Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Control (normothermic) and cold-acclimated (environmental temperature gradually reduced from 20 to 5 degrees C for 4 weeks) groups of male rats and hamsters were compared to elucidate the nature of angiogenesis in oxidative and glycolytic muscles of these species during progressive cold exposure. Skeletal muscle capillarity and fibre cross-sectional area were measured in the tonic soleus (SOL) and phasic tibialis anterior (TA). Cryostat sections were stained for alkaline phosphatase (ALP) activity to identify all capillaries, and proliferating cell nuclear antigen (PCNA) to localise the site of cellular proliferation. Cold-induced angiogenesis, indicated by an increase in capillary to fibre ratio (C:F), occurred in SOL of rats (approximately 20 % increase, P < 0.05) but not hamsters (approximately 9.5 % increase, n.s.), and in TA of hamsters (approximately 22 % increase, P < 0.01) but not rats (approximately 1 % increase, n.s.). The change in C:F was highest in the glycolytic cortex region of TA where fibre size is larger than in the oxidative core. Capillary-specific cell proliferation (co-localised ALP and PCNA labelling) increased in parallel with C:F. The total PCNA label density within the interstitium was some 5-fold higher than that co-localised with capillaries, but where angiogenesis occurred the relative increase in capillary labelling was 2-fold greater than for other cells of the interstitium. These data suggest a significant role for endothelial cell proliferation in the angiogenic response, indicative of the sprouting form of angiogenesis. There was a tendency for fibre hypertrophy in both SOL and TA of rats, especially in the core region of TA (P < 0.01), such that capillary density (CD) and intramuscular diffusion distances (DD) were largely unchanged following cold exposure. In contrast, fibre size was maintained in hamsters, DD reduced and CD increased compared to control TA (P < 0.01). In conclusion, cold acclimation stimulated angiogenesis in muscle of hamsters more than in rats, possibly due to a higher metabolic rate in the smaller species. Angiogenesis was also seen in SOL of rat, where oxidative capacity and muscle activity is higher than the TA. Thus, a combination of oxidative capacity, muscle activity, and fibre size may determine the degree of angiogenesis in response to low environmental temperature.
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PMID:Cold exposure differentially stimulates angiogenesis in glycolytic and oxidative muscles of rats and hamsters. 1460 72

We have reported that microwave cell death is a unique cell death preserving not only cell and nuclear shapes but also immunohistochemical antigenicity. However, their enzyme activity was lost, which indicated cell dysfunction and death. This peculiar observation implies that the microwave effect is likely an 'in situ' tissue fixation and that this type of cell death is morphologically different from cell death, by either oncosis or apoptosis, as previously known. To confirm whether this peculiar cell death was observed also in human tissue samples, we examined clinical samples from patients with metastatic liver cancer, which were treated with microwave irradiation. They were examined immunohistochemically for human Ki-67 antigen and proliferating cell nuclear antigen and enzyme histochemically for alkaline phosphatase, and the same morphological changes that were observed in microwave-treated rat liver were found. In conclusion, we believe that routine hematoxylin-eosin stain alone is not a suitable method to evaluate microwave treatment for cancer because microwave coagulation therapy-treated cells preserved their nuclei and cellular architectures, even after 3 months. For microwave-treated tumors, enzyme histochemistry is helpful to determine its effectiveness.
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PMID:Microwave cell death: Enzyme histochemical evaluation for metastatic carcinoma of the liver. 1462 49

Retinoic acid has been shown to adversely affect craniofacial development. Cleft palate and craniosynostosis are two examples of craniofacial defects associated with prenatal exposure to this agent. Although the effects of retinoic acid on cephalic neural crest-derived tissues have previously been studied, the specific effects of retinoic acid on the cellular biology of osteoblasts remain unclear. The purpose of this study was to analyze in detail the effects of pharmacologic doses of retinoic acid on the differentiation and proliferation of osteoblasts derived from an intramembranous source. Primary rat calvarial osteoblasts were established in culture and treated with 1 or 10 microM all-trans-retinoic acid. Retinoic acid treatment markedly increased expression of osteopontin up to 48 h after stimulation. Consistent with this early stage of differentiation, both mRNA and protein analysis of FGF receptor isoforms demonstrated a switch in predominance from fibroblast growth factor receptor 2 (fgfr2) to fgfr1. Analysis of PCNA protein confirmed inhibition of proliferation by retinoic acid. To determine whether these alterations in osteoblast biology would lead to increased differentiation, we examined short term [alkaline phosphatase (AP) activity] and long term (von Kossa staining) surrogates of bone formation in vitro. These assays confirmed that retinoic acid increased osteogenesis, with a 4-fold increase in bone nodule formation in cells treated with 10 microM retinoic acid after 28 days. Overall, our results demonstrated that pharmacologic doses of all-trans-retinoic acid decreased osteoblast proliferation and increased differentiation, suggesting that retinoic acid may effect craniofacial development by pathologically enhancing osteogenesis.
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PMID:High-dose retinoic acid modulates rat calvarial osteoblast biology. 1538 22

The hypertrophic chondrocytes lack the ability to proliferate, thus permitting matrix mineralization as well as vascular invasion from the bone in both the mandibular condyle and the epiphyseal cartilage. This study attempted to verify whether the histological appearance of the hypertrophic chondrocytes is in a steady state during postnatal development of the mouse mandibular condyle. Type X collagen immunohistochemistry apparently distinguished the fibrous layer described previously as the "articular zone," "articular layer," and "resting zone" from the hypertrophic zone. Interestingly, the ratio of the type X collagen-positive hypertrophic zone in the entire condyle seemed higher in the early stages but decreased in the later stages. Some apparently compacted cells in the hypertrophic zone showed proliferating cell nuclear antigen (PCNA) immunoreaction, indicating the potential for cell proliferation at the early stages. As the mice matured, in contrast, they further enlarged and assumed typical features of hypertrophic chondrocytes. Apoptotic cells were also discernible in the hypertrophic zone at the early but not later stages. Consistent with morphological configurations of hypertrophic chondrocytes, immunoreactions for alkaline phosphatase, osteopontin, and type I collagen were prominent at the later stage, but not the early stage. Cartilaginous matrices demonstrated scattered patches of mineralization at the early stage, but increased in their volume and connectivity at the later stage. Thus, the spatial and temporal occurrence of these immunoreactions as well as apoptosis likely reflect the prematurity of hypertrophying cells at the early stage, and imply a physiological relevance during the early development of the mandibular condyles.
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PMID:Histochemical evidences on the chronological alterations of the hypertrophic zone of mandibular condylar cartilage. 1617 89

Membranous (M) cells of the follicle-associated epithelium (FAE) are believed to sample antigens from the gut lumen. However, the origin, differentiation mechanism, and cell death of M cells are still a matter of controversy. Therefore, we investigated the process of M cell differentiation and determined their fate in the intestine of three-way crossbred female pigs. We used anti-cytokeratin 18 and anti-PCNA antibodies to distinguish M cells and proliferative cells and performed immunohistochemistry, enzyme histochemistry, and scanning electron microscopy on fresh ileal Peyer's patches. Cell migration and apoptotic cells were detected by BrdU labeling and the TUNEL method, respectively. The turnover of the FAE was similar to that of the villi. M cells were mostly observed from the FAE crypt to the FAE periphery, but not in the FAE apex. As proliferative M cells (cytokeratin 18(+)/PCNA(+) cells) have previously been detected in the FAE crypt, porcine M cells may be directly derived from intestinal epithelial stem cells and committed as a distinct cell lineage in the crypts. M cells from the FAE periphery were unstained or only weakly stained for alkaline phosphatase, whereas cytokeratin 18(+)/alkaline phosphatase(+) cells lying near to the FAE apex showed a columnar shape similar to that of adjacent enterocytes. These data suggest that the committed M cells differentiate to mature M cells by contact with lymphocytes at the FAE periphery, and that they trans-differentiate to enterocytes and are finally excluded near the FAE apex.
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PMID:Apoptotic process of porcine intestinal M cells. 1628 91

The alveolar epithelium may function as a barrier for airborne xenobiotics, and in vitro models mimicking this barrier are useful for metabolism and toxicity studies. To gain insight into the metabolic competence of alveolar epithelial cells (AECs), we investigated transcript expression of 10 different cytochrome P450 monooxygenases as well as expression of surfactant proteins A to D. We also investigated gene expression of the transcription factors PCNA, TTF-1, HNF3beta , GATA-6, C/EBPalpha and C/EBPdelta which drive, at least in part, development and differentiation of alveolar epithelium. We further studied the metabolism of testosterone, a substrate for cytochrome P450 (CYP) monooxygenases, in cultures of AECs. Essentially, medium supplementation with 5% rat serum, as opposed to 10% FCS, promoted a high level of differentiation, as judged by the mRNA expression of CYP monooxygenases, e.g. 1A1, 1A2, 2B1 and 2J3, the expression of the surfactant proteins A, B, and C, the immunohistochemical staining for surfactant protein C, and staining for alkaline phosphatase activity. Further, AECs, when cultured in the presence of 5% rat serum, promoted metabolic competence, as evidenced by the fingerprinting of individual testosterone metabolites. We thus characterized AECs in culture and found these respiratory epithelial cells to express an array of differentiation markers and showed these cultures to be metabolically competent under optimized culture conditions.
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PMID:Primary rat alveolar epithelial cells for use in biotransformation and toxicity studies. 1632 67

Hepatitis C-associated osteosclerosis (HCAO) is a rare syndrome characterized by severe, acquired, generalized osteosclerosis and hyperostosis in adults who are infected with the hepatitis C virus. However, the detail of the pathogenesis of HCAO is still unknown. We examined the effects of serum of the HCAO patient on the proliferation, alkaline phosphatase (ALP) activity and transforming growth factor (TGF)-beta-Smad signaling in mouse osteoblastic cells. The patient was compatible with HCAO, characterized by high bone mass, bone thickening and bone pain with normal lamelar bone. The serum from the HCAO patient increased the levels of TGF-beta and Smad3 expression in osteoblastic MC3T3-E1 cells, compared with the control subject. Moreover, the serum from the HCAO patient significantly augmented TGF-beta-induced transcriptional activity with luciferase assay using 3TP-Lux with a Smad3-specific responsive element. In addition, the serum from the HCAO patient significantly stimulated the MTT intensity, the level of proliferating cell nuclear antigen expression, a proliferation marker, and ALP activity in MC3T3-E1 cells, compared with that from the control subject. In conclusion, the present study indicated that the serum from the HCAO patient stimulated TGF-beta-Smad signaling, as well as the proliferation and ALP activity in osteoblastic cells. Some soluble factors other than parathyroid hormone might be related to the pathogenesis of HCAO.
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PMID:Serum soluble factors induce the proliferation, alkaline phosphatase activity and transforming growth factor-beta signal in osteoblastic cells in the patient with hepatitis C-associated osteosclerosis. 1717 44

Craniofacial anomalies resulting from impaired growth of mandibular condyles require multidisciplinary interventions, which impose a substantial burden on patients and their families. So far, correcting such deformities with an alternative strategy - gene therapy - is still an uncharted territory. Here, we established an effective in vivo gene delivery system with recombinant adeno-associated virus (rAAV)-mediated vascular endothelial growth factor (VEGF) to enhance mandibular condylar growth. With in situ hybridization, RT-PCR, immunostaining and Western blot, transgene expression was clearly detected in the mandibular condyles during the whole experiment periods. At defined time points, specific osteogenetic markers (alkaline phosphatase and osteocalcin) and chondrogenetic markers (collagen type II and collagen type X) were assessed by means of biochemical analysis and their expression significantly changed from day 30. Proliferation index by proliferating cell nuclear antigen staining showed also a significant increase in cell proliferation. Morphological measurement identified that the size of mandibular condyle significantly increased from day 30. Taken together, rAAV-VEGF was successfully established as an efficient delivery system to induce mandibular condylar growth, which provides the basis for future gene therapy to treat patients with craniofacial deformities.
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PMID:Recombinant AAV-mediated VEGF gene therapy induces mandibular condylar growth. 1746 Jul 22

One of the focuses in current cancer chemoprevention studies is the search for nontoxic chemopreventive agents that inhibit the initiation of malignant transformation. Cancer biomarkers are quantifiable molecules involved in the physiologic or pathologic events occurring between exposure to carcinogens and the development, progression of cancer. Biomarkers may be the consequence of a continuous process, such as increased cell mass, or a discrete event, such as genetic mutation. Analysis of tumor markers can be used as an indicator of tumor response to therapy. Gallic acid is a naturally available polyphenol, possess strong antioxidant activity with a capacity to inhibit the formation of tumors in several cancer models. In the present study, we investigated the antiproliferative effect of gallic acid during diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC) in male wistar albino rats. DEN treatment resulted in increased levels of aspartate transaminase, alanine transaminase, alkaline phosphatase, acid phosphatase, lactate dehydrogenase, gamma-glutamyltransferase, 5'-nucleotidase, bilirubin, alpha-fetoprotein, carcinoembryonic antigen, argyophillic nucleolar organizing regions, and proliferating cell nuclear antigen. Gallic acid treatment significantly attenuated these alterations and decreased the levels of AgNORs and PCNA. These finding suggests that gallic acid is a potent antiproliferative agent against DEN-induced HCC.
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PMID:Antiproliferative potential of gallic acid against diethylnitrosamine-induced rat hepatocellular carcinoma. 1862 14

Published and original data indicating evolutionary conservation of the morphofunctional organization of reserve stem cells providing for asexual and sexual reproduction of invertebrates are reviewed. Stem cells were studied in representatives of five animal types: archeocytes in sponge Oscarella malakhovi (Porifera), large interstitial cells in colonial hydroid Obelia longissima (Cnidaria), neoblasts in an asexual race of planarian Girardia tigrina (Platyhelmintes), stem cells in colonial rhizocephalans Peltogasterella gracilis, Polyascus polygenea, and Thylacoplethus isaevae (Arthropoda), and colonial ascidian Botryllus tuberatus (Chordata). Stem cells in animals of such diverse taxa feature the presence of germinal granules, are positive for proliferating cell nuclear antigen, demonstrate alkaline phosphatase activity (at marker of embryonic stem cells and primary germ cells in vertebrates), and rhizocephalan stem cells express the vasa-like gene (such genes are expressed in germline cells of different metazoans). The self-renewing pool of stem cells is the cellular basis of the reproductive strategy including sexual and asexual reproduction.
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PMID:[Morphofunctional organization of reserve stem cells providing for asexual and sexual reproduction of invertebrates]. 1940 44


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