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91.
To engineer an a non-islet cell capable of glucose-stimulated insulin secretion, a chinese hamster ovary cell line (CHO) was transfected with a mammalian expression vector carrying the human insulin cDNA (pCB/hINS). More proinsulin than insulin was released daily by the stably transformed cell line (CHO-INS). Examination of acid-ethanol extracts confirmed that both insulin and proinsulin were stored. Immunohistochemical analysis of the cells also showed that (pro)insulin was stored. Unlike beta cells, CHO-INS cells did not secrete insulin in response to glucose. To investigate this lack of effect, we examined whether transfection of GLUT2 cDNA, which is ordinarily not expressed in CHO-INS cells, would confer glucose-stimulated insulin secretion. Consequently, we have demonstrated that glucose regulated insulin release occurs in the CHO-INS-GLUT2 cell line and that glucose potentiates the insulin secretory response to non-glucose secretagogues.  相似文献   
92.
Monolithic columns for capillary electrochromatography have been prepared within the confines of untreated fused-silica capillaries in a single step by a simple copolymerization of mixtures of butyl methacrylate, ethylene dimethacrylate, and 2-acrylamido-2-methyl-1-propane-sulfonic acid (AMPS) in the presence of a porogenic solvent. The use of these novel macroporous monoliths eliminates the need for frits, the difficulties encountered with packed capillaries, and capillary surface functionalization. Since the porous properties of the monolithic materials can be easily tailored through changes in the composition of the ternary porogenic solvent, the effects of both pore size and the percentage of sulfonic acid monomer on the efficiency and the electroosmotic flow velocity of the capillary columns could be studied independently over a broad range. A simple increase in the content of charged functionalities within the monolith leads to an expected acceleration of the flow velocity. However, increasing the pore size leads to a substantial deterioration of the efficiency of the separation. In contrast, monoliths with increasing levels of AMPS in which the pore size remains fixed due to adjustments in the composition of the porogenic solvent show no deterioration in efficiency while maintaining the same increase in flow velocity, thus producing a significant reduction in separation time. Additionally, measurements on monoliths with constant levels of AMPS but different pore sizes suggest that flow velocity may be affected by the flow resistance within the capillary column.  相似文献   
93.
Newly developed software additions to the three-dimensional dynamic image analysis system, 3D-DIAS, are described for simultaneously reconstructing and motion analyzing in three dimensions the outer surface, nucleus and pseudopods of living, crawling cells. This new system is then used to describe for the first time a nuclear behavior cycle in translocating Dictyostelium discoideum amoebae and to investigate the role of pseudopod extension in this process. The nuclear behavior cycle is tuned to the two phases of the general cell behavior cycle [Wessels et al., 1994], and includes nuclear migration both in the z- and in the x,y-axes from the proximal border of the prior anterior pseudopod to the proximal border of a newly expanding anterior pseudopod. Nuclear migration is cued by pseudopod-substratum contact, achieves velocities in excess of 50 microm/min, and is accompanied by characteristic changes in nuclear shape. The rules and characteristics of nuclear behavior are demonstrated to be intact in two mutants affecting pseudopod formation, a myosin IB null mutant (myoB-) and a myosin II heavy chain phosphorylation mutant (3XALA). The rules and characteristics of nuclear migration, however, are disrupted upon dissolution of microtubules by colcemid. Together the above results demonstrate that the newly developed 3D-DIAS system can be used to gain new insights into the dynamic changes in the intracellular 3D architecture associated with cellular translocation.  相似文献   
94.
To gain access to information on energy use in office buildings, the German Federal Ministry for Economy launched an intensive research and demonstration programme in 1995. In advance of the 2002 EU energy performance directive a limited primary energy coefficient of about 100 kW h m−2 a−1 as a goal for the complete building services technology was postulated (HVAC + lighting) for all demonstration buildings to be supported. A further condition was that active cooling be avoided. Techniques such as natural or mechanical night ventilation or heat removal by slab cooling with vertical ground pipes as well as earth-to-air heat exchangers in the ventilation system were applied. An accompanying research was established to keep track of the results and the lessons learned from about 22 demonstration buildings realized and monitored until the end of 2005. As one outcome this paper summarises the energy performance of a selection of characteristic buildings together with an overview on the summer thermal comfort situations achieved. The research program will proceed during the next five years. Detailed reports and future results may be downloaded from the internet: www.enbau-monitor.de.  相似文献   
95.
Metaphor comprehension was studied in schizophrenics as compared to psychiatric and nonpsychiatric controls. Subjects were asked in a two-condition forced-choice response task to detect the metaphor similar or contrary to a proverb with an abstract meaning. Schizophrenic patients were impaired in both conditions. However, the paranoid patients' performances did not differ from psychiatric and nonpsychiatric controls. These results are consistent with the hypothesis that the ability of metaphorizing is preserved in some schizophrenic patients.  相似文献   
96.
Murine embryonic stem (ES) cells have been a useful model system for the study of various aspects of hematopoietic differentiation. Because we had observed a sharp peak of expression of the receptor tyrosine phosphatase gamma (Ptp gamma) gene between 14 and 18 days of ES-derived embryoid body differentiation, we investigated the effect of perturbation of expression of the Ptp gamma gene on ES cell differentiation, first by analyzing the effect of Ptp gamma overexpression. The murine full-length Ptp gamma cDNA in an expression vector was transfected into ES-D3 cells and stably transfected clones were isolated. Ptp gamma was expressed as an approximately 230-kD cell surface protein, and differentiating ES clones that overexpressed Ptp gamma gave rise to a normal number of hematopoietic colonies, approximately 1 CFU per 100 cells. There was, however, a significant increase of expression of early hematopoietic markers in colonies from Ptp gamma overexpressing ES cells. To confirm that the pertubation of hematopoietic differentiation was a result of Ptp gamma overexpression, we isolated ES stem cell clones expressing Ptp gamma antisense constructs and assayed embryoid bodies for the presence of hematopoietic precursors. We observed a complete absence of methylcellulose colonies, indicating absence of hematopoietic lineages. Results of these experiments point to an essential role for Ptp gamma in hematopoietic differentiation.  相似文献   
97.
PURPOSE: To determine medical students' experiences in working with diabetic patients, their cognitive competence and their confidence in meeting diabetic patients' educational needs, and changes in their expertise with advancing education. METHOD: The study was conducted in March 1994 at the University of Illinois College of Medicine, where students work with ambulatory patients in a longitudinal primary care experience from their second year through graduation. A combined questionnaire and objective quiz was developed and distributed to 138 second-, third-, and fourth-year students. The students were asked whether they had had experience in seeing a newly diagnosed diabetic patient, in what setting, and whether they felt confident in educating such patients on various common topics. They were also asked about their interests in and preferences for learning about diabetic patients' education. A 50-item objective quiz on the management of diabetes followed the questionnaire. Data were analyzed with several statistical methods. RESULTS: In all, 65 of the students (47%) responded; the majority were third- or fourth-year students. The percentages of students who felt confident ranged from 83% who felt confident about counseling about exercise to only 16% who felt confident about pre-conception counseling. All of the students but one felt that diabetic patients' education was an important topic, and all but two were interested in it. The students' quiz scores showed no significant difference by year of training; for all the students the average score was 39.6% correct, with no increase in score for increased training. Also, the students who were more confident about their abilities to provide patient education on a certain topic were no more likely to have accurate information about that topic than were those who were not confident. CONCLUSION: Although they were clearly interested in diabetic patients' education, the students did not feel confident in providing it, nor did they have the cognitive competence to do so. Medical schools have a responsibility to students and to their patients to formally address the issues of patient education.  相似文献   
98.
OBJECTIVE: Menopause is associated with critical changes in the cardiovascular system, and the possible effect of hormonal replacement therapy (HRT) on these changes is under investigation. The aim of our study was to evaluate in postmenopausal women the effects of HRT and clonidine on the response of plasma calcitonin gene-related peptide (CGRP) and plasma atrial natriuretic peptide (ANP) to the upright posture test and the saline infusion test respectively. METHODS: CGRP and ANP levels were measured with specific radioimmunological assays and expressed in pmol/l (means +/- S.E.M). DESIGN: Postmenopausal women (age 46-53 years) (n = 18) were studied before and after 3 months of HRT (n = 13) or clonidine treatment (n = 5). RESULTS: After HRT or clonidine treatment plasma CGRP levels (14.9 +/- 1.6 and 15.9 +/- 3.8 pmol/l) were significantly higher than before (9.8 +/- 0.6 and 10.5 +/- 1.6 pmol/l) (P < 0.01). The assumption of upright posture caused no change in plasma CGRP levels before treatment, while after HRT, but not after clonidine treatment, an increase in plasma CGRP levels was observed (P < 0.01 at 5 and 20 min). Basal plasma ANP levels significantly decreased after both HRT and clonidine treatment (P < 0.01). In untreated women the saline infusion test did not induce any change in plasma ANP levels; a significant response to the test was restored after HRT but not after clonidine treatment (P < 0.01 at 90 and 120 min). CONCLUSIONS: The results show that some of the adaptive responses modified by menopausal changes are restored by HRT but not clonidine treatment, suggesting a modulatory role for sex steroid hormones in cardiovascular function and salt and water balance.  相似文献   
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