Insulin receptors and insulin action in dissociated brain cells

The present study was conducted to characterize insulin receptors and insulin action in rat brain cells. Binding of [125I]insulin to cells obtained by mechanically dissociating rat brains was 86% specific, time-dependent and reached equilibrium within 90 min. The t1/2 of association was 14 min and t1/2 of dissociation was 8 min. Scatchard analysis demonstrated the typical curvilinear plot providing high affinity (0.03 nM) and low affinity (6.6 nM) binding sites. The total number of binding sites were 0.15 pmol/mg protein. Crosslinking of [125I]insulin to its receptors on dissociated brain cells followed by SDS-PAGE and autoradiography showed that the alpha-subunit of the receptor had a molecular weight of 122,000. This was in contrast with a molecular weight of 134,000 for the liver alpha-subunit. Incubation of dissociated brain cells with insulin resulted in a concentration-dependent inhibition of total [3H]norepinephrine (NE) uptake. This inhibitory effect of insulin on [3H]NE uptake was sodium ion-dependent suggesting that 80-90% of the sodium ion-dependent uptake was insulin-sensitive. Incubation of lectin-purified insulin receptors with insulin resulted in a time- and concentration-dependent stimulation of phosphorylation of the tyrosine residue of an exogenous substrate poly (Glu, Tyr) (4:1). In addition, insulin also stimulated the autophosphorylation of the beta-subunit of the insulin receptors. These observations corroborate our contention that insulin exerts neuromodulatory effects mediated by the specific insulin receptors in the brain.     

Rapidly growing nontuberculous mycobacteria: a new enemy of the cardiac surgeon

A review of atypical mycobacterial infections complicating cardiac operations is presented. Proven sources of infections at different institutions include contaminated porcine valves and municipal water supply, but the mode of transmission in the great majority of patients remains unclear. There are two principal clinical forms of atypical mycobacterial infections after cardiac operations--endocarditis and sternal osteomyelitis. The latter has characteristics resembling tuberculotic "cold abscess." Specialized laboratory testing is necessary to confirm the diagnosis, and surgeons may have to take the initiative to request special microbiological investigation in cases where infection is clinically suspected but routine cultures are reported as "negative." The prognosis for patients who have any atypical mycobacterial infection after a heart operation is severe. Those infected with the strain chelonei and those whose cardiac chambers were entered during operation fare worse. This dim clinical prognosis may be improved by appropriate and aggressive antibiotic and surgical therapy. Awareness of the urgency of special bacteriological studies is the key to successful management.     

Multiple sclerosis: serial study of gadolinium-enhanced MR imaging

Thirteen patients with definite multiple sclerosis (MS), studied 16-24 months previously with magnetic resonance (MR) imaging with and without enhancement by intravenously administered gadolinium diethylenetriaminepentaacetic acid (DTPA) dimeglumine, were reexamined with a similar protocol. Assessment of enhancement and clinical activity in both studies revealed that enhancement was observed in 13 of 14 cases in which clinical activity had changed within 4 weeks of the study and thus appeared more sensitive than clinical examination in determining active disease. The 3-minute postinjection, short repetition time image (TR) was the most efficient for depicting enhancement. Enhancing lesions (active plaques) arose from previously hyper- or isointense regions on long TR images. Previously active lesions reverted to areas of iso- or hyperintensity on long TR images. Serial comparison of long TR images in this population reveals a decrease in high-intensity lesions on long TR images in some cases and an increase in others. The findings of high-intensity regions on long TR images and previously enhancing lesions both becoming isointense suggests that transient inflammatory changes with concomitant edema without demyelination and/or with significant remyelination may occur in some MS lesions. MS lesions are dynamic; both active and inactive lesions may show dramatic change on longitudinal MR imaging studies.     

Shear Bond Strengths of One-Bottle Adhesives to Oil-Contaminated Enamel

Purpose: To evaluate resin bond strength to enamel contaminated with handpiece oil. Materials and Methods: Bovine teeth were randomly assigned to six groups of 20 teeth each for treatment with one of six different bonding systems (five one‐bottle and one multibottle). For each system, 10 enamel specimens were contaminated with handpiece oil before acid‐etching and 10 were contaminated after acid‐etching. The enamel was etched for 15 seconds using 35% phosphoric acid. Following adhesive application, composite resin was bonded using a gelatin capsule technique. Shear bond strengths from the two contaminated groups were compared to bond strengths to uncontaminated enamel obtained from a previous study that was performed by the same group of investigators, using the same facility, materials, and methods. Results: Two‐way analysis of variance (ANOVA) showed that the factor “surface contamination” did not have a significant effect on bond strength (p > .542). The type of adhesive and the interaction of adhesive and surface contamination were significant (p <.0001 and p <.003, respectively). When oil was applied before etching, mean bond strengths ranged from 18.0 ± 4.8 MPa for OptiBond SOLO (Kerr Corp., Orange, California) to 25.3 ± 5.6 MPa for Tenure Quik with Fluoride (Den‐Mat Corp., Santa Maria, California). With oil applied after etching, bond strengths ranged from 18.4 ± 8.0 MPa for Tenure Quik with Fluoride to 27.4 ± 5.4 MPa for Single Bond (3M Dental Products, St. Paul, Minnesota). For the same adhesive, comparing uncontaminated and “oil‐before‐etch” contaminated groups, the only statistically significant difference in bond strengths was for OptiBond SOLO: 21.8 ± 4.0 MPa (uncontaminated) versus 18.0 ± 4.8 MPa (oil before etch). Comparing uncontaminated and “oil‐after‐etch” groups, the only statistically significant difference was for Tenure: 24.5 ± 5.7 MPa (uncontaminated) and 18.4 ± 8.0 MPa (oil after etch). CLINICAL SIGNIFICANCE Surface contamination with handpiece lubricant oil does not impair appreciably the efficacy of the adhesives used in this study when bonding composite resin to enamel.     

力竭运动大鼠心室肌蛋白质组表达特征

目的:采用蛋白质组学技术,建立安静和递增运动负荷训练后力竭大鼠心室肌蛋白质组的差异性表达谱,初步筛选出心室肌对力竭运动产生反应的目标蛋白质。方法:实验于2007-03在湖南师范大学生命科学学院蛋白质化学与蛋白质组学国家教育部重点实验室和省级运动人体科学实验室完成。①实验分组:10只SD雄性大鼠随机分为对照组和运动组,每组5只。②实验方法:运动组经过7周的大强度递增运动负荷训练后(最后一次力竭),对两组心室肌组织的全蛋白进行双向凝胶电泳分离。结果:经图像分析,在运动组的电泳图谱上共展现蛋白质点(338±17)个,对照组展现蛋白质点(352±17)个。运动后差异表达的蛋白质点共有99个。对其中差异表达的9个蛋白质点进行质谱鉴定,共鉴定出7个蛋白质,Stress-70protein,NADH-ubiquinone oxidoreductase Mr75000subnunit,Long-chain specific acyl-CoA dehydrogenase,Tropomyosin-1alphachain在运动后"缺失",Nitrilase family,member2在运动后表达上调在5倍以上,一个相对分子质量为21000的未知蛋白在运动后表达下调在5倍以上,另外有两个点经鉴定均为Myosin-6,在运动后表达量相反。这些蛋白质属于收缩蛋白、能量代谢酶、分子伴侣等。结论:递增运动负荷训练后力竭时,大鼠心室肌蛋白质组明显地发生了反应。运动后"缺失"和下调的蛋白质点与心肌收缩的调控和能量代谢的方式转变以及细胞的应激反应有关,其中,成功筛选出6种在运动医学领域尚未涉足的、具有运动应激特点的目标蛋白质。