VapC12 ribonuclease toxin modulates host immune response during Mycobacterium tuberculosis infection.

Mechanistic understanding of antibiotic persistence is a prerequisite in controlling the emergence of MDR cases in Tuberculosis (TB). We have reported that the cholesterol-induced activation of VapC12 ribonuclease is critical for disease persistence in TB. In this study, we observed that relative to the wild type, mice infected with ΔvapC12 induced a pro-inflammatory response, had a higher pathogen load, and responded better to the anti-TB treatment. In a high-dose infection model, all the mice infected with ΔvapC12 succumbed early to the disease. Finally, we reported that the above phenotype of ΔvapC12 was dependent on the presence of the TLR4 receptor. Overall, the data suggests that failure of a timely resolution of the early inflammation by the ΔvapC12 infected mice led to hyperinflammation, altered T-cell response and high bacterial load. In conclusion, our findings suggest the role of the VapC12 toxin in modulating the innate immune response of the host in ways that favor the long-term survival of the pathogen inside the host.

Rv0495c regulates redox homeostasis in Mycobacterium tuberculosis.

Mycobacterium tuberculosis (Mtb) has evolved sophisticated surveillance mechanisms to neutralize the ROS-induces toxicity which otherwise would degrade a variety of biological molecules including proteins, nucleic acids and lipids. In the present study, we find that Mtb lacking the Rv0495c gene (ΔRv0495c) is presented with a highly oxidized cytosolic environment. The superoxide-induced lipid peroxidation resulted in altered colony morphology and loss of membrane integrity in ΔRv0495c. As a consequence, ΔRv0495c demonstrated enhanced susceptibility when exposed to various host-induced stress conditions. Further, as expected, we observed a mutant-specific increase in the abundance of transcripts that encode proteins involved in antioxidant defence. Surprisingly, despite showing a growth defect phenotype in macrophages, the absence of the Rv0495c enhanced the pathogenicity and augmented the ability of the Mtb to grow inside the host. Additionally, our study revealed that Rv0495c-mediated immunomodulation by the pathogen helps create a favorable niche for long-term survival of Mtb inside the host. In summary, the current study underscores the fact that the truce in the war between the host and the pathogen favours long-term disease persistence in tuberculosis. We believe targeting Rv0495c could potentially be explored as a strategy to potentiate the current anti-TB regimen.

Use of Off-Label Drugs in COVID-19: Clinicians' Perceptions Based on a Cross-Sectional Observational Study.

Introduction The absence of a common National Treatment Guideline during the second wave of the COVID-19 pandemic in India resulted in different treatment strategies, and the use of "off-label drugs" (OfLDs) was one of them. Aims This study aimed to assess the proportion of doctors who prescribed OfLDs, their perceived appropriateness, and the factors leading to their use. Settings and design This is an undergraduate student research project, in which a web-based cross-sectional survey was conducted on doctors who delivered care to COVID-19 patients during the second wave of the pandemic in Uttarakhand, India. Materials and methods The minimum sample size was 370 (for a 95% confidence level, an alpha error of 0.5, and a power of 80%). Data were collected electronically using a validated questionnaire after institutional ethical clearance and the participants' consent. Statistical analysis This is a descriptive-analytical study. Results We received 419 completed responses; all specialties had seen COVID-19 patients, and 91.4% (383) of the doctors had provided care to COVID-19 patients in some way or the other. About 90.7% (380) of the doctors used OfLDs; 62.5% (262) agreed that OfLDs were beneficial, and 78.9% (331) disagreed on universal steroid use. Only 34.1% (143) felt that using OfLDs was ethical. About 16.9% (71) of the doctors believed that alternative medicine was a useful treatment adjunct, and 20% (84) of doctors prescribed OfLDs under duress. About 21.2% (89) believed that Remdesivir was the main treatment for the disease, and 18.6% (78) believed that Tocilizumab was the main treatment for the disease. Personal experience, conviction, or advice from peers were among the various reasons that were put forward for using OfLDs. Conclusions The use of OfLDs during the COVID-19 pandemic in India was extensive. It was done sometimes under pressure and was largely based on confusion (multiplicity of guidelines, many times at variance with each other) as well as on a personal or low level of scientific evidence forwarded to support the use.

Perspective on nanochannels as cellular mediators in different disease conditions.

Tunnelling nanotubes (TNTs), also known as membrane nanochannels, are actin-based structures that facilitate cytoplasmic connections for rapid intercellular transfer of signals, organelles and membrane components. These dynamic TNTs can form de novo in animal cells and establish complex intercellular networks between distant cells up to 150 µm apart. Within the last decade, TNTs have been discovered in different cell types including tumor cells, macrophages, monocytes, endothelial cells and T cells. It has also been further elucidated that these nanotubes play a vital role in diseased conditions such as cancer, where TNT formation occurs at a higher pace and is used for rapid intercellular modulation of chemo-resistance. Viruses such as HIV, HSV and prions also hijack the existing TNT connections between host cells for rapid transmission and evasion of the host immune responses. The following review aims to describe the heterogeneity of TNTs, their role in different tissues and disease conditions in order to enhance our understanding on how these nanotubes can be used as a target for therapies.

Perspectives on the role of brain cellular players in cancer-associated brain metastasis: translational approach to understand molecular mechanism of tumor progression.

Brain metastasis is one of the leading causes of death among cancer patients. Cancer cells migrate to various sites and harbor different niche in the body which help cancer cells in their survival. The brain is one of the safest place where cancer cells are protected from immune cells. Breast, lung, and melanoma cancer cells have high propensity to migrate towards the brain. To enter the brain, cancer cells have to cross the blood brain barrier. Survival and finding new niche in the brain are directed by several mechanisms in which different cellular players take part such as astrocytes, microglia, Schwann cells, satellite cells, oligodendrocytes, and ependymal cells. Usually, cancer cells highjack the machinery of brain cellular players to survive in the brain environment. It has been shown that co-culture of M2 macrophage with cancer cells leads to increased proliferation and survival of cancer cells. One of the challenges of understanding brain metastasis is appropriate model system to understand dynamic interaction of cancer cells and brain cellular players. To meet this challenge, microfluidic-based devices are employed which can mimic the dynamic conditions as well as can be used for culturing human cells for personalized therapy. In this review, we have systematically reviewed the current status of the role of cellular players in brain metastasis along with explaining how translational approach of microfluidics can be employed for finding new drug target as well as biomarker for brain metastasis. Finally, we have also commented on the mechanism of action of drugs against brain metastasis.

Melioration in Anti-staphylococcal Activity of Conventional Antibiotic(s) by Organic Acids Present in the Cell Free Supernatant of Lactobacillus paraplantarum.

In view of emerging drug resistance in pathogens, there is a need to explore alternative strategies to combat infections. Use of probiotics is one such option. In this regard, efficacy of Lactobacillus plantarum has been reported against Staphylococcus aureus. Here, we propose that cell free supernatant (CFS) of Lactobacillus paraplantarum when used in combination with conventional antibiotics viz. ampicillin and oxacillin [to which the methicillin resistant Staphylococcus aureus (MRSA) strains were originally resistant] reduce the minimum inhibitory concentrations of these antibiotics, rendering the combination either synergistic or additive against the tested MRSA strain. The anti-staphylococcal activity was observed to be due to organic acids (acetic acid and lactic acid as confirmed by HPLC analysis) present in the CFS, as neutralization of the CFS with an alkali, sodium hydroxide (NaOH), caused the complete abrogation of its activity. The role of H2O2 and bacteriocin present in the CFS was also ruled out. The findings of this study suggest that cell free supernatant and ampicillin/oxacillin combination(s) might help in rejuvenating the use of conventional anti-staphylococcal antibiotics for the treatment of multi-drug resistant strains.

Role of concept map in teaching general awareness and pharmacotherapy of HIV/AIDS among second professional medical students.

OBJECTIVE: Medical students as future doctors will play an important role in caring for HIV-infected patients. This study assessed and evaluated the existing level of knowledge of MBBS students about HIV/AIDS given through lecture delivery methods and by use of concept map (CM). MATERIALS AND METHODS: This study was carried out on 150 professional MBBS students of tertiary care hospital. A pretest was conducted by giving 10 multiple choice questions (MCQ) of general awareness and 15 questions regarding pharmacotherapy of HIV/AIDS. In between pre- and post-test, a session of 1-week integrated teaching module was organized. After completion of integrated teaching, 2 h session of CM on general awareness and pharmacotherapy of HIV/AIDS was taken. A posttest was conducted using MCQs and problem-based question (PBQ) to assess the effect of integrated teaching and CM on their knowledge about HIV/AIDS. Feedback was also taken from the students to regarding their views about CM. RESULTS: There was a significant increase in student's score in MCQ test after integrated teaching than pretest (P < 0.05). There was also significant improvement in PBQ score after CM than that of after integrated teaching (P < 0.05). Students' perception about the effectiveness of CM was positive. CONCLUSION: CM can make a significant improvement in the knowledge of medical students and were motivated and developed interest in the subject.

Introducing structured viva voce examination in medical undergraduate pharmacology: A pilot study.

OBJECTIVE: Viva voce examination is an important tool of evaluation in medical examinations marred by high subjectivity. Gross subjectivity in viva voce assessment can be reduced by structuring it. MATERIALS AND METHODS: The marks obtained in theory and viva voce (traditional viva voce examination [TVVE]) of I sessional, II MBBS students were compared and a huge disparity was identified. A structured viva voce examination (SVVE) was then proposed and experimented as an objective and standardized alternative. Sets of equitable question cards for SVVE were prepared, each having eight questions with two parts each, arranged successively with increasing difficulty, domains of learning, and appropriate marks. The percentage variation in scoring in viva versus theory marks was calculated for both TVVE and SVVE, and students were grouped as Group I (+100 to +51%); Group II (+50 to -50%); Group III (-51 to -100%); Group IV (-101 to -150%); Group V (-151 to -200%); and Group VI (< -200%) variation, as? inappropriate, appropriate, inappropriate, erroneous, more erroneous and most erroneous respectively. Student's feedback on the SVVE was also obtained. RESULTS: In TVVE (n = 128), the students distributed were:none,17.2%, 23.4%, 22.7%, 11.7% and 25% in Group I, II, III, IV, V, and VI in contrast to SVVE (n = 107) as 7.5%, 57.9%, 19.6%, 6.5%, 5.6%, and 2.8%, respectively. Marked disparity of TVVE was annulled with SVVE. Student's feedback was quite encouraging with 83% overall acceptability and almost 66% preferred SVVE. CONCLUSION: SVVE was more realistic as compared to TVVE. Most of the students favored this approach.

Poetry in teaching pharmacology: Exploring the possibilities.

OBJECTIVES: To explore poetry as a tool for active learning in linking knowledge and affective domains and to find if correlating learning with imagination can be used in "assessment for learning." MATERIALS AND METHODS: After taking a conventional lecture on Asthma, a creative writing assignment in the form of poetry writing was given to the students. Different triggers were given to the students to channelize their thought pattern in a given direction that was linked to specific areas of academic relevance. Students were asked to reflect on this learning experience and the faculty was asked to evaluate the student assignment on a 5-point Likert scale. RESULTS: Most student groups scored well in the "overall assessment" of creative assignments and were rated as good or fair by the faculty. Students reflections were very informative and revealed that more than 90% of the students liked the exercise and many were too exuberant and liberal with emotional reactions that breathed positive. Around 5% students found the exercise average and another 5% found it very childish. CONCLUSION: Poetry writing turned out to be like a simulation exercise that linked academic knowledge, creativity, and the affective domain in an assumed scenario, rehearsed in free locales of mind. The metaphorical transition embedded in its subtle creation helped assess deeper understanding of the subject and the logical sequence of thought pattern.

A double-blind, randomized, controlled trial to compare the efficacy and tolerability of fixed doses of ropinirole, bupropion, and iron in treatment of restless legs syndrome (Willis-Ekbom disease).

BACKGROUND: We aimed to compare the efficacy of fixed doses of bupropion and ropinirole and iron alone for the treatment of restless legs syndrome (RLS) and to look for the tolerability of these medications. MATERIALS AND METHODS: Patients diagnosed with RLS were randomly divided into three groups with thirty patients in each group (Group A: Bupropion [300 mg/day], Group B: Ropinirole [0.25-0.5 mg/day], and Group C: Oral iron [150 mg elemental iron] along with folic acid [500 µg]). Each participant was then assessed for severity of RLS, as well as RLS-related quality at the baseline, and thereafter, every 14th day till 6 weeks based on the International Restless Legs Scale (IRLS) severity rating scale and Restless Legs Syndrome Quality of Life (RLSQoL) Questionnaire, respectively. RESULTS: IRLS scores differed significantly from baseline visit to last (F = 4.85; P = 0.01). The interaction between the time x treatment group was significant (F = 10.37; P < 0.001) showing an improvement with the therapy in all the groups. Pair-wise comparison depicted that ropinirole group differed from other two groups in IRLS score (F = 7.06; P = 0.001), which were comparable to each other. Regarding quality of life of these cases, within each group scores differed among all the four visits (F = 5.12; P = 0.002). Unlike IRLS, there was no significant difference among the RLSQOL scores between groups at any point of time (F = 1.2; P = 0.28). CONCLUSION: RLS severity decreased across time in all three groups; however, the ropinirole treatment was better than the bupropion and iron-folate therapy. Moreover, RLS-related quality of life although improved among all groups, it was comparable among three groups.

Translation and validation of restless leg syndrome quality of life questionnaire in Hindi language.

BACKGROUND: Restless legs syndrome (RLS) itself as well as the problems that are secondary to RLS may worsen the quality of life. AIM: The aim of this study is to translate and validate the Restless Legs Syndrome Quality of Life (RLS-QOL) questionnaire in Hindi language. SETTINGS AND DESIGNS: Patients attending psychiatry OPD and sleep clinic in a tertiary care teaching hospital. MATERIALS AND METHODS: Thirty four consecutive patients of RLS and twenty nine control subjects were included in the study. Permission for translation and validation of RLS-QOL questionnaire scale was obtained. Translation was done according to the guidelines provided by the publisher. After translation, the final version of the scale was applied in both the groups to find the reliability and validity. STATISTICAL ANALYSIS: SPSS, version 17.0 was used for the analysis. Independent sample t test was used to compare age. Chi-square test was applied to compare non-parametric variables. Pearson's and Spearman's correlations were used to find out the correlation between parametric and non-parametric items, respectively. Reliability analysis was done by using Cronbach's alpha. RESULTS: Among the RLS subjects, mean Insomnia Severity Index (ISI) score, International Restless Legs Syndrome Severity Rating Scale (IRLS) score and Computed Score of RLS-QOL questionnaire were 25.43 (7.39), 12.7 (8.34) and 29.8 (8.39), respectively. A statistically significant difference was observed between both the groups on all these scores (ISI: t= -11.2, P<0.001, 95% CI= -22.62 to -15.76; IRLS: t = -8.1, P< 0.001, 95% CI =-15.81 to -9.58; RLS-QOL: t =-19.07, P<0.001 with 95% CI = -32.69,- 28.83). A significant correlation between ISI and RLS-QOL (r² = 0.59; P< 0.001) was seen. Most of the items also showed good correlation with each other. Internal consistency done by Cronbach's alpha showed good correlation (0.85). CONCLUSION: Hindi version of the Restless Legs Syndrome Quality of Life (RLS-QOL) questionnaire is a valid and reliable tool for the assessment of the quality of life in patients with RLS.