Search

Purpose: To curtail the potential of donor corneal tissue disseminating fungi to the recipient's eye, we evaluated the addition of amphotericin B to McCarey-Kaufman (M-K)—corneal storage medium supplemented with colistin. Methods: Amphotericin B was examined for its ability to inhibit the growth of Candida albicans and Aspergillus flavus using a microbroth dilution test and checkerboard assay in combination with only gentamicin and a combination of colistin, gentamicin, and amphotericin B. The safety on epithelium and endothelium was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Results: The minimal inhibitory concentration of gentamicin was found to be >256 μg/ml against both C. albicans and A. flavus, whereas that of amphotericin B was found to be in a range of 0.25–0.5 and 1–2 μg/ml for C. albicans and A. flavus, respectively. According to the checkerboard assay, 80% (4/5) of C. albicans isolates and 100% (5/5) of A. flavus isolates responded synergistically to the combination of amphotericin B and gentamicin, but only 20% (1/5) of C. albicans isolates showed an additive effect. None of the tested isolates displayed antagonism. The combined effect of the three drugs also did not display any antagonistic effect. Additionally, the MTT assay reveals no toxic effect of the antimicrobials used on corneal epithelial and endothelial cells. Conclusion: In vitro experiments demonstrate that amphotericin B is not toxic to either epithelium or endothelium and is a promising additive to the M-K medium supplemented with colistin.

Abstract Background Previous research exploring the role of race on prostate cancer (PCa) outcomes has demonstrated greater rates of disease progression and poorer overall survival for African American (AA) compared to Caucasian American (CA) men. The current study examines self‐reported race as a predictor of long‐term PCa outcomes in patients with low and favorable‐intermediate risk disease treated with external beam radiation therapy (EBRT). Methods This retrospective cohort study examined patients who were consented to enrollment in the Center for Prostate Disease Research Multicenter National Database between January 01, 1990 and December 31, 2017. Men self‐reporting as AA or CA who underwent EBRT for newly diagnosed National Comprehensive Cancer Network‐defined low or favorable‐intermediate risk PCa were included. Dependent study outcomes included: biochemical recurrence‐free survival, (ii) distant metastasis‐free survival, and (iii) overall survival. Each outcome was modeled as a time‐to‐event endpoint using race‐stratified Kaplan–Meier estimation curves and multivariable Cox proportional hazards analysis. Results Of 840 men included in this study, 268 (32%) were AA and 572 (68%) were CA. The frequency of biochemical recurrence, distant metastasis, and deaths from any cause was 151 (18.7%), 29 (3.5%), and 333 (39.6%), respectively. AA men had a significantly younger median age at time of EBRT and slightly higher biopsy Gleason scores. Multivariable Cox proportional hazards analyses demonstrated no racial differences in any of the study endpoints. Conclusions These findings reveal no racial disparity in PCa outcomes for AA compared to CA men, in a long‐standing, longitudinal cohort of patients with comparable access to cancer care.

Purpose: To assess the efficacy of the addition of polymyxin E (colistin) in the McCarey-Kaufman (MK) corneal storage solution against multi-drug resistant strains of Enterobacteriaceae, Staphylococcus aureus, and Candida spp. Methods: A standard micro broth dilution test and a checkerboard assay were performed for five multi-drug resistant (MDR) clinical strains of P. aeruginosa and five clinical strains of methicillin-resistant S. aureus (MRSA) and C. albicans against colistin and gentamicin alone and in combination. The minimum inhibitory concentration (MIC) and the fractional inhibitory concentration index (FICI) were calculated to assess the efficacy of each combination. Results: The MIC of colistin was in the range of 1–2 μg/mL for P. aeruginosa, whereas it was 256–1024 μg/mL against S. aureus. In comparison, the MIC of gentamicin was found to be 0.5–512 μg/mL and 0.5–8 μg/mL against P. aeruginosa and S. aureus, respectively. All five isolates of C. albicans did not exhibit any susceptibility to either colistin or gentamicin even at a concentration of ≥ 512 μg/mL each. The checkerboard assay was performed to evaluate the nature of the interaction of the combination of colistin and gentamicin. Based on the FICI, it was observed that the colistin and gentamicin combination has a maximum synergistic effect (FIC 0.5–4) was 100% (5/5) for P. aeruginosa and the minimum additive effect was 20% (1/5) for S. aureus isolates. Antagonism (FIC ≥ 4) was not observed in any combination between the strains used in the study. Both colistin and gentamicin alone or in combination were, however, ineffective against Candida spp. Conclusion: The addition of colistin has an inhibitory effect on bacterial contamination that could be possibly caused by MDR strains and could potentially be considered as an additional additive in corneal storage media.

oronavirus (COVID-19) has caused marked impact on graduate medical education for all medical specialties. Radiation Oncology and the American Board of Radiology have also had to rapidly adapt to converting education and examinations to virtual platforms. We describe our small pilot experience in transitioning our in-person mock oral examinations to a virtual platform. Survey-based assessment revealed excellent feedback regarding ease of use and educational usefulness. Our mock oral examinations pilot experience adds to evidence that virtual mock oral examinations are an important considerationfor Radiation Oncology education and a feasible alternative to an in-person oral examination.

Purpose: The aim of this study is to quantify and describe the feasibility, clinical outcomes, and patient-reported outcomes of reduced planning target volume (PTV) margins for prostate cancer treatment using real-time, continuous, intrafraction monitoring with implanted radiation frequency transponder beacons. Methods and materials: For this prospective, nonrandomized trial, the Calypso localization system was used for intrafraction target localization in 31 patients with a PTV margin reduced to 2 mm in all directions. A total of 1333 fractions were analyzed with respect to movement of the prostate, pauses and interruptions, and dosimetric data. Pre- and posttreatment quality-of-life scores were tracked at baseline, during treatment, and up to 24 months after treatment. Results: The mean time of daily treatment was 10 minutes, with 96.1% of all treatments falling within a 20-minute treatment window standard. On average, beacon motion exceeded 3 mm during active treatment only 1.76% of the time. The average length of treatment interruption was 34.2 seconds, with an average of 1 interruption every 3.39 fractions. The displacement or excursion of the prostate was the greatest in the superior or inferior dimension (0.11 mm and 0.09 mm, respectively) and anterior or posterior dimension (0.07 mm and 0.13 mm, respectively), followed by the left or right dimension (0.05 mm and 0.06 mm, respectively). At 6 months, patients demonstrated a smaller change in Expanded Prostate Cancer Index Composite scores than the ProtecT comparator group (decreased short-term morbidity). However, in the Bowel and Urinary domains at 12 and 24 months, there was no significant difference. Conclusions: Our data confirm and support that the use of Calypso tracking with intensity modulated radiation therapy reliably provides minimal disruption to daily treatments and overall time of treatment, with the PTV only moving outside of a 3-mm margin < 2% of the time. The use of a 3-mm PTV margin provides adequate dosimetric coverage while minimizing genitourinary and gastrointestinal toxicity.

Fagonia arabica (FA) possesses a thrombolytic property which has been earlier reported in our laboratory. Current study was undertaken to investigate the effect of aqueous extract of FA on thrombin-induced tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) release from cultured human umbilical vein endothelial cell line (HUVE) for studying its clot lytic activity. For this, establishment of cell line model has been done by isolating the cells from human umbilical cord. Cell toxicity was evaluated using XTT assay. Estimation of t-PA and PAI-1 t-PA complex were done using ELISA technique. Thrombin treatment induces the t-PA and PAI-1 release from HUVE cell line, and FA treatment was found to antagonize the thrombin induced t-PA and PAI-1 release. Our preliminary results suggest that FA may be used as an alternative to thrombolytic drug. However, study demands further experiments using animal model of thrombosis to establish the role of FA as a novel thrombolytic drug.

Poor dietary intake or unhealthy lifestyle contributes to various health disorders, including postprandial hyperglycemia, leading to type-2 diabetes mellitus (T2DM). Reduction of postprandial glucose levels through diet is a key strategy for preventing and managing T2DM. Thus, it is essential to understand how dietary components affect glycemic regulation. Dietary polyphenols (DPs), such as anthocyanins and other phenolics found in various fruits and vegetables, are often recommended for their potential health benefits, though their systemic effectiveness is subject to ongoing debate. Therefore, this review assesses the current and historical evidence of DPs bioactivities, which regulate crucial metabolic markers to lower postprandial hyperglycemia. Significant bioactivities such as modulation of glucose transporters, activation of 5' adenosine monophosphate-activated protein kinase (AMPK), and regulation of incretins are discussed, along with prospects for diet-induced therapeutics to prevent the onset of T2DM. STATEMENT OF SIGNIFICANCE: This review uniquely synthesizes both current and historical data on the metabolic bioactivities of dietary polyphenols (DPs), particularly focusing on their mechanisms; such as modulation of glucose transporters, AMPK activation, and incretin regulation that may help lower postprandial hyperglycemia, offering a more integrated understanding of diet-induced strategies for T2DM prevention., Competing Interests: Declaration of Competing Interest ☒ The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

Leptospira borgpetersenii commonly causes human leptospirosis, including severe disease. The first published analysis of L. borgpetersenii , performed on two strains of serovar Hardjo (L550 and JB197), concluded that the L. borgpetersenii genome is in the process of genome decay with functional consequences leading to a more obligately host-dependent life cycle. Yet whole genome analysis has only been carried out on few strains of L. borgpetersenii , with limited closed genomes and comprehensive analysis. Herein we report the complete, circularized genomes of seven non-Hardjo Leptospira borgpetersenii isolates from human leptospirosis patients in Sri Lanka. These isolates (all ST144) were found to be nearly identical by whole genome analysis; serotyping showed they are a novel serovar. We show that the L. borgpetersenii isolated from humans in Sri Lanka are less genomically decayed than previously reported isolates: fewer pseudogenes (N=141) and Insertion Sequence (IS) elements (N=46) compared to N=248, N=270, and N=400 pseudogenes, and N=121 and N=116 IS elements in published L. borgpetersenii Hardjo genomes (L550, JB197 and TC112). Compared to previously published L. borgpetersenii whole genome analyses showing two to three VM proteins in L. borgpetersenii isolates from cattle, rats and humans, we found that all of the human L. borgpetersenii isolates from Sri Lanka, including previously reported serovar Piyasena, have 4 encoded VM proteins, one ortholog of L. interrogans Copenhageni LIC12339 and 3 orthologs of LIC12844. Our findings of fewer pseudogenes, IS elements and expansion of the LIC12844 homologs of the PF07598 family in these human isolates suggests that this newly identified L. borgpetersenii serovar from Sri Lanka has unique pathogenicity. Comparative genome analysis and experimental studies of these L. borgpetersenii isolates will enable deeper insights into the molecular and cellular mechanisms of leptospirosis pathogenesis., Competing Interests: Declaration of Competing Interest Some of work reported here has been filed in patent applications from Yale University. JMV and spouse have an equity interest in Luna Bioscience, Inc, which may have a future interest in licensing this work. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The field of tissue engineering has witnessed significant advancements with the advent of hydrogel nanocomposites (HNC), emerging as a highly promising platform for regenerative medicine. HNCs provide a versatile platform that significantly enhances the differentiation of stem cells into specific cell lineages, making them highly suitable for tissue engineering applications. By incorporating nanoparticles, the mechanical properties of hydrogels, such as elasticity, porosity, and stiffness, are improved, addressing common challenges such as short-term stability, cytotoxicity, and scalability. These nanocomposites also exhibit enhanced biocompatibility and bioavailability, which are crucial to their effectiveness in clinical applications. Furthermore, HNCs are responsive to various triggers, allowing for precise control over their chemical properties, which is beneficial in creating 3D microenvironments, promoting wound healing, and enabling controlled drug delivery systems. This review provides a comprehensive overview of the production methods of HNCs and the factors influencing their physicochemical and biological properties, particularly in relation to stem cell differentiation and tissue repair. Additionally, it discusses the challenges in developing HNCs and highlights their potential to transform the field of regenerative medicine through improved mechanotransduction and controlled release systems.

Background and Objectives: Regulatory requirement of fixed holding time (6 h) of whole blood (WB) at room temperature, that is, 22-24°C (RT) results in sub-optimal component separation. The aim was to evaluate the platelet concentrates (PC) prepared by both platelet rich plasma (PRP) and buffy coat (BC) methods after overnight hold (18-24 h) at RT., Materials and Methods: A prospective experimental study was performed. A total of 48 WB units collected were divided into four groups (12 each) control-1 (C1) and test-1 (T1) for PRP and control-2 (C2) and test-2 (T2) for the BC method. Control groups were processed within 6 h, and in test groups, components were prepared after overnight hold, followed by evaluation of quality parameters., Results: Irrespective of the method used, all PCs had similar volume, platelet yield, swirling, no bacterial contamination, RBC contamination, PaO 2 and PaCO 2 levels. PCs in the T1 group had significant differences in glucose and MPV values on d1, which were resolved by d5 of storage. PCs in T2 has significant differences in pH, glucose, and MPV levels throughout storage. PRBC in test and control groups had similar quality parameters till d42 of storage. FFPs in all tests were noninferior to the concurrent control groups till 3 months of storage., Conclusion: Overnight holding of WB had no lasting deleterious changes. Though a few biochemical parameters in the test groups were significantly different, they can be accepted to improve the logistics of component separation. Overall PRP method seemed to have a better result than the BC method after an overnight hold., (© 2024 British Blood Transfusion Society.)

Vector-borne diseases are a leading cause of death worldwide and pose a substantial unmet medical need. Pathogens binding to host extracellular proteins (the "exoproteome") represents a crucial interface in the etiology of vector-borne disease. Here, we used bacterial selection to elucidate host-microbe interactions in high throughput (BASEHIT)-a technique enabling interrogation of microbial interactions with 3,324 human exoproteins-to profile the interactomes of 82 human-pathogen samples, including 30 strains of arthropod-borne pathogens and 8 strains of related non-vector-borne pathogens. The resulting atlas revealed 1,303 putative interactions, including hundreds of pairings with potential roles in pathogenesis, including cell invasion, tissue colonization, immune evasion, and host sensing. Subsequent functional investigations uncovered that Lyme disease spirochetes recognize epidermal growth factor as an environmental cue of transcriptional regulation and that conserved interactions between intracellular pathogens and thioredoxins facilitate cell invasion. In summary, this interactome atlas provides molecular-level insights into microbial pathogenesis and reveals potential host-directed targets for next-generation therapeutics., Competing Interests: Declaration of interests N.W.P. and A.M.R. are inventors of a patent describing the BASEHIT technique., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Tuberculosis is a serious life-threatening disease among the top global health challenges and rapid and effective diagnostic biomarkers are vital for early diagnosis especially given the increasing prevalence of multidrug resistance., Methods: Two human whole blood microarray datasets, GSE42826 and GSE42830 were retrieved from publicly available gene expression omnibus (GEO) database. Deregulated genes (DEGs) were identified using GEO2R online tool and Gene Ontology (GO), protein-protein interaction (PPI) network analysis was performed using Metascape and STRING databases. Significant genes (n = 8) were identified using T-test/ANOVA and Molecular Complex Detection (MCODE) score ≥10, which was validated in GSE34608 dataset. The diagnostic potential of three biomarkers was assessed using Area Under Curve (AUC) of Receiver Operating Characteristic (ROC) plot. The transcriptional levels of these genes were also examined in a separate dataset GSE31348, to monitor the patterns of variation during tuberculosis treatment., Results: A total of 62 common DEGs (57 upregulated, 7 downregulated genes) were identified in two discovery datasets. GO functions and pathway enrichment analysis shed light on the functional roles of these DEGs in immune response and type-II interferon signaling. The genes in Module-1 (n = 18) were linked to innate immune response, interferon-gamma signaling. The common genes (n = 8) were validated in GSE34608 dataset, that corroborates the results obtained from discovery sets. The gene expression levels demonstrated responsiveness to Mtb infection during anti-TB therapy in GSE31348 dataset. In GSE34608 dataset, the expression levels of three specific genes, GBP5, IFITM3, and EPSTI1, emerged as potential diagnostic makers. In combination, these genes scored remarkable diagnostic performance with 100% sensitivity and 89% specificity, resulting in an impressive Area Under Curve (AUC) of 0.958. However, GBP5 alone showed the highest AUC of 0.986 with 100% sensitivity and 89% specificity., Conclusions: The study presents valuable insights into the critical gene network perturbed during tuberculosis. These genes are determinants for assessing the effectiveness of an anti-TB response and distinguishing between active TB and healthy individuals. GBP5, IFITM3 and EPSTI1 emerged as candidate core genes in TB and holds potential as novel molecular targets for the development of interventions in the treatment of TB., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Arya et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background and Objectives: Notifying blood donors of their reactive status for transfusion-transmitted infections (TTIs) plays a vital role in enabling early diagnosis and management while also preventing these donors from making future donation and transmission of the infectious agent. Given the limited data on donor notification processes in India, a narrative review was conducted to assess the existing notification process and identify areas requiring enhancement., Materials and Methods: We conducted literature searches using PubMed, Google Scholar and Scopus, employing various keywords. The review included data on the year of the study, study design, donor numbers, TTI screening methods, sero-reactive donor confirmation, notification frequency and methods, donor responses, post-test counselling and risk factor assessment., Results: Out of the 29 identified articles, 16 studies were included in the analysis. Repeat testing for initially reactive results was conducted in nine studies for 24.3% reactive donors. Phone calls were the primary notification method in most studies (8; 50%), with letters sent in cases of no response. Only 12 studies provided data on notified donors, revealing a notification rate of 71.2%. Of all initially reactive donors, 33.3% sought post-test counselling. Data from six studies indicated that 74.3% of responsive donors had identifiable TTI risk factors., Conclusion: Our review revealed significant variability in the notification processes across different studies. To enhance the management of TTI-reactive donor notifications and responses, we recommend the establishment of universal protocols encompassing pre-donation counselling, repeat/confirmatory testing, notification methods and comprehensive follow-up and treatment., (© 2023 International Society of Blood Transfusion.)

Introduction: Acute trauma resuscitation is challenging and disorganized in low- and middle-income countries as there is lack of evidence-based transfusion practices with respect to ratio, volume, trigger, cutoff points for various triggers. This study aims to study the patterns of blood use in various mechanisms of injury, by interfacility transfer status and define triggers for transfusion of various blood components. Prospective observational study, emergency department of level 1 trauma center (August 2017-August 2018)., Methods: Demographic, injury, clinical, laboratory, transfusion characteristics, and patient outcomes were collected from electronic hospital records and transfusion service records. We used multivariate logistic regression to identify triggers of transfusion of red blood cells, plasma, platelets (PLTs), cryoprecipitate, and mortality predictors., Results: Among 986 severely injured patients 80% were males, 92% had blunt injuries and commonest trauma was Head. The median length of intensive care unit stay and hospital stay were more in a referred group. Patients brought to center directly received more massive transfusions (56, 13.05% vs. 48, 8.62%). Railway track injuries received the most total median packed red blood cells (PRBCs) units in both groups. The triggers for various blood product transfusions include: Hematocrit < 32.08 for PRBC, PT >16.9 s for plasma, PLT count <130 lakhs for platelets, MAP <89.7 mmHg and PT >18.2 s for cryoprecipitate., Conclusion: Railway track injuries with inferior extremity injuries required maximum transfusion requirements and were a trigger for PRBC, plasma, PLTs, and cryoprecipitate. Smartphone-based apps and transfusion prediction models can be framed based on the triggers and cut points., Competing Interests: There are no conflicts of interest., (Copyright: © 2024 Journal of Emergencies, Trauma, and Shock.)

Tuberculosis (TB) remains a significant global health concern, necessitating accurate diagnosis and treatment monitoring. Extracellular vesicles (EVs), including exosomes, play crucial roles in disease progression, with their associated genes serving as potential biomarkers and therapeutic targets. Leveraging publicly available RNA-Seq datasets of TB patients and healthy controls (HCs), to identify differentially expressed genes (DEGs) and their associated protein-protein interaction networks and immune cell profiles, the common EV-related DEGs were identified and validated in the GSE42830 and GSE40553 datasets. We have identified nine common EV-related DEGs (SERPINA1, TNFAIP6, MAPK14, STAT1, ITGA2B, VAMP5, CTSL, CEACAM1, and PLAUR) upregulated in TB patients. Immune cell infiltration analysis revealed significant differences between TB patients and HCs, highlighting increased proportions of various immune cells in TB patients. These DEGs are involved in crucial cellular processes and pathways related to exocytosis and immune response regulation. Notably, VAMP5 exhibited excellent diagnostic performance (AUC-0.993, sensitivity-93.8%, specificity-100%), with potential as a novel biomarker for TB. The EV-related genes can serve as novel potential biomarkers that can distinguish between TB and HCs. VAMP5, which functions in exosome biogenesis and showed significant upregulation in TB, can be targeted for therapeutic interventions and treatment outcomes., Competing Interests: The authors declare no conflicts of interest.

Background: Lower limb trauma in emergency settings often leads to pain management challenges. Traditional methods, primarily non-steroidal anti-inflammatory drugs (NSAIDs) and opioids, have limitations. This study explores the efficacy of epidural analgesia in emergency for lower limb trauma patients (ELETRA) as an alternative for managing pain in lower limb trauma patients upon arrival in emergency care., Objectives:  This study primarily focuses on determining ELETRA's effectiveness in reducing pain for patients with lower limb trauma upon arrival. It also aims to evaluate ELETRA's influence on decreasing stress and shortening hospital stays compared to traditional pain management approaches., Methods: This study was executed as a prospective, parallel-design, randomized controlled trial in the emergency department of a tertiary care teaching hospital. The interventions were performed in a dedicated induction room adjacent to the emergency operating theater. The trial was registered in the Clinical Trial Registry of India with registration number CTRI/2022/08/044699 dated 16/08/2022. Participants were 18- to 50-year-old patients with lower limb injuries, classified under American Society of Anesthesiologists (ASA) class 1 or 2. Participants were randomized into two groups: one receiving ELETRA and the other standard pain control treatment. The effectiveness of pain relief was measured through a visual analog scale (VAS), and hemodynamic parameters, adverse effects, levels of acute phase reactants, and stress hormones were also measured along with patient satisfaction., Results: The study enrolled 356 participants with lower limb injuries. After excluding participants lost following the intervention, data from 157 individuals in Group A and 160 in Group B were analyzed. Group A's success rate for analgesia (VAS < 2) was 92.35% (n = 145), significantly higher than Group B's 75.62% (n = 121) (p < 0.001). The commonest side effect was hypotension (5.73%) in Group A and nausea in Group B (7.5%). C-reactive protein (CRP) levels rose to 104.71 ± 8.99 mg/dL in Group A and 192.58 ± 9.23 mg/dL in Group B; the difference was statistically significant. Serum cortisol levels were also higher in Group B (67.18 ± 9.21) compared to Group A (44.72 ± 6.14) at one week. Group B had a longer hospital stay, averaging 12.24 ± 4.81 days, against Group A's 10.19 ± 4.91 days., Conclusion: ELETRA is a safe and effective alternative for pain management in lower limb trauma patients in emergency settings. It reduces pain, improves patient satisfaction, and has a favorable impact on stress responses., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Jain et al.)

Dopaminergic (DAergic) neurodegeneration in the substantia nigra pars compacta of the human brain is the pathological feature associated with Parkinson's disease (PD). Drosophila also exhibits mobility defects and diminished levels of brain dopamine on exposure to neurotoxicants mimicking PD. Our laboratory demonstrated in a Drosophila model of sporadic PD that there is no decrease in DAergic neuronal number; instead, there is a significant reduction in tyrosine hydroxylase (TH) fluorescence intensity (FI). Here, we present a sensitive assay based on the quantification of FI of the secondary antibody (ab). As the FI is directly proportional to the amount of TH synthesis, its reduction under PD conditions denotes the decrease in the TH synthesis, suggesting DAergic neuronal dysfunction. Therefore, FI quantification is a refined and sensitive method to understand the early stages of DAergic neurodegeneration. FI quantification is performed using the ZEN 2012 SP2 single-user software; a license must be acquired to utilize the imaging system to interactively control image acquisition, image processing, and analysis. This method will be of good use to biologists, as it can also be used with little modification to characterize the extent of degeneration and changes in the level of degeneration in response to drugs in different cell types. Unlike the expensive and cumbersome confocal microscopy, the present method will be an affordable option for fund-constrained neurobiology laboratories. Key features • Allows characterizing the incipient DAergic and other catecholaminergic neurodegeneration, even in the absence of loss of neuronal cell body. • Great alternative for the fund-constrained neurobiology laboratories in developing countries to utilize this method in different cell types and their response to drugs/nutraceuticals., Competing Interests: Competing interestsThe author declares no conflicts of interest., (©Copyright : © 2024 The Authors; This is an open access article under the CC BY-NC license.)

Introduction: Vasovagal reaction (VVR) is a frequently encountered generalised donor adverse reaction, associated with donor deterrence towards future donation. Several mitigation strategies for prevention of VVR were tried but still not standardised. This quadri-armed randomised study evaluated the utility of water ingestion, applied muscle tension (AMT) and combination of both in preventing the VVR among blood donors., Methods: A quadri-armed randomised controlled trial was performed on 4320 whole blood donors. Blood donors of 18-65 years of age were randomised into four groups based on the interventions performed i.e., control with no intervention (Group 1, n = 1081), water ingestion (Group 2, n = 1082), AMT (Group 3, n = 1070) and combined intervention (Group 4, n = 1087). VVR during and immediately after blood donation were observed along with assessment of risk factors in blood donors and the effectiveness of interventions were analysed., Results: The incidence of VVR observed 1.6% in our study, with the highest occurrence in the control group (2.5%) and the lowest in the combined intervention group (0.9%). Multivariable logistic regression revealed that the control group donors faced a 1.38-fold greater risk of VVR compared to those receiving interventions (OR: 1.38, 95% CI: 1.10-1.75). Other risk factors included younger age (OR: 1.5, 95% CI: 1.05-2.17), first-time donation (OR: 5.7, 95% CI: 1.66-5.74), prior history of VVR (OR: 2.5, 95% CI: 10.4-101.52)., Discussion/conclusion: The combined approach of water ingestion and AMT proved significantly more effective in VVR prevention compared to individual interventions., (© 2023 British Blood Transfusion Society.)

Competing Interests: Declaration of Competing Interest Authors have no conflict of Interest.

To compare and evaluation of surgical operability with and without induction chemotherapy in locally advanced head and neck squamous cell carcinoma. Head and neck malignancy grossly refers to squamous cell carcinomas of head and neck (HNSCC) have multiple treatment modalities and strategies, when opted in an appropriate manner renders tumours curable. The aim of this study is to compare and evaluation of surgical operability with and without induction chemotherapy in locally advanced head and neck squamous cell carcinoma. A prospective observational study involving 50 patients of histologically proven squamous cell carcinoma of head and neck region. Patients were categorized into two major groups, group-1 patients included resectable tumour stage and group-2 included unresectable tumour stage. Both groups were compared after appropriate chemotherapy and surgical intervention. There were a total of 78% males and 22% females with majority of patients in age group of 41-60 years. 54% patients had ulcerative type of growth pattern and most patients had primary site of lesion in oral cavity. 50% patients had moderately differentiated squamous cell carcinoma. Induction chemotherapy was considered in 70% of patients, while majority of patients were belonging to T4N2M0 stage. In this study, we recommend that the borderline category of patients who are initially in an unresectable tumour stage can undergo induction chemotherapy to downstage and shrink the tumour to a resectable stage following which the appropriate surgical intervention should be done with a close monitoring and sustained follow up to prevent recurrence., Competing Interests: Competing interestsThe authors have no competing interests to declare that are relevant to the content of this article. All authors certify that they have no affiliations with or involvement in any organization or entity with any financial interest or non-financial interest in the subject matter or materials discussed in this manuscript. The authors have no financial or proprietary interests in any material discussed in this article. Authors are responsible for correctness of the statements provided in the manuscript., (© Association of Otolaryngologists of India 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Here, we investigate the structural and electrical properties on SrIrO 3 films grown on LaAlO 3 (100) substrate with varying thickness (18, 25 and 40 nm). The x-ray diffraction shows good quality epitaxial films without any chemical impurity. The out-of-plane lattice parameter increases with the film thickness. All the films show a semiconducting behavior where the resistivity increases with increasing thickness. Our analysis shows at high temperature the charge conduction mechanism follows Mott's two-dimensional variable-range-hopping model. Detailed current-voltage ( I - V ) measurements show a linear Ohmic behavior at room temperature, however, a prominent deviation from linearity has been observed at low temperatures where the exponent n (I∝Vn) increases with decreasing temperature, reachingn ∼1.5 at low temperature. Analysis of I - V data indicates that the charge conduction has dominant contribution of Poole-Frenkel mechanism rather than Schottky behavior. This evolution of charge transport with temperature is quite intriguing which may be related to the development of low temperature magnetism in films of SrIrO 3 ., (© 2023 IOP Publishing Ltd.)

Background: Coronavirus disease 2019 (COVID-19), is known for its variable severity and high infectivity. Though fewer than 15% of infected cases develop severe disease, a major proportion had prolonged stay in the intensive care unit (ICU). Prolonged ICU stay is known to have a long-term impact on behavior and quality of life. 8 Therefore, it is likely that patients discharged after severe COVID-19 have issues that persist for long term. The current study aimed to assess the long-term impact of severe COVID-19 on the Quality of life (QOL), sleep pattern, behavior, and workability., Methods: The current multicenter study adopted a cross-sectional design to analyze data from two tertiary care COVID-19 dedicated hospitals. All experimental procedures were approved by the ethics committee of the M.L.B Medical College. Participants were 20-60 age group who had been admitted to the ICU because of severe COVID-19 and had elapsed at least one and a half year since their discharge. After informed written consent the participants were assessed for: EUROHIS-QOL 8-item index; Workability Score; Quality of sleep; The major depression inventory (MDI) questionnaire; Generalized anxiety disorder 7 item scale (GAD-7); Current global health status score: an innovative subjective scale (1 -10) to determine the current global health status when 5 is the status before COVID-19., Findings: 491 participants were assessed, the median follow-up time after discharge from the hospital was 561·0 days (range, 548-580 days). The mean duration of ICU stay was 8.72 ± 2.85 days. There was significant reduction in the prevalence of obesity, diabetes, and hypertension as compared with discharge time. The mean of EUROHIS-QOL score, workability score, current global health status score was 3.28 ± 0.98, 6.87 ± 0.85, 4.53 ± 1.36 respectively. The mean MDI and anxiety scores were 4.12 ± 1.45 and 18.63 ± 3.28, respectively., Interpretation: Severe COVID-19 survivors have new-onset psychological disorders and sleep disturbances. Long term quality of life and work ability remains poor after prolong ICU admission secondary to severe COVID-19., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Recombinant fluorescent fusion proteins are fundamental to advancing many aspects of protein science. Such proteins are typically used to enable the visualization of functional proteins in experimental systems, particularly cell biology. An important problem in biotechnology is the production of functional, soluble proteins. Here we report the use of mCherry-fusions of soluble, cysteine-rich, Leptospira-secreted exotoxins in the PF07598 gene family, the so-called virulence modifying (VM) proteins. The mCherry fusion proteins facilitated the visual detection of pink colonies of the VM proteins (LA3490 and LA1402) and following them through lysis and sequential chromatography steps. CD-spectroscopy analysis confirmed the stability and robustness of the mCherry-fusion protein, with a structure comparable to AlphaFold structural predictions. LA0591, a unique member of the PF07598 gene family that lacks N-terminal ricin B-like domains, was produced without mCherry tag that strengthens the recombinant protein production protocol without fusion protein as well. The current study provides the approaches for the synthesis of 50-125 kDa soluble, cysteine-rich, high-quality fast protein liquid chromatography (FPLC)-purified protein, with and without a mCherry tag. The use of mCherry-fusion proteins enables a streamlined, efficient process of protein production and qualitative and quantitative downstream analytical and functional studies. Approaches for troubleshooting and optimization were evaluated to overcome difficulties in recombinant protein expression and purification, demonstrating biotechnology utility in accelerating recombinant protein production., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The field of injectable hydrogels has demonstrated a paramount headway in the myriad of biomedical applications and paved a path toward clinical advancements. The innate superiority of hydrogels emerging from organic constitution has exhibited dominance in overcoming the bottlenecks associated with inorganic-based hydrogels in the biological milieu. Inorganic hydrogels demonstrate various disadvantages, including limited biocompatibility, degradability, a cumbersome synthesis process, high cost, and ecotoxicity. The excellent biocompatibility, eco-friendliness, and manufacturing convenience of organo-hydrogels have demonstrated to be promising in therapizing biomedical complexities with low toxicity and augmented bioavailability. This report manifests the realization of biomimetic organo-hydrogels with the development of bioresponsive and self-healing injectable organo-hydrogels in the emerging pharmaceutical revolution. Furthermore, the influence of click chemistry in this regime as a backbone in the pharmaceutical conveyor belt has been suggested to scale up production. Moreover, we propose an avant-garde design stratagem of developing a hyaluronic acid (HA)-based injectable organo-hydrogel via click chemistry to be realized for its pharmaceutical edge. Ultimately, injectable organo-hydrogels that materialize from academia or industry are required to follow the standard set of rules established by global governing bodies, which has been delineated to comprehend their marketability. Thence, this perspective narrates the development of injectable organo-hydrogels via click chemistry as a prospective elixir to have in the arsenal of pharmaceuticals.

Living organisms are exposed to exogenous and endogenous agents that affect genomic integrity by creating DNA double strand breaks (DSBs). These breaks are repaired by DNA repair proteins to maintain homeostasis. Defects in DNA repair pathways also affect lymphocyte development and maturation, as DSB sites are critical intermediates for rearrangements required for V(D)J recombination. Recent classifications for inborn errors of immunity (IEIs) have listed DNA repair defect genes in a separate group, which suggests the importance of these genes for adaptive and innate immunity. We report an interesting case of a young female (index P1) with mutations in two different genes, DCLRE1C and FANCA, involved in DNA repair pathways. She presented with clinical manifestations attributed to both defects. With the advent of NGS, more than one defect is increasingly identified in patients with IEIs. Familial segregation studies and appropriate functional assays help ascertain the pathogenicity of these mutations and provide appropriate management and genetic counseling., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Competing Interests: Declaration of Competing Interest Authors have no conflict of Interest.

Background: Asymptomatic/presymptomatic COVID-19 affected individuals who may appear healthy during blood donor screening can donate blood despite being infective. Most blood donors in India are relatives/friends/acquaintances of patients, who under peer pressure overlook the donor selection process, which can significantly impact the transfusion safety., Aims: The prevalence of severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) antibodies among blood donors was assessed, along with the possible transmissibility of SARS-CoV-2 virus in transfusion recipients of blood components prepared from sero-reactive blood donors., Settings and Design: A prospective cross-sectional study was conducted among eligible blood donors from November-2020 to April 2021., Methods: 1500 blood donors were tested for SARS-CoV-2 IgG antibodies. Sero-reactive donors were followed-up telephonically to inquire about risk factors prior to donation or appearance of COVID-19 related symptoms postdonation. Patients transfused with blood components from seroreactive donors were also followed up for posttransfusion symptoms suggestive for COVID-19. Descriptive analysis was done for the donor and patient follow-up data., Results: A total of 452 (30.1%) donor were reactive, with median S/CO ratio of 2.8 (interquartile range 1.5-5.5). Risk factors such as travel, contact, or quarantine were significantly higher among reactive donors. History of diabetes and/or hypertension was associated with seroreactivity. Total 516 patients were transfused with blood components from these seroreactive donors. Three patients developed fever after transfusion, one of which was found to be PCR positive after 4 days of transfusion., Conclusion: Sero-reactivity rate among blood donors was lower than the general population. Optimum blood donor screening strategies can help decrease the possibility of blood collection from infected blood donors., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Asian Journal of Transfusion Science.)

During COVID-19 pandemic, many doctors were redirected from various disciplines for care of COVID-19 patients. A survey was conducted among doctors involved, to assess transfusion practices during the pandemic. To assess the knowledge, attitude, and practices of blood transfusion among doctors involved in care of COVID-19 patients. A cross-sectional survey using an online questionnaire (Google form) was used to assess knowledge and need of transfusion, attitude towards modifications in transfusion process, and practices during pandemic. Analysis was done among subgroups based on experience (designation), user type (speciality) and frequency of blood usage in parent department. Of 1900 invitations, 437 responses were received from resident doctors and faculty members across various disciplines. Of these, 354(81%) participants were included in analysis. Mean knowledge score was 6.2, majority 297(83.9%) had adequate knowledge scores (≥ 5 of total 12). Knowledge levels were higher among frequent blood users. Positive attitude towards changes in transfusion process was observed in 72.9% participants with similar scores in subgroups. Practice was assessed in 222(62.7%) participants. Mean practice score was 6.9, wherein 57.7% participants had optimal scores (≥ 7 of total 14). Positive correlation was observed between attitude and practice, unlike knowledge and practice. Although most participants had demonstrated adequate transfusion knowledge levels and positive attitude, transfusion practices during pandemic were affected mainly due to shortage of blood components and modifications in transfusion requisition process due to stringent COVID-19 containment measures. Thus, indicating need for improvement in the basic understanding of the transfusion process., Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-022-01613-2., Competing Interests: Conflict of InterestThe authors have no conflicts of interest to declare., (© The Author(s), under exclusive licence to Indian Society of Hematology and Blood Transfusion 2022. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Death of dopaminergic (DAergic) neurons in the substantia nigra pars compacta of the human brain is the characteristic pathological feature of Parkinson's disease (PD). On exposure to neurotoxicants, Drosophila too exhibits mobility defects and diminished levels of brain dopamine. In the fly model of sporadic PD, our laboratory has demonstrated that there is no loss of DAergic neuronal number, however, a significant reduction in fluorescence intensity (FI) of secondary antibodies that target the primary antibody-anti-tyrosine hydroxylase (TH). Here, we present a sensitive, economical, and repeatable assay to characterize neurodegeneration based on the quantification of FI of the secondary antibody. As the intensity of fluorescence correlates with the amount of TH synthesis, its reduction under PD conditions denotes the depletion in the TH synthesis, suggesting DAergic neuronal dysfunction. Reduction in TH protein synthesis is further confirmed through Bio-Rad Stain-Free Western Blotting. Quantification of brain DA and its metabolites (DOPAC and HVA) using HPLC-ECD further demonstrated the depleted DA level and altered DA metabolism as evident from enhanced DA turnover rate. Together all these PD marker studies suggest that FI quantification is a refined and sensitive method to understand the early stages of DAergic neurodegeneration. FI quantification is performed using ZEN 2012 SP2, a licensed software from Carl Zeiss, Germany. This method will be of good use to biologists, as it with few modifications, can also be implemented to characterize the extent of degeneration of different cell types. Unlike the expensive and cumbersome confocal microscopy, the present method using fluorescence microscopy will be a feasible option for fund-constrained neurobiology laboratories in developing countries., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ayajuddin, Chaurasia, Das, Modi, Phom, Koza and Yenisetti.)

Introduction: Neurosurgery training lacks uniformity across the world. Differences in the methods of training adopted during the training period is one of the major problems encountered in the field of neurosurgery all over the world. Moreover, neurosurgery is not "one neurosurgery"; in other words, it is not a unified whole., Material and Methods: In this study, we attempt to evaluate the present conditions of neurosurgery training in Nepal by exploring different institutions providing the training., Results: Due to various factors and challenges, the neurosurgery training provided in Nepal varies in different institutions. Many travel abroad for training due to a lack of sufficient seats in training institutions., Discussion and Conclusions: Despite the challenges, the future is bright for neurosurgery training in Nepal. With continued investment in education and training, and the adoption of new technologies and techniques, it is likely that the field of neurosurgery will continue to thrive and make a positive impact on the health and well-being of the Nepali population., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Chaurasia, Raut, Chaurasia and Thapa.)

Background and Objectives: A critical appraisal of the literature helps to assess the strength and weakness of the research and suggests ways to improve future research. Our aim was to critically appraise the knowledge, attitude, and practice (KAP) studies conducted in India for blood donation., Materials and Methods: Of 70 articles identified in our search on PubMed, Scopus and Google Scholar, 32 were assessed for quality using an appraisal tool for cross-sectional studies (AXIS) and questionnaire items., Results: Quality assessment revealed that only 6 of 32 studies had acceptable reporting (≥80% score on the AXIS tool). The most frequently identified shortcomings were failure to address the non-responders, lack of justification for sample size, assessment of outcome variables and demographic results for the survey. Our evaluation of questionnaires revealed that knowledge for need for blood donation, its benefits and site/place for blood donation were assessed by very few studies. With this, issues such as parental/family consent, religious beliefs, and indifference to blood donation process were amongst the common reasons for non-donation. Many studies also failed to ask questions related to procedural information/instructions, which are necessary for promoting voluntary blood donations., Conclusion: Most published KAP studies for blood donation in India were not appropriately described, especially the methodology and result section. These deficiencies could have led to suboptimal interpretation of the prevalent issues. Use of an open-ended and validated KAP questionnaire with a problem-based approach and inclusion of participants from various socio-cultural backgrounds is required for good quality of evidence., (© 2023 British Blood Transfusion Society.)

The journey into the field of stem cell biology has been an endeavor of paramount advancement in biomedicine, establishing new horizons in the avenue of materiobiology. The creative drive of the scientific community focuses on ameliorating the utilization of stem cells, which is currently untapped on a large scale. With similar motivation, we present a nascent strategy of maneuvering biogenic carbon quantum dots (CQDs) to eclipse the toxic hurdles of chemical synthesis of carbon allotropes to serve as a biocompatible trident in stem cell biology employing a three-prong action of stem cell differentiation, imaging, and migration. The derivation of CQDs from garlic peels as a biogenic precursor abets in realizing the optophysical features of CQDs to image mesenchymal stem cells without hampering the biological systems with cytotoxicity. We report the versatility of biogenic CQDs to generate reactive oxygen species (ROS) to robustly influence stem cell migration and concomitantly chondrocyte differentiation from human Wharton's jelly mesenchymal stem cells (hWJ-MSCs). This was orchestrated without the use of chondrogenic induction factors, which was confirmed from the expression of chondrogenic markers (Col II, Col X, ACAN). Even the collagen content of cells incubated with CQDs was quite comparable with that of chondrocyte-induced cells. Thus, we empirically propose garlic peel-derived CQDs as a tangible advancement in stem cell biology from a materiobiological frame of reference to hone significant development in this arena.

Sexual dysfunction (SD) is one of the most common non-motor symptoms of Parkinson's disease (PD) and remains the most neglected, under-reported, and under-recognized aspect of PD. Studies have shown that Dopamine (DA) in the hypothalamus plays a role in regulating sexual behavior. But the detailed mechanism of SD in PD is not known. Drosophila melanogaster shares several genes and signaling pathways with humans which makes it an ideal model for the study of a neurodegenerative disorder such as PD. Courtship behavior of Drosophila is one such behavior that is closely related to human sexual behavior and so plays an important role in understanding sexual behavior in diseased conditions as well. In the present study, a sporadic SD model of PD using Drosophila was developed and SD phenotype was observed based on abnormalities in courtship behavior markers. The Drosophila SD model was developed in such a way that at the window of neurotoxin paraquat (PQ) treatment [PQ is considered a crucial risk factor for PD due to its structural similarity with 1-methyl-4-phenyl pyridinium (MPP+), the active form of PD-inducing agent, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)], it does not exhibit mobility defects but shows SD. The whole brain tyrosine hydroxylase immunostaining showed no observable dopaminergic (DAergic) degeneration (number of DA neurons and fluorescence intensity of fluorescently labeled secondary antibodies that target anti-TH primary antibody) of the SD model. Similarly, there was no significant depletion of brain DA and its metabolite levels (HVA and DOPAC) as determined using HPLC-ECD (High-Performance Liquid Chromatography using Electrochemical Detector). The present study illustrates that the traits associated with courtship and sexual activity provide sensitive markers at the earlier stage of PD onset. This PQ-induced SD fly model throws an opportunity to decipher the molecular basis of SD under PD conditions and to screen nutraceuticals/potential therapeutic molecules to rescue SD phenotype and further to DAergic neuroprotection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Koza, Ayajuddin, Das, Chaurasia, Phom and Yenisetti.)

-Enhancing the productivity of rainfed crops, especially rice, while coping with climate adversities and saving critical natural resources is essential for ensuring the food and nutrition security of a growing population. With this context, the present study was undertaken to validate promising farm innovation and adaptation practices used by small-medium landholding farmers for rice cultivation in eastern Uttar Pradesh (UP), north India, as well as to examine the sustainability of innovative practices for large-scale adoption. For this, a 3-year study comprising extensive field surveys and experiments was undertaken to compare single transplantation (ST) and double transplantation (DT) in rice along with organic addition (farm-yard manure, FYM) on crop growth, yield, climate resilience, soil quality, and overall sustainability i.e., social (women involvements and labour productivity), environmental (water productivity and nutrient use efficiency), and economic (benefit:cost ratio) dimensions of sustainability. Field experiments were conducted in triplicate using two local rice varieties (MotiNP-360 and Sampurna Kaveri) in two agroclimatic zones, namely the middle Gangetic plains and the Vindhyan zone, in the Mirzapur district of eastern Uttar Pradesh. The DT practices of rice with and without farm yard manure (FYM) (replacing at a dose of 25% NPK) were evaluated over conventional methods of rice cultivation (i.e., ST, as control) and analysis was done periodically. The DT practice improved growth (p < 0.05), percent fertile tiller and grain (p < 0.05), and rice yield (15-20% higher than ST), while also improving soil quality, yield indices, water and labour productivity, and the benefit-cost ratio. The DT practice also resulted in early maturity (10-15 days earlier than ST), created more labour days for women, decreased lodging and pest/disease incidence, as well as a subsequent reduction in the use of synthetic chemical pesticides and associated environmental costs. Importantly, the residual effects of FYM application significantly improved (p < 0.05) the grain yield in subsequent years of cropping. Optimizing DT cultivation practices, preferably with FYM input for various agro-climatic regions, is essential for large-scale sustainable rice production under changing climatic conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Mechanisms of leptospirosis pathogenesis remain unclear despite the identification of a number of potential leptospiral virulence factors. We recently demonstrated potential mechanisms by which the virulence-modifying (VM) proteins-defined as containing a Domain of Unknown function (DUF1561), encoded by the PF07598 gene family-found only in group 1 pathogenic Leptospira -might mediate the clinical pathogenesis of leptospirosis. VM proteins belongs to classical AB toxin paradigm though have a unique AB domain architecture, unlike other AB toxins such as diphtheria toxin, pertussis toxin, shiga toxin, or ricin toxin which are typically encoded by two or more genes and self-assembled into a multi-domain holotoxin. Leptospiral VM proteins are secreted R-type lectin domain-containing exotoxins with discrete N-terminal ricin B-like domains involved in host cell surface binding, and a C-terminal DNase/toxin domain. Here we use the artificial intelligence-based AlphaFold algorithm and other computational tools to predict and elaborate on details of the VM protein structure-function relationship. Comparative AlphaFold and CD-spectroscopy defined the consistent secondary structure (Helix and ß-sheet) content, and the stability of the functional domains were further supported by molecular dynamics simulation. VM proteins comprises distinctive lectic family (QxW) 3 motifs, the Mycoplasma CARDS toxin (D3 domain, aromatic patches), C-terminal similarity with mammalian DNase I. In-silico study proposed that Gln412, Gln523, His533, Thr59 are the high binding energy or ligand binding residues plausibly anticipates in the functional activities. Divalent cation (Mg +2 -Gln412) and phosphate ion (PO 4 ] -3 -Arg615) interaction further supports the functional activities driven by C-terminal domain. Computation-driven structure-function studies of VM proteins will guide experimentation towards mechanistic understandings of leptospirosis pathogenesis, which underlie development of new therapeutic and preventive measures for this devastating disease., Competing Interests: JV and spouse have an equity interest in LeptoX, which may have a future interest in licensing this work. The work reported here has been filed in patent applications from Yale University., (Copyright © 2023 Chaurasia and Vinetz.)

Leptospirosis is an emerging infectious disease, with increasing frequency and severity of outbreaks, a changing epidemiology of populations at risk, and the emergence of new strains, serovars, serogroups, and species. Virulence-modifying (VM) proteins encoded by the PF07598 gene family are hypothesized to be Leptospira -secreted exotoxins that mediate the molecular and cellular pathogenesis of severe and fatal leptospirosis. If confirmed experimentally, this concept could revolutionize the treatment, diagnosis, prognosis, and vaccine-mediated prevention of leptospirosis by enabling a novel array of targeted interventions. VM proteins, as with other bacterial-secreted protein exotoxins, mediate their virulence effects by attaching to eukaryotic cells, competing with other microorganisms for limited resources in environmental niches, directly intoxicating target cells, and disrupting their function in the mammalian host. In contrast with the most pathogenic group of Lept ospira , particularly L. interrogans , whose genomes contain 12-15 PF07598 paralogs, strains of the livestock and human pathogen L. borgpetersenii have two PF07598 paralogs. Given the possible non-environmentally mediated transmission of some L. borgpetersenii strains and the much smaller number of VM proteins in this species, their role in infection and disease may well differ from other leptospiral species. Comparison of VM proteins from different clades of pathogenic Leptospira may deepen our understanding of leptospirosis's pathogenesis, leading to novel approaches to ameliorating Leptospira infection in humans and animals.