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181.
Human liver microsomes are capable of oxidizing lauric acid (laurate), a model medium-chain fatty acid, at both the omega- and omega-1 positions to form 12- and 11-hydroxylaurate, respectively. These laurate hydroxylation reactions are apparently catalyzed by distinct P450 enzymes. While the P450 responsible for microsomal laurate omega-1 hydroxylation in human liver has been identified as CYP2E1, the enzyme catalyzing omega-hydroxylation remains poorly defined. To that end, we employed conventional purification and immunochemical techniques to characterize the major hepatic laurate omega-hydroxylase in humans. Western blotting with rat CYP4A1 antibodies was used to monitor a cross-reactive P450 protein (M(r) = 52 kDa) during its isolation from human liver microsomes. The purified enzyme (7.4 nmol P450/mg protein) had an NH2-terminal amino acid sequence identical to that predicted from the human CYP4A11 cDNA over the first 20 residues found. Upon reconstitution with P450 reductase and cytochrome b5, CYP4A11 proved to be a potent laurate omega-hydroxylase, exhibiting a turnover rate of 45.7 nmol 12-hydroxylaurate formed/min/nmol P450 (12-fold greater than intact microsomes), while catalyzing the omega-1 hydroxylation reaction at much lower rates (5.4 nmol 11-hydroxylaurate formed/min/nmol P450). Analysis of the laurate omega-hydroxylation reaction in human liver microsomes revealed kinetic parameters (a lone Km of 48.9 microM with a VMAX of 3.72 nmol 12-hydroxylaurate formed/min/nmol P450) consistent with catalysis by CYP4A11. In fact, incubation of human liver microsomes with antibodies raised to CYP4A11 resulted in nearly 85% inhibition of laurate omega-hydroxylase activity while omega-1 hydroxylase activity remained unaffected. Furthermore, a strong correlation (r = 0.89; P < 0.001) was found between immunochemically determined CYP4A11 content and laurate omega-hydroxylase activity in liver samples from 11 different subjects. From the foregoing, it appears that CYP4A11 is the principle laurate omega-hydroxylating enzyme expressed in human liver. 相似文献
182.
In order to investigate radiation risks associated with low dose and low dose-rates, pregnant Swiss albino mice were exposed to gamma rays, 0.80 Gy from a cobalt-60 source at two different dose-rates (0.0795 and 0.0012 Gy/min) on 18 day post conception. In females exposed to lower dose-rate (0.0012 Gy/min), litter size was found to be decreased, while those exposed to higher dose-rate (0.0795 Gy/min), it remained unaltered. In both groups, appearance of fur and development of complete fur were delayed, whereas gait was delayed only in higher dose-rate group. Male offspring exhibited a biphasic mode of weight loss, while female offspring after an initial weight loss at 1 week, displayed a continuous recovery, but could not attain the normal weight till 12 weeks of age. It appears that higher dose-rate is more effective in delaying the appearance of physiological markers and weight loss, while in terms of litter size lower dose-rate (0.0012 Gy/min) is more effective. 相似文献
183.
M Radmacher M Fritz CM Kacher JP Cleveland PK Hansma 《Canadian Metallurgical Quarterly》1996,70(1):556-567
We have measured force curves as a function of the lateral position on top of human platelets with the atomic force microscope. These force curves show the indentation of the cell as the tip loads the sample. By analyzing these force curves we were able to determine the elastic modulus of the platelet with a lateral resolution of approximately 100 nm. The elastic moduli were in a range of 1-50 kPa measured in the frequency range of 1-50 Hz. Loading forces could be controlled with a resolution of 80 pN and indentations of the platelet could be determined with a resolution of 20 nm. 相似文献
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PK Yen 《Canadian Metallurgical Quarterly》1996,17(6):293-294
186.
AC Beekman PK Wierenga HJ Woerdenbag W Van Uden N Pras AW Konings FS el-Feraly AM Galal HV Wikstr?m 《Canadian Metallurgical Quarterly》1998,64(7):615-619
Epidemiologic and public health researchers frequently include several dependent variables, repeated assessments, or subgroup analyses in their investigations. These factors result in multiple tests of statistical significance and may produce type 1 experimental errors. This study examined the type 1 error rate in a sample of public health and epidemiologic research. A total of 173 articles chosen at random from 1996 issues of the American Journal of Public Health and the American Journal of Epidemiology were examined to determine the incidence of type 1 errors. Three different methods of computing type 1 error rates were used: experiment-wise error rate, error rate per experiment, and percent error rate. The results indicate a type 1 error rate substantially higher than the traditionally assumed level of 5% (p < 0.05). No practical or statistically significant difference was found between type 1 error rates across the two journals. Methods to determine and correct type 1 errors should be reported in epidemiologic and public health research investigations that include multiple statistical tests. 相似文献
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189.
PK Downes 《Canadian Metallurgical Quarterly》1998,185(10):511-518
This section explains how to make most of your time on-line by providing tips on how to reduce the cost of using the internet. Suggestions are given on how to get the best from your existing hardware and software. There are also recommendations for some additional hardware and software. 相似文献
190.
This study documents the fate of nitric oxide neurons following small bowel transplantation. Heterotopic, syngeneic small bowel transplants were performed in five male Lewis rats. The grafts were harvested 7, 36, 55, 60, and 65 days postoperatively, together with the native bowel for comparison. NADPH diaphorase staining of cryostat sections was used to identify the nitric oxide-producing neurons. NADPH diaphorase activity was concentrated in the myenteric plexus, deep muscular, and submucous plexus. No differences were detected between the transplanted and native bowel at any time after transplantation. This suggests that nitric-oxide-producing neurons are well preserved after transplantation and that posttransplant dysmotility is probably not related to interrupted nitric oxide innervation. 相似文献