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

Toward the PSTN/Internet Inter-Networking--Pre-PINT Implementations

This document contains the information relevant to the development of the inter-networking interfaces underway in the Public Switched Telephone Network (PSTN)/Internet Inter-Networking (PINT) Working Group. It addresses technologies, architectures, and several (but by no means all) existing pre-PINT implementations of the arrangements through which Internet applications can request and enrich PSTN telecommunications services. The common denominator of the enriched services (a.k.a. PINT services) is that they combine the Internet and PSTN services in such a way that the Internet is used for non-voice interactions, while the voice (and fax) are carried entirely over the PSTN. One key observation is that the pre-PINT implementations, being developed independently, do not inter-operate. It is a task of the PINT Working Group to define the inter-networking interfaces that will support inter-operation of the future implementations of PINT services

Techniques of Using Peripheral Blood Mononuclear Cells as the Cellular System to Investigate How of the Bovine Species (Indian Zebu-Jersey Crossbreds) Responds to <em>in vitro</em> Thermal Stress Stimulation (Thermal Assault/Heat Shock)

Animal production is negatively impacted by global warming and is subject to serious consequences for livestock production systems. In order to understand how PBMCs of Indian Zebu-Jersey crossbreds respond to various levels and durations of thermal assault and heat shock, in this chapter we will discuss techniques involving in vitro thermal stress stimulation (TSS) to stimulate bovine peripheral blood mononuclear cells (PBMCs) under various thermal assault conditions (TACs), including normal to extreme temperatures and varying durations of thermal exposure (DTEs). The consequences of thermal stress on bovine species can be lessened and managed with an understanding of how PBMCs as a cellular system respond to in vitro heat shock and thermal assault. To learn more about how Indian Zebu-Jersey crossbreds respond to in vitro thermal conditions, it may also be possible to explore the relationship between the decrease in PBMCs count during in vitro TSS and the expression of the heat shock protein genes (HSPs) such as HSPs 70 and 90 genes. This will be exploited to discover how Indian Zebu-Jersey crossbreds respond in vivo to diverse environmental thermal conditions and will further enable in vivo understanding of the potential for thermotolerance in bovine species for better adaptability, survival, and production performance

Emphysematous prostatitis in renal transplant

Urinary tract infections are common following renal transplant. The spectrum varies from asymptomatic bacteriuria to septicemia. Gas-producing infections of the urinary tract are rare but tend to have a grave prognosis when they do occur. We report a 57-year-old gentleman who underwent a renal transplant 20 months earlier. He presented to us with fever and dysuria. Clinical examination revealed a febrile and ill-looking patient with severe graft tenderness. An emergency pelvic CT scan revealed presence of emphysematous prostatitis, cystitis and pyelitis. Urine and blood cultures grew E. coli. Endoscopic abscess drainage was done and antibiotics given but he succumbed to his illness due to multiorgan failure within 48h. This is the first reported case of emphysematous prostatitis in a renal allograft recipient