Utility of morphologic assessment of bone marrow biopsy in diagnosis of lysosomal storage disorders.

Introduction: Lysosomal storage disorders (LSDs) are rare disorders and pose a diagnostic challenge for clinicians owing to their generalized symptomatology. In this study, we aim to classify LSDs into two broad categories, namely, Gaucher disease (GD) and Niemann-Pick/Niemann-Pick-like diseases (NP/NP-like diseases) based on the morphology of the storage cells in the bone marrow (BM) aspiration smears and trephine biopsy sections. Materials and Method: This retrospective study includes 32 BM specimens morphologically diagnosed as LSDs at our institute, in the last 10 years. Subsequently, they were subclassified into GD and NP/NP-like diseases. Further, we have compared and analyzed the clinical, hematological, and biochemical parameters for the two groups of LSDs. Results: Based on BM morphology, 59.4% (n = 19) cases were diagnosed as NP/NP-like diseases and 40.6% (n = 13) cases as GD. Abdominal distension and failure to thrive were the most common clinical manifestations in both groups of LSDs. Anemia and thrombocytopenia were frequently seen in either of the LSDs. On the assessment of metabolic profile, elevated total/direct bilirubin and liver enzymes were more commonly seen in NP/NP-like diseases when compared with GD. Conclusion: We have classified LSDs into GD and NP/NP-like diseases based on the morphology of the storage cells in the BM specimen. The hallmark findings on BM biopsy annexed with the comparative features of the two proposed categories can aid the clinician in clinching the diagnosis. Formulation of such a methodology will prove instrumental for patient care in an underresourced setting.

Effects of Menopause on Sexual Function in Indian Women: A McCoy's Questionnaire-Based Assessment.

BACKGROUND: Menopause is associated with physical, physiological, psychological changes and may lead to sexual dysfunction (SD) effecting woman's health and well-being. Scientific research in the area of female sexuality in India is scant. Therefore, this study aimed to investigate female sexual function at perimenopause and menopause and determine the association between sociodemographic and physiological factors with sexual function. MATERIALS AND METHODS: This was a cross-sectional hospital-based study carried out in perimenopausal and menopausal women. Study participant's details were collected by gynecologists and clinical research professionals following the participant's informed consent. The case report and McCoy female sexuality questionnaire were used. The association between sociodemographic status and sexual function was determined. Data were summarized using descriptive statistics for portraying profile of the participants and t-test for comparison. RESULTS: A total of 129 women in the menopausal (SD - 3.26) and 112 in the premenopausal group (SD - 6.01) were enrolled. The sociodemographic parameters did not significantly affect the sexual function scores in both groups. In terms of vaginal atrophy, a significant increase in urgency was noted in the postmenopause group. The general domain of sexual function was significantly lower in menopausal than and perimenopausal with a P < 0.001. Looking at individual domains of sexual function, for sexual interest, satisfaction, vaginal lubrication, and orgasm, the mean value of perimenopausal participants was significantly higher when compared to menopausal women; for a primary partner domain, no significant differences between the two groups were noted. CONCLUSION: Overall, the sociodemographic profile did not impact sexual function in this study. Compared with menopausal women, perimenopausal women showed better, more complete sexual function based on McCoy's score except partner-related domain that is constant from perimenopause to menopause in a monogamous relationship.

Mutant specific anti calreticulin antibody (CAL2) immunohistochemistry as a screening test for calreticulin (CALR) mutation testing.

BACKGROUND: About 50 different CALR frameshift mutations have been identified in BCR-ABL1 negative MPN, all leading to the development of common new protein C terminus. Antibody targeting this terminal epitope can be useful to identify this driver mutation using immunohistochemistry. MATERIALS AND METHODS: CALR mutation analysis was carried out in 51 JAK2V617F negative cases, PMF (n = 22) and ET (n = 29). PCR followed by fragment analysis was performed for molecular detection of CALR mutation. Bone marrow biopsy specimens of corresponding patients were subjected to IHC using mutation specific antibody CAL2. Staining pattern and intensity were observed. Staining of <2% of background nonmegakaryocytic (non- MK) cells were regarded as Pattern A, while staining of more than 2% of background nonmegakaryocytic (non-MK) was regarded as pattern B. RESULTS: CALR mutation was noted in 40.9% (9/22) and 41.4% (12/29) of JAK2V617F negative PMF and ET, respectively. All CALR mutated cases, irrespective of the mutation type, showed a positive IHC staining in the megakaryocytes with moderate to bright intensity. All CALR wild-type cases were negative on IHC. (Concordance rate- 100%). Pattern A was noted in 40% cases, while pattern B was noted in 60% cases. Pattern A staining had significantly higher chances of having type 1 mutation as compared to pattern B. In contrast, pattern B had a nonsignificant trend toward higher bone marrow cellularity and marrow fibrosis. CONCLUSION: CAL2 IHC detects all types of CALR mutation. This can act as a sensitive, specific, rapid, and cost-effective screening test for CALR mutation analysis.

Utility of thromboelastography versus routine coagulation tests for assessment of hypocoagulable state in patients undergoing cardiac bypass surgery.

INTRODUCTION: Peri-operative monitoring of coagulation is important to diagnose potential cause of hemorrhage, to manage coagulopathy and guide treatment with blood products in patients undergoing cardiac surgery with cardiopulmonary bypass. This study was done to evaluate usefulness of Thromboelastography (TEG) and routine coagulation tests (RCT) in assessing hemostatic changes and predicting postoperative bleeding in patients undergoing cardiac surgery with cardiopulmonary bypass. METHODS: Fifty adult patients undergoing cardiac surgery with cardiopulmonary bypass were enrolled in this prospective study. Preoperative and post-operative samples were collected for routine coagulation tests and TEG. Regression analysis and test of significance using Pearson's correlation coefficient was performed to assess correlation between routine coagulation tests and corresponding TEG parameters .Regression analysis was done to study relation between blood loss at 24 hours and various coagulation parameters. RESULTS:: The Routine coagulation test i.e. PT, INR, APTT showed no significant correlation with corresponding TEG parameters in pre-operative samples. However platelet count significantly correlated (p = 0.004) with MA values in postoperative samples. A significant correlation (p = 0.001) was seen between fibrinogen levels and alpha angles as well as with MA in both baseline preoperative and postoperative samples. TEG parameters R time and MA in postoperative samples were the only parameters that predicted bleeders with fair accuracy. CONCLUSION: Though the techniques of RCT and TEG are different, a few RCT e.g. platelet count and fibrinogen correlated with corresponding TEG parameters i.e. MA and Alpha angle. TEG parameters (R time and MA in postoperative samples) were able to predict blood loss better than RCT.

Formative assessment in postgraduate medical education - Perceptions of students and teachers.

CONTEXT: One of the most important factors of medical education that can revolutionize the learning process in postgraduate students (PGs) is assessment for learning by means of formative assessment (FA). FA is directed at steering and fostering learning of the students by providing feedback to the learner. However, though theoretically well suited to postgraduate training, evidence are emerging that engaging stakeholders in FA in daily clinical practice is quite complex. AIMS: To explore perceptions of PGs and teachers (Ts) about factors that determines active engagement in FA. SUBJECTS AND METHODS: It was a descriptive qualitative study involving focus group discussions (FGDs) with PGs and Ts from Departments of Pediatrics and Orthopedics. FGDs data were processed through points/remarks, data reduction, data display, coding followed by theme generation for content analysis. RESULTS: Four higher order themes emerged: Harsh reality of present summative assessment structure, individual perspectives on feedback, supportiveness of the learning environment, and the credibility of feedback and/or feedback giver. CONCLUSIONS: Engaging in FA with a genuine impact on learning is complex and quite a challenge to both students and Ts. Increased acceptability along with the effective implementation of FA structure, individual perspectives on feedback, a supportive learning environment and credibility of feedback are all important in this process. Every one of these should be taken into account when the utility of FA in postgraduate medical training is evaluated.

Effective risk assessment tools for osteoporosis in the Indian menopausal female.

OBJECTIVES: This study was designed to assess the effectiveness of self-administered osteoporosis risk score sheet, body mass index (BMI), and bone mineral density (BMD) (ultrasound) in screening females with low bone mass, and how the results of the tools correlate with each other. MATERIALS AND METHODS: The study was conducted on 536 postmenopausal women, who attended public awareness camps on Midlife Women Health held at the Menopause Research Unit, MGMCH, Sitapura, Jaipur. At these camps, in addition to several informational sessions on issues related to menopause, ultrasonic measurement of BMD was conducted on each participant. A broad questionnaire to identify midlife health problems was developed, and osteoporosis specific score sheet was designed to be self-administered. Patients were required to complete the osteoporosis specific risk score sheet and women health questionnaire (WHQ). BMI was determined. Statistical analysis was carried out to find the correlation between various variables. Sensitivity and specificity of the each risk score ascertained and cutoff risk score for identifying osteopenia was derived by comparing area under curve of each risk score on drawing receiver operational curve (ROC). RESULTS: Sensitivity of risk score system was calculated to be 78.33% with 95% confidence interval being 73.24-82.86% and specificity was 27.12% with 95% confidence interval being 21.56-33.27%, keeping the cutoff point at nine. There was statistically significant inverse relationship between risk score and BMD values with Pearson correlation coefficient of (-) 0.22 and positive relationship between BMD and BMI with correlation coefficient of 0.192. CONCLUSION: By noting down the risk factors and BMI, we can screen out the women who require further evaluation and management, thus, it is an effective tool, particularly in developing countries like India, where most of the patients cannot afford expensive DEXA scans, although considered as the gold standard for BMD assessment. With the help of such scoring systems, health resources can be judiciously utilized.

Construct optimization for protein NMR structure analysis using amide hydrogen/deuterium exchange mass spectrometry.

Disordered or unstructured regions of proteins, while often very important biologically, can pose significant challenges for resonance assignment and three-dimensional structure determination of the ordered regions of proteins by NMR methods. In this article, we demonstrate the application of (1)H/(2)H exchange mass spectrometry (DXMS) for the rapid identification of disordered segments of proteins and design of protein constructs that are more suitable for structural analysis by NMR. In this benchmark study, DXMS is applied to five NMR protein targets chosen from the Northeast Structural Genomics project. These data were then used to design optimized constructs for three partially disordered proteins. Truncated proteins obtained by deletion of disordered N- and C-terminal tails were evaluated using (1)H-(15)N HSQC and (1)H-(15)N heteronuclear NOE NMR experiments to assess their structural integrity. These constructs provide significantly improved NMR spectra, with minimal structural perturbations to the ordered regions of the protein structure. As a representative example, we compare the solution structures of the full length and DXMS-based truncated construct for a 77-residue partially disordered DUF896 family protein YnzC from Bacillus subtilis, where deletion of the disordered residues (ca. 40% of the protein) does not affect the native structure. In addition, we demonstrate that throughput of the DXMS process can be increased by analyzing mixtures of up to four proteins without reducing the sequence coverage for each protein. Our results demonstrate that DXMS can serve as a central component of a process for optimizing protein constructs for NMR structure determination.