular cells decreased from 55.9 +/- 5.6% to 25.5 +/- 4.3% (P < .05). In two patients who initially had acute tubular necrosis and then rejected their transplants, the expression of ICAM-1 on renal tubular cells tended to increase (from 27.5 +/- 2.5% to 60.0 +/- 20.0%, P = .12). In eight patients with acute tubular necrosis who never rejected their transplants, ICAM-1 expression remained low (23.1 +/- 3.8%). Immunocytology correlated well with immunohistology and the clinical diagnosis. Our findings suggest that urine immunocytology may be useful in monitoring adhesion molecule expression by renal tubular cells.
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PMID:Analysis of adhesion molecule expression by tubular epithelial cells using urine immunocytology. 776 29

The neuropeptide hormones arginine-vasopressin (AVP) and oxytocin (OT) have been found in the ovarian follicles and corpora lutea (CL) of many eutherian mammals. In ruminants, there is persuasive evidence that luteal OT is involved in luteolysis via stimulation of uterine prostaglandins. However, based on scant evidence, the marsupial ovary has been viewed as being devoid of OT-like and AVP-like peptides. In this study, corpora lutea from the brushtail possum were examined for OT, AVP, and mesotocin (MT) by a combination of reverse phase HPLC, radioimmunoassay, and immunohistochemistry (IHC). Peptides extracted from each of five CL were separated by HPLC and each fraction was assayed for AVP, MT, and OT. Two immunoreactive peaks were found, corresponding to AVP and MT standards. The amount of each peptide was 8.7 +/- 2.22 pmol MT/g (mean +/- SEM) and 5.7 +/- 1.0 pmol AVP/g, respectively. The mean MT/AVP ratio was 1.55 compared to 0.26 for the pituitary. IHC (streptavidin-peroxidase method) of Bouin's-fixed CL showed staining for MT in the cytoplasm of luteal cells which was absent in stromal tissue and nonluteal ovarian tissue. Not all luteal cells were immunopositive and no topographical distribution of stained cells was observed. IHC localization of AVP was not attempted. It was concluded that the CL of the brushtail possum contains low quantities of MT and AVP, which in the case of MT is probably synthesized by the immunochemically staining cells of the CL.
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PMID:Mesotocin and arginine-vasopressin in the corpus luteum of an Australian marsupial, the brushtail possum (Trichosurus vulpecula). 817 26

Although the pituitary is known to produce several growth factors, their effects on pituitary cell growth and differentiation are still unclear, particularly in normal tissue. Using primary cultures of aged ewe pituitaries cultured in serum-free conditions, we studied the effects of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), transforming growth factor-beta (TGF beta 1), insulin, and 17 beta-estradiol (E2) on the growth, differentiation, and expression of gonadotropin subunit genes. After 72-h incubation of the monolayer (day 5) with optimal concentrations of each factor, [3H]thymidine incorporation was increased significantly (P < 0.01) over the control values by 33 +/- 8% (mean +/- SEM; n = 3; 10 nM E2), 36 +/- 10% (1 ng/ml TGF beta 1), 83 +/- 12% (10 ng/ml bFGF), and 118 +/- 12% (1 nM EGF). Insulin showed a two-phase dose-response curve, increasing [3H]thymidine uptake by 34 +/- 9% at 10 ng/ml and by 63 +/- 13% at 10 micrograms/ml. Cell counting using a Coulter counter confirmed these results. Characterization of cell types by immunocytochemistry (avidin-biotin-peroxidase complex technique) revealed that the cell cultures were predominantly gonadotrophs. However, the cultures contained cells that did not stain with any specific ovine or human antiserum against LH beta, FSH, TSH beta, PRL, GH, ACTH, or glial fibrillary acidic protein, but were of epithelial cell lineage, as shown by positive keratin staining. Treatment with EGF and bFGF increased the proportion of these undifferentiated pituitary cells and induced changes in their morphology to large cuboidal cells containing large nuclei. After treatment with E2, insulin, and TGF beta 1, pituitary cells remained differentiated, although with E2, staining for gonadotrophs was much reduced. Northern blot analysis revealed that E2 treatment for 0-48 h progressively reduced the mRNA for FSH beta (16 +/- 4.5% of control values) and LH beta (12.4 +/- 2.5%), but had little effect on the common alpha-subunit (88.4 +/- 4.6%). TGF beta 1, however, stimulated the expression of FSH beta subunit gene by 142 +/- 4.6% (P < 0.01) of the control value, but had no significant effect on LH beta and common alpha-subunit genes. Insulin, EGF, and bFGF showed no significant effect on the expression of these three subunit genes. The data define the direct effects of growth factors and E2 on the growth and differentiation of normal sheep pituitary cells and gonadotrophs in particular, which may be of relevance to the pathophysiology of the pituitary and in the multistage process of pituitary tumorigenesis.
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PMID:Role of growth factors and estrogen as modulators of growth, differentiation, and expression of gonadotropin subunit genes in primary cultured sheep pituitary cells. 829 88

An immunohistochemical method, based on an indirect peroxidase method for staining of capillaries containing collagen type IV, is presented. Biopsies from the vastus lateralis muscle of eight endurance trained runners were stained and counted for capillaries with the collagen type IV method and with the widely used amylase-PAS method. The mean number +/- SEM of capillaries per fibre was 2.48 +/- 0.09 with the collagen type IV method and 2.41 +/- 0.09 with the collagen type IV method and 2.41 +/- 0.08 with the amylase method. The 95% confidence intervals for the differences between the two methods were - 0.03 to 0.15. We conclude that the collagen type IV immunohistochemical method for capillary staining is comparable with the amylase method under optimum conditions, but that it is likely to be more dependable. It also has the advantage that it reveals fibre outlines.
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PMID:Capillary density measurements in skeletal muscle using immunohistochemical staining with anti-collagen type IV antibodies. 856 82

The tube repair method was used to study peripheral nerve regeneration in five different inbred mouse strains. The sciatic nerve of male adult mice of the C57BL/6J, DBA/1J, C3H/HeJ, BALB/cJ and A/J strains (N = 3) was cut and both proximal and distal nerve stumps were inserted into a polyethylene tube leaving a 4-mm nerve gap. After 6 weeks the tubes containing the regenerated nerve cables were processed for total myelinated axon counts. C57BL/6J mice regenerated significantly fewer myelinated axons (1024 +/- 178, mean +/- SEM) compared to the BALB/cJ (1618 +/- 64), A/J (1788 +/- 95), DBA/1J (2168 +/- 296) or C3H/HeJ (3468 +/- 36) strains. Horseradish peroxidase was applied 3 mm distal to the tube 4 and 40 weeks after tube implantation to further characterize the reduced regenerative response of C57BL/6J mice. Labeled sensory and somatic motor neurons were counted in the spinal cord and L4,5,6 dorsal root ganglia (DRG), respectively. Sciatic nerves from four intact C57BL/6J mice were processed in the same fashion and used as normal controls. No significant difference in the number of motor neurons was detected between the experimental (4 weeks = 663 +/- 74; 40 weeks = 770 +/- 35) and control non-operated (844 +/- 13) animals. However, there were fewer labeled neurons in the DRG of the operated group (4 weeks = 1163 +/- 167; 40 weeks = 2574 +/- 104) compared to the control group (4211 +/- 96). These results indicate that sensory neurons are responsible for the diminished regenerative response in C57BL/6J mice after peripheral nerve transection.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Reduced sensory neuron regeneration by C57BL/6J mice. 858 Aug 70

The inability to separate irreversible lesions of tubular epithelia from reversible tubular atrophy constitutes a major problem in histopathology and in decisions for revascularization of shrunken kidneys with renal artery stenosis. In order to characterize reversible tubular atrophy ('kidney hibernation') we studied the physiological and biochemical parameters and morphology including histochemistry in rat kidneys made atrophic by renal artery stenosis and treatment with the angiotensin-converting enzyme inhibitor, enalapril. Renal artery stenosis was induced by a 0.2-mm clip around the left renal artery. Following 7 weeks of clipping and 2 concomitant weeks of enalapril treatment, the kidney length decreased from 17.8 +/- 0.3 to 13.7 +/- 0.7 mm (mean +/- SEM). Renal blood flow and glomerular filtration rate decreased to 39 +/- 3% and to approximately 3% of control values, respectively. The activities of the intracellular proteolytic enzymes cathepsin B and L and of Na-K-ATPase in microdissected proximal tubular segments decreased to values below 50 and 10%, respectively. All changes were significant (p < 0.05). Histochemical staining for ATPase activity in the distal tubule segments remained unchanged. Tubular cells were atrophic but not necrotic. Histochemical staining of alkaline phosphatase in the tubular brush border and of acid phosphatase and peroxidase in lysosomes was greatly reduced. All observed changes were reversible within 2-3 weeks following removal of the clip and withdrawal of enalapril either with or without contralateral nephrectomy. Thus, a form of kidney hibernation with readily reversible tubular atrophy has been described. Based on this description it may be possible in consecutive experiments to differentiate between reversible and irreversible tubular atrophy.
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PMID:Characteristics of renal tubular atrophy in experimental renovascular hypertension: a model of kidney hibernation. 868 34

An imbalance between oxidative stress and antioxidative capacity is thought to play an important role in the development and progression of chronic obstructive pulmonary disease (COPD). To assess the lung oxidative status in patients with COPD, we studied whether exhaled hydrogen peroxide (H2O2) is increased in breath condensate of patients with stable COPD (n = 12, mean FEV1 51% pred) and in patients with exacerbated COPD (n = 19, actual FEV1 36% pred) compared with a healthy control group (n = 10, FEV1 108% pred). Expired breath condensate during 15 min of tidal breathing was collected by cooling. The concentration of H2O2 was measured spectrophotometrically by means of horse radish peroxidase-catalyzed oxidation of tetramethylbenzidine. Concentrations of H2O2 (mean +/- SEM) were significantly elevated at 0.205 +/- 0.054 microM in patients with stable COPD compared with 0.029 +/- 0.012 microM in the control group (p < 0.05) and were further increased to 0.600 +/- 0.075 microM in patients with acutely exacerbated COPD (p < 0.001 compared with patients with stable COPD). Patients with pulmonary infiltrates on chest radiograph showed similar values compared with patients without obvious infiltrates. These findings demonstrate that patients with stable COPD exhibit increased oxidant production in the airways and that oxidant production increases further during exacerbations.
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PMID:Increased exhalation of hydrogen peroxide in patients with stable and unstable chronic obstructive pulmonary disease. 881 Jun 24


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