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941.
BD Abbott C Lau AR Buckalew TR Logsdon W Setzer RM Zucker KH Elstein RJ Kavlock 《Canadian Metallurgical Quarterly》1993,47(6):541-554
5-Fluorouracil (5-FU) inhibits the enzyme thymidylate synthetase (TS) which results in inhibition of DNA synthesis. 5-FU is teratogenic in many species, inducing cleft palate, limb, and tail defects. In the present study, gestation day (GD) 14 embryonic rat craniofacial explants were exposed to 5-FU in organ culture with increasing concentrations and durations of exposure. Palates exposed to 5-FU were morphologically abnormal and craniofacial shape, size, and palatal fusion pattern were affected with the severity of effects dependent on concentration and duration of exposure. Cleft palate was induced in vitro as opposing palates overlapped in a narrowed oral cavity. Palates exposed to higher levels of 5-FU were growth inhibited, but fused even though proliferation ceased and few cells were available to participate in elevation and fusion. This was demonstrated as a biphasic concentration-response profile for palatal fusion in which 0.05 to 0.15 micrograms 5-FU/ml produced decreasing rates of palatal fusion, while exposure to 0.15 to 3.0 micrograms/ml resulted in progressively increasing rates of fusion. The effects of 5-FU were detected biochemically as a reduction in TS activity which was concentration and time dependent during the first 12 hours, with a return to control levels by 24 hours. During the first day, 5-FU did not alter protein levels, but DNA levels significantly decreased at the high concentration, 2.0 micrograms/ml. After 5 days in culture, both DNA and protein decreased with increasing 5-FU concentration and duration of exposure. Also by the end of the culture period, 3H-TdR incorporation had decreased in a concentration dependent manner. It is concluded that progressive inhibition of proliferation and growth in organ culture results in two different morphological outcomes: cleft palate resulting from a narrowed oral cavity and increased incidence of anterior palatal fusion under conditions of strong growth reduction. This study demonstrates that elevation and fusion can occur in the absence of growth and proliferation. Based on these observations, severe inhibition of growth or proliferation would not necessarily be sufficient to induce cleft palate. 相似文献
942.
Pentobarbital alone, pentobarbital plus 1% lidocaine solution, pentobarbital plus 2% lidocaine solution, and pentobarbital plus 3% lidocaine solution were each used to euthanatize 6 dogs. For each dog, time between the beginning of injection of the euthanasia solution and each of the following events was recorded: collapse, onset of apnea, flat-line electrocardiogram, flat-line electroencephalogram, loss of palpable heartbeat, and loss of palpable pulse. Any signs of pain or discomfort were also recorded. There were no significant differences among groups except for time to flat-line electrocardiogram. Dogs euthanatized with pentobarbital alone had significantly longer times than did dogs euthanatized with pentobarbital in combination with any of the lidocaine concentrations. We concluded that pentobarbital in combination with lidocaine was a reasonable alternative to pentobarbital alone when euthanatizing dogs. 相似文献
943.
RF Kropman JA van Oostayen KH Zwinderman GA Lycklama A Nijehold J Schipper W Meinhardt 《Canadian Metallurgical Quarterly》1994,6(3):125-135
In 37 patients 25 to 69 years old (mean age 49.6 years) with suspected 'vasculogenic erectile dysfunction', diagnosed on the basis of repeated negative reactions to intracavernous pharmacological stimulation, duplex Doppler measurements and pharmacocavernosometry were performed in one session under simultaneous pressure registration. A statistically significant difference between patients with and without veno-occlusive dysfunction was found by combining end-diastolic velocity and pressure in time. We conclude that color Doppler scanning in combination with simultaneous pressure registration can predict veno-occlusive dysfunction in 83-95% of the patients. 相似文献
944.
B Millauer LK Shawver KH Plate W Risau A Ullrich 《Canadian Metallurgical Quarterly》1994,367(6463):576-579
Angiogenesis, the sprouting of capillaries from pre-existing blood vessels, is a fundamental process in the formation of the vascular system during embryonic development. In adulthood, angiogenesis takes place during corpus luteum formation and in pathological conditions such as wound healing, diabetic retinopathy, and tumor-igenesis. Vascularization is essential for solid tumour growth and is thought to be regulated by tumour cell-produced factors, which have a chemotactic and mitogenic effect on endothelial cells. Vascular endothelial growth factor (VEGF), a homodimeric glycoprotein of relative molecular mass 45,000, is the only mitogen, however, that specifically acts on endothelial cells, and it may be a major regulator of tumour angiogenesis in vivo. Its expression has been shown to be upregulated by hypoxia, and its cell-surface receptor, Flk-1, is exclusively expressed in endothelial cells. Here we investigate the biological relevance of the VEGF/Flk-1 receptor/ligand system for angiogenesis using a retrovirus encoding a dominant-negative mutant of the Flk-1/VEGF receptor to infect endothelial target cells in vivo, and find that tumour growth is prevented in nude mice. Our results emphasize the central role of the Flk-1/VEGF system in angiogenesis in general and in the development of solid tumours in particular. 相似文献
945.
gp93 was first described in growth cones from fetal rat brain as a 90-97-kDa glycoprotein family that binds wheat-germ agglutinin and consists of at least 12 different isoelectric variants (pl range approximately 4.9-6.4). Of particular interest is that different sets of gp93 variants are expressed in growth cones isolated from different brain regions. The preparation of a polyclonal antibody to gp93 allowed further characterization of this glycoprotein. The carbohydrate groups of gp93 were partially characterized by digestion with different glycosidases. The results indicate that most or all oligosaccharide units are N-linked (asparagine-linked) and contain sialic acid. Two-dimensional polyacrylamide gel electrophoresis and western blot with anti-gp93 show that deglycosylated gp93 is an only slightly heterogeneous polypeptide of 66 kDa, indicating that gp93 heterogeneity is due, primarily or exclusively, to differential glycosylation. Analysis of the tissue distribution in fetal rat showed gp93 to be highly enriched in the brain. Immunoblots and immunostaining of cross sections of developing cerebellum revealed that gp93 is developmentally regulated in this tissue, associated primarily with growing parallel fibers and Purkinje dendrites. Immunostaining of neurons in culture shows significant amounts of gp93 in elongating neurites and growth cones. Our results indicate that gp93 is a developmentally regulated glycoprotein of the brain that is most prominent in growth cones and growing neurites and that appears to be glycosylated differentially by different neurons. 相似文献
946.
Previous studies have shown that exogenous free n-3 polyunsaturated fatty acids (PUFA) can prevent tachyarrhythmias caused by specific agents in isolated cardiac myocytes. However, the question as to whether incorporation of the n-3 PUFA into membrane phospholipids has the same immediate protective effects remained to be answered. To answer this question, we increased the content of n-3 PUFA in the phospholipids of cultured neonatal rat myocytes by growing them 2-3 d in a culture to which eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) in 15 microM concentration was added. Analysis of the fatty acid composition of membrane phospholipids revealed a significantly higher level of EPA and DHA (from 0.2 to 7.6% and from 1.2 to 6.5%) in cells supplemented with EPA or DHA, respectively. The responses of the myocytes grown in normal media or in media enriched with the PUFA to arrhythmogenic agents were examined after free fatty acids were removed from the medium and the cells. The arrhythmogenic agents used were the beta-adrenergic agonist isoproterenol or an elevated extracellular concentration of calcium. The results showed that there was no significant difference in the induction of tachyarrhythmias by isoproterenol or by elevated [Ca2+]o in cells grown in media enriched with PUFA, as compared with cells grown in normal media in the absence of the free PUFA. Under the conditions of this study, only the unesterified PUFA were able to protect the cardiomyocytes against induced arrhythmias. There was no antiarrhythmic effect due to an increased fraction of EPA or DHA in membrane phospholipids. 相似文献
947.
The prevalence of vaginal yeast species has been studied in 140 women (41 pregnant, 66 infertile and 33 healthy controls) attending a gynaecological private clinic in Amman, Jordan. Yeast species were isolated from pregnant (68.2%), infertile (51.5%) and healthy control (48.4%) women. Patients manifesting one, two or three symptoms of vulvovaginitis were 22.1%, 26.8% or 24.2% respectively. Asymptomatic cases and cases with more than three symptoms were 22.4% and 4.5% respectively. Candida albicans was the dominant species (in 51.3% of the patients) followed by C. glabrata (17.9%). The percentage occurrence as well as the pattern of Candida species differed among the different groups of patients. Candida kefyr was found to be significantly higher in the infertile women. In vitro sensitivity tests using amphotericin B, nystatin, miconazole nitrate and chlorhexidine were carried out; amphotericin B was the most effective and miconazole nitrate the least. 相似文献
948.
KH Link G Leder J Pillasch U Butzer L Staib M Kornmann U Bruckner HG Beger 《Canadian Metallurgical Quarterly》1998,14(3):189-201
Two iron-regulated genes with deduced homology to TonB-dependent ferric siderophore receptors were cloned from Bordetella bronchiseptica by screening a library of TnphoA insertion mutants. bfrB and bfrC were iron-repressed in B. bronchiseptica by a Fur-dependent mechanism, and were expressed from promoters overlapped by potential Fur-binding sites. Both genes were highly conserved among Bordetella species and were also iron-regulated in Bordetella pertussis. bfrB and bfrC mutants of both species and a bfrB-bfrC double mutant of B. bronchiseptica had no discernible defects in utilization of known iron sources for Bordetella. 相似文献
949.
950.
K Krüger C Jochum KH Glüsenkamp C Krüsemann P Lorenz G Eberle-Adamkiewicz W Drosdziok DW Beelen HH Coenen MF Rajewsky 《Canadian Metallurgical Quarterly》1998,77(4):610-619
We have developed a new modular affinity system for the 2-step delivery of functional molecules to target cells. The system is based on the tautomer-specific monoclonal antibody (MAb) EM-6-47, which binds to 3- and 3,8-substituted adenines with high affinity (Ka > 10(9) l/mol) without cross-reacting with naturally occurring purine derivatives. This MAb serves as the hapten-specific fusion partner to produce bispecific MAbs (bs-MAbs) recognizing a target cell antigen and a low-m.w. hapten as carrier molecule for, e.g., radionuclides. Either the C-8 or the N-3 position of adenines can be used for conjugation with effector molecules; the remaining position may be substituted with different moieties to modulate the pharmacokinetics of the haptens. Different 3- and 3,8-substituted adenines conjugated to the chelates DOTA and DTPA or to the drug daunomycin were synthesized. Adenine-chelate derivatives were efficiently labeled with (111)In and 90Y, while high-affinity binding of 3-substituted adenines to MAb EM-6-47 remained almost unaffected by the conjugation to radiochelates. To confirm the validity of the delivery system, a prototype bs-MAb, EM-168-47, was generated by somatic cell fusion of MAb EM-6-47 and MAb EM-168-2, the latter recognizing a surface antigen on canine hematopoietic cells. Two-step targeting assays in vitro verified the bs-MAb-mediated, dose-dependent delivery of (111)In-labeled adenine-chelate derivatives to myeloid cells. This system represents a powerful tool for new pre-targeting approaches relying on bs-MAbs and low-m.w. haptens. Suitable cellular antigens can be targeted by fusing the appropriate MAbs with hapten-specific MAb EM-6-47, and tailor-made 3-substituted adenines may be labeled with diagnostic or therapeutic radionuclides, cytotoxic drugs or other functional molecules. 相似文献