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1.
Antisense oligonucleotide inhibition of hepatitis C virus gene expression in transformed hepatocytes
R Hanecak V Brown-Driver MC Fox RF Azad S Furusako C Nozaki C Ford H Sasmor KP Anderson 《Canadian Metallurgical Quarterly》1996,70(8):5203-5212
Genetic and biochemical studies have provided convincing evidence that the 5' noncoding region (5' NCR) of hepatitis C virus (HCV) is highly conserved among viral isolates worldwide and that translation of HCV is directed by an internal ribosome entry site (IRES) located within the 5' NCR. We have investigated inhibition of HCV gene expression using antisense oligonucleotides complementary to the 5' NCR, translation initiation codon, and core protein coding sequences. Oligonucleotides were evaluated for activity after treatment of a human hepatocyte cell line expressing the HCV 5' NCR, core protein coding sequences, and the majority of the envelope gene (E1). More than 50 oligonucleotides were evaluated for inhibition of HCV RNA and protein expression. Two oligonucleotides, ISIS 6095, targeted to a stem-loop structure within the 5' NCR known to be important for IRES function, and ISIS 6547, targeted to sequences spanning the AUG used for initiation of HCV polyprotein translation, were found to be the most effective at inhibiting HCV gene expression. ISIS 6095 and 6547 caused concentration-dependent reductions in HCV RNA and protein levels, with 50% inhibitory concentrations of 0.1 to 0.2 microM. Reduction of RNA levels, and subsequently protein levels, by these phosphorothioate oligonucleotides was consistent with RNase H cleavage of RNA at the site of oligonucleotide hybridization. Chemically modified HCV antisense phosphodiester oligonucleotides were designed and evaluated for inhibition of core protein expression to identify oligonucleotides and HCV target sequences that do not require RNase H activity to inhibit expression. A uniformly modified 2'-methoxyethoxy phosphodiester antisense oligonucleotide complementary to the initiator AUG reduced HCV core protein levels as effectively as phosphorothioate oligonucleotide ISIS 6095 but without reducing HCV RNA levels. Results of our studies show that HCV gene expression is reduced by antisense oligonucleotides and demonstrate that it is feasible to design antisense oligonucleotide inhibitors of translation that do not require RNase H activation. The data demonstrate that chemically modified antisense oligonucleotides can be used as tools to identify important regulatory sequences and/or structures important for efficient translation of HCV. 相似文献
2.
J Westhuyzen AD Cochrane PJ Tesar T Mau DB Cross MP Frenneaux FA Khafagi SJ Fleming 《Canadian Metallurgical Quarterly》1997,113(5):942-948
Augmentation of antioxidant defenses may help protect tissues against ischemia-reperfusion injury associated with operations involving cardiopulmonary bypass. In this study we examined the effect of pretreating patients with alpha-tocopherol (vitamin E) and ascorbic acid (vitamin C) or placebo on injury to the myocardium. Seventy-six subjects undergoing elective coronary artery bypass grafting participated in a prospective, double-blind, placebo-controlled randomized trial, receiving either placebo or both 750 IU dl-alpha-tocopherol per day for 7 to 10 days and 1 gm ascorbic acid 12 hours before the operation. Plasma alpha-tocopherol concentrations, raised fourfold by supplementation, fell by 70% after the operation in the supplemented group and to negligible levels in the placebo group. There were no significant differences between the groups with respect to release of creatine kinase MB isoenzyme over 72 hours, nor in the reduction of the myocardial perfusion defect determined by thallium 201 uptake. Electrocardiography provided no evidence of a benefit from antioxidant supplementation. Thus the supplementation regimen prevented the depletion of the primary lipid soluble antioxidant in plasma, but provided no measurable reduction in myocardial injury after the operation. 相似文献
3.
AD Santin PL Hermonat JC Hiserodt J Fruehauf V Schranz D Barclay S Pecorelli GP Parham 《Canadian Metallurgical Quarterly》1997,64(3):477-480
Tumor cells from eight freshly isolated cervical cancers (i.e., four adenocarcinomas and four squamous carcinomas) were analyzed for their production of the immune-inhibitory cytokine transforming growth factor-beta (TGF-beta) in vitro. All fresh adenocarcinomas secreted significant levels of TGF-beta (mean 397, range between 207 and 782 pg/ml/10(5) cells/48 hr). In contrast, no detectable TGF-beta was present in the supernatants from the four fresh squamous carcinoma cultures (P < 0.001). These data suggest that major differences in the secretion of the immunoinhibitory cytokine TGF-beta exist between squamous cell carcinomas and adenocarcinomas of the uterine cervix. Furthermore, these findings suggest that at least some of the differences in the natural biologic behavior, as well as in the response to radiation treatment, between these two histologic types of cervical cancer could be related to differences in secretion of this immune-inhibitory cytokine. 相似文献
4.
5.
In this study the authors have examined the effects of transluminal angioplasty on cerebral blood flow (CBF) in the management of intractable vasospasm following aneurysmal subarachnoid hemorrhage (SAH). Fourteen consecutively enrolled patients underwent attempted angioplasty with or without intraarterial infusion of papaverine. Twelve patients underwent pre- and postangioplasty xenon-enhanced computerized tomography (Xe-CT) scanning to measure regional CBF in 55 to 65 regions of interest (ROIs) per patient. Angioplasty was possible in 13 (93%) of 14 patients, with angiographically demonstrated improvement in all 13. Twelve (92%) of the 13 patients were neurologically improved following angioplasty; seven (58%) of the 12 patients who improved had a complete reversal of all delayed ischemic deficits. Angioplasty significantly decreased the mean number of ROIs at risk (11.4 ROIs pre- and 0.9 ROIs postangioplasty) (p < 0.00005, t-test). All patients had a reduction in the number of ROIs at risk after angioplasty; six (50%) of 12 no longer had any ROIs remaining at risk after angioplasty. Angioplasty significantly increased the mean CBF within at-risk ROIs (13 ml/100 g/minute pre- and 44 ml/100 g/minute postangioplasty) (p < 0.00005, t-test). All patients experienced an improvement in mean CBF in at-risk ROIs after angioplasty, with the mean CBF improving to above 20 ml/100 g/minute in all cases. No differences in the degree of improvement were found in patients who received intraarterial papaverine compared with those who did not. In the majority of patients with refractory vasospasm following SAH, angioplasty effectively dilated spastic arteries, reversed delayed neurological deficits, and significantly improved CBF in areas of brain at risk of infarction. 相似文献
6.
SC Langley GW Rickett A Hunt FJ Kelly AD Postle DA York 《Canadian Metallurgical Quarterly》1993,45(3):543-551
The biochemical maturation of the lung in late gestation and in the young animal is regulated by glucocorticoids. The present study was aimed at dissociating the different glucocorticoid receptor sites involved in these regulatory functions. The obese Zucker rat was selected as a model for this study as it exhibits hypersensitivity to glucocorticoid hormone action by virtue of its elevated receptor numbers and activity. Two synthetic steroid analogues were administered to obese animals; RU28362, a specific type II receptor agonist, and the type II antagonist RU486. RU28362 promoted a strong catabolic effect, which was associated with reduced food intake and the abolition of growth in the rats. The agonist, RU28362, attenuated developmental increases in antioxidant enzyme activities, and altered the growth of the tissue. At the age studied, development of the lung phosphatidylcholine (PC) system was almost complete, but RU28362 increased disaturated PC 16:0/16:0 concentrations by almost 2-fold, and altered the molecular composition of total pulmonary PC. RU486 attenuated the growth of the rats and reduced their food intake. Treatment with the type II antagonist attenuated lung growth and increased the activities of pulmonary copper zinc (Cu/Zn) and manganese (Mn) superoxide dismutases. RU486 had no effect on lung PC concentrations and molecular composition. The data suggest a role for type I glucocorticoid receptors in the regulation of the antioxidant enzyme system in the lung, as type II antagonism will channel endogenous glucocorticoid binding to the type I site. Type II receptor binding would appear to play a role in regulating the lung PC content. 相似文献
7.
Dynamic mechanical properties have been investigated for interpenetrating-network systems based on polyol-cured polyurethanes (PU) and 2 to 1 n-butyl acrylate-n-butyl methacrylate (Ac) networks. The systems were formed simultaneously (SIN) from all of the precursors and reactants for both networks in the same vessel, and sequentially (SIPN) by swelling a precured PU with the reactants that will form the Ac network. If the Ac network is formed after gelation of the PU, the IPNs are transparent and appear to have single T (tan δmax) between those of the homonetworks; visible-phase separation occurs if the Ac is intentionally polymerized prior to PU gelation. Damping curves were lower and broader and the T (tan δmax) and rubber moduli were higher for the SIN than for the SIPN systems. Up to 65 percent Ac, the T (tan δmax) data for both SIN and SIPN fit the Gordon-Taylor equation if a T (tan δmax) for the Ac homonetwork 7°C higher than observed is used, suggesting a higher crosslink density for the Ac network under these conditions. The differences in properties of the SIN and SIPN are assumed to be dependent on sample homogeneity and upon the presence of a tin catalyst in the SIN preparation. This can result in limited Ac-network formation and consequent phase separation before PU gelation has occurred, and the catalyst may also increase the extent of interaction, such as grafting or hydrogen-bond formation between the networks. 相似文献
8.
In this article, “Life time of influence” refers to Robert K. Merton's impact broadly, and emblematically, to his influence
upon my work. The article discusses 1) the scope and influence of Merton's ideas about social structure and explanations of
social processes; 2) his vast scholarship establishing the study of science as a social institution, with implications for
theory and research; and 3) his fostering of the social study of science through immense published work, and through impact
upon an inter-generational network of scholars.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
9.
Garey A. Fox John W. Fuchs Victor F. Medina Kaumudi Atapattu 《Canadian Metallurgical Quarterly》2007,133(3):255-262
Recent research has proposed the use of asphalt and tall-oil-pitch emulsions for stabilizing radioactive contamination deposited on surfaces in urban areas. The objective of this project was to investigate whether surface applied emulsions could capture airborne radioactive particulate. Laboratory experiments included wind-blown particulate capture studies using an acrylic column and particulate retainment experiments using a wind box capable of producing wind speeds of 96?km/h. A probe methodology was developed to relate particulate retainment to a tack force on the emulsion surface. Experiments were also performed to determine the potential for such emulsions to absorb particulate matter into their emulsion matrix. Tall-oil-pitch emulsions outperformed asphalt emulsions in terms of particulate retention, tack force, and the ability to absorb magnesium silicate. Both tall-oil-pitch and asphalt emulsions were capable of extracting 22–24?g?m?2 of powder from particulate-laden airflow. Tall-oil-pitch emulsions were capable of retaining as much as 5–10% of magnesium silicate powder applied (i.e., retainment densities of 10–20?g?m?2) even after seven?days of curing and after applying 96.5?km/h (60?mph) wind. Tall-oil-pitch emulsions were able to absorb surface-applied magnesium silicate (approximately 0.1–0.2?g of magnesium silicate per 1.0?g of emulsion within three?days) into their emulsion matrix, preventing the magnesium silicate from being exposed to the external environment. Initial results with these five different emulsion formulations suggested particulate capture was feasible. Future emulsion formulations (i.e., longer curing times with greater acid concentrations) should be tested to optimize this postdetonation response strategy. 相似文献
10.
MH Maslowska AD Sniderman LD MacLean K Cianflone 《Canadian Metallurgical Quarterly》1993,34(2):219-228
The initial suspicion that obesity increases coronary risk has been much sharpened with the demonstration that risk is more tightly linked to abdominal than to peripheral obesity, and tighter yet again when the mass of omental adipose tissue is taken into account. These data suggest that important metabolic differences might exist between adipocytes from different regions, and indeed, it has long been appreciated that triacylglycerol hydrolysis can be stimulated to a greater extent in omental than in subcutaneous adipocytes. The present study focuses on triacylglycerol synthesis in human subcutaneous and omental adipocytes, a process which, by contrast, has received relatively little attention. Experiments were done on adipose tissue removed at laparotomy and on cultured preadipocytes. With the former, triacylglycerol synthesis was measured in the presence and absence of oleate added to the medium using radiolabeled glucose and oleate as tracers. The results demonstrate that under all conditions examined triacylglycerol synthesis in subcutaneous adipose tissue exceeded that in deep omental adipose tissue. To study the cells in more detail, preadipocytes were cultured and triacylglycerol synthesis was examined again under basal conditions and with stimulation with insulin and acylation stimulating protein (ASP). Under basal conditions, particularly when oleate was added to the medium, clear differences were present such that triacylglycerol synthesis was substantially greater in subcutaneous preadipocytes than in omentally derived preadipocytes. These differences were more pronounced when the cells were stimulated with either insulin or acylation stimulating protein. Overall, triacylglycerol synthetic capacity in subcutaneous tissue exceeded that in omental tissue. As a consequence, omental tissue as compared to subcutaneous adipose tissue would have a limited capacity to prevent fatty acids from reaching the liver and stimulating hepatic lipoprotein synthesis. 相似文献