Establishing proof of concept for utility of Trueprep®-extracted DNA in line-probe assay testing.

BACKGROUND AND OBJECTIVES: With an increased demand for rapid, diagnostic tools for TB and drug resistance detection, Truenat® MTB-RIF assay has proven to be a rapid point of care molecular test. The present study aimed to establish a proof of concept of using Trueprep-extracted DNA for line-probe assay (LPA) testing.METHODS: A total of 150 sputum samples (MTB-positive at Truenat sites) were divided into two aliquots. One aliquot was used for DNA extraction using the Trueprep device and MTB testing. The second aliquot of the sample was subjected to GenoLyse® DNA extraction. DNA from both the Trueprep and GenoLyse methods was subjected to first-line (FL) and second-line (SL) LPA testing.RESULTS: Of 139 Trueprep-extracted DNA, respectively 135 (97%) and 105 (75%) had interpretable results by FL and SL-LPA testing. Of 128 GenoLyse-extracted DNA, all 128 (100%) had interpretable FL-LPA results and 114 (89%) had interpretable SL-LPA results.CONCLUSION: The results obtained in this study indicate that Trueprep-extracted DNA can be used in obtaining valid LPA results. However, the study needs to be conducted on a larger sample size before our recommendations can be used for policy-making decisions.

A descriptive study of onychoscopic features in various subtypes of onychomycosis.

Background: The diagnosis of onychomycosis is usually clinical and is confirmed by 40% KOH examination. A diagnostic dilemma occurs when KOH examination is negative despite strong clinical suspicion. Dermoscopic evaluation of the nail is referred to as onychoscopy. We attempted to assess the dermoscopic findings in Onychomycosis positive with KOH examination. Methods: A cross sectional study was conducted in a tertiary care center including 122 patients with clinical suspicion of onychomycosis with KOH positivity. After assessment of risk factors and gross nail examination, onychoscopic examination was done to identify the presence of the specific features. Results: Primary findings of onychoscopic examination were 'spiked pattern' in 80.3% subjects, of which 95 were distal lateral subungual onychomycosis (DLSO), 8 of total dystrophic onychomycosis (TDO). True leukonychia was seen in the single patient of proximal subungual onychomycosis (PSO) and pseudoleukonychia in the single patient of white superficial onychomycosis (WSO). Distal irregular termination was observed in 23% of subjects - 8 from DLSO and in all 20 patients of TDO. 'Ruin appearance' was observed in all 20 patients of TDO, 56 patients with DLSO and not seen in other types of onychomycosis (OM). Presence of spiked pattern, Longitudinal striae, Distal irregular termination and Ruin appearance were found to be statistically significant (p < 0.001). Conclusion: In suspected onychomycosis, specific onychoscopic findings such as Spiked pattern, Longitudinal striae, Ruin appearance and Distal irregular termination can be used as supporting evidence for diagnosing onychomycosis clinically and initiating antifungal therapy if mycological testing is unavailable or negative.

VUV photoabsorption of thermally processed carbon disulfide and ammonia ice mixtures - Implications for icy objects in the solar system.

Many icy bodies in the solar system have been found to contain a rich mixture of simple molecules on their surfaces. Similarly, comets are now known to be a reservoir of molecules ranging from water to amides. The processing of planetary/cometary ices leads to the synthesis of more complex molecules some of which may be the harbingers of life. Carbon disulphide (CS2) and ammonia (NH3) are known to be present on many icy satellites and comets. Reactions involving CS2 and NH3 may lead to the formation of larger molecules that are stable under space conditions. In this paper we present temperature dependent VUV spectra of pure CS2 in the ice phase, and of CS2 and NH3 ices deposited as (i) layered, and (ii) mixed ices at 10 K and warmed to higher temperatures until their sublimation. Pure CS2 ice is found to have a broad absorption in the VUV region, which is unique for a small molecule in the ice phase. In layered and mixed ices, the molecules tend to affect the phase change and sublimation temperature of each other and also leave behind a form of CS2-NH3 complex after thermal annealing. This study of CS2-NH3 ice systems in layered and mixed configurations would support the detection of these species/complexes in mixed molecular ices analogous to that on planetary and cometary surfaces.

Covid-19 vaccination and possible link to Herpes zoster.

PURPOSE: To report 3 otherwise healthy patients with Herpes zoster reactivation shortly after administration of a mRNA vaccine against the novel COVID-19 virus. OBSERVATIONS: Patient 1 is a 54 year old who presented with Herpes zoster meningitis complicated by enhancing nodular leptomeningeal lesions of the spinal cord. The subsequent two patients had Herpes zoster ophthalmicus of the cornea (Case 2) and eyelid (Case 3). All three presented within 2 weeks of receiving the Pfizer/BioNTech COVID-19 vaccine. CONCLUSIONS: Herpes zoster may be a side effect of m RNA vaccination against the Sars-CoV2 vaccine and requires further investigation.

Vacuum ultraviolet photoabsorption spectra of icy isoprene and its oligomers.

Isoprene and its oligomers, terpenes, are expected to be present, along with other complex organic molecules in the diverse environments of the ISM and in our solar system. Due to insufficient spectral information of these molecules at low temperature, detection and understanding the importance of these molecules has been rather incomplete. For this purpose, we have carried out the vacuum ultraviolet (VUV) photoabsorption measurements on pure molecular ices of isoprene and a few simple terpenes: limonene, α-pinene and ß-pinene by forming icy mantles on cold dust analogs. From these experiments, we report the first low temperature (10 K) VUV spectra of isoprene and its oligomers limonene, α-pinene and ß-pinene. VUV photoabsorption spectra of all the molecules reported here reveal similarities in the ice and gas phase as expected, with an exception of isoprene where a prominent red shift is observed in the ice phase absorption. This unqiue property of isoprene along with distinctive absorption at longer wavelengths supports its candidature for detection on icy bodies.

Lax skin and blurring of vision- A case report of pseudoxanthoma elasticum.

Increased laxity of the skin can be caused by aging, significant weight loss, or defects in the elastic tissue. A 38-year-old female presented with increased laxity of the skin over the neck, thighs, and abdomen for 6 years, associated with headache and blurring of vision for a week. On cutaneous examination, prominent skin folds, laxity, and wrinkles were noted over the neck, abdomen, thighs, and groin, with yellowish papules along the neck creases. Ocular examination revealed features suggestive of angioid streaks. Skin biopsy showed fragmented elastic fibers and intervening calcium deposits on Verhoeff Van Gieson and Von Kossa stains. Based on these findings, a diagnosis of pseudoxanthoma elasticum (PXE) was made. The patient was started on oral and topical sunscreens and eye protection and advised regular follow-up. Diagnosing the condition early based on skin findings can help prevent further multi-system manifestations by taking appropriate preventive measures as this condition is progressive and has no cure.

A descriptive study of the clinical and etiological profile of balanoposthitis.

BACKGROUND: Balanoposthitis is defined as an inflammatory condition of glans penis and prepuce. There are wide variety of etiologies including both infectious and noninfectious conditions. This study attempts to throw light on information regarding clinical and microbiological aspects of balanoposthitis. OBJECTIVES: To study various clinical patterns, etiologies, and predisposing factors of balanoposthitis. METHODOLOGY: A descriptive study was undertaken on 106 cases who presented to sexually transmitted disease (STD) clinic with balanoposthitis between November 2017 and April 2019. A detailed history, physical examination, and investigations like KOH mount, leishman staining, gram staining, dark field microscopy, cultures, and other investigations were done wherever indicated. The data collected was tabulated and analyzed. RESULTS: In our study, infectious etiology was the most common and was found in 77.36% cases. About 13.41% of cases with infectious balanoposthitis had multiple etiological agents. Noninfectious etiology was found in 22.64% cases. The most common infectious cause of balanoposthitis was candida, noted in 59.76% cases, followed by herpes simplex virus (19.51%), human papilloma virus (13.41%), and scabies (8.54%). Among noninfectious etiologies, adverse drug reaction (4.72% of total cases) was the most common, followed by lichen planus (3.77%) and psoriasis (3.77%). There was significantly higher incidence of phimosis in diabetic patients with candidal balanoposthitis. CONCLUSION: Identifying the etiology facilitates early treatment and hence reduces the infectivity and transmission of disease and also the disease complications like phimosis. In addition, multiple infectious etiologies should always be kept in mind while evaluating STDs.

Named entity recognition on bio-medical literature documents using hybrid based approach.

There have been many changes in the medical field due to technological advances. The progression in technologies provides lot of opportunities to extract valuable insights from huge amount of unstructured data. The literature documents published by the researchers in medical domain consists enormous amount of knowledge. Many organizations are involving in retrieving the hidden information from the literature documents. Extracting the drug names, diseases, symptoms, route of administration, species and dosage forms from the textual document is an easy task due to the innovation of technologies in the Natural Language Processing. In this article, a new hybrid based approach is proposed to identify named entity from the medical literature documents. New dictionary has been built for route of administration, dosage forms and symptoms to annotate the entities in the medical documents. The annotated entities are trained by the blank Spacy machine learning model. The trained model provide a decent accuracy when compared with the existing model. The hybrid model is validated with the dictionary and human (optional)to calculate the confusion matrix. It is able to identify more entities than the prevailing model. The average F1 score for five entities of the proposed hybrid based approach 73.79%.

Spatiotemporal and functional characterisation of transient receptor potential vanilloid 4 (TRPV4) in the murine intervertebral disc.

The molecular regulators of mechano-transduction in intervertebral disc (IVD) cells are not well understood. The aim of the present study was to characterise the expression and function of the mechano-sensitive ion channel TRPV4 in the IVD. A novel transgenic reporter mouse, in which the endogenous Trpv4 locus drove the expression of LacZ, was used to localise Trpv4 expression at specific stages of spine development [embryonic day (E) 8.5, 12.5, 17.5, postnatal day 1] and time points following skeletal maturity (2.5, 6, 9 and 12 months). The TRPV4-specific agonist GSK1016790A and antagonist GSK2193874 were used to assess the functional response of annulus fibrosus (AF) cells using epifluorescence imaging with Ca2+-sensitive Fura-2 dye and F-actin staining. The effects of TRPV4 agonism and antagonism in mechanically stimulated AF cells were quantified by gene expression analysis. Trpv4 expression was specific to the developing notochord and intervertebral mesenchyme at E12.5. At 2.5, 6 and 9 months, Trpv4 expression was detected in the nucleus pulposus, inner AF, cartilage endplate and vertebral growth plate. AF cells treated with GSK1016790A demonstrated heterogeneity in TRPV4-dependent Ca2+ responses (no response, calcium oscillation or sustained response). TRPV4-induced Ca2+ signalling was associated with Rho/ROCK-dependent actin cytoskeleton remodelling and stress-fibre formation. In AF cells, cyclic-tensile-strain-induced changes in Acan and Prg4 expression were mediated by TRPV4 channel activation. These data establish TRPV4 as an important mechano- sensor regulating IVD mechano-biology.

Strengthened capacity of India´s bedaquiline Conditional Access Programme for introducing new drugs and regimens.

BACKGROUND: Addressing TB in India is critical to meeting global targets. With the scale-up of diagnostic networks and the availability of new TB drugs, India had the opportunity to improve the detection and treatment outcomes in drug-resistant TB (DR-TB).OBJECTIVE: To document how the introduction of new drugs and regimens is helping India improve the care of DR-TB patients.DESIGN: In 2016, India´s National TB Programme (NTP) introduced bedaquiline (BDQ) under a Conditional Access Programme (BDQ-CAP) at six sites after providing extensive training and strengthening laboratory testing, pre-treatment evaluation, active drug safety monitoring and management (aDSM) and follow-up systems.RESULTS: An interim analysis reflected earlier and better culture conversion rates: 83% of the 620 patients converted within a median time of 60 days. However, 248 serious adverse events were reported, including 73 deaths (12%) and 100 cardiotoxicity events (16.3%). Encouraged by the evidence of safety and efficacy of BDQ, the NTP took steps to systematically expand its access to cover the entire population by 2018.CONCLUSION: The cautious yet focused approach used to introduce BDQ under BDQ-CAP paved the way for the rapid introduction of delamanid, as well as the shorter treatment regimen and the all-oral regimen for DR-TB.

Detection of estrous biomarkers in the body exudates of Kangayam cattle (Bos indicus) from interplay of hormones and behavioral expressions.

Behavioral expressions and biochemical composition of body exudates are significantly altered in concert with the endocrine status, which are all clear indicators of physiological conditions of animals. In this study, we sought to infer about the reproductive physiological status of Kangayam cattle (Bos indicus) by analyzing behaviors, endocrine pattern, and body exudates and further to discover estrous biomarkers so as to facilitate timely artificial insemination/mating and to aid in aspects of conservation of the species. Therefore, in this study, we followed Kangayam cows through pre-estrous to post-estrous phases to correlate the endocrine dependence of biochemical constituents in urine and cervical mucus and sought to identify estrous biomarkers. Behavioral estrus was confirmed in 10 cows, from which urine samples were collected and subjected to determination of LH, FSH, estrogens, progesterone, proteins, and lipids. Furthermore, urinary fatty acids and proteins were profiled using gas chromatography and SDS-PAGE, respectively. The volatile compounds in the urine and cervical mucus were identified by gas chromatography-mass spectrometry analysis. The data revealed that LH, FSH, and estrogen levels increased significantly in estrous urine compared with nonestrous urine, whereas progesterone status was vice versa (P < 0.05). The lipid content was also significantly higher in estrous urine than in pre- and post-estrous urines (P < 0.05). There were also cyclical variations of volatiles and fatty acid profiles across phases of the estrous cycle. More acidic compounds were present in estrous urine, rendering it more acidic, than in pre- and post-estrous urines. Interestingly, oleic acid, which was present as a fatty acid in estrous and post-estrous urines, appeared to be a volatile in post-estrous urine and estrous cervical mucus. In addition, octanoic and butanoic acids were specific to both estrous urine and cervical mucus, indicating their possible candidature as estrous biomarkers. SDS-PAGE analysis showed pronounced expression of a 98 kDa protein in post-estrous urine, which in matrix-assisted laser desorption ionization-time of flight mass spectrometry was identified as albumin. Our results demonstrate multiple biomarkers in estrous urine and specific volatiles in cervical mucus that offer scope to develop viable estrus detection kits for Kangayam cows.

Collisional excitation of NH(3Σ-) by Ar: A new ab initio 3D potential energy surface and scattering calculations.

Collisional excitation of light hydrides is important to fully understand the complex chemical and physical processes of atmospheric and astrophysical environments. Here, we focus on the NH(X3Σ-)-Ar van der Waals system. First, we have calculated a new three-dimensional Potential Energy Surface (PES), which explicitly includes the NH bond vibration. We have carried out the ab initio calculations of the PES employing the open-shell single- and double-excitation couple cluster method with noniterative perturbational treatment of the triple excitations. To achieve a better accuracy, we have first obtained the energies using the augmented correlation-consistent aug-cc-pVXZ (X = T, Q, 5) basis sets and then we have extrapolated the final values to the complete basis set limit. We have also studied the collisional excitation of NH(X3Σ-)-Ar at the close-coupling level, employing our new PES. We calculated collisional excitation cross sections of the fine-structure levels of NH by Ar for energies up to 3000 cm-1. After thermal average of the cross sections, we have then obtained the rate coefficients for temperatures up to 350 K. The propensity rules between the fine-structure levels are in good agreement with those of similar collisional systems, even though they are not as strong and pronounced as for lighter systems, such as NH-He. The final theoretical values are also compared with the few available experimental data.

Loss of Subpodocytic Space Predicts Poor Response to Tacrolimus in Steroid-Resistant Calcineurin Inhibitor-Naïve Adult-Onset Primary Focal Segmental Glomerulosclerosis.

Focal segmental glomerulosclerosis (FSGS) is the most common cause of adult-onset nephrotic syndrome, but its pathophysiology is poorly understood. The question as to why only a subset of patients responds to treatment in unanswered. In the past few years, change of podocytic phenotype from stationary type in health to migratory type in disease has been described, of which loss of subpodocytic space is a surrogate marker. Diagnostic biopsies of adult-onset steroid-resistant calcineurin inhibitor-naïve primary FSGS cases, which were subsequently treated with tacrolimus were included in this retrospective study conducted from 2011 to 2013. The ultrastructure of all cases was studied in detail, especially in context to the presence or absence of subpodocytic space. In the present study, we have compared presence or absence of subpodocytic space in tacrolimus-responsive versus tacrolimus-resistant cases to identify potential electron microscopic features predictive of response to treatment, of which loss of subpodocytic space indicating migratory phenotype is the most important and consistent feature. The present series included 7 tacrolimus responsive cases (includes two cases with partial response) and seven tacrolimus-resistant cases. The tacrolimus-resistant patients were of older age, had a longer duration of illness, and a lower eGFR as compared to tacrolimus responsive cases. The subpodocytic space was preserved in patients on tacrolimus with complete remission and lost in patients with partial response and tacrolimus-resistant cases.

Intestinal Tuberculosis: A Rare Case of Massive Gastrointestinal Bleed in a Post-Renal Transplant Recipient.

Massive rectal bleeding is an uncommon presentation of ileal tuberculosis (TB). We report an uncommon cause of anemia in a post-renal transplant patient due to massive lower gastrointestinal (LGI) bleed. The index case had a normal upper and LGI endoscopy, but the 99technetium labeled red blood cell scan showed active bleeding from terminal ileum and caecum. Microscopic examination of the resected specimen revealed tubercular granuloma with acid-fast bacilli. Intestinal TB should be a differential diagnosis for massive LGI bleed in immunosuppressed patients in developing country.

Missed Monoclonal Disease Manifesting in Early Post-renal Transplant Period.

A 63-year-old diabetic gentleman with microvascular complications presented with advanced azotemia and anemia. He was stabilized with blood transfusion and hemodialysis. With the probable diagnosis of diabetic nephropathy-related end-stage renal disease, he underwent kidney transplantation. He had delayed graft function. Graft biopsy done on the 2nd postoperative day showed acute tubular necrosis. Graft biopsy repeated after 2 weeks for persistent graft dysfunction showed myeloma cast nephropathy (MCN) and light chain proximal tubulopathy. Work-up for multiple myeloma was positive. He was started on plasmapheresis and chemotherapy. However, he suffered sudden cardiac death during dialysis after 1 week. The presence of MCN in the early graft biopsy implies that it must have been the cause for his native kidney failure. Thus, renal failure in a diabetic should not always be presumed to be due to diabetic nephropathy, and kidney biopsy should be done in diabetics with atypical features.

Epidemiology of chikungunya based on laboratory surveillance data-India, 2016-2018.

BACKGROUND: We analysed the laboratory surveillance data generated by the network of virology laboratories from January 2016 to July 2018 to describe the epidemiology of chikungunya in India. METHODS: Suspected patients reporting to virology laboratories are investigated for the presence of immunoglobulin M (IgM) antibodies against chikungunya virus (CHIKV) as evidence of recent infection. We analysed the data to describe the distribution of seropositive individuals by time, place and demographic characteristics. RESULTS: Of the 49 380 sera tested from suspected patients, 20.5% were seropositive. CHIKV IgM positivity was seen throughout the year, with a peak between September and December. The laboratories diagnosed 28 outbreaks of chikungunya during the study period. CONCLUSIONS: Laboratory surveillance data indicate continued transmission of CHIKV in many regions of India.

Assessment of urinary volatile compounds and proteins in the female goat Capra hircus: A pilot study to reveal potential indicators of oestrus.

Urine samples of female goats in pro-oestrus, oestrus and post-oestrus phases were analysed for finding oestrus-specific volatile compounds using gas chromatograph-mass spectrometry (GC-MS), and proteins using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS). Fourteen urinary volatile were identified covering all three phases among which four compounds, 1-Tetradecanol, n-Pentadecanol, 3-Methylene tridecane and 2-Ethyl-1-dodecene, were unique to oestrus. Also, oestrus urine contained a 25 kDa protein, which was totally absent in pro-oestrus urine, and less-expressed in post-oestrus urine. This protein revealed to be complement C3 fragment. This pilot study, for the first time, reveals the difference in urinary volatile compounds and proteins in the female goat during the different phases of oestrous cycle. The four unique volatile compounds and a 25 kDa protein that appeared as oestrus-specific in this study warrant further investigation to consider them as urinary biomarkers of oestrus in goats.

Epidemiology of dengue fever in India, based on laboratory surveillance data, 2014-2017.

OBJECTIVES: The Indian Council of Medical Research and the Department of Health Research have established a network of Virus Research and Diagnostic Laboratories (VRDL) to strengthen laboratory capacity in India. We analyzed the data generated by the 52 VRDLs during 2014-2017 to describe the epidemiology of dengue fever (DF) in India. METHODS: As per the laboratory protocol, suspected DF patients reporting to various hospitals where VRDLs are located, or samples from suspected DF outbreaks are investigated for the presence of NS1 antigen or IgM antibodies against dengue. The data were analyzed to describe the distribution of DF by time (month and year), place (district and state) and person (age and sex) characteristics. RESULTS: Between 2014-2017, VRDLs investigated 211,432 suspected DF patients, 28.4% of whom were serologically confirmed. The median age of dengue positive patients was 25 years (IQR: 16-36). Dengue positivity was significantly higher among males. The mean monthly dengue positivity ranged from 7.7% to 37%, with higher positivity reported during September and October months. VRDLs provided diagnosis to 190 suspected outbreaks. CONCLUSIONS: The data from the VRDL network indicate dengue was the etiology in one-fourth of AFI cases. Dengue was the second commonest etiology of suspected outbreaks diagnosed by VRDLs.