Direct Microbial Identification using An Automated Microbial Identification System to Facilitate the EUCAST RAST Method Without Mass Spectrometry.

Gram-negative (GN) sepsis is a medical emergency where management in resource-limited settings relies on conventional microbiological culture techniques providing results in 3-4 days. Recognizing this delay in turnaround time (TAT), both EUCAST and CLSI have developed protocols for determining AST results directly from positively flagged automated blood culture bottles (+aBCs). EUCAST rapid AST (RAST) protocol was first introduced in 2018, where zone diameter breakpoints for four common etiological agents of GN sepsis, i.e., Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii complex can be reported. However, those clinical laboratories that have implemented this method in their routine workflow rely on mass spectrometry-based microbial identification, which is not easily available, thus precluding its implementation in resource-limited settings. To circumvent it, we evaluated a direct inoculum protocol (DIP) using a commercial automated microbial identification and antimicrobial susceptibility testing system (aMIAST) to enable early microbial identification within 8 h of positive flagging of aBC. We evaluated this protocol from January to October 2023 to identify the four RAST reportable GN (RR-GN) in the positively flagged aBC. The microbial identification results in DIP were compared with the standard inoculum preparation protocol (SIP) in aMIAST. Of 204 +aBCs with monomorphic GN (+naBC), one of the 4 RR-GN was identified in 105 +naBCs by SIP (E. coli: 50, K. pneumoniae: 20, P. aeruginosa: 9 and A. baumannii complex: 26). Of these, 94% (98/105) were correctly identified by DIP whereas major error and very major error rates were 6% (7/105) and 1.7% (4/240), respectively. When DIP for microbial identification is done using the EUCAST RAST method, provisional clinical reports can be provided within 24 h of receiving the sample. This approach has the potential to significantly reduce the TAT, enabling early institution of appropriate antimicrobial therapy.

Hit or Miss: The Dilemma of Specimen Selection for Microbiological Diagnosis of Rhino-Orbital-Cerebral- COVID-Associated Mucormycosis (ROCM-CAM).

BACKGROUND: We diagnosed various cases of rhino-orbital-cerebral- COVID-associated Mucormycosis (ROCM-CAM) during India's second wave of COVID-19. This helped formulate novel suggestions for improving laboratory output, applicable anywhere in the world. METHODS: To diagnose ROCM-CAM by microbiological methods, we used direct microscopy and conventional culture on various clinical samples within the shortest turn-around time. DESIGN: Prospective single-center observational study. PARTICIPANTS: Patients with ROCM-CAM. RESULTS: Of 113 suspected cases of ROCM-CAM during May 2021, direct microscopy and culture could confirm the disease in 87.61% and 44.25% of patients, respectively. The highest pathogen isolation was seen from maxillary bone fragments, FESS-guided biopsy from pterygopalatine fossae, nasal turbinates and nasal mucosal biopsy. Direct microscopy could diagnose the disease in almost 40% of patients within 24 hours and 60% within two days. Conventional cultures yielded Rhizopus spp. (86%) as the commonest fungal pathogen followed by Mucor spp. (12%) within 7 days. Deep tissue biopsies are more useful for rapid diagnosis than superficial specimens. Routine fungal cultures can supplement case detection and help prognosticate survivors. CONCLUSION: The management of ROCM is a surgical emergency. The diagnosis of the condition must therefore be prompt and precise. Despite ongoing antifungal therapy, nasal mucosal tissue, FESSguided, and intra-operative tissue biopsies showed the pathogen's highest diagnostic yield. The diagnostic index improved further when multiple (4-5) high-quality specimens were collected. Nasal swabs and crusts, among the most commonly requested specimens worldwide, were found to have an overall low diagnostic potential.

Anti-SARS-CoV-2 IgG status among frontline health workers involved in COVID-19 survey at Jodhpur, Rajasthan, India.

Background: Asymptomatic carriers are responsible for the consistent spread of coronavirus disease 2019 (COVID-19) in the community. The Government of India has deputed house-to-house survey teams to aid in identifying asymptomatic individuals and their susceptible contacts. We selected door-to-door survey teams of a COVID-19 red zone in western India and determined their infectioncontrol practices and anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) immunoglobin G (IgG) status. Materials and Methods: This single-day prospective cross-sectional study was conducted by the Department of Microbiology of a tertiary care hospital of Jodhpur, in collaboration with the Rajasthan State Health Services. Participants were asked to fill out a questionnaire regarding personal protective equipment (PPE) use after written informed consent. Venous blood samples were collected and Kavach enzyme-linked immunosorbent assay (ELISA) (J Mitra and Co.) was performed to determine anti-SARS-CoV-2 IgG status. Results: Out of the total 39 participants, IgG antibody was detected in four. Of them, three reported mild symptoms in the past. Out of two previously real-time polymerase chain reaction (RT-PCR) SARS-CoV-2-positive participants, only one had detectable IgG antibodies (Ab) in serum. Cloth mask was used by 24, N95 mask by 11, and surgical masks by four. Conclusion: Anti-SARS-CoV-2 IgG Abs were detected among four members of house-to-house COVID-19 survey teams in Jodhpur. Most of the team members used cloth masks, whereas the Government of India guidelines has recommended triple-layered surgical masks as minimum essential PPE for healthcare workers in India. More such studies should be conducted to ascertain infection prevention and control practices among such vulnerable frontline workers in our country.

Retrospection of heatwave and heat index.

The frequency and intensity of extreme events especially heat waves (HW) are growing all around the world which ultimately poses a serious threat to the health of individuals. To quantify the effects of extreme temperature, appropriate information, and the importance of HW and heat index (HI) are carefully discussed for different parts of the world. Varied definitions of the HW and HI formula proposed and used by different countries are carried out systematically continent-wise. Different studies highlighted the number of definitions of HW; however, mostly used Steadman's formulae, which was developed in the late 1970s, for the calculation of HI that uses surface air temperature and relative humidity as climatic fields. Since then, dramatic changes in climatic conditions have been observed as evident from the ERA5 datasets which need to be addressed; likewise, the definition of HW, which is modified by the researchers as per the geographic conditions. It is evident from the ERA5 data that the temperature has increased by 1-2 °C as compared to the 1980s. There is a threefold increase in the number of heatwave days over most of the continents in the last 40 years. This study will help the researcher community to understand the importance of HW and HI. Furthermore, it opens the scope to develop an equation based on the present scenario keeping in mind the basics of an index as considered by Steadman.

The usefulness of crossword puzzle as a self-learning tool in pharmacology.

INTRODUCTION: Pharmacology is perceived as a volatile subject as it's difficult to recall and recite the core of the subject. Enriching the learning environment through incorporation of a variety of teaching and learning strategies and methods yields enhanced learning. Crossword puzzles provide expansion of vocabulary, stimulate thinking capacity, boost confidence, and fasten up the learning capacity; hence, the present study was conducted to investigate the usefulness of crossword puzzle as an innovative self-learning tool in pharmacology. METHODS: This prospective study was conducted among 5th semester students of the second professional MBBS course. A total of 139 students participated in this study and were evaluated with formative examination and feedback questionnaire. Permission was taken from Institutional Ethics Committee for the study. A crossword puzzle consisting of 32 questions on endocrine pharmacology was prepared and divided into two sections: the across section had 17 questions and the down section contained 15 questions. The data were analyzed, using Graph Pad Software and presented as percentage of the responses. RESULTS: On average, out of 32 questions, one mark each, the students scored 52.69% and all students responded correctly on questions on the topic of hormonal contraceptives. 75.5% of the students had an enjoyable experience and the majority of them agreed that it helped them enhance their knowledge of drugs, remember diseases and drug names, and overall learning about the topic. They were also of the opinion that this should be inculcated in pharmacology curriculum. CONCLUSION: Incorporation of crossword puzzles, as an adjunct tool, was useful as majority of the students reported that this improved their attitude of learning, thereby improving their performance.

Introducing objective structured practical examination as a method of learning and evaluation for undergraduate pharmacology.

OBJECTIVE: Assessment method can influence student learning. Use of objective structured practical examination (OSPE) has been reported in various institutes with great benefits. We evaluated OSPE for the assessment of practical skills in pharmacology examination for undergraduate medical students and compared it with conventional practical examination (CPE). MATERIALS AND METHODS: After sensitizing the 2nd year MBBS students to OSPE, the students were divided into four batches with twenty students in each batch. Students were assessed by attending five OSPE stations, each for duration of 5 min. The effectiveness was assessed through a student's feedback questionnaire and was checked for its reliability by Cronbach's alpha. The result of OSPE was compared with that of CPE of the same batch. RESULTS: Cronbach's alpha of the feedback questionnaire was 0.71, with high internal consistency. The feedback given was categorized into three domains: cognitive, psychomotor, and affective, and an assessment was also done for its further use. In cognitive domain, 74% of the students felt that the questions asked and the syllabus taught were well correlated. In psychomotor domain, 81% agreed that it is excellent for assessing the applied part of the subject. Seventy percent of students opined that it was associated with lesser stress than CPE. On overall assessment, 76% rated this methodology as good/satisfactory and 23% as excellent in terms of better scoring. There was a significant difference in the mean score between the results of OSPE and CPE (P < 0.001, df = 158, confidence interval = 95%). CONCLUSION: OSPE is a feasible and skill enhancing tool for the assessment in pharmacology examinations for undergraduate students.

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.