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91.
Isolation and characterization of adenovirus type 2 vertex capsomer (penton base). 总被引:11,自引:0,他引:11
Using two temperature-sensitive mutants of human adenovirus 2, H2 ts-115 and H2 ts-125, both defective in fiber production at 39.5°, it was possible to isolate appreciable amounts of vertex capsomer (penton base) devoid of fiber projection. The penton base was purified by Freon extraction, ammonium sulfate precipitation, DEAE-Sephadex, and hydroxyapatite chromatography. The final product was homogeneous by two-dimensional immunoelectrophoresis, analytical ultracentrifugation, SDS-polyacrylamide gel electrophoresis, and electron microscopy. The penton base thus isolated had a sedimentation coefficient of 9.1 S and an apparent molecular weight of about 500, 000. Stoichiomeric analysis of the penton base and fiber components of adenovirion, performed by biochemical and immunological methods, suggested that there are five subunits of 85,000 daltons per penton base structure. This finding and the sedimentation data implied an elongated shape for the vertex capsomer. The penton base had an isoelectric point of 5.8. No free N-terminal amino acid was detectable and the amino acid composition showed a relatively high content of dicarboxylic amino acids, leucine, and significant amounts of cysteine. The penton base had a cell-detaching effect, but did not seem to induce neutralizing antibodies. No endonuclease activity was found associated with the purified penton base obtained after hydroxyapatite chromatography, or even after the DEAE-Sephadex step. 相似文献
92.
Boulanger Henri; Haymann Jean Philippe; Fouqueray Bruno; Mansouri Rafik; Metivier Fabien; Sarfati Emile; Glotz Denis 《Nephrology, dialysis, transplantation》2005,20(12):2865
Sir, In our experience, 10% of renal allograft recipients developsustained hyperparathyroidism and hypercalcaemia during thefirst year following renal transplantation. Persistent hypercalcaemiausually requires parathyroidectomy, which represents the onlydefinitive treatment currently available. Cinacalcet, a calcimimeticdrug, represents from now on an alternative 相似文献
93.
Circulating endothelial microparticles are associated with vascular dysfunction in patients with end-stage renal failure 总被引:13,自引:0,他引:13
Amabile N Guérin AP Leroyer A Mallat Z Nguyen C Boddaert J London GM Tedgui A Boulanger CM 《Journal of the American Society of Nephrology : JASN》2005,16(11):3381-3388
Endothelial dysfunction and arterial stiffness are major determinants of cardiovascular risk in patients with end-stage renal failure (ESRF). Microparticles are membrane fragments shed from damaged or activated cells. Because microparticles can affect endothelial cells, this study investigated the relationship between circulating microparticles and arterial dysfunction in patients with ESRF and identified the cellular origin of microparticles associated with these alterations. Flow cytometry analysis of platelet-free plasma from 44 patients with ESRF indicated that circulating levels of Annexin V+ microparticles were increased compared with 32 healthy subjects, as were levels of microparticles derived from endothelial cells (three-fold), platelets (16.5-fold), and erythrocytes (1.6-fold). However, when arterial function was evaluated noninvasively in patients with ESRF, only endothelial microparticle levels correlated highly with loss of flow-mediated dilation (r = -0.543; P = 0.004), increased aortic pulse wave velocity (r = 0.642, P < 0.0001), and increased common carotid artery augmentation index (r = 0.463, P = 0.0017), whereas platelet-derived, erythrocyte-derived, and Annexin V+ microparticle levels did not. In vitro, microparticles from patients with ESRF impaired endothelium-dependent relaxations and cyclic guanosine monophosphate generation, whereas microparticles from healthy subjects did not. Moreover, in vitro endothelial dysfunction correlated with endothelial-derived (r = 0.891; P = 0.003) but not platelet-derived microparticle concentrations. In fact, endothelial microparticles alone decreased endothelial nitric oxide release by 59 +/- 7% (P = 0.025). This study suggests that circulating microparticles of endothelial origin are tightly associated with endothelial dysfunction and arterial dysfunction in ESRF. 相似文献
94.
We report on the synthesis of various acridine (Acr)-spacer-nuclear localization signal (NLS) peptide conjugates and explore whether their use as NLS-labeling agent of plasmidic DNA could improve gene nuclear import and expression into cells when mediated by synthetic DNA complexes. As the conditions of successful use of the NLS properties to enhance gene transfer are not clear, and with the aim of detecting and defining the requirements of NLS-enhanced transfection, we investigated gene delivery and expression into various cell lines with various DNA complexes (lipoplexes or polyplexes) that were formulated for various N/P ratios from various preformed Acr-spacer-NLS/DNA complexes (1:1, 5:1 and 10:1 molar ratio). For the in vitro transfection assays, the lipoplexes and polyplexes were formulated from the preformed Acr-spacer-NLS/DNA complexes and dioctadecylamidoglycylspermine (DOGS)/dioleylphosphatidylethanolamine (DOPE) 1:1 mol and branched polyethyleneimine (PEI) 25 kDa, respectively, which are very efficient in vitro gene transfer systems. We show by fluorescence experiments that part of the acridine-NLS-conjugates remains intercalated within the plasmid for most of the N/P lipoplexes and polyplexes investigated. We show that, as several other studies performed with NLS-conjugates that are not covalently linked to DNA, the expression of the transgene is in most cases not improved upon complexation of plasmidic DNA with NLS-intercalating conjugates prior to its formulation as lipoplexes or polyplexes. 相似文献
95.
Multiple forms of autosomal ataxia exist which can be identified by genetic testing. Due to their wide variety, the identification of the appropriate genetic test is difficult but could be aided by magnetic resonance data. In this study, magnetic resonance spectroscopy (MRS) and imaging (MRI) data were recorded for 20 ataxia patients of six different types and compared to 20 normal subjects. Spectra were acquired in the pons, left frontal lobe, left basal ganglia, left cerebellar hemisphere and vermis. Both metabolite spectra and absolute metabolite concentrations were determined. Differences in metabolite levels were observed between ataxia patients and control subjects and between ataxia patients of different types. A number of correlations were found between metabolite ratios, atrophy levels, number of repeats on the small and large allele, age at examination, symptoms duration and age at symptoms onset for ataxia patients. These MR characteristics are expected to be useful for the identification of the ataxia type. 相似文献
96.
97.
The use of laparostomy in damage control surgery and uncontrolled intra-abdominal infection has been well described. We examined 71 patients who required laparostomy to see if trends in management and outcome could be identified based on the underlying disease state. The underlying etiology included gastrointestinal sepsis (n = 25), pancreatitis (n = 21), or trauma (n = 25). Pancreatitis patients required more operations per patient (P < 0.05). The likelihood and type of closure (fascial, mesh, or none) was related to the underlying etiology: trauma patients were more likely to have fascial closure (P < 0.02), patients with GI sepsis were more likely to require mesh closure, and pancreatitis patients were more likely to have no formal closure (P < 0.02). Only 29 per cent of patients achieved definitive fascial closure. Mortality in trauma patients was 20 per cent, 36 per cent for GI sepsis, and 43 per cent in patients with pancreatitis. Complications of laparostomy included enterocutaneous fistula (16.9%) and abscess formation (7%). Though the use of laparostomy has become more prevalent, it is still associated with significant hospital stay, morbidity, and mortality. In our study, the number of operations and likelihood of abdominal closure appears to correlate with the etiology of the underlying disease requiring use of laparostomy. 相似文献
98.
99.
100.
Questioning the cardiocirculatory excitatory effects of opioids under volatile anaesthesia 总被引:1,自引:0,他引:1
Docquier MA Lavand'homme P Boulanger V Collet V De Kock M 《British journal of anaesthesia》2004,93(3):408-413
Background. Opioid-induced hyperalgesia has been demonstratedin awake animals. We observed an increased haemodynamic reactivityin response to noxious stimuli in rats under sevoflurane anaesthesiatreated with a very low dose of sufentanil. The aim of thisinvestigation was to determine whether the two phenomena sharea common origin: an opioid-induced excitatory reaction. To addressthis, we administered several drugs with proven efficacy inopioid hyperalgesia to rats presenting with haemodynamic hyper-reactivity. Methods. The MACbar of sevoflurane was measured in controlsand in animals treated with sufentanil 0.005 µg kg1min1 before and after administration of i.v. (0.25, 0.5mg kg1) and intrathecal (i.t.) (250 µg) ketamine,i.v. (0.5, 1 mg kg1) and i.t. (30 µg) MK-801(NMDAantagonist), i.v. (0.1, 0.5 mg kg1) naloxone, i.v. (10mg kg1) and i.t. (50, 100 µg) ketorolac or i.t.(100, 150 µg) meloxicam (COX-2 inhibitor). Results. Sufentanil 0.005 µg kg1 min1 significantlyincreased MACbar (3.2 (SD 0.3) versus 1.9 (0.3) vol%). Withthe exception of naloxone, all drugs displayed a significantMACbar-sparing effect (>50%) in controls. Naloxone completelyprevented haemodynamic hyperactivity. Two patterns of reactionwere recorded for the other drugs: either hyper-reactivity wassuppressed and the MACbar-sparing effect was maintained (i.t.ketamine, i.t. MK-801, i.t. ketorolac [100 µg], i.t. meloxicam[150 µg]) or hyper-reactivity was blocked but MACbar-sparingeffect was lost (i.v. ketamine [0.5 mg kg1], i.v. MK-801[0.5, 1 mg kg1], i.v. ketorolac [10 µg kg1],i.t. ketorolac [50 µg], i.t. meloxicam [100 µg]). Conclusions. We have demonstrated that low-dose sufentanil-inducedhaemodynamic hyper-reactivity is an excitatory µ-opiate-relatedphenomenon. This effect is reversed by drugs effective in treatingopiate-induced hyperalgesia. 相似文献