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Extraction and concentration of pure nucleic acid from complex biofluids is the prerequisite for nucleic acid amplification test (NAAT) applications in pathogen detection, biowarfare prevention, and genetic diseases. However, conventional spin-column mediated nucleic acid extraction is constricted by the requirement for costly power-intensive centralized lab infrastructure, making them unsuitable for limited-resource settings. Significant progress on lab-on-a-chip devices or cartridges (e.g., Cepheid GeneXpert®) that integrate nucleic acid extraction and amplification have been made, but these approaches either require additional equipment or are costly. Similarly, their complexities make them difficult to fabricate in low-resource settings by the end-user themselves. The application of magnetic particles such as silica-coated iron oxide beads for nucleic acid extraction is relatively instrument-free, rapid, user-friendly, and amenable to automation. But, they rely on hazardous chaotropic salt chemistry and ethanol desalting that could limit their efficacy for downstream NAATs. Recent advances in several types of novel material (e.g., polyamine) coated magnetic bead-based chaotropic salt-free extraction methods offer a possible solution to this problem. However, these materials also involve multistep synthesis impermissible in limited-resource settings. To offer a possible instrument-free magnetic particle-based nucleic acid extraction doable at limited-resource settings, we investigated the nucleic acid capture ability of two chitosan-coated magnetic particle syntheses that are preparable by minimally trained personnel using only a water bath and magnetic stirrer within 6 – 8 h. We quantitatively probed the efficiency of the passive (without any electrical shaking or vortex-aided) DNA magnetocapture (i.e., binding to chitosan magnetic particles, physical separation from its sample of origin, and release from the particles) using UV260. To explore their suitability towards clinically relevant sensitive downstream NAATs, 100 - 1000 copies (i.e., in the order of zeptomole) of Escherichia coli (E. coli) or human genomic DNA from aqueous solution, crude cell lysate, and fetal bovine serum were extracted by them and then successfully detected using quantitative real-time loop-mediated isothermal amplification (LAMP) or real-time polymerase chain reaction (PCR). Alongside, their suitability with gel-based LAMP, colorimetric LAMP, and in situ (on beads) LAMP was also probed. The required optimization of the amplification methods has been discussed. Overall, the turnaround time for the magnetocapture combined with NAAT was 1.5 – 2 h and is thus expected to aid rapid clinical decision making. With the ease of preparation, reproducibility, and compatibility with downstream NAATs, we anticipate that these magnetic particles would facilitate the expansion and decentralization of nucleic acid-based diagnosis for limited-resource settings.

IntroductionScreening for G6PD deficiency can inform disease management including malaria. Treatment with the antimalarial drugs primaquine and tafenoquine can be guided by point-of-care testing for G6PD deficiency.Methods and FindingsData from similar clinical studies evaluating the performance of the STANDARD™G6PD Test (SD Biosensor, South Korea) conducted in Bangladesh, Brazil, Ethiopia, India, Thailand, the United Kingdom, and the United States were pooled. Test performance was assessed in a retrospective analysis on capillary and venous specimens. All study sites used spectrophotometry for reference G6PD testing, and either the HemoCue or complete blood count for reference hemoglobin measurement.The sensitivity of the STANDARD™G6PD Test using the manufacturer thresholds for G6PD deficient and intermediate cases in capillary specimens from 4212 study participants was 100% (95% Confidence Interval (CI): 97.5%–100%) for G6PD deficient cases with 60% on the reference assay. Negative predictive values for females with G6PD activity >60% was 99.6% (95% CI 99.1%–99.8%) on capillary specimens. Test sensitivity among 396P. vivaxmalaria cases was 100% (69.2%–100.0%) for both deficient and intermediate cases. In the study population, a high proportion of those classified as G6PD deficient or intermediate resulted from true normal cases. Despite this, the majority cases would receive the correct medication and no true G6PD deficient cases would be treated inappropriately.ConclusionsThe STANDARD G6PD Test enables safe access to drugs which are contraindicated for individuals with G6PD deficiency. Operational considerations will inform test uptake in specific settings.

Data from the National Clinical Registry for COVID-19 (NCRC) were analyzed with an aim to describe the clinical characteristics, course and outcomes of patients hospitalized with COVID-19 in the third wave of the pandemic and compare them with patients admitted earlier.The NCRC, launched in September 2020, is a multicentre observational initiative, which provided the platform for the current investigation. Demographic, clinical, treatment and outcome data of hospitalized COVID-19 patients were captured in an electronic data portal from 38 hospitals across India. Patients enrolled during December 16, 2021 to January 17, 2022 were considered representative of the third wave of COVID-19 and compared with those registered during November 15 to December 15, 2021, representative of the tail end of the second wave.Between November 15, 2021 and January 17, 2022, 3230 patients were recruited in NCRC. Patients admitted in the third wave were significantly younger than those admitted earlier (46.7±20.5 vs. 54.6±18 yr). The patients admitted in the third wave had a lower requirement of drugs including steroids, interleukin (IL)-6 inhibitors and remdesivir as well as lower oxygen supplementation and mechanical ventilation. They had improved hospital outcomes with significantly lower in-hospital mortality (11.2 vs. 15.1%). The outcomes were better among the fully vaccinated when compared to the unvaccinated or partially vaccinated.The pattern of illness and outcomes were observed to be different in the third wave compared to the last wave. Hospitalized patients were younger with fewer comorbidities, decreased symptoms and improved outcomes, with fully vaccinated patients faring better than the unvaccinated and partially vaccinated ones.

ObjectivesThis study aims to describe the demographic and clinical profile and ascertain the determinants of outcome among hospitalized coronavirus disease 2019 (COVID-19) adult patients enrolled in the National Clinical Registry for COVID-19 (NCRC).MethodsNCRC is an on-going data collection platform operational in 42 hospitals across India. Data of hospitalized COVID-19 patients enrolled in NCRC between 1st September 2020 to 26th October 2021 were examined.ResultsAnalysis of 29 509 hospitalized, adult COVID-19 patients [mean (SD) age: 51.1 (16.2) year; male: 18 752 (63.6%)] showed that 15 678 (53.1%) had at least one comorbidity. Among 25 715 (87.1%) symptomatic patients, fever was the commonest symptom (72.3%) followed by shortness of breath (48.9%) and dry cough (45.5%). In-hospital mortality was 14.5% (n = 3957). Adjusted odds of dying were significantly higher in age group ≥60 years, males, with diabetes, chronic kidney diseases, chronic liver disease, malignancy and tuberculosis, presenting with dyspnoea and neurological symptoms. WHO ordinal scale 4 or above at admission carried the highest odds of dying [5.6 (95% CI: 4.6–7.0)]. Patients receiving one [OR: 0.5 (95% CI: 0.4–0.7)] or two doses of anti-SARS CoV-2 vaccine [OR: 0.4 (95% CI: 0.3–0.7)] were protected from in-hospital mortality.ConclusionsWHO ordinal scale at admission is the most important independent predictor for in-hospital death in COVID-19 patients. Anti-SARS-CoV2 vaccination provides significant protection against mortality.

ObjectivesThis study aims to describe the demographic and clinical profile and ascertain the determinants of outcome among hospitalised COVID-19 adult patients enrolled in the National Clinical Registry for COVID-19 (NCRC).MethodsNCRC is an on-going data collection platform operational in 42 hospitals across India. Data of hospitalized COVID-19 patients enrolled in NCRC between 1st September 2020 to 26th October 2021 were examined.ResultsAnalysis of 29,509 hospitalised, adult COVID-19 patients [mean (SD) age: 51.1 (16.2) year; male: 18752 (63.6%)] showed that 15678 (53.1%) had at least one comorbidity. Among 25715 (87.1%) symptomatic patients, fever was the commonest symptom (72.3%) followed by shortness of breath (48.9%) and dry cough (45.5%). In-hospital mortality was 14.5% (n=3957). Adjusted odds of dying were significantly higher in age-group ≥60 years, males, with diabetes, chronic kidney diseases, chronic liver disease, malignancy, and tuberculosis, presenting with dyspnea and neurological symptoms. WHO ordinal scale 4 or above at admission carried the highest odds of dying [5.6 (95% CI: 4.6, 7.0)]. Patients receiving one [OR: 0.5 (95% CI: 0.4, 0.7)] or two doses of anti-SARS CoV-2 vaccine [OR: 0.4 (95% CI: 0.3, 0.7)] were protected from in-hospital mortality.ConclusionsWHO ordinal scale at admission is the most important independent predictor for in-hospital death in COVID-19 patients. Anti-SARS-CoV2 vaccination provides significant protection against mortality.

BackgroundSevere Corona virus disease (COVID-19) is associated with high mortality. Although single centre intensive care units (ICU) have reported clinical characteristics and outcomes, no large scale multicentric study from India has been published. The present retrospective, multi-centre study was aimed to describe the predictors and outcomes of COVID-19 patients requiring ICU admission from COVID-19 Registry of Indian council of Medical Research (ICMR), India.MethodsProspectively collected data from multiple participating institutions was entered in the electronic National Clinical Registry of COVID 19. We enrolled patients aged>18 years with COVID-19 pneumonia requiring ICU admission between March 2020 and August 2021. Exclusion criteria were negative RT PCR, death within 24 hours of ICU admission, or patients with incomplete data in the registry Their demographic characteristics, laboratory variables, ICU severity indices, treatment strategies and outcomes were analysed.ResultsA total of 5865 patients, with mean age 56±15 years, with 3840/5865 (65.4%) men, were enrolled in the ICMR registry.. Overall mortality was 2535/5865 (43.5%). Non-survivors were older than survivors (58.2±15.4 years vs 53.6 ±14.7 years; P=0.001). Non-survivors had multiple comorbidities (n=1951, 52.9%) with hypertension (47.2%) and diabetes (45.6%) being the most common, higher creatinine (1.6 ± P=0.001, high D-dimer (1.56 vs 1.37, P=0.001), higher CT severity index (16.8±5.2 vs 13.5 ±5.47 ) compared to survivors. Non survivors had longer hospital and ICU stay (P=0.001). On multivariate regression analysis, high NLR (HR 1.017, 95% CI 1.005- 1.029, P=0.001), high CRP (HR 1.008, 95% CI 1.006- 1.010, P=0.001), high D dimer ((HR 1.089, 95% CI 1.065- 1.113, P=0.001) were associated with mechanical ventilation while younger age, (HR 0.974, CI 0.965-0.983, p=0.001), high D dimer (HR-1.014, CI 1.001-1.027, P=0.035) and use of prophylactic LMWH (HR 0.647, CI 0.527-0.794, p=0.001) were independently associated with mortality. ConclusionIn this large retrospective study of 5865 critically ill COVID 19 patients admitted to ICU, overall mortality was 2535/5865 (43.5%). Age, high D dimer, CT Severity score and use of prophylactic LMWH were independently associated with mortality.

Community engagement in relation to HIV/AIDS has so often been a topic of discussion in India, substantial involvement of the community has rarely been a mainstream practice The general concept of community engagement is letting research projects achieve success by involving the participants directly into the research work, and implementing the results and the inferences obtained from those projects for the betterment of participants. We tried our level best to put this theory in practice at Medical College and Hospital, Kolkata, India during year 2015-2016. Apart from some teething problems that cropped up at the initial stage of the study, which was difficult and time taking to solve. The advantages of community involvement, as the experience showed, outweighed such minor difficulties. The final developed model for community engagement among marginalized population was successful and we can recommend this model for robust data collection and interpretation expecting a fruitful end result. Keywords: HIV/AIDS, Community engagement, Community involvement.

Nucleic acid amplification tests (NAATs) such as quantitative real-time reverse transcriptase PCR (qRT-PCR) or isothermal NAATs (iNAATs) such as loop-mediated isothermal amplification (LAMP) require pure nucleic acid free of any polymerase inhibitors as its substrate. This in turn, warrants the use of spin-column mediated extraction with centralized high-speed centrifuges. Additionally, the utilization of centralized real-time fluorescence readout and TaqMan-like molecular probes in qRT-PCR and real-time LAMP add cost and restrict their deployment. To circumvent these disadvantages, we report a novel sample-to-answer workflow comprising an indirect sequence-specific magneto-extraction (also referred to as magnetocapture, magneto-preconcentration, or magneto-enrichment) for detecting SARS-CoV-2 nucleic acid. It was followed by in situ fluorescence or electrochemical LAMP. After in silico validation of the approach's sequence selectivity against SARS-CoV-2 variants of concern, the comparative performance of indirect and direct magnetocapture in detecting SARS-CoV-2 nucleic acid in the presence of excess host nucleic acid or serum was probed. After proven superior, the sensitivity of the indirect sequence-specific magnetocapture in conjunction with electrochemical LAMP was investigated. In each case, its sensitivity was assessed through the detection of clinically relevant 10

Context: Malaria is an inflammatory condition triggered by the infection of parasite Plasmodium on erythrocytes. It is characterised by periodic fever with chills due to rupture of erythrocytes to release the progeny parasites. It is marked by release of variety of cytokines like IL-6, IL-12, IFN-Ɣ, TNF—α etc. Uric acid is one of the emerging inflammatory markers in malaria that demands attention. One of the proposed mechanisms is that there is accumulation of uric acid and its precursor, hypoxanthine in the infected erythrocytes. These are released into the blood on rupture of the schizont. Aim: To find association of serum uric acid level with severity of malaria.Settings and Design: This was a hospital based longitudinal study conducted at a tertiary care centre in Kolkata, India. Materials and Methods: We measured the plasma levels of uric acid and various inflammatory markers {Ferritin, C-reactive protein(CRP), LDH, C3, C4} in eighty eight patients admitted with microscopically proven malaria (severe or non-severe type). The levels of uric acid were compared with the disease severity and the inflammatory markers stated above. Statistical Analysis used: The data was analyzed using MedCalc software and Microsoft Excel 2010 and further graphically plotted. Results: The serum uric acid levels were raised in 18.51% of patients with severe malaria compared to only 4.91% with non-severe variety (p= 0.04). The uric acid levels demonstrated a positive correlation with CRP (r=0.3334, p=0.0015); procalcitonin & ferritn (r=0.3701, p

Since the beginning of the SARS-CoV-2 pandemic, nucleic acid amplification test (NAAT) such as quantitative real-time reverse transcriptase PCR (qRT-PCR) has remained the primary intervention for diagnostics and containment of SARS-CoV-2. Despite its remarkable clinical as well as analytical specificity and sensitivity, qRT-PCR necessitates pure nucleic acid free of any polymerase inhibitors (from complex biological matrices) as its substrate. Similarly, isothermal NAATs (iNAATs), despite their advantage over qRT-PCR in terms of thermal cycler independence, still require pure nucleic acid as a template. The requirement of pure nucleic acid in turn warrants the use of spin-column mediated extraction with centralized high-speed centrifuges. Additionally, utilization of centralized real-time fluorescence readout and use of sequence-specific molecular probes like TaqMan further prevent their deployment in decentralized locations. To circumvent these disadvantages, we have envisioned a sample-to-answer workflow comprising of indirect sequence-specific magneto-extraction (also referred to as magnetocapture, magneto-preconcentration, or magneto-enrichment in this manuscript) followed by in situ fluorescence or electrochemical LAMP. This study, using SARS-CoV-2 nucleic acid as the analyte, compared the analytical effectiveness of indirect and direct sequence-specific magneto-extraction followed by LAMP. Since contamination carryover may affect the efficacy of sequence-specific indirect magnetocapture, its performance in presence of excess host nucleic acid or serum was probed. Through these experiments, we have established a comprehensive limited resource-adoptable and highly specific nucleic acid detection method with the limit of detection of 2.5 copies/L. Its advantage lies in the flexibility of using either centralized real-time SYBR-based fluorescence LAMP or portable electrochemical LAMP as the readout. Simultaneously, the performance with magneto-capture aided fluorescence and electrochemical LAMP readouts were weighed against each other in terms of analytical sensitivity, specificity, and turnaround time. Additionally, the analytical efficacy of the magnetocapture-LAMP workflow was also checked against that of LAMP using pure nucleic acid as a template. Besides being the first report utilizing electrochemical LAMP to detect SARS-CoV-2 nucleic acid, this would probably be the first study to make the analytical comparative assessment of magnetic preconcentration combined with in situ fluorescence and electrochemical LAMP. It is probably also the first study (to the best of our knowledge) to compare the analytical efficacy of a sequence-specific magnetic target enrichment-LAMP (fluorescence and electrochemical) to that of a LAMP assay using pure nucleic acid as a template.

Background Point-of-care glucose-6-phosphate dehydrogenase (G6PD) testing has the potential to make the use of radical treatment for vivax malaria safer and more effective. Widespread use of G6PD tests as part of malaria case management has been limited, in part due to due concerns regarding product usability, user training, and supervision. This study seeks to assess how well end users can understand the Standard™ G6PD Test (SD Biosensor, Suwon, South Korea) workflow, result output, and label after training. This will ultimately help inform test registration and introduction. Methods Potential G6PD test users who provide malaria case management at three sites in Brazil, Ethiopia, and India were trained on the use of the SD Biosensor Standard G6PD Test and assessed based on their ability to understand the test workflow and interpret results. The assessment was done through a questionnaire, designed to assess product usability against key technical product specifications and fulfill regulatory evidence requirements. Any participant who obtained 85% or above correct responses to the questionnaire was considered to adequately comprehend how to use and interpret the test. Results Forty-five participants, including malaria microscopists, laboratory staff, nurses, and community health workers took part in the study. Seventy-eight percent of all participants in the study (35/45) obtained passing scores on the assessment with minimal training. Responses to the multiple-choice questions indicate that most participants understood well the test intended use, safety claims, and warnings. The greatest source of error regarding the test was around the correct operating temperature. Most test results were also read and interpreted correctly, with the haemoglobin measurement being a more problematic output to interpret than the G6PD measurement. Conclusions These data results show how a standardized tool can be used to assess a user’s ability to run a point-of-care diagnostic and interpret results. When applied to the SD Biosensor Standard G6PD Test, this tool demonstrates that a range of users across multiple contexts can use the test and suggests improvements to the test instructions and training that can improve product usability, increase user comprehension, and ultimately contribute to more widespread effective use of point-of-care G6PD tests. Trial registration: NCT04033640

Layman Summary: Nucleic acid extraction is a key prerequisite for any nucleic acid amplification test (NAAT) or isothermal NAAT (iNAAT) based molecular diagnosis assays. Existing methods utilizes spin column system for nucleic acid extraction which are unsuitable for limited resource settings. Our work explores two methods for chitosan coated magnetic particle preparation that can be executed within 6 h from commonly available chemicals with nothing but a magnetic stirrer and water bath and doable by a minimally trained person. We will also investigated the compatibility of the extracted nucleic acid with downstream NAATs such as real time LAMP, colorimetric LAMP, and real time PCR. In the process, we established the analytical sensitivity of the overall method.Characterization methods: SEM, XRD, EDX, FT-IRBioanalytical methods: Real time LAMP, Colorimetric LAMP, Real time PCR

Management of severe malaria remains a critical global challenge. In this study, using a multiplexed quantitative proteomics pipeline we systematically investigated the plasma proteome alterations in non-severe and severe malaria patients. We identified a few parasite proteins in severe malaria patients, which could be promising from a diagnostic perspective. Further, from host proteome analysis we observed substantial modulations in many crucial physiological pathways, including lipid metabolism, cytokine signaling, complement, and coagulation cascades in severe malaria. We propose that severe manifestations of malaria are possibly underpinned by modulations of the host physiology and defense machinery, which is evidently reflected in the plasma proteome alterations. Importantly, we identified multiple blood markers that can effectively define different complications of severe falciparum malaria, including cerebral syndromes and severe anemia. The ability of our identified blood markers to distinguish different severe complications of malaria may aid in developing new clinical tests for monitoring malaria severity., Kumar et al. use proteomes of patients with falciparum and vivax malaria and dengue fever, and identify differentially abundant proteins and their associated functions in the different severities and diseases. This study offers mechanistic pathways to malaria severity, and adopts a machine learning approach for predictive proteins for the malarial types and severities.

Context: Parasitic opportunistic infections (POIs) frequently occur in HIV/AIDS patients and affect the quality of life. Aims: This study assessing the standard organisms in the stool of HIV-positive patients, their comparison with HIV-negative controls, their relation with various factors, is the first of its kind in the eastern part of India. Settings and Design: hospital-based case-control study. Materials and Methods: A total of 194 antiretroviral therapy naïve HIV-positive patients (18-60 years) were taken as cases and 98 age- and sex-matched HIV-negative family members as controls. Demographical, clinical, biochemical, and microbiological parameters were studied. Statistical Analysis Used: Odds ratio, 95% confidence interval, and P (< 0.05 is to be significant) were calculated using Epi Info 7 software. Results: POI was significantly higher among HIV-seropositive cases (61.86%) (P < 0.001). Cryptosporidium was the most common POI in HIV-seropositive patients overall and without diarrhea; Entameba was the most common POI in patients with acute diarrhea, and Isospora was the most common POI in the patients having chronic diarrhea. Entameba was the most common POI in CD4 count 350 cells/μl Cryptosporidium was the most common POI. Mean CD4 count was significantly (P < 0.001) lower among people having multiple infections. Male sex, hemoglobin

Background Early detection of endothelial dysfunction in patients with systemic lupus erythematosus (SLE) is important since they show accelerated atherosclerosis and an increased risk of cardiovascular mortality. Aim The aim of the study was to describe the correlation of flow-mediated dilation (FMD) with common inflammatory and immunological markers in patients with SLE. An attempt was made to predict a model of FMD with the help of these markers. Material and methods The study included 44 treatment-naive SLE patients as per American College of Rheumatology (ACR) criteria (1982) and 44 age- and sex-matched healthy controls. Each group consists of 42 females and 2 males. FMD of the brachial artery by B-mode ultrasonography was performed in both groups to compare the FMD value. Serum hsCRP, fibrinogen, uric acid, fasting insulin, serum ferritin, total cholesterol, triglyceride, VLDL, LDL, and HDL levels were measured as markers of inflammation in SLE patients to determine whether there was any correlation with FMD. Results There were no significant differences in age and gender between the SLE and control groups. The mean FMD was 16.85 ± 10.64 and 21.89 ± 4.6 among cases and controls, respectively. FMD was significantly less among SLE patients (p = 0.005). The hsCRP, uric acid, fibrinogen, insulin resistance, and ferritin values were found to have significant correlations with FMD values among treatment-naive SLE patients. Multiple linear regression by the ENTER method was used to predict a model of FMD: FMD = 48.252 − 1.565 hsCRP − 0.143 fibrinogen − 0.217 uric acid + 0.001 ferritin + 7.658 IR. Conclusion Inflammatory markers of SLE, such as hsCRP, fibrinogen, insulin resistance, and uric acid positively correlate with FMD values. FMD can be predicted from the value of these biochemical parameters.

Photoplethysmographic (PPG) measurements are susceptible to motion artifacts (MA) due to movement of the peripheral body parts. In this paper, we present a new approach to identify the MA corrupted PPG beats and then rectify the beat morphology using artificial neural network (ANN). Initially, beat quality assessment was done to identify the clean PPG beats by a pretrained feedback ANN to generate a reference beat template for each person. The PPG data were decomposed using principal component analysis (PCA) and reconstructed using fixed energy retention. A weight coefficient was assigned for each PPG sample in such a way that when they are multiplied, the modified beat morphology matches the reference template. A particle swarm optimization-based technique was utilized to select the best weight vector coefficients to tune another feedback ANN, fed with a set of significant features generated by an auto-encoder from PCA reconstructed data. For real-time implementation, this pretrained ANN was operated in feed-forward mode to directly generate the weight vectors for any subsequent measurements of PPG. The method was validated with PPG data collected from 55 human subjects. An average root-mean-square error of 0.28 and signal-to-noise ratio improvement of 14.54 dB were obtained, with an average improvement of 36% and 47% measurement accuracies on crest time and systolic to diastolic peak height ratio, respectively. With the IEEE Signal Processing Cup 2015 challenge database, Pearson’s correlation coefficient between PPG-estimated and ECG-derived heart rates was 0.990. The proposed method can be useful for personal health monitoring applications.

India significantly contributes to the global malaria burden and has the largest population in the world at risk of malaria. This study aims to analyze alterations in the human serum proteome as a consequence of non-severe and severe infections by the malaria parasite Plasmodium falciparum to identify markers related to disease severity and to obtain mechanistic insights about disease pathogenesis and host immune responses. In discovery phase of the study, a comprehensive quantitative proteomic analysis was performed using gel-based (2D-DIGE) and gel-free (iTRAQ) techniques on two independent mass spectrometry platforms (ESI-Q-TOF and Q-Exactive mass spectrometry), and selected targets were validated by ELISA. Proteins showing altered serum abundance in falciparum malaria patients revealed the modulation of different physiological pathways including chemokine and cytokine signaling, IL-12 signaling and production in macrophages, complement cascades, blood coagulation, and protein ubiquitination pathways. Some muscle related and cytoskeletal proteins such as titin and galectin-3-binding protein were found to be up-regulated in severe malaria patients. Hemoglobin levels and platelet counts were also found to be drastically lower in severe malaria patients. Identified proteins including serum amyloid A, C-reactive protein, apolipoprotein E and haptoglobin, which exhibited sequential alterations in their serum abundance in different severity levels of malaria, could serve as potential predictive markers for disease severity. To the best of our information, we report here the first comprehensive analysis describing the serum proteomic alterations observed in severe P. falciparum infected patients from different malaria endemic regions of India. This article is part of a Special Issue entitled: Proteomics in India. (C) 2015 Elsevier B.V. All rights reserved.

Dengue fever (DF) is a major global health burden with a pathophysiology that is still incompletely understood. Biomarkers that predict and explain susceptibility to DF and its progression to its more severe hemorrhagic form are much needed. DF is endemic in tropical and subtropical regions of the world, with a rapidly increasing incidence of disease severity. We conducted a clinical biomarker discovery study using both a case-control and longitudinal study design. Plasma proteome alterations in patients with DF (n = 12) and dengue hemorrhagic fever (DHF, n = 24) were analyzed in comparison to healthy controls (HCs, n = 16), using the isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomics methodology (false discovery rate of 1%, ≥2 peptides). Several proteins such as the alpha-2 macroglobulin, angiotensinogen, apolipoprotein B-100, serotransferrin, and ceruloplasmin were upregulated (fold change1.2) in all DHF cases, and downregulated in DF (fold change0.83), compared with HCs. Plasma cytokine profiling (8 DF, 8 DHF, and 8 HC) on two consecutive time points, at day 0 (day of admission) and days 5-7, found significant elevation in IL-1RA, IL-7, TNF-α, MCP1-MCAF, and MIP-1β levels, but only in the DHF cases, which is the severe disease, and not in DF, compared with HCs (p 0.05). These new observations on changes in the plasma proteome and cytokine profiles in patients with dengue infection identify several putative molecular leads for future biomarker development and precision medicine in relation to forecasting DF disease severity.

SCOPE Haptoglobin (Hp), an acute phase inflammatory protein is associated with malaria pathogenesis in several proteomics and genomics studies. The Hp gene has two co-dominant alleles: Hp1 and Hp2 that produce three genotypes: Hp1/Hp1, Hp1/Hp2 and Hp2/Hp2. EXPERIMENTAL DESIGN In this study, validation of the proteomics data with Multiple Reaction Monitoring Mass Spectroscopy (MRM-MS) is performed and the association of the Hp gene variants with severe, non-severe malaria and community (healthy) controls using genotyping PCRs and DNA sequencing is analysed. RESULTS Highly significant values of Hp is observed in the MRM assay that show a correlation with severity of malaria and is clearly distinguished from another febrile disease, dengue. Moreover, the Hp2/Hp2 genotype is seen in high percentages in non-severe malaria patients (74%) and community controls (72%) whereas patients diagnosed with severe malaria show only (31%) of this genotype. Sequencing of the Hp promoter region reveals three SNPs along with 10 unique haplotypes, out of which five are associated with non-severe and three with severe malaria populations (χ2 = 130; df = 18; p

Most urethral strictures arise from iatrogenic, traumatic, or inflammatory causes. We report the familial occurrence of a congenital bulbar urethral stricture in two brothers. Retrograde and voiding cystourethrography was performed. A Cobb′s collar was diagnosed after radiological and endoscopic evaluation in both cases and was successfully managed with urethroplasty. Cobb′s collar is a rarely recognized cause of a membranous stricture of bulbar urethra that can lead to several urinary problems. In cases of adolescent and young adults presenting with symptoms of progressive urinary obstruction and enuresis with or without urinary tract infection, Cobb′s collar can be seen as a minor constriction in the bulbar urethra, but is not frequently symptomatic, and the familial occurrence of such a stricture is even rarer.

Background: Detection of HIV among elderly is increasing globally and this is going to be a major public health issue in recent future. In India, data regarding demographical and clinical picture of HIV among elderly are sparse more so for elderly women. Elderly individuals especially women are still largely outside the light of HIV awareness programs and represent a major portion of undiagnosed HIV infected population with a poor quality of life. Methods and Materials: This cross-sectional study was conducted at HIV clinic of a tertiary care hospital in eastern India. 567 cases (men 457, women 110) that were detected to be HIV-positive at or after age of 50 years from 2008 to 2012 were analyzed on socio-demographical and clinical basis. Results: Detection of HIV was found to be increasing (5.7% in 2008 to 12.8% in 2012) among the elderly over last five years. Illiteracy (p < 0.01), unemployment (p < 0.001), seroconcordance of spouse (p = 0.02) and widowhood (p = 0.017) were seen more among elderly women. Frailty was the most common clinical presentation among elderly men (31%) whereas most of the elderly women were asymptomatic (27%) at the time of diagnosis. Mean CD4 count among HIV infected elderly men and women were 112 cells/dl and 137 cells/dl respectively at the time of diagnosis. Conclusions: HIV in elderly is an emerging entity in India and posing elderly women a worse socioeconomic burden despite a better clinical profile at the time of presentation. Clinicians, social workers and policymakers should come forward with wider awareness program targeted for elderly HIV patients especially women.

Background: Kolkata, one of the major metropolitan cities of India, is also the capital of the state West Bengal, contributes largest number of malaria cases reported from West Bengal. The present study was undertaken to assess the anti-malarial prescribing pattern in a tertiary care teaching hospital in Kolkata. Methods: This was an observational, prospective, cross-sectional study for a period of one year (from March 2017 to February 2018) in which prescriptions of diagnosed pediatric and adult malaria patients were scanned and reviewed for anti-malarial use pattern. Core drug use indicators were also analyzed to assess the rational prescribing pattern. Results: During one-year study period, 122 adult and 24 child malaria patient encounters were screened. Among adult patients, 48(39.3%) patients had P. falciparum and 74(60.7%) patients had P. vivax malaria; in children, 9(37.5%) patients had P. falciparum and 15(62.5%) patients had P. vivax malaria. All adult and pediatric P. vivax malaria patients were treated with chloroquine. Artemisinin derivatives were prescribed to 91.67% of adult and 88.88% of pediatric falciparum malaria patients, 77.09% of adults and 66.67% of children received ACT. Artemether- lumefantrine was the most commonly prescribed ACT (33.34% in adults and 55.56% in children). Prescriptions were usually in generic name and from National EDL. Percentage of encounters with antibiotics was high in both age group but percentage of encounters with injections was low in adults and children. Conclusion: Chloroquine was used rationally for treatment of P. vivax malaria patients. Artemether-lumefantrine was the most common ACT used for treatment of P.falciparum malaria cases though the National guideline for treatment of malaria does not recommend Artemether-lumefantrine for this state and region for treatment of falciprum cases.

Tuberculosis (TB) remains one of the most important causes of death from an infectious disease, and it poses formidable challenges to global health at the public health, scientific, and political level. Miliary TB is a potentially fatal form of TB that results from massive lymphohematogenous dissemination of Mycobacterium tuberculosis bacilli. The epidemiology of miliary TB has been altered by the emergence of the human immunodeficiency virus (HIV) infection and widespread use of immunosuppressive drugs. Diagnosis of miliary TB is a challenge that can perplex even the most experienced clinicians. There are nonspecific clinical symptoms, and the chest radiographs do not always reveal classical miliary changes. Atypical presentations like cryptic miliary TB and acute respiratory distress syndrome often lead to delayed diagnosis. High-resolution computed tomography (HRCT) is relatively more sensitive and shows randomly distributed miliary nodules. In extrapulmonary locations, ultrasonography, CT, and magnetic resonance imaging are useful in discerning the extent of organ involvement by lesions of miliary TB. Recently, positron-emission tomographic CT has been investigated as a promising tool for evaluation of suspected TB. Fundus examination for choroid tubercles, histopathological examination of tissue biopsy specimens, and rapid culture methods for isolation of M. tuberculosis in sputum, body fluids, and other body tissues aid in confirming the diagnosis. Several novel diagnostic tests have recently become available for detecting active TB disease, screening for latent M. tuberculosis infection, and identifying drug-resistant strains of M. tuberculosis. However, progress toward a robust point-of-care test has been limited, and novel biomarker discovery remains challenging. A high index of clinical suspicion and early diagnosis and timely institution of antituberculosis treatment can be lifesaving. Response to first-line antituberculosis drugs is good, but drug-induced hepatotoxicity and drug–drug interactions in HIV/TB coinfected patients create significant problems during treatment. Data available from randomized controlled trials are insufficient to define the optimum regimen and duration of treatment in patients with drug-sensitive as well as drug-resistant miliary TB, including those with HIV/AIDS, and the role of adjunctive corticosteroid treatment has not been properly studied. Research is going on worldwide in an attempt to provide a more effective vaccine than bacille Calmette–Guerin. This review highlights the epidemiology and clinical manifestation of miliary TB, challenges, recent advances, needs, and opportunities related to TB diagnostics and treatment.

Cerebral venous sinus thrombosis (CVST) is not an uncommon cause of stroke but very often unrecognized at initial presentation due to lack of clinical suspicion and thus frequently left untreated. CVST is a potentially serious condition which manifests with diverse clinical manifestations, from isolated headache to focal neurological signs and even coma. CVST usually takes place either an inherited thrombophilia or any acquired hyperviscosity state and thus prompting anticoagulation was regimen as is the cornerstone of successful treatment. We describe a 47-year-old woman who presented with recurrent bouts of vomiting in the post-operative period and later developed cortical blindness and asymmetric limb weakness. Magnetic resonance imaging (MRI) showed hyperintensity involving bilateral parieto-occipital corticomedullary junction. MR venography showed signal void in the superior sagittal sinus. She was diagnosed as CVST and achieved complete recovery with anticoagulation therapy. Bilateral occipital infarction as a consequence of cerebral venous thrombosis is a rare cause of visual loss. Thrombosis in the superior sagittal sinus was related to her cortical blindness and weakness. This case illustrates that cerebral venous thrombosis should be considered in cases of occipital vascular lesions leading to acute painless loss of vision prompting anticoagulation therapy which can improve the outcome significantly. Dehydration could be considered as a risk factor for development of CVST in appropriate situations.

In Plasmodium vivax malaria, mechanisms that trigger transition from uncomplicated to fatal severe infections are obscure. In this multi-disciplinary study we have performed a comprehensive analysis of clinicopathological parameters and serum proteome profiles of vivax malaria patients with different severity levels of infection to investigate pathogenesis of severe malaria and identify surrogate markers of severity. Clinicopathological analysis and proteomics profiling has provided evidences for the modulation of diverse physiological pathways including oxidative stress, cytoskeletal regulation, lipid metabolism and complement cascades in severe malaria. Strikingly, unlike severe falciparum malaria the blood coagulation cascade was not found to be affected adversely in acute P. vivax infection. To the best of our knowledge, this is the first comprehensive proteomics study, which identified some possible cues for severe P. vivax infection. Our results suggest that Superoxide dismutase, Vitronectin, Titin, Apolipoprotein E, Serum amyloid A, and Haptoglobin are potential predictive markers for malaria severity.

AIM: To assess the uptake of an HIV screening program and therapeutic intervention to minimize the risk of mother to child transmission. METHODS: This was a cross-sectional observational study with retrospective data obtained from patient medical records. Pregnant women presenting to a tertiary care center in Kolkata India from 1 January 2004 - 31 December 2007 underwent HIV serology by rapid test after receiving group counseling. Care was administered using a standard national protocol by a multi-disciplinary team of health-care personnel. Main outcome measures were: acceptance of pretest counseling and HIV testing by pregnant women attending antenatal clinic services post-test counseling rate coverage rate of nevirapine to mother-child pairs and averted HIV infection in children. RESULTS: Of the 52 127 new antenatal booking visits 49 580 (95.11%) women attended pretest counseling and 47 506 (91.13%) women accepted HIV testing. Eighty-six women were found to be seropositive. The seroprevalence rate of HIV infection was 0.17%. Thirty-seven mothers (88%) and all newborn infants (100%) had received nevirapine prophylaxis. Overall nevirapine coverage rate was 48%. Twenty-four of the mother-infant pairs that we assessed had infants who were aged over 18 months by June 2008. Eleven (45.83%) of these women turned up with their babies for a blood test at 18 months. Three (27.27%) babies tested reactive. CONCLUSION: As uptake of testing is high and detection of HIV-infected women in pregnancy remains very low a radical rethinking of policies on therapeutic intervention and their implementation now needs to be undertaken.

Objective: To compare the risk of HIV and sexually transmitted diseases (STD) among homeless Muslim (circumcised) and Hindu (uncircumcised) men in Kolkata, India. Background: Many observational studies and clinical trials in Africa have demonstrated that male circumcision provides protection against HIV acquisition, but there are sparse data on circumcision and HIV in India, which has the largest number of HIV cases in the world. Methods: Using a two-stage probability proportionate to size cluster design among homeless men aged 18–49 years in Kolkata, India, data were obtained on religion, behavioral risk factors, and HIV/STD prevalence, by administering an anonymous questionnaire. Rapid HIV tests and testing for syphilis were performed on blood, and urine samples were obtained to test for gonorrhea. Results: The odds ratio for HIV among Muslims (circumcised) compared to Hindus (uncircumcised) was 0.43 (95% confidence interval 0.29–0.67). Despite Muslims having more partners and visits to commercial sex workers, the rates of syphilis and gonorrhea were similar. The results suggest that a biological effect of circumcision protects against HIV infection. Conclusion: The beneficial effect of circumcision should be communicated to high-risk groups, as well as to the general population.

Filarial infection can have varied manifestations, but hydropneumothorax at presentation has not yet been reported. A 28-year-old man presented to our hospital with heaviness of the left chest for the past 10 days, which was preceded by a sudden, short stabbing pain in the left chest after straining. Chest X-ray revealed left-sided hydropneumothorax. A peripheral blood picture revealed significant eosinophilia. A pleural fluid report also showed eosinophilia and a few motile microfilaria of Wuchereria bancrofti. Microfilaria was also documented in peripheral blood. There was no evidence of other organ system involvement. The patient was diagnosed with ‘Filarial Hydropneumothorax’. After treatment with a temporary chest drain and oral diethylcarbamazine citrate, there was dramatic relief of symptoms and radiological improvement. The patient has been symptom free with no features of recurrence through 8 months of follow-up.

Tuberculosis (TB) remains one of the most important causes of death from an infectious disease, and it poses formidable challenges to global health at the public health, scientific, and political level. Miliary TB is a potentially fatal form of TB that results from massive lymphohematogenous dissemination of Mycobacterium tuberculosis bacilli. The epidemiology of miliary TB has been altered by the emergence of the human immunodeficiency virus (HIV) infection and widespread use of immunosuppressive drugs. Diagnosis of miliary TB is a challenge that can perplex even the most experienced clinicians. There are nonspecific clinical symptoms, and the chest radiographs do not always reveal classical miliary changes. Atypical presentations like cryptic miliary TB and acute respiratory distress syndrome often lead to delayed diagnosis. High-resolution computed tomography (HRCT) is relatively more sensitive and shows randomly distributed miliary nodules. In extrapulmonary locations, ultrasonography, CT, and magnetic resonance imaging are useful in discerning the extent of organ involvement by lesions of miliary TB. Recently, positron-emission tomographic CT has been investigated as a promising tool for evaluation of suspected TB. Fundus examination for choroid tubercles, histopathological examination of tissue biopsy specimens, and rapid culture methods for isolation of M. tuberculosis in sputum, body fluids, and other body tissues aid in confirming the diagnosis. Several novel diagnostic tests have recently become available for detecting active TB disease, screening for latent M. tuberculosis infection, and identifying drug-resistant strains of M. tuberculosis. However, progress toward a robust point-of-care test has been limited, and novel biomarker discovery remains challenging. A high index of clinical suspicion and early diagnosis and timely institution of antituberculosis treatment can be lifesaving. Response to first-line antituberculosis drugs is good, but drug-induced hepatotoxicity and drug-drug interactions in HIV/TB coinfected patients create significant problems during treatment. Data available from randomized controlled trials are insufficient to define the optimum regimen and duration of treatment in patients with drug-sensitive as well as drug-resistant miliary TB, including those with HIV/AIDS, and the role of adjunctive corticosteroid treatment has not been properly studied. Research is going on worldwide in an attempt to provide a more effective vaccine than bacille Calmette-Guérin. This review highlights the epidemiology and clinical manifestation of miliary TB, challenges, recent advances, needs, and opportunities related to TB diagnostics and treatment.

Cobb syndrome is an exceedingly rare clinical condition defined by the presence of a vascular skin nevus and an angioma in the spinal canal at the same metamere. We report the case of a 14-year-old boy who presented with sudden onset paraplegia. Physical examination showed port-wine stains over buttock and thigh. Magnetic resonance (MR) angiogram of the dorso-lumbar spine revealed a large arteriovenous malformation (AVM) at D11-D12 to L2-L3 levels. These concurrent findings led to the diagnosis of Cobb’s syndrome. The patient received orally administered prednisolone therapy and underwent endovascular embolization of spinal angioma. Cobb’s syndrome is a rare disease entity and literature search revealed only a few case reports and series mentioning this condition to date. The importance lies in the recognition that cutaneous vascular lesions may clue to an associated spinal cord angioma or AVM that may lead to weakness or paralysis.

Background: Alcoholism is a health problem not only in developed countries but also in developing countries. Cirrhosis due to alcohol is a common cause of death among individuals abusing alcohol. A better knowledge of the spectrum of alcoholic liver diseases, its clinical, biochemical and histopathological features could result in early detection and prevention of alcoholic liver diseases before it’s catastrophic and life threatening effects. Materials and Methods: A total of 200 patients with alcoholic liver diseases were studied with respect to alcohol consumption, clinical features, biochemical and histopathological changes. The clinical features, biochemical parameters, and histopathology of liver including Ishak’s modified histological activity index (HAI) were correlated with the amount and duration of alcohol consumed. Result: Majority of the patients were in the age group of 40-49 years and all the cases were males. Majority consumed alcohol of about 75-90 grams per day for a duration of 10–12 years. Anorexia and jaundice were the most common symptom and clinical finding respectively. Hyperbilirubinemia and hypoalbuminemia were the most common abnormalities observed in liver function tests. Advanced HAI stages with features of cirrhosis were most frequent histo-pathological finding noted in this study. Clinico-biochemical profile was significantly correlated with degree of alcohol ingestion as well as with liver histopathology. Conclusion: The wide prevalence of alcoholic liver disease including cirrhosis among Indian males was noted with significantly lower quantity and duration of alcohol ingestion. The severity of liver damage is directly proportional to the quantity and duration of alcohol consumed. Clinical features and biochemical changes may forecast the liver histopathology among the patients of alcoholic liver disease.

Although mental retardation is generally associated with Bardet-Biedl (BBS) syndrome, a rare autosomal recessive disorder with multisystem involvement, autism is an unusual comorbidity. An 8-year-old boy presented to our psychiatry department with poor social skills and night blindness. On further assessment autism, mild mental retardation, retinitis pigmentosa, polydactyly and syndactyly, obesity, micropenis, maldescended testis, hypodontia and high-arched palate were noted and subsequently a diagnosis of BBS was made. To the best of our knowledge, this is the first reported case of BBS with autism from eastern India; it also emphasises the importance of thorough physical examination even in a patient presenting with pure psychiatric symptoms.

Abrus precatorius poisoning leading to hemorrhagic gastroduodenitis– a rare experience Abrus precatorius, popularly known as ‘kuch’ or ‘rati’, is used in weighing gold, making jewelry and children’s play. One to two crushed seeds can be fatal and very often used as cattle poison which mimics viper bite. Human poisoning with ‘kuch’ seeds is rarely reported in literature and almost always lifethreatening without prompt identification and aggressive supportive therapy. The authors here present a case of acute hemorrhagic gastroduodenitis following accidental ingestion of some beans with bright red and black color which was found to be Abrus precatorius. Patient himself identified those seeds as well as corroborated by forensic medicine and toxicology department. Moreover with vigilant monitoring the patient survived without any residual illness.

Lambert–Eaton myasthenic syndrome (LEMS) is a rare autoimmune disorder of neuromuscular transmission characterized clinically by muscle weakness, hyporeflexia or areflexia, and autonomic dysfunction. LEMS can occur as a remote effect of a neoplasm, in the paraneoplastic form (PLEMS), usually in association with small cell lung cancer (SCLC), or in the non-paraneoplastic form (NPLEMS). To denote a case as a patient with non-paraneoplastic LEMS needs a long-term follow up as in paraneoplastic cases where neurological symptoms can precede the diagnosis of primary neoplasm. Brain metastases from small cell lung cancer (SCLC) are found in approximately 10–14% of cases at presentation, with 65–95% of these cases also having disease outside the central nervous system (CNS) on diagnosis or shortly after. A solitary brain metastasis with an undetected primary is a rare presentation in cases of SCLC. Sometimes combinations of paraneoplastic syndromes (PS) and metastatic features might be the only presentation of the underlying occult primary SCLC. Certain clinical features together with relevant investigation help to clinch the diagnosis which has significant therapeutic implications in patients with occult primary SCLC. This chapter presents varied clinical features, diagnostic modalities, and therapeutic options in patients with LEMS and brain metastasis arising from occult primary SCLC. The authors also share their experience of a case of PLEMS and solitary cerebellar metastasis occurring simultaneously in the absence of a florid picture of SCLC.

Tuberculosis (TB) involving the pancreas are uncommon, especially when present in immunocompetent hosts. Pancreatic TB is more frequently associated with miliary TB or widely disseminated disease. Pancreatic TB may present as cystic or solid pancreatic masses, pancreatic abscess or acute or chronic pancreatitis. Majority of the cases are diagnosed after surgical exploration for presumed pancreatic malignancy and pre-operative diagnosis is quite difficult. However, improvement in imaging techniques and the resulting image-guided interventions gradually can obviate the need for more invasive diagnostic surgical procedures and expedite the planning of therapy. Herein, we report a rare case of isolated pancreatic TB which presented with pancreatic mass lesion in an immunocompetent host. Diagnosis was made by contrast enhanced computed tomography and guided fine needle aspiration of the pancreatic mass which revealed acid-fast bacilli on Ziehl-Neelsen stain. The case was treated successfully with antituberculous drugs. Pancreatic tuberculosis should be considered in the differential diagnosis of a pancreatic mass when the patient is young, residing in the endemic zone of tuberculosis. Every attempt should be made to diagnose the cases to prevent unnecessary operation.