Antimicrobial efficiency against fish pathogens on the green synthesized silver nanoparticles.

Fish-borne pathogens such as A. hydrophila and F. aquidurense are the most resistant strains in pisciculture farming. Removing the aforementioned pathogens without antibiotics presents a formidable challenge. To overcome this problem, silver nanoparticles (AgNPs) are synthesized using silver nitrate, water medium, and as an AzadirachtaIndica leaf extract via the green synthesis route. X-ray diffraction (XRD) pattern results authenticate the synthesized material is the face-centered cubic structure of silver. The optical absorption edge of the synthesized product was found at the wavelength of 440 nm from the UV-visible spectra, which is confirmed to relate to the Surface Plasmon Resonance peaks of silver particles. In addition, the optical band gap value of the synthesized Ag sample is measured to be 2.81 eV from the obtained optical absorption spectra. EDX spectrum of the synthesized product also supports confirming the silver particle formation. The FT-IR spectra of the neem extract and silver nanoparticles showed their characteristic functional groups, respectively. The presence of bands between 1000 cm-1 to 500 cm-1 indicates to the formation of silver particles. Spherical particles appeared in the synthesized Ag using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The particle size of Ag NPs was measured as 40 nm and 62 ± 10 nm by TEM and Dynamic Light Scattering (DLS). The zeta potential was also measured as -12 mV showing the synthesized sample's stable nature. Using the DPPH assay, synthesized AgNPs were taken along with the various concentrations of ascorbic acid (20, 40, 60, 80, and 100 µg/mL) to examine the free radical scavenging activity (RSA). RSA value is higher (84 ± 2 %) for synthesized AgNPs at higher concentration (100 µg/mL) than 21 ± 2 % at low concentration (100 µg/mL). The antimicrobial efficacy of the AgNPs against A. hydrophila and F. aquidurense was performed through the agar diffusion method and its results showed the inhibitory zones of the F.aquidurense and A. hydrophila were measured as 25 ± 3 mm, and 28 ± 4 mm respectively. The synthesized Ag particles showed excellent antimicrobial and antioxidant properties confirmed by antimicrobial and DPPH experiments. It implies that the green synthesized silver nanoparticles could be a good alternative for antibiotics in aquaculture farms. The exposure of low concentrations of silver nanoparticles to zebrafish and brine shrimp does not affect the viability and morphology. The exposure of silver nanoparticles in the fisheries in optimized concentration and time could control the fish-borne pathogens without antibiotics.

Nailfold capillaroscopy changes associated with anti-RNP antibodies in systemic lupus erythematosus.

Anti-U1RNP antibody is associated with distinct organ involvement in patients with systemic lupus erythematosus (SLE). Nailfold capillaroscopy (NFC) allows non-invasive assessment of microvascular abnormalities in several connective tissue diseases. The objective of this study is to determine the association of anti-U1RNP antibody with microvascular changes by NFC in RNP-positive SLE patients in comparison with RNP-negative SLE patients (negative disease controls) and mixed connective tissue disease (MCTD) cases (positive disease controls). NFC examination was performed in consecutive patients with SLE with or without anti-U1RNP positivity. MCTD patients were recruited as disease controls. Abnormalities noted in the three groups were compared using non-parametric tests. Ordinal logistic or linear regression was used wherever applicable. 81 patients were studied, of whom 28 were diagnosed as RNP-positive SLE (age 30.0 ± 10.37; 26 females), 26 were RNP-negative SLE (age 29.42 ± 9.20; 25 females) and 27 had MCTD (age36.5 ± 9.70; 25 females). RNP-positive SLE patients had more frequent giant capillaries, enlarged capillaries and ramified capillaries as compared to RNP-negative SLE (p = 0.05, < 0.01 and 0.03, respectively). The capillary density was lower in patients with MCTD as compared with patients with RNP-positive SLE (5.11 ± 1.69/mm vs 7.25 ± 1.38/ mm, p < 0.01) and RNP-negative SLE (8.92 ± 1.13/mm, p < 0.01). The reduction in capillary density was less severe in patients with RNP-negative SLE as compared with RNP-positive SLE (OR = 0.1058 [95% CI = 0.02-0.546], p < 0.01) which was independent of the presence of Raynaud's phenomenon, interstitial lung disease and disease duration. Presence of anti-U1RNP antibody is associated with notable patterns of microvascular abnormalities in SLE. These NFC abnormalities are noted more profoundly in patients with MCTD and are less marked in RNP-negative SLE patients.

Deep CNN with Hybrid Binary Local Search and Particle Swarm Optimizer for Exudates Classification from Fundus Images.

Diabetic retinopathy is a chronic condition that causes vision loss if not detected early. In the early stage, it can be diagnosed with the aid of exudates which are called lesions. However, it is arduous to detect the exudate lesion due to the availability of blood vessels and other distractions. To tackle these issues, we proposed a novel exudates classification from the fundus image known as hybrid convolutional neural network (CNN)-based binary local search optimizer-based particle swarm optimization algorithm. The proposed method from this paper exploits image augmentation to enlarge the fundus image to the required size without losing any features. The features from the resized fundus images are extracted as a feature vector and fed into the feed-forward CNN as the input. Henceforth, it classifies the exudates from the fundus image. Further, the hyperparameters are optimized to reduce the computational complexities by utilization of binary local search optimizer (BLSO) and particle swarm optimization (PSO). The experimental analysis is conducted on the public ROC and real-time ARA400 datasets and compared with the state-of-art works such as support vector machine classifiers, multi-modal/multi-scale, random forest, and CNN for the performance metrics. The classification accuracy is high for the proposed work, and thus, our proposed outperforms all the other approaches.

Validity evidence for a novel instrument assessing medical student attitudes toward instruction in implicit bias recognition and management.

BACKGROUND: Implicit bias instruction is becoming more prevalent in health professions education, with calls for skills-based curricula moving from awareness and recognition to management of implicit bias. Evidence suggests that health professionals and students learning about implicit bias ("learners") have varying attitudes about instruction in implicit bias, including the concept of implicit bias itself. Assessing learner attitudes could inform curriculum development and enable instructional designs that optimize learner engagement. To date, there are no instruments with evidence for construct validity that assess learner attitudes about implicit bias instruction and its relevance to clinical care. METHODS: The authors developed a novel instrument, the Attitude Towards Implicit Bias Instrument (ATIBI) and gathered evidence for three types of construct validity- content, internal consistency, and relationship to other variables. RESULTS: Authors utilized a modified Delphi technique with an interprofessional team of experts, as well as cognitive interviews with medical students leading to item refinement to improve content validity. Seven cohorts of medical students, N = 1072 completed the ATIBI. Psychometric analysis demonstrated high internal consistency (α = 0.90). Exploratory factor analysis resulted in five factors. Analysis of a subset of 100 medical students demonstrated a moderate correlation with similar instruments, the Integrative Medicine Attitude Questionnaire (r = 0.63, 95% CI: [0.59, 0.66]) and the Internal Motivation to Respond Without Prejudice Scale (r = 0.36, 95% CI: [0.32, 0.40]), providing evidence for convergent validity. Scores on our instrument had low correlation to the External Motivation to Respond Without Prejudice Scale (r = 0.15, 95% CI: [0.09, 0.19]) and the Groningen Reflection Ability Scale (r = 0.12, 95% CI: [0.06, 0.17]) providing evidence for discriminant validity. Analysis resulted in eighteen items in the final instrument; it is easy to administer, both on paper form and online. CONCLUSION: The Attitudes Toward Implicit Bias Instrument is a novel instrument that produces reliable and valid scores and may be used to measure medical student attitudes related to implicit bias recognition and management, including attitudes toward acceptance of bias in oneself, implicit bias instruction, and its relevance to clinical care.

Creatively tiding over the crisis.

Background: A patient with a retained intraocular metallic foreign body post-trauma was taken up for vitrectomy and intraocular foreign body removal. Unfortunately, the intraocular magnet was not available at the moment on the table! How a little bit of creativity and innovative thought helped us tide over this crisis is the content of this video. Purpose: To demonstrate magnetization of a metallic surgical instrument for temporary use in the event of unavailability of the intraocular magnet for intraocular foreign body removal. Synopsis: A ferromagnetic substance can be magnetized temporarily using an existing magnet. We obtained a general-purpose magnet and wrapped it in sterile plastic, using which we magnetized normal intraocular forceps and a Micro Vitreo Retinal (MVR) blade by giving about 20-30 strokes over the magnet in a single direction. This aligned the magnetic domains in the metal in a parallel fashion. These Do It Yourself (DIY)- magnetic instruments were then effectively utilized to remove the metallic intraocular foreign body. Highlights: The video showcases effectively harnessing the available resources and tiding over the dearth of a necessary instrument, with the right use of an innovative idea and some creativity!. Video link: https://youtu.be/QtRC-AK5FLU.

A comparative study on perception and use of generic drugs between public and private health practitioners.

Context: The perception of generic drugs may vary significantly between government and private doctors because physicians in the private sector have more prescribing choices and flexibility. Hence, this study was undertaken to analyse the knowledge, attitude and perception (KAP) of government and private physicians on generic drugs. Materials and Methods: This was a questionnaire-based cross-sectional study conducted among physicians working in public and private health sectors. The questionnaire had 25 closed-ended questions related to the KAP of generic medicine. The overall scores were categorised using Bloom's cut-off point. The Chi-square or Mann-Whitney U-test was used to compare the differences between the two groups. Results: About 80% of the participants in both groups agreed that generic medicines contain the same active ingredients as brand-name drugs, are less expensive and are available in the Indian market. Nearly 84% of government physicians and only 64% of private physicians believed that generic medicines are just as effective and secure as branded medicines (P - 0.003). The majority of physicians from both groups concurred that there is a lack of quality check in generic drug manufacturing, and they require more information about bioequivalence studies. In both categories, about 75% of participants preferred generic medications for their patients. However, in both groups, more than 50% of physicians were concerned about therapeutic failure and expressed reluctance to prescribe generic medications in life-threatening situations. Conclusions: Knowledge and acceptance of generic drugs regarding efficacy, safety, bioequivalence and therapeutic failure are low among both government and private physicians.

Sociocultural and drug-related factors associated with adherence to iron-folic acid supplementation among pregnant women - A mixed-methods study.

BACKGROUND: The National Family Health Survey of India (NFHS-5) revealed a lapse in the advancement of mitigating anemia despite free supplementation of iron-folic acid tablets (IFAT) and improvement in IFAT coverage during pregnancy. The local sociocultural beliefs and community perspective toward IFAT are pivotal in reducing the gap between IFAT coverage and consumption. Hence, we proposed the study to assess adherence to IFAT among rural pregnant women and explore the associated factors. MATERIALS AND METHODS: The present study was conducted as a mixed-methods study with a sequential exploratory design in a rural setting of the Model Rural Health Research Unit (MRHRU) from October 2020 to May 2021. Ten focus group discussions (FGDs; 8 - antenatal women, 1 - mother/mother-in-law, and 1 - health care worker) were conducted, and framework analysis was done to identify themes followed by a quantitative survey with a semi-structured questionnaire among antenatal women (n = 236). Logistic regression was used to analyze factors associated with adherence. RESULTS: The major themes that emerged from FGDs were the sociocultural factors (gender norms, communal fallacies), lack of awareness, and drug-related factors (unpalatability, misperceptions, and experienced side effects). Around 57% were adherent to IFAT. Side effects experienced on IFAT intake (P = 0.001, OR = 2.33), misconceptions regarding IFAT, like weight gain in IFAT use (P = 0.001, OR = 2.86), a big baby with IFAT use (P = 0.000, OR = 5.93) negatively influenced adherence. CONCLUSIONS: The significant gaps between IFAT coverage and consumption surrounded the unpleasant odor and stench of IFAT, side effects, lack of individualized counseling, and misperceptions regarding IFAT use.

Tuning the physiochemical properties of polycaprolactone-hydroxyapatite composite films by gamma irradiation for biomedical applications.

Physiochemical properties of polycaprolactone-hydroxyapatite (PCL-HAp) composites were investigated in the pristine and after irradiation of γ rays (25, 50, 75, and 100 kGy). PCL-HAp composites were synthesized by solvent evaporation and characterized using spectroscopic methods as well as biological assays. The surface roughness (RMS) of the irradiated composite film (at 75 kGy) was 80 times higher than that of the pristine. Irradiation tailors the contact angle of the films from 77° to 90° (at 100 kGy). A decrease in particle size (at 100 kGy) of HAp nanorods in PCL-HAp composites film was observed. The XRD peak of PCL was slightly shifted from 21.2° to 21.7° (at 100 kGy) with the decrease in crystallite size. The peak intensity of the PCL and HAp altered on irradiation that was confirmed by FTIR and Raman analysis. Further, the bandgap of the irradiated film was lowered by 13 % (at 25 kGy). The luminescence intensity decreased due to the non-radiative process induced by the irradiation defects. All the samples possess hemocompatibility percentage of <10 % as per ASTM standards. At 75 kGy, fibroblast cell proliferation was higher than the pristine and other doses. The gamma-irradiated PCL-HAp composite films are potential candidates for tissue engineering applications.

Evaluation of glycemic status among hypercholesterolemic patients on atorvastatin in a tertiary care hospital - A retrospective study.

INTRODUCTION: Statins are effective in reducing low-density lipoprotein cholesterol and are favorable in primary and secondary prevention of cardiovascular disease. Recent large trials have linked the use of statins and increased incidence of new-onset diabetes mellitus, the possibility of worsening of glucose level in individuals with diabetes following statin therapy, and this possibility is increased with the use of atorvastatin. This study was undertaken to analyze the possibility of the diabetogenic potential of atorvastatin among hypercholesterolemic patients. MATERIALS AND METHODS: This retrospective cohort study was conducted in the cardiology department from July 2019 to December 2019. Patients on atorvastatin for more than 6 months with normoglycemia on commencement of therapy were included. The occurrence of prediabetes or new-onset diabetes mellitus after atorvastatin therapy is the outcome of the study. Adverse drug effects to atorvastatin were also recorded and WHO-UMC causality assessment was performed. Descriptive statistics were performed for baseline and demographic characteristics. RESULTS: Sixty study participants were included in the study. Eighteen (30%) study participants developed prediabetes with an HbA1c value of 5.97 ± 0.22 and 17 (28%) of participants developed new-onset diabetes mellitus with an HbA1c value of 7.24 ± 0.50. Atorvastatin at dose of 40 mg was found to be the most frequently prescribed dose. CONCLUSION: Atorvastatin has a dose-dependent risk of developing new-onset diabetes mellitus. Hence, the following statin therapy glycemic status should be periodically monitored especially in patients with a large dose of atorvastatin and also in patients with higher risk factors for diabetes.

Detection of the mineral constituents in human renal calculi by vibrational spectroscopic analysis combined with allied techniques Powder XRD, TGA, SEM, IR imaging and TXRF.

Detection of the mineral constituents in a batch of 310 samples of human urinary calculi (kidney stones-235 and bladder stones-75) combined with a semi-quantitative analysis has been presented on the basis of Fourier Transform based IR and Raman spectral measurements. Some of the observed characteristic IR and Raman bands have been proposed as 'Marker Bands' for the most reliable identification of the constituents. A detailed vibrational spectral analysis combined with a DFT level calculation for the functional groups in Calcium Oxalate Monohydrate (COM), Magnesium Ammonium Phosphate Hexahydrate (MAPH), Calcium Hydrogen Phosphate Dihydrate (CHPD), Penta-Calcium Hydroxy-Triphosphate (PCHT) and Uric Acid (UA) has been proposed. It has been shown that the identified mineral constituents as major or minor components can be deduced from the application of Lambert-Beer law of radiation absorption and results are in agreement with quantitative Spectral Data base. This simple method has the potential to be integrated into the management of Urolithiasis, a process of forming renal calculi in the kidney, bladder and/or urethra. Employment of powder XRD, TGA, SEM, TXRF and IR Imaging techniques has provided additional support for the proposed foolproof identification of the mineral constituents. Among the mineral constituents, Calcium Oxalate Monohydrate, Calcium Oxalate Dihydrate or their mixture account for 85% of the total number of samples; the remaining 15% and 5% samples contain Phosphate and Uric acid stones respectively.

Adapting SQUIRE 2.0 to Create a Quality Improvement Evidence-Based Medicine Critical Appraisal Tool (QI-EBM-CAT) for Graduate Medical Education Trainees.

Background: Evidence-based medicine (EBM) has long been taught to physician trainees for critical appraisal of research manuscripts. There is no parallel or similar framework to guide trainees in the appraisal of quality improvement (QI) literature. Objective: To adapt existing guidelines of QI manuscript reporting into an educational QI-EBM appraisal tool to help residents distinguish research and QI manuscripts, assess QI designs and methodologies, and evaluate QI manuscripts' strengths and weaknesses. Methods: Between 2018 and 2021, we developed a QI-EBM critical appraisal tool (QI-EBM-CAT) and performed 3 plan-do-study-act cycles to refine the tool based on JAMA and SQUIRE 2.0 guidelines. We then surveyed residents regarding the usefulness of the tool and their confidence in evaluating QI manuscripts before and after completing a QI-EBM workshop using the QI appraisal tool. Results: Sixty-six of 74 internal medicine postgraduate year (PGY)-1 to PGY-3 residents (89.2%) completed the workshop and assessment surveys in 2021. The workshop was found to be moderately to very useful by 85.1% (63 of 74) of residents as a framework for QI manuscript critical analysis. The summary confidence score in QI manuscript critical appraisal improved from a 64% rating of moderately to very confident in the pre-period to 94.6% in the post-period (P<.001) with statistical improvements in all 5 confidence areas assessed (P<.001). Conclusions: The QI-EBM-CAT, designed to teach residents how to critically assess QI manuscripts using EBM principles, resulted in subjective improvements in confidence of QI manuscript analysis.

Pharmacogenomic landscape of COVID-19 therapies from Indian population genomes.

Aim: Numerous drugs are being widely prescribed for COVID-19 treatment without any direct evidence for the drug safety/efficacy in patients across diverse ethnic populations. Materials & methods: We analyzed whole genomes of 1029 Indian individuals (IndiGen) to understand the extent of drug-gene (pharmacogenetic), drug-drug and drug-drug-gene interactions associated with COVID-19 therapy in the Indian population. Results: We identified 30 clinically significant pharmacogenetic variants and 73 predicted deleterious pharmacogenetic variants. COVID-19-associated pharmacogenes were substantially overlapped with those of metabolic disorder therapeutics. CYP3A4, ABCB1 and ALB are the most shared pharmacogenes. Fifteen COVID-19 therapeutics were predicted as likely drug-drug interaction candidates when used with four CYP inhibitor drugs. Conclusion: Our findings provide actionable insights for future validation studies and improved clinical decisions for COVID-19 therapy in Indians.

Genetic epidemiology of autoinflammatory disease variants in Indian population from 1029 whole genomes.

BACKGROUND: Autoinflammatory disorders are the group of inherited inflammatory disorders caused due to the genetic defect in the genes that regulates innate immune systems. These have been clinically characterized based on the duration and occurrence of unprovoked fever, skin rash, and patient's ancestry. There are several autoinflammatory disorders that are found to be prevalent in a specific population and whose disease genetic epidemiology within the population has been well understood. However, India has a limited number of genetic studies reported for autoinflammatory disorders till date. The whole genome sequencing and analysis of 1029 Indian individuals performed under the IndiGen project persuaded us to perform the genetic epidemiology of the autoinflammatory disorders in India. RESULTS: We have systematically annotated the genetic variants of 56 genes implicated in autoinflammatory disorder. These genetic variants were reclassified into five categories (i.e., pathogenic, likely pathogenic, benign, likely benign, and variant of uncertain significance (VUS)) according to the American College of Medical Genetics and Association of Molecular pathology (ACMG-AMP) guidelines. Our analysis revealed 20 pathogenic and likely pathogenic variants with significant differences in the allele frequency compared with the global population. We also found six causal founder variants in the IndiGen dataset belonging to different ancestry. We have performed haplotype prediction analysis for founder mutations haplotype that reveals the admixture of the South Asian population with other populations. The cumulative carrier frequency of the autoinflammatory disorder in India was found to be 3.5% which is much higher than reported. CONCLUSION: With such frequency in the Indian population, there is a great need for awareness among clinicians as well as the general public regarding the autoinflammatory disorder. To the best of our knowledge, this is the first and most comprehensive population scale genetic epidemiological study being reported from India.

Effect of Nickel Doping on the Properties of Hydroxyapatite Nanoparticles.

Herein, we studied the effect of nickel (Ni) doping on the properties of hydroxyapatite (HAp) nanoparticles synthesized by facile ultrasonication assisted wet chemical synthesis process. Various doping concentrations of nickel, i.e., 0.01 M, 0.05 M and 0.10 M, were used to dope into hydroxyapatite nanoparticles. The synthesized nanoparticles were characterized by X-ray diffraction (XRD) pattern, scanning electron spectroscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, UV-Visible spectroscopy and Raman-scattering spectroscopy. The detailed structural characterizations confirmed that the crystallite sizes of the Ni-doped hydroxyapatite nanoparticles were reduced up to 53% compared to pure hydroxyapatite upon the doping of different concentrations of Ni ions. The agglomeration in the nanoparticles was also reduced by increasing the doping concentration of Ni ions. The XRD studies revealed that the average crystallite size of the synthesized Ni-doped HAp was decreased with increasing the concentration of Ni2+ ion doping and this observation was well-consistent with the SEM results. The FTIR and Raman studies well-confirmed the formation of pure HAp and Ni-doped HAp. Further, doping with Ni creates a new level of energy between the conductive band and the valence band and hence with increasing the concentration of Ni2+, the intensity in the UV-vis spectra was enhanced.

Morphological and elemental mapping of gallstones using synchrotron microtomography and synchrotron X-ray fluorescence spectroscopy.

BACKGROUND AND AIM: Regional differences in gallstone (GS) composition are well documented in the Indian subcontinent. The reasons for the same are unknown. Etiopathogenesis of GS remains elusive despite advances in instrumentation. This was an in-depth analysis of the chemical, structural, and elemental composition of GS with special reference to synchroton studies. METHODS: We used high-end sensitive analytical complementary microscopic and spectroscopic methods techniques, such as X-ray diffraction, scanning electron microscopy, Fourier transform infrared, synchrotron X-ray fluorescence spectroscopy (SR-XRF), and 2D and 3D synchrotron microtomography (SR-µCT), to study the ultra structure and trace element composition of three major types of GS (cholesterol, mixed, and pigment). SR-XRF quantified the trace elements in GS. RESULTS: The cholesterol GS (monohydrate and anhydrate) were crystalline, with high calcium content. The pigment GS were amorphous, featureless, black, and fragile, with high calcium bilirubinate and carbonate salts. They had the highest concentration of iron (average 31.50 ppm) and copper (average 92.73 ppm), with bacterial inclusion. The mixed stones had features of both cholesterol and pigment GS with intermediate levels of copper (average 20.8 ppm) and iron (average 17.78 ppm). CONCLUSION: SR-µCT has, for the first time, provided cross-sectional computed imaging delineating the framework of GS and mineral distribution. It provided excellent mapping of cholesterol GS. SR-XRF confirmed that pigment GS had high concentrations of copper and iron with bacterial inclusions, the latter possibly serving as a nidus to the formation of these stones.

How to Make or Break Implicit Bias Instruction: Implications for Curriculum Development.

PURPOSE: To analyze faculty experiences regarding facilitating discussions as part of the institution's curriculum on racial and ethnic implicit bias recognition and management. METHOD: Between July 2014 and September 2016, the authors conducted 21 in-depth interviews with faculty who had experience teaching in implicit bias instruction or were interested in facilitating discussions related to implicit bias and the Implicit Association Test. Grounded theory methodology was used to analyze interview transcripts. RESULTS: Participants identified challenges that affect their ability to facilitate instruction in implicit bias. Faculty described the influence of their own background and identities as well as the influence of institutional values on their ability to facilitate implicit bias discussions. They noted the impact of resistant learners and faculty during discussions and made suggestions for institutional measures including the need for implementation of formalized longitudinal implicit bias curricula and faculty development. CONCLUSIONS: Faculty facilitating sessions on implicit bias must attend faculty development sessions to be equipped to deal with some of the challenges they may face. Buy-in from institutional leadership is essential for successful implementation of implicit bias teaching, and medical educators need to consider formalized longitudinal curricula addressing the recognition and management of implicit biases.

Chemical and structural analysis of gallstones from the Indian subcontinent.

Representative gallstones from north and southern parts of India were analyzed by a combination of physicochemical methods: X-ray diffraction (XRD), infrared spectroscopy (IR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), CHNS analysis, thermal analysis and Nuclear Magnetic Resonance (NMR) spectroscopy (1H and 13C). The stones from north Indian were predominantly consisting of cholesterol monohydrate and anhydrous cholesterol which was confirmed by XRD analysis. FTIR spectroscopy confirmed the presence of cholesterol and calcium bilirubinate in the south Indian gallstones. EDX spectroscopy revealed the presence of carbon, nitrogen, oxygen, calcium, sulfur, sodium and magnesium and chloride in both south Indian and north Indian gallstones. FTIR and NMR spectroscopy confirmed the occurrence of cholesterol in north Indian gallstones. The respective colour of the north Indian and south Indian gallstones was yellowish and black. The morphology of the constituent crystals of the north Indian and south Indian gallstones were platy and globular respectively. The appreciable variation in colour, morphology and composition of south and north Indian gallstones may be due to different food habit and habitat.

Younger patients with primary Sjögren's syndrome are more likely to have salivary IgG anti-muscarinic acetylcholine receptor type 3 antibodies.

Acetylcholine type 3 receptor (M3R) is recognized as an autoantigen in primary Sjögren's syndrome (pSS). Assay of anti-M3R antibody levels in serum is fraught with low sensitivity for diagnosis of pSS. Salivary assay is more likely to improve the diagnostic accuracy. Patients with pSS classified either by the American European Consensus Group (AECG) or American college of Rheumatology (ACR) criteria, attending rheumatology clinic between October 2014 and July 2015 were included. Hospital staff and lupus patients constituted healthy and disease controls, respectively. Evaluation of pSS included clinical evaluation, laboratory tests, ESSDAI and ESSPRI scoring. Unstimulated saliva was collected by the spitting method. Salivary IgG antibody against M3R (anti-M3R) was quantified by indirect ELISA. In this study, 43 patients with pSS, 34 with lupus and 42 healthy controls were recruited. The frequency of anti-M3R antibody levels was 55.81, 17.64 and 7 % for pSS, lupus and healthy controls, respectively. Area under the Receiver Operator Characteristic was 0.7791 (95 % CI,, 0.67-0.87). Sensitivity and specificity of the assay for diagnosis of pSS were 44.19 and 88.16 %, respectively. Salivary anti-M3R IgG antibody positivity was associated with lower age, shorter disease duration and higher globulin levels in our cohort. Salivary anti-M3R IgG antibody assay has high specificity in pSS; younger patients and those with hyperglobulinemia more frequently tested positive for this antibody.