Browsing by Author "Subodh Kumar"

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A clinical trial to evaluate the safety and immunogenicity of the LEISH-F1+MPL-SE vaccine for use in the prevention of visceral leishmaniasis
(2011) Jaya Chakravarty; Subodh Kumar; Sonali Trivedi; Vijay K. Rai; Anup Singh; Jill A. Ashman; Elsa M. Laughlin; Rhea N. Coler; Stuart J. Kahn; Anna Marie Beckmann; Karen D. Cowgill; Steven G. Reed; Shyam Sundar; Franco M. Piazza
Healthy Indian adult volunteers, with or without a history of leishmaniasis, were evaluated for evidence of previous infection with Leishmania donovani based on the direct agglutination test (DAT). Three cohorts of 6 DAT-negative and 6 DAT-positive subjects were enrolled in an open-label, dose-escalating, uncontrolled clinical trial and received three injections of the LEISH-F1+MPL-SE vaccine (consisting of 5μg, 10μg, or 20μg recombinant Leishmania polyprotein LEISH-F1 antigen+25μg MPL®-SE adjuvant). The study injections were given subcutaneously on days 0, 28, and 56, and the subjects were followed through day 168 for safety and immunological endpoints. The vaccine was safe and well-tolerated in DAT-negative and DAT-positive subjects and induced T-cell production of IFN-γ and other cytokines in response to stimulation with the LEISH-F1 antigen. This clinical trial shows that the LEISH-F1+MPL-SE vaccine is safe and immunogenic in healthy subjects with and without history of previous infection with Leishmania donovani. © 2011 Elsevier Ltd.
A novel 12.6-kDa protein of leishmania donovani for the diagnosis of Indian visceral leishmaniasis
(2011) Dinesh Kumar; Subodh Kumar; Jaya Chakravarty; Shyam Sundar
Background: For the diagnosis of visceral leishmaniasis (VL), rK39 antigen-based rapid test is widely used. Unfortunately, up to 32% healthy individuals from endemic region test positive with this antigen. There is an urgent need to search for a more specific antigen with sensitivity similar to rK39. Methods: We identified a Leishmania donovani-specific 12.6-kDa (BHUP3) soluble promastigote antigen through sensitive western blot technique. The identified protein was partially purified from sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the antigenic response of eluted protein was determined by western blot with different groups of individual sera. The diagnostic potential was further validated by enzyme-linked immunosorbent assay using serum of 100 VL patients, 93 nonendemic healthy control individuals, 110 endemic healthy control individuals, and 110 disease control individuals. Further, it was characterized by two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ionization-time-of-flight analysis. Results: On blotting, antibody against this protein was recognized by all (9/9) VL patient's sera, but it was absent in every control group (nonendemic healthy control and endemic healthy control). Sensitivity of the enzyme-linked immunosorbent assay was 88% (89/101), whereas the specificity for endemic healthy, nonendemic healthy, and different disease groups were 96% (106/110), 100% (93/93), and 97% (107/110), respectively. The two-dimensional gel electrophoresis showed a single spot, and matrix-assisted laser desorption/ionization-time-of-flight analysis revealed that it is a 113-amino-acid-long putative uncharacterized protein of 12.6-kDa anamorsin homolog matched completely with Leishmania major (GenBank accession number: Q4QIS1). Conclusion: Despite marginally lower sensitivity of BHUP3, excellent specificity warrants its further development as a tool for diagnosis of VL. © Copyright 2011, Mary Ann Liebert, Inc.
Antileishmanial activity of nano-amphotericin B deoxycholate
(2008) Krishna Das Manandhar; Thakur Prasad Yadav; Vijay Kumar Prajapati; Subodh Kumar; Madhukar Rai; Anuradha Dube; Onkar Nath Srivastava; Shyam Sundar
Objectives: The aim of the present study was to compare the efficacy of a nano form of amphotericin B deoxycholate with that of conventional amphotericin B deoxycholate for the treatment of visceral leishmaniasis. Methods We have formulated nanoparticles (10-20 μM) from amphotericin B deoxycholate (1-2 nM) by applying high-pressure (150 argon) milling homogenization and have tested their efficacy in a J774A cell line and in hamsters. Parasite survival and tissue burden in spleen were evaluated for nano-amphotericin B and conventional amphotericin B. Both nano-amphotericin B and conventional amphotericin B were injected intraperitoneally at 5 mg/kg per day for 5 days. Results: The inhibition of amastigotes in the splenic tissue with nano-amphotericin B was significantly more than with conventional amphotericin B (92.18% versus 74.57%, P = 0.005). Similarly, the suppression of parasite replication in the spleen was also found to be significant (99.18% versus 97.17%, P = 0.05). In a cytotoxicity test, nano-amphotericin B against the J774A cell line had a CC50 of 12.67 mg/L in comparison with 10.61 mg/L for amphotericin B, far higher than the doses used for ED50. Conclusions: Nanoparticles of amphotericin B had significantly greater efficacy than conventional amphotericin B. This formulation may have a favourable safety profile, and if production costs are low, it may prove to be a feasible alternative to conventional amphotericin B. © The Author 2008. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.
Application of essential oils for the production of dietary supplements and as traditional self-medication purposes
(Elsevier, 2023) Raghvendra Raman Mishra; Pragya Mishra; Subodh Kumar; Divya Gupta
Despite the fact that essential oils (EOs) have been studied for more than 60 years, interest in them has increased recently as more individuals look for natural remedies. Essential oils have been used in rituals and medicine for millennia because of their healing properties. This has existed since the Paleolithic era. Since they are more complex and contain a variety of volatile and naturally occurring bioactive substances, EOs are commonly used in the food sector as the finest alternatives. Despite the fact that EOs are difficult to use practically as an effective antibacterial due to their volatility, low solubility, and high instability, plant-based EOs have gained interest as a potential replacement for synthetic preservatives due to their environmental friendliness and widely acknowledged safety status. Among its traditional uses, the use of EOs in food products undoubtedly stands out. While there have been a number of recent reviews that have concentrated on the use and potential of EOs as food preservatives, there has been relatively less research on other functional roles of EOs or their components in food products, which are nevertheless generating significant public interest and a growing body of scientific evidence: their health-promoting role beyond preservation and basic nutrition. The present chapter points toward the use of EOs in the manufacturing of dietary supplements and traditional self-medication. © 2024 Elsevier Inc. All rights reserved.
Evaluation of rk39 immunochromatographic test with urine for diagnosis of visceral leishmaniasis
(2011) Jaya Chakravarty; Subodh Kumar; Rajiv Kumar; Shalini Gautam; Madhukar Rai; Shyam Sundar
This study evaluates commercially available rK39 immunochromatographic strips using urine for diagnosis of visceral leishmaniasis (VL). Freshly collected urine and serum samples of 280 parasitologically confirmed VL patients and 66 endemic healthy controls (EHC), 48 nonendemic healthy controls (NEHC) and 45 different diseases were tested with rK39 strips. The sensitivity of rK39 in urine was 96.4% while the specificity was low varying from 66.7% in EHC, 77.08% in NEHC to 62.2% in different diseases. With serum, sensitivity was 100% whereas the specificity was 100%, 92.4% and 95.55% for the respective control groups. In the present format, the immunochromatographic strips cannot be used for the diagnosis of VL using urine samples. © 2011 Royal Society of Tropical Medicine and Hygiene.
Identification and characterization of a novel Leishmania donovani antigen for serodiagnosis of visceral leishmaniasis
(2012) Subodh Kumar; Dinesh Kumar; Jaya Chakravarty; Madhukar Rai; Shyam Sundar
Despite several drawbacks, rK39-based rapid immunochromatographic test is widely used for the diagnosis of visceral leishmaniasis (VL) in the Indian subcontinent. There is an urgent need to develop a better antigen. In this study we separated crude soluble antigens of Leishmania donovani by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hybridized with pool sera from pre- and post-treated VL patients, 6 months follow-up, endemic healthy (EHC), and nonendemic healthy controls (NEHC) by Western blotting. The sensitivity of enzyme-linked immunosorbent assay with identified protein was 95% (confidence interval [CI] = 89.6-98.01%), whereas the specificity for EHC, NEHC, and different disease groups were 96.3%(CI = 89.8-98.6%), 100% (CI = 95.8-100%), and 97.4%(CI = 91.02-99.3%), respectively. This specific antigen was subjected to two-dimensional gel electrophoresis and after tryptic digestion, antigen was characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Further analysis showed that it is a member of the heat shock protein family of 70 kDa, designated as BHUP1, and has great potential in the diagnosis of VL. Copyright © 2012 by The American Society of Tropical Medicine and Hygiene.
Identification and characterization of a novel, 37-kilodalton Leishmania donovani antigen for diagnosis of indian visceral leishmaniasis
(2011) Subodh Kumar; Dinesh Kumar; Jaya Chakravarty; Shyam Sundar
The biggest challenge in the serological diagnosis of visceral leishmaniasis (VL) is to find a biomarker with a high specificity. This study was undertaken to identify novel Leishmania donovani antigens to solve the existing problem. The soluble L. donovani promastigote antigen was separated by SDS-PAGE, and a Western blot was probed with pooled sera of five subjects with confirmed VL before (n = 9 pools) and after (n = 9 pools) treatment and at the 6-month follow-up visit (n = 9 pools), healthy controls not from an area of endemicity (n = 9 pools), and healthy controls from an area of endemicity. The antibody response to the identified partially purified antigen was ascertained by an enzyme-linked immunosorbent assay (ELISA) with 70 sera from patients with parasitologically confirmed VL, 48 sera from healthy controls from an area where the disease is not endemic, 60 sera from healthy controls from an area of endemicity, and 42 sera from patients in different disease groups. The eluted protein was subjected to two-dimensional (2D) gel electrophoresis, Western blotted, and probed with sera from patients with confirmed VL and from healthy controls not from an area of endemicity. The antigenic protein was further characterized by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. The identified protein (BHUP2) corresponds to a cytochrome c-like synthesis protein of 37 kDa. ELISA results were 94% sensitive, whereas specificities with sera from healthy controls from an area of endemicity, healthy controls not from an area of endemicity, and disease controls were 98%, 100%, and 97%, respectively. The antigen identified via a proteomics-based approach has a strong potential for further development as a diagnostic tool for VL. Copyright © 2011, American Society for Microbiology. All Rights Reserved.
Laser photoacoustic and photothermal spectroscopy for defense and security
(Elsevier, 2022) Subodh Kumar; Ramesh C. Sharma; Abhishek Parmar; Surya N. Thakur
Photoacoustic and photothermal techniques provide direct as well as indirect methods to measure optical absorption and thermal characteristics of a sample. Periodic or pulsed optical absorption followed by nonradiative transitions causes change in the thermodynamic state and leads to pressure, temperature, or density fluctuations of the sample. The detection of changes in temperature, pressure, or density occurring due to optical absorption eventually generates the photoacoustic or photothermal signal. Laser photoacoustic techniques using microphone, piezoelectric transducer, and quartz crystal tuning fork as detectors have been discussed. These techniques have been used in point detection as well as standoff detection configurations for studies on hazardous chemicals, explosive, and pollutants. Photothermal technique has also been developed to measure the temperature gradient in the sample generated by a wavelength tunable laser beam, and the method involves a pump probe configuration to sense change in refractive index in samples. © 2023 Elsevier Inc. All rights reserved.
LC-MS/MS–Based Proteomics Study of Cyanobacteria
(CRC Press, 2024) Soumila Mondal; Subodh Kumar; Phulen Sarma; Bikash Medhi; Anjali Gupta; Sapna Tiwari; Pankaj K. Maurya; Shailendra P. Singh
Cyanobacteria are a monophyletic group of Gram-negative, oxygen-producing photosynthetic microorganisms that are well known for their significant contribution to global CO2 and N2 fixation (Zahra et al. 2020). Several cyanobacteria can be easily genetically modified and have simple growth requirements as compared to algae and plants (Berla et al. 2013; Singh and Sinha 2020). A cyanobacterial biomass of a wild type as well as genetically engineered strains have commercial and industrial importance for the production of various secondary metabolites such as long-chain alcohols, alkanes, fatty acids, ethylene, 2,3-butanediol, ethanol, polyhydroxybutyrate and hydrogen (Knoot et al. 2018). Due to the obligatory requirement of light for their growth, cyanobacteria are exposed to a dynamic light environment, both diurnally and seasonally in their natural habitat, as well as in large-scale cultivation systems (Ooms et al. 2016). In addition to light, several other abiotic factors such as salinity, drought, temperature, pH and nutrient availability affect their overall fitness, growth and development. Also, installation of novel metabolic pathways and associated metabolite production could alter the overall regulatory network of the organism and its response to environmental factors (Ungerer et al. 2018; Gupta et al. 2020). Therefore, transcriptomics and proteomics studies have become important tools for the identification and quantification of transcriptomes and proteomes of native as well as genetically engineered strains to understand their response to different environmental conditions. However, information obtained from transcriptomic study may not necessarily translate to a protein level. Therefore, proteomic studies have emerged as a powerful tool to understand the total protein complement of any genome and its role as a part of a complex network system (Aebersold and Mann 2003; Karpievitch et al. 2010). © 2024 selection and editorial matter, Shailendra P. Singh, Rajeshwar P. Sinha and Donat-P. Häder; individual chapters, the contributors.
Photomechanical detection of bioaerosol fluorescence free-from solar background
(Elsevier Ltd, 2022) Ramesh C. Sharma; Subodh Kumar; Abhishek Parmar; Akansha Tyagi; Kamal P. Singh; Surya N. Thakur
Detection of biological molecules in aerosol via fluorescence signature is well established. However, standoff detection of weak target fluorescence deeply submerged in the ambient bright solar background is challenging with conventional approaches (using photo-multiplier tubes, avalanche photodiodes or ICCD) due to saturation effects and overlapping spectral signatures. Here, we demonstrate a Quartz tuning fork (QTF) enhanced photomechanical detection of fluorescence spectra from bio-aerosols free from broadband solar light. The detection is based on resonant excitation of QTF at 32.78 kHz by on–off modulated laser-induced fluorescence from aerosol while the stray solar background is essentially suppressed by the QTF. A remote detection of fluorescence spectra of Bacillus Globbigi and Riboflavin aerosols was demonstrated at 5 m standoff distance using two near-UV excitation wavelengths with good signal-to-noise ratio. The fluorescence signal increases linearly with excitation power, concentration and quality factor of the QTF. Besides being low-cost and reliable, the detection range of the presented approach can be extended to large standoff distance free-from ambient sunlight noise. © 2022 Elsevier Ltd
SELSI Consensus Statement for Safe Cholecystectomy — Prevention and Management of Bile Duct Injury — Part A
(Springer, 2021) Virinder Kumar Bansal; Mahesh C. Misra; Anil K Agarwal; Jb Agrawal; Pn Agarwal; Sandeep Aggarwal; Mohammad Aslam; Asuri Krishna; Aditya Baksi; Anu Behari; Hemanga K Bhattacharjee; Rajesh Bhojwani; Jagdish Chander; Tk Chattopadhyay; Chintamani; Pradeep Chowbey; Abhay Dalvi; Nr Dash; Ik Dhawan; Shivanand Gamangatti; Pk Garg; Nm Gupta; Rajesh Gupta; Sk Gupta; Vikas Gupta; L. Kaman; Bml Kapur; Kamal Kataria; Muneer Khan; Ajay K Khanna; Rajesh Khullar; Anand Kumar; Atin Kumar; Sandeep Kumar; Subodh Kumar; Pawanindra Lal; Sd Maurya; Gs Moirangthem; Sujoy Pal; Rajesh Panwar; Rajinder Parshad; Biju Pottakkat; Om Prakash Prajapati; Shailesh Puntambekar; Piyush Ranjan; Yashwant Singh Rathore; Peush Sahni; Rathindra Sarangi; Vuthaluru Seenu; Rajeev Sharma; Vk Shukla; Dp Singh; Jaspal Singh; Rajdeep Singh; Rajeev Sinha; Sadiq S Sikora; Amit Srivastava; Anurag Srivastava; Kn Srivastava; Shaji Thomas; Gr Verma; Jd Wig; Vk Kapoor
Cholecystectomy is one of the most common general surgical operations performed worldwide. The risk of bile duct injury (BDI) during laparoscopic cholecystectomy is two to three times higher than during open cholecystectomy. The worldwide incidence of bile duct injury is 0.5% or 1 in 200 cases. BDI and its consequences result in significant morbidity and may even cause mortality; it also increases the cost of treatment and can be a common reason for medico-legal suits against the surgeons. To minimize the incidence of BDI and to manage it timely and appropriately, a set of guidelines was deemed necessary by a group of senior surgeons during a Society of Endoscopic and Laparoscopic Surgeons of India (SELSI) meeting in 2016. Guidelines for “Safe Laparoscopic Cholecystectomy” and bile duct injury management formulated by other international societies are already available. The applicability of these guidelines to Indian subjects, especially in small peripheral centers, was limited. Hence, a decision was taken to form a set of guidelines for general surgeons with basic laparoscopic skills with little or no advanced laparoscopic skills. Those working in a solo practice, nursing homes, and small private hospitals at talukas or districts should have “Safe Cholecystectomy” guidelines and management of BDI suitable to their situation. These guidelines were formed after three consensus meetings and have been approved by a SELSI Expert Group. © 2019, Association of Surgeons of India.
SELSI Consensus Statement for Safe Cholecystectomy—Prevention and Management of Bile Duct Injury—Part B
(Springer, 2021) Virinder Kumar Bansal; Mahesh C. Misra; Anil K Agarwal; Jb Agrawal; Pn Agarwal; Sandeep Aggarwal; Mohammad Aslam; Asuri Krishna; Aditya Baksi; Anu Behari; Hemanga K Bhattacharjee; Rajesh Bhojwani; Jagdish Chander; Tk Chattopadhyay; Chintamani; Pradeep Chowbey; Abhay Dalvi; Nr Dash; Ik Dhawan; Shivanand Gamangatti; Pk Garg; Nm Gupta; Rajesh Gupta; Sk Gupta; Vikas Gupta; L. Kaman; Bml Kapur; Kamal Kataria; Muneer Khan; Ajay K Khanna; Rajesh Khullar; Anand Kumar; Atin Kumar; Sandeep Kumar; Subodh Kumar; Pawanindra Lal; Sd Maurya; Gs Moirangthem; Sujoy Pal; Rajesh Panwar; Rajinder Parshad; Biju Pottakkat; Om Prakash Prajapati; Shailesh Puntambekar; Piyush Ranjan; Yashwant Singh Rathore; Peush Sahni; Rathindra Sarangi; Vuthaluru Seenu; Rajeev Sharma; Vk Shukla; Dp Singh; Jaspal Singh; Rajdeep Singh; Rajeev Sinha; Sadiq S Sikora; Amit Srivastava; Anurag Srivastava; Kn Srivastava; Shaji Thomas; Gr Verma; Jd Wig; Vk Kapoor
Cholecystectomy is one of the commonest general surgical procedures performed all over India. The risk of bile duct injury (BDI) during laparoscopic cholecystectomy is two to three times higher than during open cholecystectomy. The worldwide incidence of bile duct injury is 0.5% or 1 in 200 cases. BDI and its consequences result in significant morbidity and may even cause mortality. BDI increases the cost of treatment and is a common reason for a medicolegal suit against the surgeons. To minimize the incidence of BDI and to manage it timely and appropriately, a set of guidelines was deemed necessary by a group of senior surgeons during a Society of Endoscopic and Laparoscopic Surgeons of India (SELSI) meeting in 2016. Guidelines for “Safe Laparoscopic Cholecystectomy” and bile duct injury management formulated by other international societies are already available. The applicability of these guidelines to Indian subjects especially in small peripheral centers was limited. Hence, a decision was taken to form a set of guidelines for general surgeons with basic laparoscopic skills with little or no advanced laparoscopic skills. Those working in a solo practice, nursing homes, and small private hospitals at talukas or districts should have “Safe Cholecystectomy” guidelines and management of BDI suitable to their situation. These guidelines were formed after three consensus meetings and have been approved by a SELSI Expert Group. © 2019, Association of Surgeons of India.
Standoff pump-probe photothermal detection of hazardous chemicals
(Nature Research, 2020) Ramesh C. Sharma; Subodh Kumar; Abhishek Parmar; Mohit Mann; Satya Prakash; Surya N. Thakur
A novel pump-probe Photothermal methodology using Quartz Tuning Fork (QTF) detector has been demonstrated for the first time. A tunable mid-IR Quantum Cascade Laser (QCL) and a CW fixed wavelength visible laser have been used as the pump and probe beam respectively. The developed Photothermal (PT) technique is based on Quartz Tuning Fork (QTF) detector for the detection of hazardous/explosive molecules adsorbed on plastic surface and also in aerosols form. PT spectra of various trace molecules in the fingerprinting mid- infrared spectral band 7–9 µm from distance of 25 m have been recorded. The PT spectra of explosives RDX, TNT and Acetone have been recorded at very low quantities. Acetone is the precursor of explosive Tri-Acetone Tri-Phosphate (TATP). The experimentations using pump and probe lasers, exhibit detection sensitivity of less than 5 μg/cm2 for RDX, TNT powders and of ~ 200 nl quantity for Nitrobenzene (NB) and Acetone (in liquid form) adsorbed on surfaces, from a distance of ~ 25 m. The sensitivity of the same order achieved from a distance of 15 m by using only a mid-IR tunable pump laser coupled to QTF detector. Thus the pump-probe PT technique is more sensitive in comparison to single tunable QCL pump beam technique and it is better suited for standoff detection of hazardous chemicals for homeland security as well as for forensic applications. © 2020, The Author(s).
STAT3 Protein–Protein Interaction Analysis Finds P300 as a Regulator of STAT3 and Histone 3 Lysine 27 Acetylation in Pericytes
(Multidisciplinary Digital Publishing Institute (MDPI), 2024) Gautam Kundu; Maryam Ghasemi; Seungbin Yim; Ayanna Rohil; Cuiyan Xin; Leo Ren; Shraddha Srivastava; Akinwande Akinfolarin; Subodh Kumar; Gyan P. Srivastava; Venkata S. Sabbisetti; Gopal Murugaiyan; Amrendra K. Ajay
Background: Signal transducer and activator of transcription 3 (STAT3) is a member of the cytoplasmic inducible transcription factors and plays an important role in mediating signals from cytokines, chemokines, and growth factors. We and others have found that STAT3 directly regulates pro-fibrotic signaling in the kidney. The STAT3 protein–protein interaction plays an important role in activating its transcriptional activity. It is necessary to identify these interactions to investigate their function in kidney disease. Here, we investigated the protein–protein interaction among three species to find crucial interactions that can be targeted to alleviate kidney disease. Method: In this study, we examined common protein–protein interactions leading to the activation or downregulation of STAT3 among three different species: humans (Homo sapiens), mice (Mus musculus), and rabbits (Oryctolagus cuniculus). Further, we chose to investigate the P300 and STAT3 interaction and performed studies of the activation of STAT3 using IL-6 and inhibition of the P300 by its specific inhibitor A-485 in pericytes. Next, we performed immunoprecipitation to confirm whether A-485 inhibits the binding of P300 to STAT3. Results: Using the STRING application from ExPASy, we found that six proteins, including PIAS3, JAK1, JAK2, EGFR, SRC, and EP300, showed highly confident interactions with STAT3 in humans, mice, and rabbits. We also found that IL-6 treatment increased the acetylation of STAT3 and increased histone 3 lysine acetylation (H3K27ac). Furthermore, we found that the disruption of STAT3 and P300 interaction by the P300 inhibitor A-485 decreased STAT3 acetylation and H3K27ac. Finally, we confirmed that the P300 inhibitor A-485 inhibited the binding of STAT3 with P300, which inhibited its transcriptional activity by reducing the expression of Ccnd1 (Cyclin D1). Conclusions: Targeting the P300 protein interaction with STAT3 may alleviate STAT3-mediated fibrotic signaling in humans and other species. © 2024 by the authors.