Quantitation and characterization of glutathionyl haemoglobin as an oxidative stress marker in chronic renal failure by mass spectrometry

OBJECTIVES: Glutathionyl haemoglobin (GS-Hb) belonging to the class of glutathionylated proteins has been investigated as a possible marker of oxidative stress in different chronic diseases. The purpose of this study was to examine whether glutathionyl haemoglobin can serve as an oxidative stress marker in non-diabetic chronic renal failure patients on different renal replacement therapies (RRT) through its quantitation, and characterization of the specific binding site of glutathione in haemoglobin molecule by mass spectrometric analysis. DESIGN AND METHODS: The study group consisted of non-diabetic chronic renal failure patients on renal replacement therapy (RRT): hemodialysis (HD), continuous ambulatory peritoneal dialysis (CAPD) and renal allograft transplant (Txp) patients. Haemoglobin samples of these subjects were analyzed by liquid chromatography electrospray ionization mass spectrometry for GS-Hb quantitation. Characterization of GS-Hb was done by tandem mass spectrometry. Levels of erythrocyte glutathione (GSH) and lipid peroxidation (as thiobarbituric acid reacting substances) were measured spectrophotometrically, while glycated haemoglobin (HbA1c) was measured by HPLC. RESULTS: GS-Hb levels were markedly elevated in the dialysis group and marginally in the transplant group as compared to the controls. GS-Hb levels correlated positively with lipid peroxidation and negatively with the erythrocyte glutathione levels in RRT groups indicating enhanced oxidative stress. De novo sequencing of the chymotryptic fragment of GS-Hb established that glutathione is attached to Cys-93 of the beta globin chain. Mass spectrometric quantitation of total glycated haemoglobin showed good agreement with HbA1c estimation by conventional HPLC method. CONCLUSIONS: Glutathionyl haemoglobin can serve as a clinical marker of oxidative stress in chronic debilitating therapies like RRT. Mass spectrometry provides a reliable analytical tool for quantitation and residue level characterization of different post-translational modifications of haemoglobin.     

Modular-level alterations of single-subject gray matter networks in schizophrenia

Brain Imaging and Behavior - Schizophrenia is often regarded as a psychiatric disorder caused by disrupted connections in the brain. Evidence suggests that the gray matter of schizophrenia patients...     

Appropriate Source Control and Antifungal Therapy are Associated with Improved Survival in Critically Ill Surgical Patients with Intra-abdominal Candidiasis

World Journal of Surgery - Intra-abdominal candidiasis (IAC) is the predominant type of invasive candidiasis with high mortality in surgical intensive care patients. The purpose of this study was...     

Plasma osteopontin levels in patients with acute kidney injury requiring dialysis: a study in a tertiary care institute in South India

International Urology and Nephrology - Osteopontin (OPN) is evolving as a novel biomarker of injury, overall survival and renal outcome in critically ill patients with acute kidney injury (AKI),...     

输尿管软镜碎石术治疗上尿路结石的疗效和安全性

目的探讨输尿管软镜碎石术(retrograde intrarenal stone surgery,RIRS)治疗上尿路结石的疗效和安全性。方法回顾性分析2016年4月至2019年1月武汉大学人民医院行RIRS的640例上尿路结石患者的临床资料。男424例,女216例。年龄(46.2±12.8)岁。结石最大径(1.4±0.7)cm。肾下盏结石126例,非肾下盏结石514例。单侧输尿管上段结石196例,单侧输尿管上段结石合并肾结石118例,单侧肾结石236例,双肾结石90例。104例术前留置双J管。马蹄肾8例,孤立肾合并肾功能不全30例,盆腔异位肾合并旋转不良4例,先天性输尿管畸形6例,海绵肾2例。术前血红蛋白(133.2±5.6)g/L,血清肌酐(84.4±12.2)μmol/L。手术均采用全麻,患者取截石位。采用输尿管软镜联合钬激光碎石,软镜顺利进入肾盂后首先观察肾盂及各肾盏并寻及结石。使用200μm光纤碎石,钬激光功率为12~45 W(0.5~1.5 J/10~30 Hz),根据实际情况辅助取石网篮套取结石。术中检查各肾盂、肾盏,确保结石已完全粉末化(结石最大径<0.3 cm),留置双J管和尿管。手术均由同等资历的术者完成。结果所有手术均顺利完成,手术时间(45.6±14.6)min。术后第1天复查血清肌酐(76.0±10.6)μmol/L,与术前比较差异有统计学意义(t=64.76,P<0.05);血红蛋白(126.4±9.6)g/L,与术前比较差异无统计学意义(t=2.02,P=0.064)。术后住院时间(4.8±1.5)d。术后3例(0.9%)发生严重并发症,分别为2例脓毒血症,1例包膜下血肿。术后3个月596例获得随访,其中552例达到结石清除标准,结石清除率(stone-free rate,SFR)为92.6%;余44例未达到结石清除标准者采用体外冲击波碎石、再次输尿管软镜手术或拔除双J管后观察。结石大小(χ^2=29.569,P<0.05)和位置(χ^2=44.949,P<0.05)是SFR的影响因素。多因素回归分析结果显示结石大小不是影响SFR的独立危险因素(P=0.639),结石位置是影响SFR的独立危险因素(P=0.013)。结论RIRS对于上尿路中小结石患者是一种可靠的治疗方式,疗效确切,并发症少,安全性高。但对于大结石及肾下盏结石的治疗还存在一定的局限性。肾下盏结石是影响RIRS疗效的独立危险因素。     

Phosphorylated ganciclovir derivatives: design,synthesis and in vitro and in vivo immunomodulatory activity

Synthesis of a series of novel phosphorylated ganciclovir derivatives with bioactive amines/aminoacid esters as substituents was accomplished. These compounds were structurally characterized by IR, NMR (¹H, ¹³C, ³¹P), mass spectra and CHN analysis. The compounds (5a–m) have been evaluated for its in vitro and in vivo immunomodulatory activities. The amino acid ester-substituted ganciclovir derivatives, especially the 5g and 5i, increased the intracellular killing activity of the stimulated neutrophils. The in vivo experiment results show that the administration of compounds 5g and 5i (8 mg/kg body weight) to diet-induced immune impaired obese rats ameliorated the significant increase in immune cell counts (lymphocytes, neutrophils and monocytes). In addition, TNF-α, IL-6 and IL-1 secretions were considerably restored to normal by the compounds 5g and 5i with regulation in the release of C-reactive protein suggesting their potentiating effect on immune system dysfunction. These in vitro and in vivo results suggest that the phosphorylated ganciclovir derivatives 5g and 5i are strong immunomodulators.     

Synthesis,antitubercular and anticancer activity of new Baylis–Hillman adduct-derived N-cinnamyl-substituted isatin derivatives

Baylis–Hillman adduct-derived N-cinnamyl-substituted isatin derivatives were synthesized and evaluated for their antitubercular activity on Mycobacterium tuberculosis H₃₇Rv strain ATCC 27294 by agar dilution method. Anticancer activity for the same compounds was also screened on four different cell lines: Chinese hamster ovary (CHO cells), Colo 205 (human colon cancer), Sup-T1 (human lymphoma) and C6 glioma (rat glioma) by MTT assay method. The compounds (3j–l) have shown significant activity against Mycobacterium strain and the compound 3l has shown specific cytotoxic activity.