Objective structured practical examination (OSPE) as a tool in formative assessment of II MBBS students, in pathology

Background: Assessment drives the student learning. Regular periodical assessment not only improves learning habits, but also enhances the competence in all levels of medical education. Traditional practical examination is more subjective. It depends on examiners subjectivity, varying difficulty level of various experiments, and also internal marks variation etc. These flaws can be overcome by newer methods like OSPE. The aim of the study was to implement OSPE as a tool of internal assessment for practical skills in the II MBBS. To compare this with traditional practical examination (TPE). To obtain the students and faculty feedback regarding OSPE as a tool of assessment.Methods: A cross sectional study was carried out for 158 students in II internal pathology practical examination for six days in the second week of September 2016 at Department of Pathology, Dr. Pinnamaneni Siddhartha institute of medical sciences & Research Foundation, Chinnaoutpalli. Faculty and students were sensitized; blueprint were used to arrange twenty OSPE stations for the exercises conducted as per TPE and for the same 25 marks as per TPE. Simultaneously, all the students were subjected to both TPE and OSPE at the same time and venue. TPE was assessed by two professors and OSPE by separate eight faculty members independently without interaction with the students. The procedural stations were evaluated by using checklist and the response stations which consisted of short answers and MCQs, facilitated correction. Feedback was given to the student on their performance and feedback was obtained from the students and faculty regarding OSPE by questionnaire with Yes/No answers.Results: Performance score of students in OPSE (13.73 ±2.49) was higher as compared to TPE (9.27±1.86) which was statistically significant. Based on the response to the questionnaire, students perception towards OSPE was analyzed. Majority strongly agree OSPE to be fairer, more transparent and objective in comparison to TPE. In contrast, all the faculty members unanimously opined that OSPE was difficult to arrange, time taken and faculty versus students ratio was high for evaluation. Though, the faculty (91%) overall opined that OSPE should be included as a method of assessment.Conclusions: Present study revealed that OSPE was acceptable, feasible and reliable to the students as well as for faculty for the internal assessment in pathology. Opinions of both students and faculties strongly agreed that OPSE is more effective objective assessment tool

Contrastive Deep Encoding Enables Uncertainty-aware Machine-learning-assisted Histopathology

Deep neural network models can learn clinically relevant features from millions of histopathology images. However generating high-quality annotations to train such models for each hospital, each cancer type, and each diagnostic task is prohibitively laborious. On the other hand, terabytes of training data -- while lacking reliable annotations -- are readily available in the public domain in some cases. In this work, we explore how these large datasets can be consciously utilized to pre-train deep networks to encode informative representations. We then fine-tune our pre-trained models on a fraction of annotated training data to perform specific downstream tasks. We show that our approach can reach the state-of-the-art (SOTA) for patch-level classification with only 1-10% randomly selected annotations compared to other SOTA approaches. Moreover, we propose an uncertainty-aware loss function, to quantify the model confidence during inference. Quantified uncertainty helps experts select the best instances to label for further training. Our uncertainty-aware labeling reaches the SOTA with significantly fewer annotations compared to random labeling. Last, we demonstrate how our pre-trained encoders can surpass current SOTA for whole-slide image classification with weak supervision. Our work lays the foundation for data and task-agnostic pre-trained deep networks with quantified uncertainty.Comment: 18 pages, 8 figure

Effect of Phosphine and Methyl Bromide Fumigation of Different Life Stages of Peanut Bruchid, Caryedon serratus Olivier

To ensure seed quality, peanut seeds received for export by the Plant Quarantine Unit of the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT) were subjected to phosphine and methyl bromide fumigation. Effective control of eggs, larvae, and adults of Caryedon serratus Olivier was achieved with methyl bromide vacuum fumigation (16 g/m3 for 4 h exposure). Under normal atmospheric pressure (NAP), phosphine fumigation @ 0.5,1.0, and 2.0 g a.I./m3 for 24, and also with 0.25 g a.l. for 72 h gave effective control of eggs and larvae. Ph~phlne (1.0 g a.l./40 kg burlap bag) with bruchld Infested pods for 120 h resulted in 100% larval and 93% adult mortality. Both the fumigants did not impair the viability of peanut seeds. A dosage of 16 g/m3 of methyl bromide for 4 h under vacuum or 0.25 g a.l./m3 of phosphine for 72 h under NAP or placing 1.0 g a.l. of ph~phlne in dry pods stored in 40 kg burlap bag covered with polythene sheets for 120 h can satisfy the seed health requirements

Generation of LUMCi041-A-2: equipping a PAX3 reporter iPSC line with doxycycline inducible H2B-mTurquoise2 for live cell imaging

An induced pluripotent stem cell (iPSC) line, in which a H2B-fluorescent protein fusion is temporally expressed, is a valuable tool to track cells and study cell divisions and apoptosis. To this end we introduced a 3rd generation "all-in-one" doxycycline-inducible H2B-mTurquoise2 vector into the AAVS1 locus of PAX3-Venus iPSCs via CRISPR/Cas9. H2B-mTurquoise2 expression is absent but readily induced by doxycycline allowing quantification of cell divisions and imaging of living cells. Besides being a universal reporter in iPSC-based differentiation and toxicity assays, the generated pluripotent and genomically normal LUMCi041-A-2 line is particularly suited to study PAX3-positive stages of development.Therapeutic cell differentiatio

Extracts from Acacia catechu suppress HIV-1 replication by inhibiting the activities of the viral protease and Tat

Background: Acacia catechu (Mimosa family) stem bark extracts have been used traditionally as a dietary supplement as well as a folk medicine given its reported anti-inflammatory, immunomodulatory, hepatoprotective, antioxidant, anti-microbial and anti-tumor activities. The present study was undertaken to evaluate the anti-HIV-1 activity of the extracts from stem bark of A. catechu. Methods. The aqueous and 50% ethanolic extracts of A. catechu stem bark were prepared and 50% ethanolic extract was further fractioned by successively partitioning with petroleum ether, chloroform and n-butanol. All the extracts and fractions were evaluated for cytotoxicity and anti-HIV-1 activity using different in vitro assays. The active n-butanol fraction was evaluated for its inhibition against HIV-1 reverse transcriptase, integrase, protease, pro-viral genome integration and viral Tat protein mediated transactivation. The effect of n-butanol fraction on the induction of pro-inflammatory cytokines secretion in Vk2/E6E7 cells and transepithelial resistance in Caco-2 and HEC-1A cells was investigated. Results: The aqueous and 50% ethanolic extracts of A. catechu showed IC§ssub§50§esub§ values of 1.8 ± 0.18 μg/ml and 3.6 ± 0.31 μg/ml, respectively in cell-free virus based assay using TZM-bl cells and HIV-1§ssub§NL4.3§esub§ (X-4 tropic). In the above assay, n-butanol fraction exhibited anti-HIV-1 activity with an IC§ssub§50§esub§ of 1.7 ± 0.12 μg/ml. The n-butanol fraction showed a dose-dependent inhibition against HIV-1§ssub§NL4.3§esub§ infection of the peripheral blood lymphocytes and against HIV-1§ssub§BaL§esub§(R-5-tropic) as well as two different primary viral isolates of HIV-1 infection of TZM-bl cells. The n-butanol fraction demonstrates a potent inhibitory activity against the viral protease (IC§ssub§50§esub§ = 12.9 μg/ml), but not reverse transcriptase or integrase. Further, in Alu-PCR no effect on viral integration was observed. The n-butanol fraction interfered with the Tat-mediated Long Terminal Repeat transactivation in TZM-bl cells, mRNA quantitation (qRT-PCR) and electrophoretic mobility shift assay (EMSA). The n-butanol fraction did not cause an enhanced secretion of pro-inflammatory cytokines in Vk2/E6E7 cells. Additionally, no adverse effects were observed to the monolayer formed by the Caco-2 and HEC-1A epithelial cells. Conclusions: The results presented here show a potential anti-HIV-1 activity of A. catechu mediated by the inhibition of the functions of the viral protein and Tat. © 2013 Nutan et al.; licensee BioMed Central Ltd

Molecular Epidemiology of HIV-1 Subtypes in India: Origin and Evolutionary History of the Predominant Subtype C

This thesis describes the translational genomics of HIV-1subtype C in India from its origin to therapeutic response with the aim to improve our knowledge for better therapeutic and preventive strategies to combat HIV/AIDS. In a systemic approach, we identified the molecular phylogeny of HIV-1 subtypes circulating in India and the time to most recent common ancestors (tMRCA) of predominant HIV-1 subtype C strains. Additionally, this thesis also studied drug resistance mutations in children, adolescents and adults, the role of host factors in evolution of drug resistance, and population dynamics of viremia and viral co-receptor tropism in perinatal transmission. Finally, the long term therapeutic responses on Indian national first-line antiretroviral therapy were also studied. In Paper I, we reported an increase in the HIV-1 recombinant forms in the HIV-1 epidemiology using a robust subtyping methodology. While the study confirmed HIV- 1 subtype C as a dominant subtype, its origin was dated back to the early 1970s from a single or few genetically related strains from South Africa, whereafter, it has evolved independently. In Paper II, the lethal hypermutations due to the activity of human apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3G (hA3G) was significantly associated with antiretroviral therapy (ART) failure in Indian HIV-1 subtype C patients. The presence of M184I and M230I mutations were observed due to the editing of hA3G in the proviral compartment but stop codons were also found in the open reading frames and the same drug resistance mutations were absent in plasma virus. Therefore, it is unlikely that the viral variants which exhibit hypermutated sequences and M184I and/or M230I will mature and expand in vivo and hence are unlikely to have any clinical significance. The high concordance of drug resistance genotyping in the plasma and proviral compartments in therapy-naïve patients, gives weight to the idea of using whole blood for surveillance of drug resistance mutations which precludes logistic challenges of cold chain transport. In Papers III and IV, we identified a substantial proportion of HIV-1 subtype C perinatally-infected older children who had a high burden of plasma viremia but also had high CD4+ T-cell counts. In addition, older children with HIV-1 subtype C infection presented a high prevalence of predicted X4 and R5/X4 tropic strains which indicates that HIV-1 subtype C strains required longer duration of infection and greater disease progression to co-receptor transition from R5- to X4-tropic strains (IV). Our studies also indicate that transmitted drug resistance is low among Indian HIV-1 infected children, adolescents (III) and adults (II). In Paper V, in a longitudinal cohort study, a good long-term response to the Indian national first-line therapy for a median of nearly four years with 2.8% viral failure, indicating the overall success of the Indian ART program. Our study also showed that three immunologically well patients with virological rebound and major viral drug resistance mutations (M184V, K103N and Y181C) during one study visit had undetectable viral load at their next visit. These findings suggest that use of multiple parameters like patients’ immunological (CD4+ T-cell count), virological (viral load) and drug resistance data should all be used to optimize the treatment switch to second line therapy. In conclusion, this translational genomics study enhances our knowledge about the HIV-1 subtype C strains circulating in India which are genetically distinct from prototype African subtype C strains. Considerably more research using appropriate models need to be performed to understand the phenotypic and biological characteristics of these strains to guide efficient disease intervention and management strategies

Evolutionary Modeling of Rate Shifts Reveals Specificity Determinants in HIV-1 Subtypes

A hallmark of the human immunodeficiency virus 1 (HIV-1) is its rapid rate of evolution within and among its various subtypes. Two complementary hypotheses are suggested to explain the sequence variability among HIV-1 subtypes. The first suggests that the functional constraints at each site remain the same across all subtypes, and the differences among subtypes are a direct reflection of random substitutions, which have occurred during the time elapsed since their divergence. The alternative hypothesis suggests that the functional constraints themselves have evolved, and thus sequence differences among subtypes in some sites reflect shifts in function. To determine the contribution of each of these two alternatives to HIV-1 subtype evolution, we have developed a novel Bayesian method for testing and detecting site-specific rate shifts. The RAte Shift EstimatoR (RASER) method determines whether or not site-specific functional shifts characterize the evolution of a protein and, if so, points to the specific sites and lineages in which these shifts have most likely occurred. Applying RASER to a dataset composed of large samples of HIV-1 sequences from different group M subtypes, we reveal rampant evolutionary shifts throughout the HIV-1 proteome. Most of these rate shifts have occurred during the divergence of the major subtypes, establishing that subtype divergence occurred together with functional diversification. We report further evidence for the emergence of a new sub-subtype, characterized by abundant rate-shifting sites. When focusing on the rate-shifting sites detected, we find that many are associated with known function relating to viral life cycle and drug resistance. Finally, we discuss mechanisms of covariation of rate-shifting sites