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991.
S Gagliardi G Nadler E Consolandi C Parini M Morvan MN Legave P Belfiore A Zocchetti GD Clarke I James P Nambi M Gowen C Farina 《Canadian Metallurgical Quarterly》1998,41(10):1568-1573
The vacuolar H+-ATPase (V-ATPase), located on the ruffled border of the osteoclast, is a proton pump which is responsible for secreting the massive amounts of protons that are required for the bone resorption process. With the aim to identify new agents which are able to prevent the excessive bone resorption associated with osteoporosis, we have designed a novel class of potent and selective inhibitors of the osteoclast proton pump, starting from the structure of the specific V-ATPase inhibitor bafilomycin A1. Compounds 3a-d potently inhibited the V-ATPase in chicken osteoclast membranes (IC50 = 60-180 nM) and were able to prevent bone resorption by human osteoclasts in vitro at low-nanomolar concentrations. Notably, the EC50 of compound 3c in this assay was 45-fold lower than the concentration required for half-maximal inhibition of the V-ATPase from human kidney cortex. These results support the validity of the osteoclast proton pump as a useful molecular target to produce novel inhibitors of bone resorption, potentially useful as antiosteporotic agents. 相似文献
992.
SJ George JL Johnson GD Angelini AC Newby AH Baker 《Canadian Metallurgical Quarterly》1998,9(6):867-877
Neointimal formation involving smooth muscle cell (SMC) migration and proliferation is a common feature of atherosclerosis, restenosis after angioplasty, and vein graft intimal thickening. Extracellular matrix remodeling by metalloproteinase (MMP) enzymes is an essential component of neointimal formation and therefore MMPs are a potential target for localized gene therapy. To evaluate this concept using human tissue, we used the highly reproducible organ culture model of neointimal formation in human saphenous vein to investigate the effect of adenovirus-mediated gene transfer of tissue inhibitor of metalloproteinase 1 (TIMP-1) and the bacterial LacZ gene (RAd35) as a control. Incubating veins with 100 microl of RAd35 (1.2 x 10(10) pfu/ml) led to expression of LacZ in 39 +/- 7% of surface cells but had no effect on SMC proliferation, migration, or neointimal formation. Similar infection with RAdTIMP-1 increased explanation of TIMP-1 in surface cells and significantly inhibited neointimal formation and SMC migration after 14 days by 54% and 78%, respectively (n = 6, p < 0.05 Student's paired t test). No effect on SMC proliferation or deleterious effect on cell viability was observed. A specific MMP inhibitory effect was detected using in situ zymography. These data confirm the importance of MMPs in neointimal formation and highlight the potential for application of TIMP gene therapy. 相似文献
993.
JA Grass DJ Hei K Metchette GD Cimino GP Wiesehahn L Corash L Lin 《Canadian Metallurgical Quarterly》1998,91(6):2180-2188
A photochemical treatment (PCT) process using a novel psoralen and long wavelength ultraviolet light (UVA, 320-400 nm) has been developed to inactivate bacteria and viruses in platelet concentrates. This study evaluated the efficacy of PCT for inactivation of leukocytes that contaminate platelet preparations. Three psoralens, 8-methoxypsoralen (8-MOP), 4'-aminomethyl 4,5', 8-trimethylpsoralen (AMT), and the novel psoralen S-59, were compared using the following four independent but complementary biological and molecular assays. (1) T-cell viability: Treatment with 150 mumol/L S-59 and 1.0 to 3.0 Joules/cm2 UVA inactivated >5.4 +/- 0.3 log10 of T cells in full-sized single-donor plateletpheresis units. Using 1.0 Joule/cm2 UVA, the lowest dose of S-59, AMT and 8-MOP required to reduce the number of T cells to the limit of detection was 0.05 micromol/L, 1.0 micromol/L, and 10.0 micromol/L, respectively. (2) Cytokine synthesis: Treatment with 1.9 Joules/cm2 UVA and 150 micromol/L S-59 or AMT completely inhibited synthesis of the cytokine IL-8 by contaminating leukocytes during 5 days of platelet storage. After treatment with 75 micromol/L 8-MOP and 1.9 Joules/cm2 UVA, only low levels of IL-8 were detected. (3) Psoralen-DNA adduct formation: The combination of 1.9 Joules/cm2 UVA and 150 micromol/L S-59, AMT, or 8-MOP induced 12.0 +/- 3.0, 6.0 +/- 0. 9, and 0.7 psoralen adducts per 1,000 bp DNA, respectively. (4) Replication competence: Polymerase chain reaction (PCR) amplification of small genomic DNA sequences (242-439 bp) after PCT was inhibited. The degree of PCR amplification inhibition correlated with the level of adduct formation (S-59 > AMT > 8-MOP). In contrast, 2,500 cGy gamma radiation, a dose that inactivates >5 log10 of T cells in blood products, had minimal effect on cytokine synthesis and did not induce sufficient DNA strand breaks to inhibit PCR amplification of the same small DNA sequences. These results demonstrate that leukocytes are sensitive to PCT with psoralens and among the psoralens tested S-59 is the most effective. Therefore, PCT has the potential to reduce the incidence of leukocyte-mediated adverse immune reactions associated with platelet transfusion. 相似文献
994.
JM Winchell S Routray PW Betts HJ Van Kruiningen LK Silbart 《Canadian Metallurgical Quarterly》1998,178(3):850-853
A plasmid encoding T1-SP10MN(A), a peptide derived from immunodominant regions of human immunodeficiency virus type 1 gp120, was delivered to rabbit Peyer's patches using a helium-driven gene gun. Six weeks thereafter, 2 of 5 animals were given an intradermal booster immunization. Blood, feces, and vaginal washes were collected weekly and assayed by ELISA. High titer T1-SP10MN(A)-specific fecal and vaginal secretory IgA responses were observed, and the response appeared to be augmented following dermal booster immunizations. Specific serum IgG was also detected within 1 week of immunization and remained elevated through week 20 in the 2 animals receiving dermal boosts (titers > or = 6400). This study establishes the Peyer's patch as a promising target tissue for DNA vaccination and demonstrates the efficacy of gene gun-mediated delivery of foreign DNA to a mucosal tissue for the induction of an immune response. 相似文献
995.
FM Hughes CD Bortner GD Purdy JA Cidlowski 《Canadian Metallurgical Quarterly》1997,272(48):30567-30576
Little is known about the mechanisms of suppression of apoptosis. We have addressed the novel possibility that the level of intracellular K+ regulates the apoptotic process by controlling the activity of death enzymes. We show that K+, at normal intracellular levels, inhibits both apoptotic DNA fragmentation and caspase-3(CPP32)-like protease activation, suggesting that intracellular K+ loss must occur early during apoptosis. Direct measurement of K+ by inductively coupled plasma/mass spectrometry and flow cytometry indicates a major decrease in intracellular K+ concentration in the apoptotic cell. Flow cytometric analysis revealed that caspase and nuclease activity were restricted to the subpopulation of cells with reduced K+. Disruption of the natural K+ electrochemical gradient suppressed the activity of both caspase and nuclease independent of the mode of activation of the apoptotic inducing agent, demonstrating that a decrease in intracellular K+ concentration is a necessary, early event in programmed cell death. 相似文献
996.
997.
NA Scott GD Cipolla CE Ross B Dunn FH Martin L Simonet JN Wilcox 《Canadian Metallurgical Quarterly》1996,93(12):2178-2187
BACKGROUND: In the present series of experiments, we examined the onset of cell proliferation and growth factor expression after balloon overstretch injury to porcine coronary arteries. METHODS AND RESULTS: Domestic juvenile swine underwent balloon overstretch injury to the left anterior descending and circumflex coronary arteries with standard percutaneous transluminal coronary angioplasty balloon catheters. To identify proliferating cells, 5-bromo-2-deoxyuridine (BrDU) was administered over a period of 24 hours before the animals were killed at either 1, 3, 7, or 14 days after injury. Immunohistochemistry was performed with monoclonal antibodies to BrDU and smooth muscle cell markers. Three days after injury, a large number of proliferating cells were located in the adventitia, with significantly fewer positive cells found in the media and lumen. Seven days after injury, proliferating cells were found primarily in the neointima, extending along the luminal surface. In situ hybridization for PDGF A-chain and beta-receptor mRNAs revealed that the expression of these two genes was closely correlated with the sites of proliferation at each time point. Studies in which BrDU was injected between days 2 and 3 and the animals were killed on day 14 suggested that the proliferating adventitial cells may migrate into the neointima. CONCLUSIONS: These data suggest that adventitial myofibroblasts contribute to the process of vascular lesion formation by proliferating, synthesizing growth factors, and possibly migrating into the neointima. Increased synthesis of alpha-smooth muscle actin observed in the adventitial cells after arterial injury may constrict the injured vessel and contribute to the process of arterial remodeling and late lumen loss after angioplasty. 相似文献
998.
GD Marty EF Freiberg TR Meyers J Wilcock TB Farver DE Hinton 《Canadian Metallurgical Quarterly》1998,32(1):15-40
A recently proposed Jovanovic-Freundlich isotherm model for single component adsorption without lateral interactions on heterogeneous surfaces is extended to account for lateral interactions and for competitive adsorption. The model is tested using previously reported single component and competitive adsorption data of 2-phenylethanol and 3-phenylpropanol on ODS-silica with methanol-water as the mobile phase. A comparison is made regarding the ability of the Jovanovic-Freundlich and Langmuir-Freundlich models to predict competitive equilibria using the single component identified parameters. Fair predictions of the competitive data were obtained when using heterogeneous-surface models which do not take into account the possible interactions of phenylalcohols in the adsorbed phase via hydrogen bonding. Markedly improved predictions were obtained with models which account simultaneously for the two main sources of adsorbed phase nonideal behavior, i.e. adsorbate-adsorbate interactions and heterogeneity of the adsorbent surface. 相似文献
999.
H Wang EA Spillare QS Wang Sabourin CLK GD Stoner 《Canadian Metallurgical Quarterly》1998,12(2):325-328
The temporal correlation between adenosine outflow and changes in field excitatory post synaptic potentials (fEPSP) occurring during ischemia-like conditions was investigated in rat hippocampal slices. Five-minute long ischemia-like conditions resulted in a 100% depression of fEPSP amplitude, followed by a complete recovery after 6-7 min of reperfusion. By reducing the duration of the ischemic insult to 2 min, fEPSP was depressed by 50%. During both 5 and 2 min of ischemia-like conditions, a significant increase in adenosine outflow was detected. During reperfusion, when fEPSP amplitude recovered completely, the adenosine level in the extracellular fluid returned to basal values. The strict relationship between the increase in adenosine outflow and fEPSP inhibition supports the hypothesis that adenosine is largely responsible for the synaptic transmission depression during cerebral ischemia. 相似文献
1000.
An enzyme with alpha-L-rhamnosidase activity was purified by anion exchange chromatography from an Aspergillus niger commercial preparation. The alpha-L-rhamnosidase was shown to be N-glycosylated, and had a molecular mass of 85 kD on sodium dodecylsulfate-polyacrylamide gel electrophoresis of which approximately 12% was contributed by carbohydrate. The enzyme was optimally active at pH 4.5 and 65 degrees C. When tested towards p-nitrophenyl-alpha-L-rhamnopyranoside it showed Km and Vmax values of 2.9 mM and 20.6 U mg-1, respectively whereas it was inhibited competitively by L-rhamnose (Ki 3.5 mM). Substrate specificity studies showed alpha-L-rhamnosidase to be active both on alpha-1,2 and alpha-1,6 linkages to beta-D-glucose. Moreover, the enzyme was able to release L-rhamnose from geranyl-beta-D-rutinoside and 2-phenylethyl-beta-D-rutinoside. 相似文献