NMR study of the cataract-linked P23T mutant of human γD-crystallin shows minor changes in hydrophobic patches that reflect its retrograde solubility

The Pro23 to Thr (P23T) mutation in human γD-crystallin (HGD) shows several cataract phenotypes. We found earlier [A. Pande, O. Annunziata, N. Asherie, O. Ogun, G.B. Benedek, J. Pande, Decrease in protein solubility and cataract formation caused by the Pro23 to Thr mutation in human gamma D-crystallin, Biochemistry 44 (2005) 2491-2500] that the mutation dramatically lowers the solubility of P23T but the overall protein fold is maintained. Recently we observed that solutions of P23T showed liquid-liquid phase transition behavior similar to that of HGD but the liquid-protein crystal phase transition was altered, suggesting an asymmetric distribution of “sticky” patches on the protein surface [J.J. McManus, A. Lomakin, O. Ogun, A. Pande, M. Basan, J. Pande, G.B. Benedek, Altered phase diagram due to a single point mutation in human gammaD-crystallin, Proc. Natl. Acad. Sci. USA 104 (2007) 16856-16861]. Here we present high-resolution NMR studies of HGD and P23T in which we have made nearly complete backbone assignments. The data provide a structural basis for explaining the retrograde solubility of P23T by (a) identifying possible “sticky” patches on the surface of P23T and (b) highlighting their asymmetric distribution.     

Biological activities of the natural imidazole-containing peptidomimetics n-acetylcarnosine, carcinine and L-carnosine in ophthalmic and skin care products

Apart from genetically programmed cell aging, different external aggressors related to oxidative stress and lipid peroxidation (LPO) can accelerate the skin aging phenomenon. Oxidative stress associated with the formation of lipid peroxides is suggested to contribute to pathological processes in aging and systemic diseases known as the risk factors for cataract. Despite the fact that L-carnosine-related peptidomimetics N-acetylcarnosine (N-acetyl-beta-alanyl-L-histidine) (NAC) and carcinine (beta-alanylhistamine) are metabolically related to L-carnosine and have been demonstrated to occur in tissues of many vertebrates, including humans, these compounds were shown resistant toward enzymatic hydrolysis. A series of related biocompatible imidazole-containing peptidomimetics were synthesized in order to confer resistance to enzymatic hydrolysis and ex vivo improvement of protective antioxidative properties related to L-carnosine. The included findings revealed a greater role of N-acetylcarnosine (NAC) and carcinine ex vivo in the prolongation and potentiation of physiological responses to the therapeutical and cosmetics treatments with L-carnosine as antioxidant. 3-D molecular conformation studies proposed the antioxidant activity of peptidomimetics (carcinine, L-prolylhistamine, N-acetylcarnosine, L-carnosine) for metal ion binding, quenching of a number free radicals, and binding of hydroperoxide or aldehyde (including dialdehyde LPO products) in an imidazole-peroxide adducts. NAC can act as a time release (carrier) stable version of L-carnosine during application in ophthalmic pharmaceutical and cosmetics formulations which include lubricants. Carcinine, L-prolylhistamine show efficient deactivation of lipid hydroperoxides monitored by HPLC and protection of membrane phospholipids and water soluble proteins from the lipid peroxides-induced damages. This activity is superior over the lipophilic antioxidant vitamin E. The biologically significant applications of carnosine mimetics were patented by Dr. Babizhayev and the alliance Groups (WO 2004/028536 A1; WO 94/19325; WO 95/12581; WO 2004/064866 A1).     

超声乳化手术治疗慢性闭角型青光眼合并白内障的临床疗效分析

目的:探讨超声乳化手术治疗慢性闭角型青光眼合并白内障的临床效果。方法:慢性闭角型青光眼合并白内障64例(64眼)根据治疗方法的不同分为治疗组与对照组各32例,对照组采用传统小梁切除手术,治疗组采用超声乳化手术。结果:(1)两组术前视力对比无明显差异,治疗后视力情况都明显改善,同时组间对比有明显差异(P0.05)。(2)两组术前眼压比较无显著性差异,术后两组组间与组内对比都有明显差异(P0.05)。(3)两组术前前房深度比较无显著性差异,治疗后都有明显上升(P0.05),同时组间对比差异明显(P0.05)。(4)两组患者术中与术后都无严重并发症发生。结论:对于慢性闭角型青光眼合并白内障患者行超声乳化手术具备加深前房、控制眼压、提高视力的作用,同时安全性好,可作为标准治疗选择。     

Glutaredoxin 2 (Grx2) Gene Deletion Induces Early Onset of Age-dependent Cataracts in Mice

Glutaredoxin 2 (Grx2) is an isozyme of glutaredoxin1 (thioltransferase) present in the mitochondria and nucleus with disulfide reductase and peroxidase activities, and it controls thiol/disulfide balance in cells. In this study, we investigated whether Grx2 gene deletion could induce faster age-related cataract formation and elucidated the biochemical changes effected by Grx2 gene deletion that may contribute to lens opacity. Slit lamp was used to examine the lenses in Grx2 knock-out (KO) mice and age-matched wild-type (WT) mice ages 1 to 16 months. In the Grx2 null mice, the lens nuclear opacity began at 5 months, 3 months sooner than that of the control mice, and the progression of cataracts was also much faster than the age-matched controls. Lenses of KO mice contained lower levels of protein thiols and GSH with a significant accumulation of S-glutathionylated proteins. Actin, αA-crystallin, and βB2-crystallin were identified by Western blot and mass spectroscopy as the major S-glutathionylated proteins in the lenses of 16-month-old Grx2 KO mice. Compared with the WT control, the lens of Grx2 KO mice had only 50% of the activity in complex I and complex IV and less than 10% of the ATP pool. It was concluded that Grx2 gene deletion altered the function of lens structural proteins through S-glutathionylation and also caused severe disturbance in mitochondrial function. These combined alterations affected lens transparency.     

Defect of Mitotic Vimentin Phosphorylation Causes Microophthalmia and Cataract via Aneuploidy and Senescence in Lens Epithelial Cells

Vimentin, a type III intermediate filament (IF) protein, is phosphorylated predominantly in mitosis. The expression of a phosphorylation-compromised vimentin mutant in T24 cultured cells leads to cytokinetic failure, resulting in binucleation (multinucleation). The physiological significance of intermediate filament phosphorylation during mitosis for organogenesis and tissue homeostasis was uncertain. Here, we generated knock-in mice expressing vimentin that have had the serine sites phosphorylated during mitosis substituted by alanine residues. Homozygotic mice (VIM^SA/SA) presented with microophthalmia and cataracts in the lens, whereas heterozygotic mice (VIM^WT/SA) were indistinguishable from WT (VIM^WT/WT) mice. In VIM^SA/SA mice, lens epithelial cell number was not only reduced but the cells also exhibited chromosomal instability, including binucleation and aneuploidy. Electron microscopy revealed fiber membranes that were disorganized in the lenses of VIM^SA/SA, reminiscent of similar characteristic changes seen in age-related cataracts. Because the mRNA level of the senescence (aging)-related gene was significantly elevated in samples from VIM^SA/SA, the lens phenotype suggests a possible causal relationship between chromosomal instability and premature aging.     

不同手术联合超声乳化人工晶体植入对闭角型青光眼合并白内障临床观察

目的:对不同手术联合超声乳化人工晶体植入对闭角型青光眼合并白内障进行临床观察。方法:选择2012年3月~2014年4月在我院进行诊治的闭角型青光眼合并白内障患者109例(123眼),按照房角粘连状况和眼压大小选择不同的手术治疗方法。房角粘连≤180°、眼压≤30 mm Hg的58例(65眼)患者为A组,采用房角分离术联合超声乳化人工晶体植入。房角粘连180°、眼压30 mm Hg的51例(58眼)患者为B组,采用小梁切除术联合超声乳化人工晶体植入。术后随访半年,比较两组手术前后的眼压、视力、前房深度。结果:术后6个月,两组的矫正视力均较术前明显提高(P0.05);两组术后7 d、1、3、6个月的眼压均明显低于术前(P0.05);两组术后6个月的前房深度均较术前明显增加(P0.05)。结论:对于闭角型青光眼合并白内障患者,应按照患者房角粘连状况和眼压大小选择合适的手术治疗方法。     

三种不同手术方案治疗急性闭角型青光眼合并白内障患者的疗效比较

目的:研究比较三种不同手术方案治疗急性闭角型青光眼(AACG)合并白内障患者的疗效。方法:选择2012年10月至2015年10月在我院接受手术治疗的AACG合并白内障患者50例(50只眼)进行研究,按患者前房角的粘连关闭程度将其分成A、B、C三组,A组给予小切口囊外摘除术,B组在A组基础上给予前房角分离术,C组在A组基础上给予小梁切除术,比较各组术后疗效。结果:A、B、C三组患者术后的眼压均显著低于术前,前房中央深度和视力均显著高于术前,差异均有统计学意义(P0.05)。A、B、C三组术后视力的分布中,0.3~0.5的比例均显著高于0.3及0.5的比例,差异有统计学意义(P0.05)。A组术后的并发症总发生率为15.79%,B组为31.25%,C组为40.00%,三组间比较差异无统计学意义(P0.05)。结论:AACG合并白内障患者可通过多种手术方案进行治疗,但应根据房角关闭程度合理地选择最佳术式,从而有效改善患者的症状和视力状况。     

AC1及AC3抗白内障形成中晶状体谷胱甘肽代谢相关酶活性的变化

观察了亚硒酸钠,AC1,AC3对大鼠晶状体中谷胱甘肽过氧化物酶(GSH-Px),谷胱甘肽还原酶(GR)及谷胱甘肽硫转移酶(GST)的影响。结果表明,亚硒酸钠组大鼠的晶状体尚未混浊前已出现GSH-Px活性增高及GR和GST的活性降低。GR活性下降随白内障进展而加重。AC1及AC3均可使亚硒酸钠所致的酶活性变化逆转,但对正常晶状体的酶活性没有影响。     

正常与患白内障大鼠晶状体中脲溶性蛋白质性质的研究

本文研究了正常及三种类型白内障大鼠晶状体中脲溶性蛋白质的含量及性质的变化,发现在每种类型白内障晶状体中,水溶性蛋白质均减少,水不溶性蛋白质则都相对增加。经SephadexG-200柱层析及SDS聚丙烯酰胺凝胶电泳发现,晶状体中脲溶性蛋白质主要是由二硫键交联而成的高分子聚合物。经巯基乙醇还原后,绝大部分高分子聚合物可分解成低分子量蛋白质,其分子量与水溶性的γ晶体蛋白相同。这提示晶状体中脲溶性蛋白质的主要成分很可能是以二硫键交联而成的γ晶体蛋白聚合物。此结果与本实验室所得白内障晶状体水溶性蛋白质的变化相吻合。