A Critical Appraisal of the New Competency-Based Medical Undergraduate Curriculum in Biochemistry.

The new competency-based medical education undergraduate curriculum (CBMC) was launched for the 2019 admission batch of MBBS students. The programme is designed to create an "Indian Medical Graduate" (IMG) possessing the requisite knowledge, skills, attitudes, values and responsiveness, so that the graduate may function appropriately and effectively as a physician of first contact with the community while being globally relevant. Given that implementation of this curriculum is still in its infancy across the country, we stand to gain from a unified approach to its implementation. Phase I of the curriculum includes anatomy, physiology, and biochemistry along with professional and personal development modules. Biochemistry enjoys an enviable position in the medical curriculum as it explains the molecular basis of diseases. We present an appraisal of the curriculum in Biochemistry by reviewing the components against Harden's six themes which are considered when planning or developing a curriculum. Further, five core components of CBME are selected on the basis of three research papers to characterize underlying assumptions of CBME to suggest ways of logical implementation for achieving the competencies expected of the Indian Medical Graduate. The insight gained shall help students to be equipped with competencies which they shall be able to use in their day- to- day work, which shall ultimately help benefit patient care and the society at large. Supplementary Information: The online version contains supplementary material available at 10.1007/s12291-022-01088-y.

Perceptions of faculty toward "social obligation" at an Indian medical school.

BACKGROUND: The World Health Organization has defined social accountability of medical schools as "…obligation to direct their education, research, and service activities toward addressing the priority health concerns of the community." The current study looked at the extent to which the concept was understood in an Indian medical school, exploring how faculty perceived and were involved in directing a response to the social obligation of their medical school. METHODS: Seventeen semi-structured audio-recorded interviews were conducted by purposive sampling of faculty from different disciplines. Interviews were transcribed and analyzed through a collaborative thematic approach to gain insight into faculty knowledge of the "obligation triad" of responsibility, responsiveness, and accountability; enablers and barriers in implementation; and understanding stakeholder roles. RESULTS: Faculty were unfamiliar with the terms and were unaware of the movement towards socially accountable schools. They were, however, sensitive to their responsibilities towards students and the community. Four major themes emerged: Perceptions of social obligation, awareness of social and cultural values, the role of partnerships, and moving toward a socially accountable model. DISCUSSION: Sensitizing students towards community needs, impact of cultural and socio-economic backgrounds, importance of contextual curriculum, and stakeholder roles were some of the challenges highlighted in developing a socially accountable medical school.

Oxidative Stress Markers in Tuberculosis and HIV/TB Co-Infection.

INTRODUCTION: Dysfunction of redox homeostasis has been implicated in many pathological conditions. An imbalance of pro- and anti-oxidants have been observed in Tuberculosis (TB) and its co-morbidities especially HIV/AIDS. The pro inflammatory milieu in either condition aggravates the physiological balance of the redox mechanisms. The present study therefore focuses on assessing the redox status of patients suffering from TB and HIV-TB co-infection. AIM: To assess the oxidative stress markers in the HIV-TB and TB study cohort. MATERIALS AND METHODS: The current prospective study was conducted in Haffkine Institute, Parel, Maharashtra, India, during January 2013 to December 2015. Blood samples from 50 patients each suffering from active TB and HIV-TB co-infection were collected from Seth G.S.Medical College and KEM Hospital Mumbai and Group of Tuberculosis Hospital, Sewree Mumbai. Samples were processed and the experiments were carried out at the Department of Biochemistry, Haffkine Institute. Samples from 50 healthy volunteers were used as controls. Serum was assessed for pro-oxidant markers such as Nitric Oxide (NO), Thiobarbituric Acid Reactive Species (TBARS), C-Reactive Protein (CRP), superoxide anion. Antioxidant markers such as catalase and Superoxide Dismutase (SOD) were assessed. Total serum protein, was also assessed. RESULTS: Among the pro-oxidants, serum NO levels were decreased in TB group while no change was seen in HIV-TB group. TBARS and CRP levels showed significant increase in both groups; superoxide anion increased significantly in HIV-TB group. Catalase levels showed decreased activities in TB group. SOD activity significantly increased in HIV-TB but not in TB group. The total serum proteins were significantly increased in HIV-TB and TB groups. The values of Control cohort were with the normal reference ranges. CONCLUSION: In the present study, we found the presence of oxidative stress to be profound in the TB and HIV-TB co-infection population.

Hepatic Enzyme's Reference Intervals and Their Modulating Factors in Western Indian Population.

The reference intervals (RIs) of serum aminotransferases and Gamma-glutamyl transferase (GGT) have been established many years ago. Recent RIs are not available. The prospective study was conducted to re-evaluate the RIs of liver enzymes and the effect of demographic and anthropometric variables on them in western Indian population. A total of 1059 blood donors comprised the study population. Anthropometry and serum liver enzymes levels were measured. Subjects were categorized into normal weight and overweight by using body mass index (BMI) and waist circumference (WC). For RI determination, non-parametric methodology recommended by IFCC/CLSI was adopted. Mann-Whitney test and Spearman's rank correlation were used for statistical analysis. Upper limit of normal reference value of liver enzymes were lower in female compared to male. (ALT-23.55 F vs 36.00 M, GGT-34.58 F vs 36.20 M) When RI of liver enzymes were calculated according to body mass index, the upper limit of normal of ALT and GGT were higher in overweight group compared to normal weight group. (ALT-38.00 vs 27.00 IU/L and GGT-37.59 vs 35.26 IU/L). In both male and female, liver enzymes correlated significantly with age. WC and BMI were positively correlated with AST, ALT and GGT in both subgroups and the correlation was stronger in male. Demographic factors should be considered for making liver enzyme tests more clinically relevant. Gender based partitioning should be adopted for serum alanine aminotransferase (ALT) and GGT reference values for Western Indian population.

Steric Crowding of the Turn Region Alters the Tertiary Fold of Amyloid-ß18-35 and Makes It Soluble.

Aß self-assembles into parallel cross-ß fibrillar aggregates, which is associated with Alzheimer's disease pathology. A central hairpin turn around residues 23-29 is a defining characteristic of Aß in its aggregated state. Major biophysical properties of Aß, including this turn, remain unaltered in the central fragment Aß18-35. Here, we synthesize a single deletion mutant, ΔG25, with the aim of sterically hindering the hairpin turn in Aß18-35. We find that the solubility of the peptide goes up by more than 20-fold. Although some oligomeric structures do form, solution state NMR spectroscopy shows that they have mostly random coil conformations. Fibrils ultimately form at a much higher concentration but have widths approximately twice that of Aß18-35, suggesting an opening of the hairpin bend. Surprisingly, two-dimensional solid state NMR shows that the contact between Phe(19) and Leu(34) residues, observed in full-length Aß and Aß18-35, is still intact in these fibrils. This is possible if the monomers in the fibril are arranged in an antiparallel ß-sheet conformation. Indeed, IR measurements, supported by tyrosine cross-linking experiments, provide a characteristic signature of the antiparallel ß-sheet. We conclude that the self-assembly of Aß is critically dependent on the hairpin turn and on the contact between the Phe(19) and Leu(34) regions, making them potentially sensitive targets for Alzheimer's therapeutics. Our results show the importance of specific conformations in an aggregation process thought to be primarily driven by nonspecific hydrophobic interactions.

The rs10993994 in the proximal MSMB promoter region is a functional polymorphism in Asian Indian subjects.

BACKGROUND: The microseminoprotein gene encoding prostate secretory protein of 94 amino acids (PSP94) harbours a potential risk allele (rs10993994) for prostate cancer (PCa) in its promoter region. However, studies on rs10993994 have been sparse in Asian Indians. METHODS: The present study recruited a sample population of 44 benign prostatic hyperplasia patients, 33 PCa patients and 60 healthy participants, of which, participants without other confounding risk factors for PCa were retained. The serum PSP94 (sPSP94) levels were measured by a serum-based ELISA in an earlier study. A novel RFLP technique was developed to screen for rs10993994 which was validated with direct sequencing. RESULTS: Sequencing showed additional 4 SNPs (rs41274660, rs141211965, rs12770171, rs10669586) and 2 novel variants (GenBank accession nos. KM265191 and KM265192). In silico DNA topographical studies predicted that KM265192 would have higher cleavage intensity and more accessibility for binding of transcription factors. Even though, similar frequencies were observed for all the variants in all the three study groups, the risk allele 'T' (rs10993994) was seen to be associated with reduced PSP94 expression both at mRNA and protein level. Further, mRNA expression as studied by real-time PCR correlated positively with sPSP94 levels. Interestingly, CC genotype of rs10993994 showed highest sPSP94 levels in all the three study groups and was associated with Gleason score ≤7 in PCa patients. In contrast, TT genotype of rs10993994 was associated with lesser sPSP94 levels and with aggressiveness of PCa. CONCLUSION: rs10993994 was found to be a functional SNP in the studied Asian Indian population.

Development of an ELISA for sPSP94 and utility of the sPSP94/sPSA ratio as a diagnostic indicator to differentiate between benign prostatic hyperplasia and prostate cancer.

BACKGROUND: The serum PSA (sPSA) test has low specificity for prostate cancer (PCa), since sPSA also rises in benign prostatic hyperplasia (BPH). Serum PSP94 (sPSP94), a major secreted prostate protein, is indicated as a PCa marker. The potential of sPSP94 and sPSA in conjunction with each other to improve specificity of diagnostic test for PCa needs to be evaluated. METHODS: PCa patients (n=33), BPH patients (n=44) and healthy controls (n=50) were recruited. A serum-based sandwich ELISA was developed to measure sPSP94 concentrations. Utility of sPSP94 in improving specificity of sPSA test was evaluated by studying sPSP94/sPSA ratios of study participants. RESULTS: Considerable decrease in overlap among sPSP94/sPSA ratio values of BPH and PCa patients was observed, as compared to sPSP94 or sPSA alone. For differentiating between BPH and PCa patients, this ratio had a maximum area under the curve (AUC) of 0.859 (P=0.0132) and had a comparable sensitivity (90.91%) to sPSA with an increased specificity of 70.45%. Further, decision curve analysis (DCA) showed that sPSP94/sPSA ratio had a superior net benefit in identifying PCa, in patients opting for biopsy. CONCLUSION: The sPSP94/sPSA ratio can be a better differentiating marker between BPH and PCa, than sPSP94 or sPSA alone.

A survey validation and analysis of undergraduate medical biochemistry practical curriculum in maharashtra, India.

In order to review the strengths and weaknesses of medical biochemistry practical curriculum for undergraduates and to generate ideas to improve it, a questionnaire was sent to 50 biochemistry faculty members selected (through simple random sampling method) from 42 medical colleges of Maharashtra, India. 39 responded to the questionnaire, representing a 78% response rate. The internal consistency of the questionnaire sections was found to be satisfactory (>0.7). The respondents did not agree that the ongoing curriculum was in alignment with learning outcomes (8%), that it encouraged active learning (28%), helped to apply knowledge to clinical situations (18%) and promoted critical thinking and problem solving skills (28%). There were a number of qualitative experiments that were rated 'irrelevant'. Qualitative and quantitative experiments related to recent advances were suggested to be introduced by the respondents. Checklists for the practicals and new curriculum objectives provided in the questionnaire were also approved. The results of the curriculum evaluation suggest a need for re-structuring of practical biochemistry curriculum and introduction of a modified curriculum with more clinical relevance.

Influence of method of specimen collection on various preanalytical sample quality indicators in EDTA blood collected for cell counting.

Improving specimen quality as well as healthcare worker (HCW) safety poses significant concerns for today's laboratories. With an increasing number of diagnostic tests requested, laboratory professionals are faced with challenges to reduce laboratory errors, improve the quality of laboratory results to assure accurate diagnosis and implement initiatives to ensure healthcare worker safety and minimize risk of exposure to bloodborne pathogens. A prior study conducted in 2008 reported that variations in blood collection methods for clinical chemistry assays may affect overall specimen quality. As a follow up, the current study assessed the quality of 22563 patient specimens for cell counting in EDTA blood collection tubes that were obtained with needle and syringe collection (open) using either disposable tubes or re-washed glass vials or with an evacuated blood collection system (closed). Based on the observations, the use of the evacuated blood collection system resulted in better preanalytical specimen quality as compared with needle and syringe collection. The findings also showed an approximately 70-fold reduction in the incidence of clotting as well as fewer instrument-generated flags using the evacuated collection system. In addition, the use of an evacuated collection system for venous blood collection demonstrated lesser chance of blood exposure to healthcare workers.

Influence of blood specimen collection method on various preanalytical sample quality indicators.

Preanalytical errors contribute to a large proportion of total laboratory errors. In order to achieve continuous laboratory improvement, it is important to focus on all phases of patient specimen testing i.e. preanalytical, analytical and post-analytical. With large variations in the way venous blood specimens are collected using diverse devices in the country, the effect of such practices on specimen quality is not known. The purpose of this study was to monitor fourteen specimen preanalytical quality indicators in order to compare the usage of evacuated blood collection devices with needle and syringe open collection using either disposable tubes or re-washed glass vials. The study involved 26638 patient specimens assessed over a period of 6 months. The results demonstrated that evacuated closed blood collection resulted in an approximate 100-fold reduction in the incidence of hemolysis in samples. Similarly, there was a 200-fold reduction in incidence of insufficient specimen quantity while using evacuated collection system. It was also found that incidence of specimen contamination, improper volume of sample collected, and specimen spillage was also lower when the evacuated collection system was used. Further, it was also observed that the facility with a laboratory information system demonstrated much lower specimen identification and related errors. The observed results clearly demonstrate that the usage of the evacuated blood collection system resulted in improvement of preanalytical specimen quality as compared to needle and syringe usage.

Role of proteases in the pathophysiology of cardiac disease.

Cardiovascular disease is a major cause of death and thus a great deal of effort has been made in salvaging the diseased myocardium. Although various factors have been identified as possible causes of different cardiac diseases such as heart failure and ischemic heart disease, there is a real need to elucidate their role for the better understanding of the cardiac disease pathology and formulation of strategies for developing newer therapeutic interventions. In view of the intimate involvement of different types of proteases in maintaining cellular structure, the role of proteases in various cardiac diseases has become the focus of recent research. Proteases are present in the cytosol as well as are localized in a number of subcellular organelles in the cell. These are known to use extracellular matrix, cytoskeletal, sarcolemmal, sarcoplasmic reticular, mitochondrial and myofibrillar proteins as substrates. Work from different laboratories using a wide variety of techniques has shown that the activation of proteases causes alterations of a number of specific proteins leading to subcellular remodeling and cardiac dysfunction. Inhibition of protease action by different drugs and agents, therefore, has a clinical relevance and is expected to form a part of new treatment paradigm for improving heart function. This review examines the biochemistry and localization of some of the proteases in the cardiac tissue in addition to identification of the sites of action of some protease inhibitors.

Development and validation of an ELISA for hemoglobin-A2: a novel method for beta-thalassemia screening in developing countries.

Hemoglobin-A2 (HbA2) measurement in human hemolysates has great significance, since its level can indicate beta-thalassemia carrier status in otherwise healthy individuals. An ELISA for HbA2 using antiserum monospecific to the delta chain of HbA2 and affinity purified antirabbit gamma globulins (ARGG) conjugated to horseradish peroxidase (HRP) have been developed. The monospecific antiserum used does not cross react with other hemoglobins. Hemolysates from volunteers are used for measurement of HbA2. In a limited trial for beta-thalassemia carrier screening (n = 350), the results obtained with the developed ELISA are comparable with those obtained with a micro-column chromatography method (r > or = 0.89). The developed ELISA is simple, accurate, precise, inexpensive, and several samples can be processed simultaneously with ease, making this system a suitable candidate for transforming into a user friendly kit.

Role of proteases in the pathophysiology of cardiac disease.

Cardiovascular disease is a major cause of death and thus a great deal of effort has been made in salvaging the diseased myocardium. Although various factors have been identified as possible causes of different cardiac diseases such as heart failure and ischemic heart disease, there is a real need to elucidate their role for the better understanding of the cardiac disease pathology and formulation of strategies for developing newer therapeutic interventions. In view of the intimate involvement of different types of proteases in maintaining cellular structure, the role of proteases in various cardiac diseases has become the focus of recent research. Proteases are present in the cytosol as well as are localized in a number of subcellular organelles in the cell. These are known to use extracellular matrix, cytoskeletal, sarcolemmal, sarcoplasmic reticular, mitochondrial and myofibrillar proteins as substrates. Work from different laboratories using a wide variety of techniques has shown that the activation of proteases causes alterations of a number of specific proteins leading to subcellular remodeling and cardiac dysfunction. Inhibition of protease action by different drugs and agents, therefore, has a clinical relevance and is expected to form a part of new treatment paradigm for improving heart function. This review examines the biochemistry and localization of some of the proteases in the cardiac tissue in addition to identification of the sites of action of some protease inhibitors. (Mol Cell Biochem 263: 241-256, 2004).