Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: HUMANGGP:025734 (ANOVA)
 22,580 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In order to study human fat cell morphology and metabolism in both sexes, with different levels of fatness and physical training, 53 subjects (14 women and 39 men), 25.9 +/- 7.6 (mean +/- SD) years of age, weighing 64.8 +/- 10.8 kg and with 14.8 +/- 4.7% body fat were submitted to a biopsy of subcutaneous adipose tissue in the supra-iliac region. Average fat cell diameter (ACD) was assessed by measuring at least 500 cells per subject. Basal (BL), epinephrine sub-maximal (10(-5) M) (ESML) and epinephrine maximal (10(-4) M) (EML) stimulated lipolysis have been determined on collagenase isolated fat cells. Female subjects have larger ACD (83.4 +/- 12.3 micrometer vs. 62.5 +/- 11.9 micrometer) (P less than or equal to 0.001) and lower BL, ESML, and EML than men (P less than or equal to 0.05). After control over age and sex of subjects, there were significant and negative correlations between all fatness indicators (ACD, fat mass in kg, percent body fat, sum of 9 skinfolds) and BL, ESML and EML. A completely randomized factorial ANOVA design revealed a significant effect of fatness level on BL, ESML and EML, but a non-significant trend for the effect of physical training on the same variables. It is concluded that lipolytic activity is more related to body fatness than to the training status. However, training tends to accentuate lipolytic activity as shown by values of BL, ESML and EML found in the lean highly trained subjects of the study.
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PMID:[Morphology and metabolism of human adipose tissues: relation to sex, body fat and level of training]. 619 Apr 51

A comparison of collagen synthesis in the peripheral and central regions of the normal medial meniscus, the normal lateral meniscus, and the medial meniscus, three months after reconstruction of the anterior cruciate ligament (ACL), was performed in a canine model. Triangular-shaped explants were labeled in vitro with tritiated proline to compare collagen synthesis. Regional cellularity was also quantified. In the normal medial and lateral meniscus, cellularity was similar. Total collagen synthesis, as assessed by collagenase digestible material, was significantly elevated in the peripheral one-third of the medial meniscus versus the central region (paired t-test, p = 0.026). However, in the medial meniscus after ACL reconstruction, cellularity and total collagen synthesis were significantly higher in the peripheral one-third versus the contiguous central region. They were also elevated relative to the peripheral and central region of the normal medial and lateral meniscus (ANOVA, p < 0.05; Student-Neuman-Kuel test, p < 0.05). Thus, in the normal medial meniscus, the peripheral one-third may have an increased healing potential based on its increased ability to synthesize collagen. Furthermore, after ACL reconstruction, only the peripheral, and not the central, region of the canine meniscus responds to the altered mechanical and synovial environment by significantly increasing cellularity and collagen synthesis.
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PMID:Comparison of collagen synthesis in the peripheral and central region of the canine meniscus. 819 43

Other investigators have shown that exogenously administered transforming growth factor-beta (TGF-beta) inhibits lymphocyte adherence to vascular endothelial cells (VEC). We examined the role of TGF-beta 1 as an autocrine mediator of lymphocyte adhesion to adult human VECs. VECs were harvested from eight saphenous or cadaveric iliac veins using 0.2% collagenase. Low-passage VECs in MCDB + 0.1% BSA were pretreated for 24 hr with monoclonal anti-TGF-beta 1 antibody (5 micrograms/ml), LPS (5 micrograms/ml), or IL-1 (10 U/ml). Adherence of fluorescently labeled lymphocytes to pretreated VECs was quantitated and results were expressed as relative adhesion compared to untreated control. Total mRNA from LPS- or IL-1-treated VECs was subjected to Northern analysis to determine relative TGF-beta 1 expression. Total TGF-beta 1 protein concentration in supernatants from LPS- or IL-1-treated VECs was determined by ELISA. Data (means +/- SEM) were analyzed by ANOVA with a Newman-Keuls posttest. Neutralizing endogenous TGF-beta 1 with anti-TGF-beta 1 antibody significantly increased adhesion of lymphocytes to VEC monolayers compared to control (125 +/- 3 vs 101 +/- 2%, P < 0.01, n = 8). The level of adhesion was equivalent to that seen with IL-1 stimulation (131 +/- 6%). Spearman correlation of lymphocyte adherence to IL-1- or LPS-treated VECs vs TGF-beta 1 mRNA expression or vs relative TGF-beta 1 protein concentration showed significant inverse relationships (r = -0.82, P < 0.001, and r = -0.87, P < 0.001, respectively). Endogenous TGF-beta 1's inhibitory effect on lymphocyte adhesion was blocked by a specific neutralizing antibody. VEC TGF-beta 1 mRNA expression and TGF-beta 1 production were inversely proportional to lymphocyte adhesion, suggesting down-regulation of TGF-beta 1 in response to proinflammatory cytokines. Together, these observations support the hypothesis that TGF-beta 1 has an autocrine inhibitory role in regulation of lymphocyte adhesion to VECs.
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PMID:Transforming growth factor-beta 1 serves as an autocrine inhibitor of human endothelial cell/lymphocyte adhesion. 853 71

Prostaglandin E2 (PGE2) stimulates collagen gene promoter activity in transfected human chondrocytes though no canonical cyclic AMP (cAMP) response element has been yet identified. Human insulin-like growth factor-1 (IGF-1) induces an increase in collagen type II expression and synthesis in chondrocytes. Since our preliminary data suggested that PGE2 can stimulate IGF-1 release from human articular chondrocytes, we examined whether the eicosanoid could influence collagen synthesis and whether the effect was mediated by IGF-1. Incubation of primary cultures of human articular chondrocytes with increasing concentrations of PGE2 resulted in a dose-dependent (ANOVA, F= 51.62, P < 0.0001, n = 5) and saturable increase in the synthesis and release of IGF-1 and expression of IGF-1 mRNA. At relatively low concentrations (30 pmol/1 to 30 nmol/l), PGE2 stimulated an increase in the incorporation of [3H]proline into collagenase digestible protein (CDP) (P < 0.01, n = 5) whereas at high levels (300 nmol/l to 3 micromol/l) of the eicosanoid, incorporation diminished precipitously. Human IGF-1 mimicked the effects of low PGE2 concentrations by stimulating in a dose-dependent (ANOVA, F= 31.65, P < 0.001, n = 3) and saturable fashion the incorporation of [3H]proline into CDP although the magnitude of the response induced by IGF-1 was far greater (3.5-fold). An IGF-1 receptor blocking antibody completely abrogated the IGF-1 induced response suggesting that the effect was specifically IGF-1 receptor mediated. Furthermore, the PGE2-induced increase in [3H]proline incorporation into CDP was inhibited (63%, P < 0.001, n = 7) by the addition to the culture medium of an anti-IGF-1 antibody. We conclude that PGE2 may act as a secretagogue of IGF-1 and that the latter growth factor may mediate, via an autocrine loop and the IGF-1 receptor, at least some of the anabolic effects of the eicosanoid on cartilage metabolism.
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PMID:Prostaglandin E2 stimulates incorporation of proline into collagenase digestible proteins in human articular chondrocytes: identification of an effector autocrine loop involving insulin-like growth factor I. 891 8

The pathogenesis of tobacco-related periodontal diseases is not well understood. The purpose of this study was therefore to investigate smokeless tobacco extract (ST) and nicotine effects on prostaglandin E2 (PGE2) and interleukin-1beta (IL-1beta) secretion by peripheral blood mononuclear cells (PBMC, consisting of monocytes and lymphocytes) and gingival mononuclear cells (GMC). Both peripheral blood and gingival tissue adjacent to the alveolar crest were taken from non-smoking adult periodontitis patients. Gingival tissue was treated with collagenase and deoxyribonuclease and GMC and PBMC were isolated by Ficoll-Hypaque centrifugation. GMC and PBMC (100,000 cells/200 microl) were cultured for 24 hours in supplemented RPMI 1640 alone (control), or in supplemented RPMI 1640 containing 1% ST, 100 microg/ml nicotine, 1 microg/ml Porphyromonas gingivalis LPS, or 1 microg/ml P. gingivalis LPS and either 100 microg/ml nicotine or 1% ST. Enzyme immunoassays were used to quantify PGE2 and IL-1beta. Treatments were compared by repeated measures ANOVA. 100 microg/ml nicotine (7-fold, p<0.02) and 1% ST (3.5-fold, p<0.004) significantly increased secretion of PGE2 by PBMC relative to control cultures. 100 microg/ml nicotine and 1% ST, however, had no effect on IL-1beta secretion by PBMC. Enhanced PGE2 secretion also was seen when PBMC were treated with P. gingivalis LPS+ 100 microg/ml nicotine relative to P. gingivalis LPS alone (p<0.007). In contrast, 100 microg/ml nicotine significantly downregulated IL-1beta secretion by GMC relative to medium alone (p<0.008) and had no effect on PGE2 secretion by GMC. These data indicate that while nicotine and ST can stimulate PBMC to secrete PGE2, they cannot activate further mononuclear cells extracted from gingiva, possibly due to maximal previous stimulation in the periodontitis lesion.
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PMID:Nicotine and smokeless tobacco effects on gingival and peripheral blood mononuclear cells. 954 95

Recently developed technologies enabling the production of a reproducible, purified enzyme blend for optimal human pancreatic islet isolation has renewed interest in clinical islet transplantation. The canine model has been an ideal preclinical model for the development of islet transplantation protocols. As seen in other species, the application of crude collagenase for isolating canine islets resulted in highly variable islet yields, extensive islet fragmentation, and variable islet functionality. We compared the function of commercially available crude collagenases with that of Liberase-CI purified enzyme blend for canine islet isolation. We also compared two manufacturing runs of Liberase-CI enzyme (lots 1 and 2) to demonstrate reproducibility of islet recovery and function. We report on the improved recovery and function of islets isolated using Liberase-CI enzyme. No difference in dog age, mean body weight, or pancreas weight were observed between the experimental groups. We observed a significantly higher postpurification recovery of islet equivalent number (IE) from pancreases processed using two lots of Liberase-CI enzyme (189 +/- 20 x 10(3) IE, n = 4) and lot 2 (234 +/- 39 x 10(3) IE, n = 7) than that obtained from pancreases processed with Sigma Type V (116.8 +/- 27 x 10(3) IE, n = 5), Serva collagenase (49 +/- 11.6 x 10(3) IE, n = 5, p < 0.05) or Boehringer-Mannheim (BM) Type P collagenase (85.4 +/- 25 x 10(3) IE, n = 5, p < 0.05, ANOVA). No significant differences were observed in islet yield recovery from pancreases processed using the two production lots of Liberase-CI enzyme. Islet survival after 48 h in culture at 37 degrees C was significantly higher from islets isolated using Liberase-CI enzyme (88 +/- 3.7% survival) when compared to Sigma Type V (81.8 +/- 3.3%), Serva (71.7 +/- 2.8%), and BM Type P (77 +/- 7.2%) (p < 0.05). Islet functional testing in vitro demonstrated islets isolated using crude collagenase had an increased insulin basal release and a reduced insulin stimulated response when compared with islets isolated using the two lots of Liberase-CI enzyme. The calculated stimulation index was 7.8 +/- 1.7, 3.1 +/- 0.6, and 4.8 +/- 1.1 for Sigma Type V, Serva, and BM Type P isolated islets, respectively, compared to 15.7 +/- 1.6 and 16.2 +/- 1.9 for islets isolated with Liberase-CI enzyme production lots 1 and 2, respectively (p < 0.05). This evaluation demonstrates that a purified enzyme blend can significantly improve islet recovery and function. It also demonstrates the manufacturing reproducibility of Liberase-CI enzyme lots resulting in the isolation of canine islets with the same degree of efficacy. A blend of purified enzymes, specifically formulated for canine islet isolation, can consistently yield large numbers of islets that survive longer in culture and demonstrate an improved insulin response in vitro.
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PMID:Evaluation of a purified enzyme blend for the recovery and function of canine pancreatic islets. 971 Mar 5

Sinusoidal entry is the first obligatory process preceding intracellular drug removal in liver. Transport of the angiotensin converting enzyme inhibitor enalapril (1-750 microM with [(3)H]enalapril), a substrate of Oatp1, the sodium-independent organic anion transporting polypeptide 1 cloned from rat liver, was studied in rat hepatocytes isolated from all zones of the liver (homogeneous) and from enriched periportal (PP) and perivenous (PV) hepatocytes prepared by collagenase perfusion and zone-selective destruction with digitonin, respectively. Uptake was linear over 1 min and was concentration-dependent. Transport by the homogeneous hepatocytes (in the presence and absence of Na(+)) and PP and PV cells was described by single saturable components of similar kinetic constants (K(m) values of 344-461 microM and V(max) values of 9.5-11.6 nmol/min/10(6) cells; P >.05, ANOVA). The K(m) value for enalapril uptake in hepatocytes was of the same order of magnitude compared with that for Oatp1 expressed in HeLa cells transfected with cDNA-Oatp1 and Western blot analysis revealed similar levels of immunoreactive Oatp1 expression in PP and PV hepatocytes. However, enalapril was not taken up by Oatp2 nor by the human OATP expressed in recombinant vaccinia systems.
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PMID:Uptake of enalapril and expression of organic anion transporting polypeptide 1 in zonal, isolated rat hepatocytes. 1085 54

Both length of extra-alveolar time and type of storage media are significant factors that can affect the long-term prognosis for replanted teeth. Numerous studies have examined various media in an attempt to determine the ideal material for storage of the avulsed tooth. The purpose of this study was to compare the number of viable periodontium ligament (PDL) cells in different storage media using a collagenase assay. Thirty-three freshly extracted human teeth were divided into four experimental and two control groups. The positive and negative controls corresponded to 0 min and an 8-h dry time, respectively. The experimental teeth were stored dry for 30 min and then immersed in one of four media (Hank's balanced salt solution (HBSS), milk, saline, water) for 45 min. The teeth were then treated with dispase grade II and collagenase for 30 min. The number of viable and nonviable PDL cells was counted with a hemocytometer and analyzed. An ANOVA demonstrated no statistically significant differences in the viability of PDL cells among saline, HBSS and milk. Within the parameters of this study, it appears that milk or saline is an equally viable alternative to HBSS for storage of avulsed teeth.
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PMID:Assessment of post-traumatic PDL cells viability by a novel collagenase assay. 1244 27

Melatonin exerts a marked antiproliferative action in numerous experimentally-induced tumors in vivo as well as in both animal and human cell lines in vitro. However, the mechanisms of oncostatic action of melatonin is not clear, and the involvement of both membrane and nuclear receptors are suggested. Therefore, the aim of this study was to investigate effects of melatonin, and both agonist (CGP 52608), and antagonist (CGP 55644) of RZR/ROR nuclear receptors on the growth of diethylstilbestrol-induced rat prolactin-secreting pituitary tumor cells in vitro. Pituitary tumors were induced by subcutaneous implantation of a single silastic capsule containing 10 mg of diethylstilbestrol in 4-wk-old male Fischer 344 rats. Four months after the implantation of capsules the animals were killed by decapitation, pituitary tumors were aseptically removed, mechanically dispersed, and enzymatically digested with 0.2% collagenase and 0.2% hyaluronidase. The cells (6 x 105 cells/well) were incubated for 24 hr in the presence of melatonin, CGP 52608, CGP 55644 and CGP 55644 plus melatonin (at the concentrations of 107 and 10-9 m) at 37 degrees C in the humidified atmosphere of 95% air and 5% CO2. The group with the addition of solvent only served as control. The growth of cell was measured using the EZ4U system. Statistical analysis was performed using ANOVA followed by LSD test. Both melatonin and CGP 52608 significantly suppressed growth of tumor cells in vitro in both used concentrations. CGP 55644 stimulated growth of tumor cells and blocked the inhibitory effects of melatonin in vitro. Results of the present study as well as other experimental evidence strongly support the hypothesis that both membrane and nuclear receptors are involved in the oncostatic action of melatonin, and indicate that nuclear signalling plays an important role in this process.
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PMID:Melatonin inhibits growth of diethylstilbestrol-induced prolactin-secreting pituitary tumor in vitro: possible involvement of nuclear RZR/ROR receptors. 1266 53

In this study, we investigated the effect of inflammatory cytokines on matrix metalloproteinase (MMP-1) and TIMP-1 production in human dermal fibroblasts, which play a pivotal role in wound healing, ranging from the synthesis and remodeling of extracellular matrix (ECM) to the synthesis of growth factors. The balance of MMPs and TIMPs is crucial in directing successful wound repair. Human adult dermal fibroblasts were seeded in six well plates (7.5 x 10(4) cells/ml) in complete media. Eighty to ninety percent confluent cells were treated with interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) (10 ng/ml) for 6h in serum free media with suitable controls run in triplicate. Supernatants were assayed for pro-MMP-1 & TIMP-1. Extracted total RNA was used for reverse transcription polymerase chain reaction (RT-PCR) with sequence specific primers for MMP-1, TIMP-1 and beta-actin. Signal intensity was normalized to the internal control (beta-actin). Statistical analysis used ANOVA. MMP-1 and TIMP-1 mRNA expression were markedly increased with IL-6 and TNF-alpha treatment and remains unchanged with IL-1beta. Pro-MMP-1 protein levels are unchanged with TNF-alpha and significantly increased with IL-1beta and IL-6 treatment. However, TNF-alpha significantly increases TIMP-1 protein levels. Data suggests differential regulation of MMP-1 and TIMP-1 protein levels by the cytokines found in stimulated dermal fibroblasts. Further characterization of this response will provide an understanding of the mechanisms of pathogenesis of extracellular matrix (ECM) and the potential role of metalloproteinases in tissue remodeling after injury.
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PMID:Matrix metalloproteinase expression in cytokine stimulated human dermal fibroblasts. 1292 75


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