Prevalance of tuberculosis amongst healthcare workers, working in DOTS/sputum microscopy centre in two different districts of state of Uttarakhand & Uttar Pradesh of India.

BACKGROUND & OBJECTIVE: The Healthcare workers (HCWs) who work in DOTS/Sputum microscopy centre are exposed to higher risk of contacting tuberculosis (TB) comparatively to other health workers who are serving the other health sectors. The HCWs in DOTS are more exposed due to direct contact with patients suffering from TB or through sharing the infected air space with the infectious patients. The aim of the study is to know the prevalance of TB disease amongst the HCWs who are working in DOTS cum Sputum Microscopy Centre's under RNTCP in two different districts of state of Uttar Pradesh (UP) and Uttarakhand (UK) of india. METHODS: The prospective cross-sectional study is conducted in two districts of different states having high burden of TB disease in UP and low burden of TB disease in UK state. All 100% (130) staff i.e. Medical officers, Sputum microscopy technicians, DOTS providers of DOTS cum Sputum Microscopy centre's of both selected Ghaziabad (UP) and Dehradun (UK) districts are covered in the study. RESULTS: The 4.6% (6) healthcare workers of both the districts were taking ATT at the time of interview and 13.8% (18) HCWs had taken the ATT in past. The 62.5% (15) HCWs i.e 55.5% (5) from Dehradun district and 66.6% (10) from Ghaziabad district preferred to have a ATT from the private medical store inspite of taking DOTS with assumption of low efficacy of drugs and high toxicity. The 58.33% (14) HCWs ie 55.5% (5) staff members of DOTS/sputum microscopy centre in Dehradun & 60.0% (9) staff members of DOTS/sputum microscopy centre in Ghaziabad district had not notified about the status of their disease to the health care authority due the assumption that they may be asked to leave the job or to go on a long unpaid leave. CONCLUSION: The 18.4% (24) HCWs of both the district got TB disease during their working in DOTS/Sputum microscopy centre and 4.6% (6) HCWs of both the districts were taking the ATT at the time of interview.

BK virus associated haemorrhagic cystitis in related donor haematopoietic stem cell transplant recipients: A developing country experience.

A retrospective cross sectional study was conducted at the Virology Department, Armed Forces Institute of Pathology (AFIP) and Armed Forces Bone Marrow Transplant Centre (AFBMTC), Rawalpindi, from January 2016 to July 2018. Medical records of 193 patients were examined to determine the number of patients developing Haemorrhagic Cystitis associated with BK virus (BKV). BKV PCR testing was done on the patients' urine samples. Cytomegalovirus reactivation was also assessed weekly from day 30 to day 100, by CMV quantitative PCR testing on blood samples. Out of 193 patients, 11 (5.6%) developed haemorrhagic cystitis and all these patients were positive for BKV on urine samples. The maximum number of positive cases, i.e. 5 (2.6%) was in the age group three months to 10 years. Primary disease in seven out of 11 cases was Beta-Thalassemia Major.

Special issues related to the diagnosis and management of acquired aplastic anemia in countries with restricted resources, a report on behalf of the Eastern Mediterranean blood and marrow transplantation (EMBMT) group and severe aplastic anemia working party of the European Society for blood and marrow transplantation (SAAWP of EBMT).

Aplastic anemia is a relatively rare but potentially fatal disorder, with a reported higher incidence in developing countries in comparison to the West. There are significant variations in epidemiological as well as etiological factors of bone marrow failure syndromes in the developing countries in comparison to the developed world. Furthermore, the management of bone marrow failure syndromes in resource constraint settings has significant challenges including delayed diagnosis and referral, limited accessibility to healthcare facilities, treatment modalities as well as limitations related to patients who require allogeneic stem cell transplantation. Here we will provide a review of the available evidence related to specific issues of aplastic anemia in the developing countries and we summarize suggested recommendations from the Eastern Mediterranean blood and bone marrow transplantation (EMBMT) group and the severe aplastic anemia working party of the European Society of blood and marrow transplantation (SAAWP of EBMT) related to the diagnosis and therapeutic options in countries with restricted resources.

Structural alteration in hypochlorous acid modified antithrombin indicates generation of neo-epitopes.

Increased tendency of cancer patients to develop venous thromboembolism (VTE) is associated with high rates of mortality. Elevation of procoagulant proteins and down regulation of naturally occurring coagulation inhibitors appears to form the basis of high risk of VTE in malignancy. A reduced level of anticoagulant protein like antithrombin (AT) will influence both coagulation and angiogenesis, as its cleaved and latent conformations show potent antiangiogenic activity. We show a concentration dependent perturbation in the secondary and tertiary structures of AT conformers exposed to hypochlorous acid (HOCl). Modulated under a very narrow concentration range of HOCl, native AT undergoes oligomerization, aggregation and fragmentation based on spectroscopic, SDS and native-PAGE studies. Factor Xa inhibition assay demonstrated a progressive decrease in inhibition activity of AT on modification by HOCl. Bis-ANS result showed that hydrophobic patches were more exposed in the case of HOCl-modified AT when assessed fluorometrically. Dosage of HOCl-modified AT in experimental animals induced high titer antibodies showing more specificity towards modified forms in comparison to unmodified forms. Auto-antibodies isolated from cancer patients also showed enhanced binding with HOCl-modified AT in comparison to native counterpart. Compared to normal AT, structurally and functionally altered conformation of HOCl-modified AT showed increased immunogenic sensitivity. HOCl modified AT can contribute to prothrombotic and angiogenic environment during cancer progression/development.

Renal cell carcinoma in ectopic pelvic kidney: A rare case report with relevant surgical anatomy.

Renal Ectopia is a rare developmental anomaly, leading to failure of mature kidney to reach its normal location within the renal fossa. Most ectopic kidneys are asymphtomatic and gets diagnosed as incidental finding on radiography or at surgery. The incidence of RCC in ectopic kidney are very uncommon, presenting with atypical symptoms causes a diagnostic as well as therapeutic dilemma in front of urologists. We present a case of 65 years old gentleman, having complaints of hematuria having heterogeneously enhancing SOL in left pelvic ectopic kidney.

Identification of a novel alternatively spliced isoform of antithrombin containing an additional RCL-like loop.

Antithrombin (AT3) is one of the most important inhibitors of blood coagulation proteases that belong to the serpin family of protease inhibitors. In this study, a novel alternatively spliced isoform of AT3 was identified, both at transcript and protein level. This novel transcript contains an additional region in the continuation of exon 3b that was included in the transcript due to use of an alternate 5' splice site. The existence of the novel transcript was confirmed in human brain and liver through RT-PCR. An analysis of the complete transcript indicated that the native reactive centre loop (RCL) of AT3 is maintained; however the novel amino acid sequence projects out as an additional loop as evident from MD simulation studies. A unique amino acid sequence present in the novel isoform was used for the development of polyclonal antibody. The expression of novel isoform was confirmed in human brain and liver tissue using Western blot analysis. Interestingly an alignment of RCL like domain with other inhibitory serpins showed significant similarity with the neuroserpin RCL. To the best of our knowledge, this is the first evidence of alternatively spliced AT3 sequence containing an additional loop and could have physiological relevance.

Neo-epitopes on crotonaldehyde modified DNA preferably recognize circulating autoantibodies in cancer patients.

DNA damage is one of the leading causes of various pathological conditions including carcinogenesis. Crotonaldehyde is a 4-carbon unsaturated bifunctional aldehyde which is found ubiquitously and produced both exogenously and endogenously. It reacts with deoxyguanosine and form adducts with DNA. These adducts were detected and found involved in tumor formation in rats treated with crotonaldehyde. In the present study, structural changes in DNA by crotonaldehyde were evaluated by Fourier transform infrared (FTIR) spectroscopy, differential scanning colorimetry (DSC), dynamic light scattering (DLS), high-performance liquid chromatography (HPLC), and atomic force microscopy (AFM). Enhanced binding was observed in cancer autoantibodies with the DNA modified by crotonaldehyde than the native counterpart. Immunological studies revealed enhanced binding of cancer autoantibodies with crotonaldehyde modified DNA, compared to the native form. Furthermore, lymphocyte DNA isolated from cancer patients demonstrated considerable recognition of anti-Cro-DNA IgG as compared to the DNA from healthy individuals. Therefore, we suggest that crotonaldehyde modified DNA presents unique epitopes, that may trigger autoantibody induction in cancer patients.

Fructosylation generates neo-epitopes on human serum albumin.

Hyperglycemia is the defining feature of diabetes mellitus. The persistently high levels of reducing sugars like glucose and fructose cause glycation of various macromolecules in the body. Human serum albumin (HSA), the most abundant serum protein with a myriad of functions, is prone to glycation and consequent alteration in its structural and biological properties. This study aimed to assess the role of fructose-modified human serum albumin as a marker of diabetic pathophysiology. We carried out modification of HSA with fructose and the changes induced were studied by various physicochemical studies. Fructose modified-HSA showed hyperchromicity in UV spectrum and increased AGE-specific fluorescence as well as quenching of tryptophan fluorescence. In SDS-PAGE protein aggregation was seen. Amadori products were detected by NBT. The fructose modified HSA had higher content of carbonyls along with perturbations in secondary structure as revealed by CD and FT-IR. A greater hydrodynamic radius of fructose-modified HSA was evident by DLS measurement. The fructose-modified HSA induced high titre antibodies in experimental animals exhibiting high specificity towards the immunogen.

Preferential recognition of peroxynitrite-modified human serum albumin by circulating autoantibodies in cancer.

Peroxynitrite is a potent oxidizing and nitrating agent and has in vivo existence. Several studies have shown the damaging role of this molecule in biological system. Human serum albumin (HSA), being most abundant plasma protein, is easily targeted by different oxidizing and nitrating agents. Free radicals increase the onset of different cancers as evident by several researchers. In the present study, structural perturbations in HSA by peroxynitrite were observed by MALDI-MS, DSC and DLS. Immunological studies showed enhanced binding of peroxynitrite-modified HSA with cancer autoantibodies, compared to the native protein. A decline in the antioxidant property of peroxynitrite-modified HSA was also observed. Therefore, we may conclude that peroxynitrite exposure results in structural alteration and hence generation of neo-epitopes in HSA molecule along with the decrease in its antioxidant property. The possible role of peroxynitrite-modified HSA in carcinogenesis has been discussed.

Pathophysiological Role of Peroxynitrite Induced DNA Damage in Human Diseases: A Special Focus on Poly(ADP-ribose) Polymerase (PARP).

Peroxynitrite is formed in biological systems when nitric oxide and superoxide rapidly interact at near equimolar ratio. Peroxynitrite, though not a free radical by chemical nature, is a powerful oxidant which reacts with proteins, DNA and lipids. These reactions trigger a wide array of cellular responses ranging from subtle modulations of cell signaling to overwhelming oxidative injury, committing cells to necrosis or apoptosis. The present review outlines the various peroxynitrite-induced DNA modifications with special mention to the formation of 8-nitroguanine and 8-oxoguanine as well as the induction of DNA single strand breakage. Low concentrations of peroxynitrite cause apoptotic death, whereas higher concentrations cause necrosis with cellular energetics (ATP and NAD(+)) serving as control between the two modes of cell death. DNA damage induced by peroxynitrite triggers the activation of DNA repair systems. A DNA nick sensing enzyme, poly(ADP-ribose) polymerase-1 (PARP-1) becomes activated upon detecting DNA breakage and it cleaves NAD(+) into nicotinamide and ADP-ribose and polymerizes the latter on nuclear acceptor proteins. Over-activation of PARP induced by peroxynitrite consumes NAD(+) and consequently ATP decreases, culminating in cell dysfunction, apoptosis or necrosis. This mechanism has been implicated in the pathogenesis of various diseases like diabetes, cardiovascular diseases and neurodegenerative diseases. In this review, we have discussed the cytotoxic effects (apoptosis and necrosis) of peroxynitrite in the etiology of the mentioned diseases, focusing on the role of PARP in DNA repair in presence of peroxynitrite.

Peroxynitrite induced structural changes result in the generation of neo-epitopes on human serum albumin.

Human serum albumin (HSA), the most abundant plasma protein, is quite vulnerable to oxidizing and nitrating agents. In this study, peroxynitrite induced nitration and oxidation of HSA was assessed by various physicochemical techniques. Cross-linking of HSA was evident on polyacrylamide gel electrophoresis. The carbonyl content was markedly elevated in peroxynitrite-modified HSA as compared to the native protein. Dityrosine and 3-nitrotyrosine were present only in peroxynitrite-modified HSA. The peroxynitrite-modified HSA induced high titre antibodies in experimental animals showing high specificity towards the immunogen. Spectroscopic studies showed structural alterations in the HSA molecule upon peroxynitrite treatment which result in the generation of neo-epitopes and enhanced immunogenicity. The possible role of damaged HSA in various diseases has been suggested.

Subjective fatigue and subjective sleepiness: two independent consequences of sleep disorders?

The objective of this investigation was to evaluate subjective fatigue versus subjective sleepiness as independent consequences of sleep disorders. Furthermore, we tried to explore how these symptoms relate to alertness, depressive symptoms and illness intrusiveness. In a prospective observational study, 283 sleep-disordered patients referred to a hospital-based sleep laboratory for various indications over a 1-year period were evaluated vis-à-vis fatigue and sleepiness. All patients completed five subjective questionnaires, underwent objective sleep recording and attended a clinical interview with a sleep specialist. The subjective questionnaires included the Epworth Sleepiness Scale, the Fatigue Severity Scale, the Toronto Hospital Alertness Test, the Illness Intrusiveness Rating Scale and the Center for Epidemiologic Studies-Depression Scale. Only 4% of the total sample was referred to the sleep clinic due to a complaint of excessive fatigue compared with 17% for excessive daytime sleepiness. However, during the assessment, 64% of referred patients reported pathological fatigue without overlap of sleepiness and only 4% reported pathological sleepiness without overlap of fatigue. Pearson's correlation analysis indicated a weak association (r=0.18) between subjective fatigue and sleepiness in the total sample. Analysis of variance testing showed strong association between those patients with prominent fatigue and depressive symptoms (P<0.01) and illness intrusiveness (P<0.001). The findings support the notion that subjective fatigue and sleepiness can be independent manifestations of sleep disorders. Furthermore, predominantly fatigued individuals with sleep disorders seem vulnerable to additional negative consequences due to possible interplay between amplified fatigue and psychological distress.