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The cytoskeletal components of hamster oocytes, zygotes, and spontaneously activated parthogenotes were examined after immunocytochemical labeling. Microtubules were found only in the anastral, tangentially arranged second meiotic spindle of unfertilized oocytes. Taxol treatment of unfertilized oocytes greatly augmented astral microtubules in both the metaphase II spindle and the cortex. Disruption of the meiotic spindle microtubules with nocodazole resulted in cortical chromosomal scattering. During hamster sperm incorporation and pronuclear formation, no sperm aster was detected in association with the male DNA. Instead, a large overlapping array of microtubules assembled in the cortex. By mitosis, this interphase array disassembled and an anastral metaphase spindle formed. Microtubule and chromatin configurations were also imaged in hamster oocytes injected with human sperm. Astral microtubules were absent from the sperm centrosome. The implications of these results are discussed in relation to the hamster oocyte penetration assay, a test commonly used by in vitro fertilization clinics to demonstrate the fertilizing ability of human sperm. We conclude that since hamsters and humans follow different methods of centrosome inheritance, maternal and paternal, respectively, the hamster may be an inappropriate model for exploring microtubule and centrosomal defects in humans or for assaying postinsemination forms of human male fertility defects. 相似文献
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JM Flanagan M Kataoka T Fujisawa DM Engelman 《Canadian Metallurgical Quarterly》1993,32(39):10359-10370
Deletion of 13 amino acids from the carboxyl terminus of staphylococcal nuclease (WTSNase delta) results in a denatured, partially unfolded molecule that lacks significant persistent secondary structure but is relatively compact and monomeric under physiological conditions [Shortle & Meeker (1989) Biochemistry 28, 936-944; Flanagan et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 748-752]. Because of these and other properties of the SNase delta polypeptide, it is a useful model system for investigating the conformation of the denatured state of a protein without using extreme temperature or solvent conditions. Moreover, since the modification is a carboxyl-terminal deletion, SNase delta may also resemble a transient state of the polypeptide chain as it emerges from a ribosome prior to its folding. In the present study, we have examined the sizes and conformations of mutated forms of SNase delta, using small-angle X-ray scattering and circular dichroism spectroscopy. Seven mutated forms were studied: four with single substitutions, two with double substitutions, and one triple substitution. When present in the full-length SNase, each of these mutated forms exhibited unusual behavior upon solvent or thermal denaturation. In the case of the truncated form (SNase delta), the small-angle scattering curves of the mutated forms fall into two classes: one resembling the scattering curve of compact native nuclease and the other having features consistent with those expected for an expanded coil-like polymer. In contrast, the scattering curve of WT SNase delta exhibits features intermediate between those observed for globular proteins and random polymers. The amino acid substitutions that gave rise to compact, native-like versions of SNase delta were all of the m--type (m-substitutions are predicted to decrease the size of the denatured state). Those which gave rise to versions of SNase delta that were more extended and coil-like than WT SNase delta were of the m+ type (m+ substitutions are predicted to increase the size of the denatured state). Estimates of the residual secondary structure present in WT SNase delta, as well as both the m+ and m-substituted versions of SNase delta, as determined by CD, suggest that the formation of secondary structure and compaction of the polypeptide chain occur concurrently. Our results show that single amino acid substitutions can radically alter the conformational distribution of a partially condensed polypeptide chain.(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
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DM Wyatt 《Canadian Metallurgical Quarterly》1993,23(7):32U, 32Y-32Z passim
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Ockene Judith K.; Kristeller Jean; Pbert Lori; Hebert James R.; Luippold Rose; Goldberg Robert J.; Landon Joan; Kalan Kathryn 《Canadian Metallurgical Quarterly》1994,13(3):278
Patterns of smoking cessation using 6- and 12-mo follow-up data are reported for 1,261 primary care patients randomized to 3 physician-delivered smoking interventions: advice only (AO), counseling (CI), and counseling plus availability of nicotine-containing gum (CI?+?NCG). One-week point-prevalence cessation rates at 12 mo did not differ among the interventions: AO (15.2%), CI (12.9%) and CI?+?NCG (16.7%). However, maintained cessation rates (abstinent at both 6 and 12 mo) increased with intervention intensity: AO (6.0%), CI (7.8%), and CI?+?NCG (10.0%): Test of trend χ–2?=?5.06, p?=?.02. CI?+?NCG was significantly higher than AO (p?=?.02). The findings support the following conclusions: Brief physician delivered intervention with availability of nicotine-containing gum can have a beneficial long-term effect on smoking cessation, and cohort data as well as point-prevalence rates are important when assessing the long-term impact of lifestyle interventions. (PsycINFO Database Record (c) 2010 APA, all rights reserved) 相似文献
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