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991.
Ku, a heterodimer of polypeptides of approximately 70 kDa and 80 kDa (Ku70 and Ku80, respectively), binds avidly to DNA double-strand breaks (DSBs). Mammalian cells defective in Ku are hypersensitive to ionizing radiation due to a deficiency in DSB repair. Here, we show that the simple inactivation of the Saccharomyces cerevisiae Ku70 homologue (Yku70p), does not lead to increased radiosensitivity. However, yku70 mutations enhance the radiosensitivity of rad52 strains, which are deficient in homologous recombination. Through establishing a rapid and reproducible in vivo plasmid rejoining assay, we show that Yku70p plays a crucial role in the repair of DSBs bearing cohesive termini. Whereas this damage is repaired accurately in YKU70 backgrounds, in yku70 mutant strains terminal deletions of up to several hundred bp occur before ligation ensues. Interestingly, this error-prone DNA repair pathway utilizes short homologies between the two recombining molecules and is thus highly reminiscent of a predominant form of DSB repair that operates in vertebrates. These data therefore provide evidence for two distinct and evolutionarily conserved illegitimate recombination pathways. One of these is accurate and Yku70p-dependent, whereas the other is error-prone and Yku70-independent. Furthermore, our studies suggest that Yku70 promotes genomic stability both by promoting accurate DNA repair and by serving as a barrier to error-prone repair processes. 相似文献
992.
RR de Ruyter van Steveninck GD Lewen SP Strong R Koberle W Bialek 《Canadian Metallurgical Quarterly》1997,275(5307):1805-1808
To provide information about dynamic sensory stimuli, the pattern of action potentials in spiking neurons must be variable. To ensure reliability these variations must be related, reproducibly, to the stimulus. For H1, a motion-sensitive neuron in the fly's visual system, constant-velocity motion produces irregular spike firing patterns, and spike counts typically have a variance comparable to the mean, for cells in the mammalian cortex. But more natural, time-dependent input signals yield patterns of spikes that are much more reproducible, both in terms of timing and of counting precision. Variability and reproducibility are quantified with ideas from information theory, and measured spike sequences in H1 carry more than twice the amount of information they would if they followed the variance-mean relation seen with constant inputs. Thus, models that may accurately account for the neural response to static stimuli can significantly underestimate the reliability of signal transfer under more natural conditions. 相似文献
993.
N Patel R Padera GH Sanders SM Cannizzaro MC Davies R Langer CJ Roberts SJ Tendler PM Williams KM Shakesheff 《Canadian Metallurgical Quarterly》1998,12(14):1447-1454
Controlling receptor-mediated interactions between cells and template surfaces is a central principle in many tissue engineering procedures (1-3). Biomaterial surfaces engineered to present cell adhesion ligands undergo integrin-mediated molecular interactions with cells (1, 4, 5), stimulating cell spreading, and differentiation (6-8). This provides a mechanism for mimicking natural cell-to-matrix interactions. Further sophistication in the control of cell interactions can be achieved by fabricating surfaces on which the spatial distribution of ligands is restricted to micron-scale pattern features (9-14). Patterning technology promises to facilitate spatially controlled tissue engineering with applications in the regeneration of highly organized tissues. These new applications require the formation of ligand patterns on biocompatible and biodegradable templates, which control tissue regeneration processes, before removal by metabolism. We have developed a method of generating micron-scale patterns of any biotinylated ligand on the surface of a biodegradable block copolymer, polylactide-poly(ethylene glycol). The technique achieves control of biomolecule deposition with nanometer precision. Spatial control over cell development has been observed when using these templates to culture bovine aortic endothelial cells and PC12 nerve cells. Furthermore, neurite extension on the biodegradable polymer surface is directed by pattern features composed of peptides containing the IKVAV sequence (15, 16), suggesting that directional control over nerve regeneration on biodegradable biomaterials can be achieved. 相似文献
994.
SP Kamwendo GA Ingram FL Musisi AJ Trees DH Molyneux 《Canadian Metallurgical Quarterly》1993,87(5):525-535
Incubation of fluorescein-conjugated and biotinylated lectins with Rhipicephalus appendiculatus salivary glands revealed differences between the basal laminae of the haemocoelic surfaces of the three acinal types. There were some similarities in the lectin binding characteristics of the surfaces of type II and type III acini when Con A, PNA and TVA were applied, indicating the presence of galactose, mannose and glucose moieties on the acinal surfaces. The binding of BPA, HPA and HAA lectins, specific for galactosyl and glycosyl ligands, which occurred only on the surface of type III acini, was moderate to intense. The remaining galactose-reactive lectins (GMA, DBA and VVA) also bound only to type III acini and the level of binding was weak to moderate. Although the relationship between the haemocoelic surface of the R. appendiculatus salivary gland acini and Theileria kinetes is not known, the consistent differences in the surface carbohydrate composition of the acini confirm the existence of differences in the basic physiology of the acini which may correlate with the specific susceptibility of the type III acinus to Theileria parva infection. 相似文献
995.
A series of 228 eyes implanted with one-piece all poly(methyl methacrylate) (PMMA) biconvex posterior chamber intraocular lenses was examined for posterior capsule opacification. One hundred forty-one eyes (61.8%) had opacification at an average postoperative period of 19.7 months. Seventy eyes (30.7%) developed an unusual form of early central posterior capsular fibrosis (ECPCF), which was confined to the capsulorhexis opening, sparing the peripheral aspect of the anterior and posterior capsules. Risk factors for developing this form of opacification were close apposition of peripheral anterior and posterior capsules caused by placing a posteriorly vaulted biconvex optic anterior to a capsulorhexis opening smaller than the optic diameter. This opacification occurred most often in cases of haptic fixation in the ciliary sulcus. The cumulative capsulotomy rate in this series was 5.26% at three months, 9.1% at 12 months, and 13.2% at 20 months. Of the ECPCF cases, 34.3% eventually required neodymium: YAG (Nd:YAG) laser capsulotomy; the capsulotomy rate for ECPCF was 4.8 times higher than that for Elschnig pearls. Early onset of ECPCF (average onset = 19.4 weeks) resulted in early Nd:YAG capsulotomy (average = 8.0 months after surgery). One-piece all-PMMA biconvex intraocular lenses may promote early central fibrosis of the posterior capsule if the lens optic is anterior to a capsulorhexis opening smaller than the optic diameter. The early onset of this form of opacification predisposes to earlier Nd:YAG capsulotomy with a higher risk of complications. 相似文献
996.
997.
M Stubbs SP Robinson LM Rodrigues CS Parkins DR Collingridge JR Griffiths 《Canadian Metallurgical Quarterly》1998,78(11):1449-1456
Characteristics of the tumour metabolic profile play a role in both the tumour-host interaction and in resistance to treatment. Because carbogen (95% oxygen/5% carbon dioxide) breathing can both increase sensitivity to radiation and improve chemotherapeutic efficacy, we have studied its effects on the metabolic characteristics of Morris hepatoma 9618a. Host carbogen breathing increased both arterial blood pCO2 and pO2, but decreased blood pH. A fourfold increase in tumour pO2 (measured polarographically) and a twofold increase in image intensity [measured by gradient recalled echo magnetic resonance (MR) imaging sensitive to changes in oxy/deoxyhaemoglobin] were observed. No changes were seen in blood flow measured by laser Doppler flowmetry. Tumour intracellular pH remained neutral, whereas extracellular pH decreased significantly (P < 0.01). Nucleoside triphosphate/inorganic phosphate (NTP/Pi), tissue and plasma glucose increased twofold and lactate decreased in both intra- and extracellular compartments, suggesting a change to a more oxidative metabolism. The improvement in energy status of the tumour was reflected in changes in tissue ions, including Na+, through ionic equilibria. The findings suggest that the metabolic profile of hepatoma 9618a is defined partly by intrinsic tumour properties caused by transformation and partly by tissue hypoxia, but that it can respond to environmental changes induced by carbogen with implications for improvements in therapeutic efficacy. 相似文献
998.
PJ Blair LH Boise SP Perfetto BL Levine G McCrary KF Wagner DC St Louis CB Thompson JN Siegel CH June 《Canadian Metallurgical Quarterly》1997,17(3):234-246
Progression to AIDS in asymptomatic HIV-infected individuals is characterized by a gradual but progressive loss of CD4+ T cells. While the mechanisms underlying this decline are currently unknown, recent evidence suggests that these cells are abnormally sensitive to apoptosis in response to activation signals. Recent work has implicated downregulation of Bcl-2 with the increased spontaneous apoptosis in lymphocytes from HIV-infected patients. We have evaluated the roles of the apoptosis-protective proteins Bcl-2 and Bcl-x in stimulated PBMC from asymptomatic HIV-infected and HIV-uninfected individuals. We found that Bcl-2 was constitutively expressed in PBMC from both HIV-infected and uninfected samples. However, Bcl-x induction was delayed and responses were decreased in stimulated HIV-infected samples. Additionally, single-cell intracellular staining of Bcl-x revealed a significant inverse correlation between PWM-induced Bcl-x expression and apoptosis (r = -0.695, P = 0.05). This was confirmed at the single-cell level in direct experiments when stimulated cells were sorted based on Bcl-x induction and then measured for apoptosis. Furthermore, low Bcl-x expression was not due to reduced lymphocyte activation following PWM stimulation. Our data indicate that the induction of Bcl-x is markedly impaired in asymptomatic HIV-infected patients and that stimuli which induce inadequate expression of Bcl-x are associated with increased levels of apoptosis in these cells. 相似文献
999.
Fucan, a sulfated polysaccharide extracted from brown seaweeds, inhibits smooth muscle cell (SMC) proliferation with a higher antiproliferative activity than heparin (Logeart et al., Eur. J. Cell Biol. 74, 1997, this issue). In order to investigate the structure-activity relationship of fucan on SMC growth, we have prepared by size exclusion chromatography fucan fractions of various molecular masses ranging from 5.5 to 556 kDa. Our experiments showed that the antiproliferative activity is dependent on the molecular weight of the polysaccharide. The molecular weight threshold indicated that about 30 saccharidic units on fucan were necessary to give the antiproliferative activity on SMCs. A kinetics study of DNA synthesis using tritiated thymidine uptake was also performed with different molecular weight fucan fractions. Although all tested fractions acted as soon as the cells enter the first cell cycle, the duration and potency of action varied. Moreover, displacement experiments of iodinated fucan revealed that the low molecular fucan fraction interacted weakly with the binding sites. Finally, gel permeation chromatography of internalized radiolabeled heparin and fucans was performed with SMCs. A rapid degradation of internalized heparin was observed, whereas only low molecular weight fucan fractions were partially degraded by SMCs. Together, these results indicate the significance of molecular weight on the antiproliferative activity of fucans on SMCs, and might help to understand their mechanism of action. In addition, the degradation experiments with internalized heparin and fucans ruled out a direct link between polysaccharide degradation and the antiproliferative effect on SMCs. 相似文献
1000.