Molecular Characterization and Analysis of Risk Factors Associated with Brucellosis in Central Indian and Meghalaya Population.

Objective: In this study we evaluated the utility of Abortus Melitensis Ovis Suis Brucella PCR (AMOS PCR) for the molecular characterization of Brucella species and analyzed the associated risk factors for brucellosis in Central Indian and Meghalayan population. Methods: AMOS PCR was carried out in a total of 160 BSCP-31 PCR-positive DNA samples isolated previously from the blood of Central Indian (n = 90) and Meghalayan cohorts (n = 70). Clinical and associated risk factors recorded earlier were used to establish strain-specific disease outcomes in study cohorts. Results: Brucella melitensis was found to be the dominant strain in both Central Indian and Meghalayan cohorts (57.7% and 54.28%, respectively) followed by Brucella abortus (42.22% and 38.57%). Although rare, brucellosis cases in the Meghalayan population also showed the presence of Brucella suis (7.14%) and Brucella ovis (2.85%). Febrile illness was a major clinical risk factor in both study cohorts, while occupational risk factors like exposure to animals and raw milk consumption were major mediating factors for brucellosis in Central Indian cohorts. On the contrary, meat consumption was found to be significant predisposing factor for brucellosis in Meghalaya. Conclusion: Molecular characterization of Brucella species provides important public health data for mitigation, advocacy, and antimicrobial stewardship.

K-Wire-Based External Fixator for Management of Salter-Harris Type-II Distal Femur Physeal Injury.

We present a case of a 16-year-old male with a Salter-Harris type II physeal slip of the distal femur managed with closed reduction and K wire and clamp-based external fixator. Knee range of motion exercises were initiated after one week. The union was observed at 10 weeks, and implant removal was done on an outpatient basis. At one year follow-up, the patient had good clinical and radiological outcomes. The K-wire-based external fixator frame is an effective fixation method for distal femur physeal slips in older children, providing favorable radiological and functional outcomes.

Glass transition phenomena and dielectric relaxations in supercooled d-lyxose aqueous solutions.

Differential scanning calorimeter and broadband dielectric spectroscopy in a broad range of temperatures (150-300 K) were employed to study the d-lyxose aqueous mixture at different hydration levels. Two relaxation processes were observed in all investigated d-lyxose aqueous mixtures. A relaxation process (process-I) usually known as the primary relaxation mode which is accountable for the collective motion of d-lyxose aqueous solution, was observed above the glass transition temperature (Tg). Below Tg, another process designated as process-II was found which is mainly related to the water molecule relaxation inside the d-lyxose matrix. The average relaxation times as a function of temperature and dielectric strengths of both observed relaxation processes (I & II) were analyzed for all hydration levels in d-lyxose. It was identified that the relaxation amplitude of process-II in the d-lyxose aqueous mixture was increased drastically and their activation energies were found to be approximately independent of the content of water above critical concentration, xc = 0.28. This suggests that the dynamical process observed above xc was dominated by the presence of water clusters. In the current aqueous mixture, the critical content of water (xc) is slightly higher as compared to previously reported aqueous mixtures, indicating a more cooperative nature of water molecules with a d-lyxose matrix. Additionally, the Tg of pure water was estimated at 128 ± 5.8 K from the extrapolation of DSC Tg data of the d-lyxose aqueous solution by using the well-known Gordon-Taylor equation. Our current result gives further support to the well-accepted glass transition (Tg) of pure water.

Investigation of the Contents of the Stack Emissions of Iron Ore Sinter Plants With and Without Bag Filter.

This investigation aims to identify the reasons for the plumes' visibility, compare the stacks with other sinter plant stacks worldwide, and suggest countermeasures to completely stop the visibility of emissions. The appearance of the sinter plant stack emission changes with time and the background color of the sky due to the scattering effect of the sunlight and incidence angle. The flue gas samples were collected at the outlet of the emission control equipment and observed under optical and scanning electron microscopes. The characterization was performed with the help of an electron dispersive spectroscope and mapping technique. The contents of the stack of a sinter plant without a bag filter had much higher levels of PM10, SO2, and NOx. The emissions from all the sinter plants were invariably found to have particulates of SO2 and NOx of size less than 2.5 microns. It is suggested to opt for state-of-the-art fabric filter technology to eliminate PM2.5 emissions also.

Diagnostic utility of LAMP PCR targeting bcsp-31 gene for human brucellosis infection.

PURPOSE: Human brucellosis is a neglected zoonotic disease of significant public health concern. Molecular diagnosis of brucella remains challenging in low resource settings, due to the high infrastructure and cost involved. Loop-mediated isothermal amplification (LAMP) is a rapid point of care polymerase chain reaction (PCR) with the utility of on-field molecular diagnosis and offers a convenient alternative to conventional PCR. In the present study, we developed and evaluated the diagnostic utility of in house LAMP PCR targeting the Brucella genus-specific bcsp-31 gene in patients having febrile illness. MATERIALS AND METHODS: The analytical sensitivity and specificity of bcsp-31 LAMP PCR was first evaluated using brucella (n â€‹= â€‹8) and non-brucella cultures (n â€‹= â€‹5), along with spiked clinical samples. The overall diagnostic utility of developed LAMP PCR was then further evaluated in 393 human samples suspected of brucellosis. RESULTS: The developed LAMP PCR could detect as low as 8 â€‹fg of DNA by visual detection within 35min. We report sensitivity and specificity of the developed LAMP PCR as 90.91% and 99.37%.The accuracy of the developed test assay was found to be 98.60%. In clinical samples, LAMP gave positivity of 20% with the concordance of 89% with conventional PCR. CONCLUSION: To conclude, a rapid, efficacious, sensitive LAMP PCR targeting the bcsp 31 gene was developed. The existing LAMP PCR can be used as a point of care screening test in various low resource endemic setting in lieu of conventional PCR for estimation of prevalence data, diagnosis and treatment of brucellosis.

Membrane Tucking Technique using Ligature Clips with Intermembranous Placement of Oxidized Regenerated Cellulose in Multilayered Chronic Subdural Hematoma: Description of Novel Technique and our Initial Experience.

Chronic subdural hematoma (cSDH) is an encapsulated collection of blood and blood degradation products between dural border cell layers, the pathophysiology of formation and expansion of which is still debatable. It is usually seen in the elderly population, and surgical evacuation is the primary mode of treatment. The main hurdle in the treatment of cSDH is postoperative recurrences and the need for repeat operations. A few authors have classified cSDH into homogenous, gradation, separated, trabecular, and laminar types based on the internal architecture of hematoma and proposed that separated, laminar, and gradation types of cSDH have a high propensity of recurrence after surgery. A similar problem was described with multi-layered or multi-membrane cSDH. Based on the most accepted theory of formation and expansion of cSDH that suggests a complex and vicious process of membrane formation, chronic inflammation, neoangiogenesis, rebleeding from fragile capillaries, and increased fibrinolysis, we propose our theory of intermembranous placement of oxidized regenerated cellulose and membrane tucking using ligature clips to prevent recurrence in multi membranous cSDH by ceasing the ongoing cascade in hematoma's internal milieu and thus preventing recurrence and reoperation in such cases. This is the first in the world literature report describing such a technique for treating multi-layered cSDH and in our series, the reoperation and postoperative recurrence rates were 0% in patients treated by the described technique.

Design and Analysis of Planar Waveguide-Based SPR Sensor for Formalin Detection Using Ag-Chloride-BP Structure.

Nowadays, Food additives and preservatives have become a hot topic especially formalin, which is a chemical substance used to preserve food. Chronic cancer is caused by swallowing the formalin-contaminated food on a regular basis. As a result, detection of the formalin in food ingredients is a critical need, and this requirement is becoming increasingly important in new terrains. Therefore, a new approach, planar waveguide-based surface plasmon resonance (SPR) sensor is presented in this paper employing silver-chloride materials-black phosphorus structure to detect the formalin. In this study, the optimization of metal thickness and optimal performance of chloride material are analyzed by observing the transmittance power and resonance shift for conventional and chloride-based sensors, respectively. Moreover, the performance parameters, such as sensitivity of 344°/ RIU, quality-factor of [Formula: see text], detection-accuracy of 3.34 and figure-of-merit of 164.74, are observed for the proposed sensor. Furthermore, the performance comparative study is conducted and found that the proposed sensor highlighted the enhanced the SPR sensor and highlighted the performance of FOM. Finally, the electromagnetic filed distributions of the proposed WG-based SPR sensor are shown using the Opti-FDTD software.

Development of multiple reaction monitoring (MRM) assays to identify Brucella abortus proteins in the serum of humans and livestock.

In the present study, a targeted multiple reaction monitoring-mass spectrometry (MRM-MS) approach was developed to screen and identify protein biomarkers for brucellosis in humans and livestock. The selection of proteotypic peptides was carried out by generating in silico tryptic peptides of the Brucella proteome. Using bioinformatics analysis, 30 synthetic peptides corresponding to 10 immunodominant Brucella abortus proteins were generated. MRM-MS assays for the accurate detection of these peptides were optimized using 117 serum samples of human and livestock stratified as clinically confirmed (45), suspected (62), and control (10). Using high throughput MRM assays, transitions for four peptides were identified in several clinically confirmed and suspected human and livestock serum samples. Of these, peptide NAIYDVVTR corresponding to B. abortus proteins: BruAb2_0537 was consistently detected in the clinically confirmed serum samples of both humans and livestock with 100% specificity. To conclude, a high throughput MRM-MS-based protocol for detecting endogenous B. abortus peptides in serum samples of humans and livestock was developed. The developed protocol will help design sensitive assays to accurately diagnose brucellosis in humans and livestock. The data associated with this study are deposited in Panorama Public (https://panoramaweb.org/rNOZCy.url with ProteomeXchange ID: PXD034407).

Comprehensive Proteomic Analysis of Brucella melitensis ATCC23457 Strain Reveals Metabolic Adaptations in Response to Nutrient Stress.

In the present study, a comprehensive proteomic analysis of Brucella melitensis (B. melitensis) strain ATCC23457 was carried out to investigate proteome alterations in response to in vitro-induced nutrient stress. Our analysis resulted in the identification of 2440 proteins, including 365 hypothetical proteins and 850 potentially secretory proteins representing ~77.8% of the B. melitensis proteome. Utilizing a proteogenomics approach, we provide translational evidence for eight novel putative protein-coding genes and confirmed the coding potential of 31 putatively annotated pseudogenes, thus refining the existing genome annotation. Further, using a label-free quantitative proteomic approach, new insights into the cellular processes governed by nutrient stress, including enrichment of amino acid metabolism (E), transcription (K), energy production and conversion (C), and biogenesis (J) processes were obtained. Pathway analysis revealed the enrichment of survival and homeostasis maintenance pathways, including type IV secretion system, nitrogen metabolism, and urease pathways in response to nutrient limitation. To conclude, our analysis demonstrates the utility of in-depth proteomic analysis in enabling improved annotation of the B. melitensis genome. Further, our results indicate that B. melitensis undergoes metabolic adaptations during nutrient stress similar to other Brucella. sp, and adapts itself for long-term persistence and survival.

Seroepidemiological study of human brucellosis in the Northeast region of Meghalaya, India.

Introduction: Brucellosis is a neglected zoonotic disease of major public health concern. In India, the incidence of brucellosis remains vastly underreported due to its non-specific clinical presentation and sub-optimal sensitivity of existing gold standard tests. Studies in Northeast India have shown high incidences of brucellosis in livestock, but the region lacks data on human brucellosis despite its high associated risk. In the present study, we report the seroprevalence of human brucellosis and its associated risk factors in Meghalaya, Northeast India. Materials and Methods: A prospective observational study was conducted in East Khasi Hills and Ri.Bhoi districts of Meghalaya, from July 2018 to July 2020. A total of 1046 suspected patients with febrile illness along with associated risk factors were recruited through camps and various diagnostic laboratories in the defined region as per the pre.specified inclusion and exclusion criteria. Baseline, demographics, and clinical characteristics were recorded of all the consenting participants. Blood samples were analyzed for brucellosis-specific IgM antibodies through enzyme-linked immunosorbent assay (ELISA). Results and discussion: The overall seroprevalence of brucellosis was found to be 11.37% in Meghalaya. Among recruited participants, females were found to be more susceptible than males. Risk factors such as consumption of meat were found to be more significantly associated with brucellosis disease in the study region. Among the clinical presentations, pyrexia of unknown origin, myalgia, and chronic fatigue syndrome were found to be significantly associated with brucellosis disease in IgM.positive cases. Conclusion: Our result suggests further epidemiological investigations for human brucellosis in Northeast India toward improved advocacy for accurate diagnosis, and development of proper response mechanism in areas of high endemicity.

Breeding More Crops in Less Time: A Perspective on Speed Breeding.

Breeding crops in a conventional way demands considerable time, space, inputs for selection, and the subsequent crossing of desirable plants. The duration of the seed-to-seed cycle is one of the crucial bottlenecks in the progress of plant research and breeding. In this context, speed breeding (SB), relying mainly on photoperiod extension, temperature control, and early seed harvest, has the potential to accelerate the rate of plant improvement. Well demonstrated in the case of long-day plants, the SB protocols are being extended to short-day plants to reduce the generation interval time. Flexibility in SB protocols allows them to align and integrate with diverse research purposes including population development, genomic selection, phenotyping, and genomic editing. In this review, we discuss the different SB methodologies and their application to hasten future plant improvement. Though SB has been extensively used in plant phenotyping and the pyramiding of multiple traits for the development of new crop varieties, certain challenges and limitations hamper its widespread application across diverse crops. However, the existing constraints can be resolved by further optimization of the SB protocols for critical food crops and their efficient integration in plant breeding pipelines.

A Plus Shaped Cavity in Optical Fiber Based Refractive Index Sensor.

The optical fiber grating sensors have strong potential for the detection of biological samples. However, a careful effort is still in demand to enhance the performance of existing grating sensors especially in biological sensing. Therefore, in this work, we have introduced a novel plus shaped cavity (PSC) in optical fiber model and used it for the detection of haemoglobin (Hb) refractive index (RI). The numerical analysis of designed model is done by the testing of single and double vertical slots cavity in optical fiber core structure. The testing of designed sensor model is done at the wavelength of 800 nm at which the RI of oxygenated and deoxygenated Hb is 1.392 and 1.389, respectively. The analysis of reported PSC sensor model is done in the wide range of Hb RI from 1.333 to 1.392. The tested range of RI corresponds to the Hb concentration from 0 to 140 gl-1. The obtained results states that for the tested range of RI, the autocorrelation coefficient of R2 = 99.51 % is achieved. The analysis of projected work is done by using finite difference time domain (FDTD) method. The introduction of PSC can increase in sensitivity. In proposed PSC, the length and width of created slots are [Formula: see text] and [Formula: see text], respectively, which is quite enough to observe the response of analytes RI. This can minimize the creation of multiple gratings required for observing the analyte response.

Determination of Etiology and Epidemiology of Viral Central Nervous System Infections by Quantitative Real-Time Polymerase Chain Reaction in Central India Population.

INTRODUCTION: Viral infections of the central nervous system (CNS) are the most common cause of hospital admission in worldwide and remain a challenging disease for diagnosis and treatment. The most common infectious agents associated with viral CNS infections are cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), Japanese encephalitis virus (JEV), Dengue virus (DENV),West Nile virus(WNV), and Chandipura virus(CHPV). The aim of the present work was to find the etiology of CNS viral infection in the Central India population by transcriptase PCR (RT-PCR) comparing real-time polymerase chain reaction (PCR) method [one-step and two-step reverse transcriptase (RT-PCR)] in cerebrospinal fluid (CSF) and blood samples of CNS viral infections patients. MATERIALS AND METHODS: One-step and two-step real-time PCR assays were evaluated in CSF and parallel blood samples from patients with viral CNS infections for detection of DNA and RNA viruses. A comparative analysis was also done between gDNA, gRNA, cDNA, and plasmid-based real-time PCR methods for an efficient quantitation of viral particles in clinical samples for determination of viral etiology. RESULT: On evaluation of 150 CSF and 50 parallel blood samples from suspected cases of viral CNS infections, a viral etiology was confirmed in 21 (14%) cases, including 3% for EBV, 1% of CMV, and 5% for VZV and JEV. The one-step RT-PCR has a higher detection limit for detection and quantitation of viral RNA in comparison to two-step RT-PCR. CONCLUSION: Our result reveals that VZV and JEV are the most usual cases of CNS viral infection in hospitalized patients in the Central India population and one-step RT-PCR shows higher viral load detection limits for quantitation of viral genome and more sensitivity in comparison to two-step RT-PCR.

The Aftermath of COVID-19 Lockdown- Why and How Should We Be Ready?

INTRODUCTION: Severe acute respiratory syndrome, coronavirus 2 (SARS-COV 2) has inexplicably and irreversibly changed the way of neurosurgery practice. There has been a substantial reduction in neurosurgical operations during the period of lockdown. The lockdown might be the most effective measure to curtail viral transmission. Once we return to the normalization of the lifestyle, there will be a backlog of unoperated pending cases along with the possibility of further spread of the coronavirus. METHODS: We reviewed the available literature and protocols for neurosurgical practice in different geographic locations. We drafted a consensus statement based on the literature and protocols suggested by the World Health Organization (WHO) and various professional societies to prevent the spread of SARS-COV2 while streamlining the neurosurgical practice. RESULTS: The consensus statement suggests the patient triage, workflow, resource distribution, and operational efficacy for care providers at different stages of management. The priority is set at personal protection while ensuring patients' safety, timely management, and capacity building. We performed a detailed subsection analysis for the management of trauma and set up for COVID-free hospitals for simultaneous management of routine neurosurgical indications. In this time of medicolegal upheaval, special consent from the patients should be taken in view of the chances of delay in management and the added risk of corona infection. The consensus statements are applicable to neurosurgical setups of all capacities. CONCLUSION: Along with the glaring problem of infection, there is another threat of neurosurgery emergency building up. This wave may overwhelm the already stretched systems to the hilt. We need to flatten this curve while avoiding contagion. These measures may guide neurosurgery practitioners to effectively manage patients ensuring the safety of caregivers and care seekers both.

Detection of Collagen-IV Using Highly Reflective Metal Nanoparticles-Immobilized Photosensitive Optical Fiber-Based MZI Structure.

In this work, a photosensitive (PS) optical fiber-based Mach-Zehnder interferometer (MZI) structure is developed to diagnose the presence of collagen-IV in human bodies. The MZI is fabricated by sequentially splicing the single mode-multimode-photosensitive-multimode-single mode (SMPMS) fiber segments. The sensing region in MZI structure is created by partially removing the cladding of photosensitive fiber by using 40% hydrofluoric (HF) acid and depositing the layers of highly reflective metal nanoparticles (NPs) over it. The used NPs are polyvinyl alcohol stabilized silver nanoparticles (PVA-AgNPs), gold nanoparticles (AuNPs), and zinc oxide nanoparticles (ZnO-NPs). The size of AuNPs, PVA-AgNPs, and ZnO-NPs are 10 ± 0.2 nm,  âˆ¼  4 -5 nm, and < 50 nm, respectively. In order to avoid the interference of other biomolecules in the detection of collagen-IV, the sensing region is functionalized with a collagenase enzyme. The sensing ability of the probe is ascertained by sensing a wide concentration of collagen solution ranging from 0 ng/ml to [Formula: see text]/ml. It is observed that sensing performance of probe is much better on immobilizing it with PVA-AgNPs and ZnO-NPs.

Diagnostic Challenges and Prospects Associated With Zoonotic Tuberculosis of Central Nervous System.

INTRODUCTION: The diagnosis of Tuberculous Meningitis (TBM) has remained a challenge due to its insidious onset and the failure of conventional diagnostic tests. The present study aimed to identify the mycobacterial pathogen in the CSF of patients with TBM and a poor prognosis. METHODS: We retrospectively recruited 224 TBM and 34 non-TBM patients admitted to the Central India Institute of Medical Sciences, Nagpur, India, in 2014. The CSF samples of these patients were subjected to a duplex PCR assay for the species-specific identification of the causative pathogen. RESULTS: M. bovis and infection with M.tuberculosis were detected in 7% (18) and 32.9% (85) of the patients, respectively. Moreover, 14% (36) of the study samples were culture positive; however, the mycobacterial pathogens could not be differentiated to the species level. CONCLUSION: The present study findings emphasized the potentially vital importance of M. bovis identification for appropriate patient management. The obtained data also demonstrated the persistent significance of M. bovis, as a zoonotic pathogen.

Development of an Immunodiagnostic Test for Screening Human Brucellosis Cases Using the Whole-Cell Antigens of Brucella abortus.

BACKGROUND: Human brucellosis is an important zoonotic disease of public health and often remains neglected owing to lack of sensitive and efficient diagnostic methods. This study evaluates diagnostic utility of in-house designed enzyme-linked immunosorbent assay (ELISA) using whole-cell antigens of Brucella abortus (B. abortus) S19 against the commercially available kits. METHODS: A prospective cohort study involving different populations within the Vidarbha regions of Maharashtra, India was conducted through camps organised from May 2009 to October 2015. A total of 568 serum samples were collected from high-risk people recruited as study cohorts based on inclusion criteria, additional risk factors and clinical symptoms. Samples were evaluated by indirect ELISA using the whole-cell antigens of B. abortus. The results were compared with the commercially available IgG detection ELISA kit to ascertain the specificity and sensitivity of the developed test. RESULTS: Fever, body ache, joint pain, lower back pain, loss of appetite and weight loss were major symptoms associated with the disease. With the cut-off of > 0.8, the positivity of brucellosis infection was at 12.32% (70/568) compared to 9.33% (53/568) as detected by the commercial kit. The in-house developed ELISA method yielded a sensitivity of 87.5% and specificity of 99.18% as compared to the commercial kits (sensitivity -80.30% and specificity -99.6%). DISCUSSION: The B. abortus S19-derived whole-cell protein-based ELISA is rapid and cost-effective and can be used for screening brucellosis infection in lieu of the commercially available ELISA kits.

Draft Genome Sequence of Listeria monocytogenes CIIMS-NV-3, a Strain Isolated from Vaginal Discharge of a Woman from Central India.

We present here the draft genome sequence of Listeria monocytogenes CIIMS-NV-3, a serovar 4b strain isolated from the vaginal swab of a female patient from central India. The availability of this genome may provide useful information on virulence characteristics for comparative genomic analysis.

Differential proteome analysis of rat plasma after diisopropyl fluorophosphate (DFP) intoxication, a surrogate of nerve agent sarin.

Diisopropyl fluorophosphate (DFP), a surrogate of nerve agent sarin, is an organophosphorus (OP) compound which inhibits neuronal enzyme acetylcholinesterase (AChE). Exposure of this compound leads to a wide range of toxic symptoms and survivors may exhibit long term neurotoxicity related to cognitive and memory defects. Due to ease of availability and similar mechanism of action to other highly toxic nerve agent, DFP is widely used as model compound to trace changes associated with nerve agent exposures. Proximal fluids are widely used for the elucidation of biomarkers for exposure to toxic substances and to study the mechanism of toxicity. Using a rat model of OP intoxication, the present study was carried out to elucidate proteomic changes in plasma associated with DFP intoxication. Rats were exposed to a single dose (0.5 LD50) of DFP and their plasma proteome was studied, one day post exposure by two dimensional gel electrophoresis - mass spectrometry (2DE-MS). Some of the milestone changes were validated by Western blot analysis. A total 15 proteins showed significant fold changes in expression with respect to control after 1 day of DFP intoxication. Most of the proteins showing changes in expression at initial stages were related to immunogenic function, acute phase response, blood coagulation, and stress response. Experiments reported here demonstrate that 0.5 LD50 DFP intoxication leads to AChE inhibition, modulation of immunogenic function, and generation of stress at an early stage. Although, some proteins and their putative functional ramifications indicated similarity with those observed in our previous plasma proteome study, neurodegenerative changes were not observed in plasma of 0.5 LD50 DFP treated animals.