Browsing by Author "Aditya Agrawal"

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Budesonide: A promising candidate therapeutic for early COVID-19
(Elsevier Ltd, 2021) Shailesh Kumar Patel; Rakesh Kumar; Jigyasa Rana; Aditya Agrawal; Alok Singh; Adesh Kumar
[No abstract available]
Detection of porcine enteric picornaviruses from faecal samples of Indian pigs
(Springer, 2022) Shailesh Kumar Patel; Aditya Agrawal; Mamta Pathak; Alok Singh; Rajat Varshney; Jigyasa Rana; G. Saikumar
Porcine enteric picornaviruses often consequence diarrhoea and nervous complications in pig and pose enormous loss to pig farming. The present study expands the limited Indian data of porcine enteric picornaviruses which is needed for the early implementation of control measures and to check further outbreaks. A total of 398 porcine faecal samples from Uttar Pradesh, Madhya Pradesh, Chhattisgarh and Jharkhand state of India were screened for porcine teschovirus (PTV), porcine sapelovirus (PSV) and enterovirus G (EV-G) by reverse transcriptase-polymerase chain reaction (RT-PCR) using 5′UTR-specific primers. The prevalence of PTV, PSV and EV-G was found to be 12.81% (51/398), 5.77% (23/398) and 24.37% (97/398), respectively. EV-G was relatively higher in circulation in Indian pigs among all the included enteric picornaviruses. Conversely, the concurrent infection of more than one enteric picornavirus was also frequent. © 2022, The Author(s), under exclusive licence to Indian Virological Society.
Development of Hemagglutinin–Neuraminidase Homologous Peptides as Novel Promising Therapeutic Agents Against Peste des Petits Ruminants Virus
(Springer, 2023) Aditya Agrawal; Rajat Varshney; Anil Gattani; Mahvash Hira Khan; Rohini Gupta; Khushal Singh Solanki; Shailesh Kumar Patel; R.P. Singh; Praveen Singh
The lack of specific antiviral therapy and complications associated with the existing peste des petits ruminants (PPR) vaccines accentuates the search of novel antiviral blocking agents in order to curtail the PPR infection at initial level. The synthetic hemagglutinin–neuraminidase (HN) homologous peptides may compete with the natural HN protein of PPR virus for binding to signaling lymphocytic activation molecule (SLAM) receptor, consequently, may disrupt peste des petits ruminants virus (PPRV) at entry level. Therefore, insilico analysis, synthesis, purification and subsequent characterization of HN homologous peptides were conducted in this study. The HN homologous peptides were synthesized by means of solid phase chemistry and were purified by reversed-phase-high performance liquid chromatography. The mass as well as sequence of HN homologous peptides were assessed by mass spectroscopy while its secondary structure was elucidated by circular dichroism spectroscopy. The binding (interaction) efficacy of HN homologous peptides with PPRV antibodies was assessed via indirect enzyme linked immunosorbent assay, visual detection test (red wine to purple), bathochromic shift under UV–Vis spectrophotometry and lateral flow immunochromatographic strip test. The antiviral properties and cytotoxicity of these peptides were also assessed in B95a cell line with changes in cytopathic effect and titer of PPRV (Sungri/96). The presence of green fluorescein isothiocyanate over the B95a cell surface pointed towards the binding of HN homologous peptides with surface SLAM receptor. Moreover, the intact beta sheet configuration in water and lower cytotoxicity [cytotoxic concentration 50 (CC50) > 1000 µg/ml] of these peptides signifies its in vivo use. Among HN homologous peptides, the binding efficacy and antiviral properties of pep A was relatively high in comparison to pep B and Pep ppr peptides. The prerequisite concentration of HN homologous peptides (pep A = 12.5 µg/ml; pep B = 25 µg/ml; pep ppr = 25 µg/ml) to exemplify its antiviral effect was much lower than its CC50 level. Hence, this study signifies the therapeutic potential of synthetic HN homologous peptides. © 2023, The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.
Exploration of antigenic determinants in spike glycoprotein of SARS-CoV2 and identification of five salient potential epitopes
(Springer, 2021) Aditya Agrawal; Rajat Varshney; Mamta Pathak; Shailesh Kumar Patel; Vishal Rai; Sourabh Sulabh; Rohini Gupta; Khushal Singh Solanki; Ritu Varshney; Ramadevi Nimmanapalli
Emerging pathogens have been an eternal threat to mankind. In a series of pandemics caused by notorious coronaviruses, a newly emerged SARS-CoV2 virus is creating panic among the world population. The unavailability of reliable theranostics insists the exploration of antigenic determinants in spike glycoprotein of SARS-CoV2. The four novel inserts (‘70VSGTNGT76’, ‘150KSWM153’, 247SYLTPG252 and 674QTQTNSPRR682) in SARS-CoV2 spike protein were unraveled via multiple sequence alignment of spike proteins of SARS-CoV2, SARS-CoV, and MERS-CoV. The three-dimension (3D) modeling of the spike protein of the SARS-CoV2 and their interaction with the ACE2 receptor was delineated with the help of SWISS-MODEL and 3DLigandSite web servers. The predicted 3D model of SARS-CoV2 was further verified by SAVES, RAMPAGE, and ProSA-web tools. The potential B-cell immunogenic epitopes of SARS-CoV2 were predicted out by using various software viz. IEDB B-cell epitopes prediction tool, BepiPred linear epitope prediction tool, Emini Surface Accessibility Prediction tool, and Kolaskar-Tongaonkar antigenicity web tool. The five epitopes (i.e. ‘71SGTNGTKRFDN81, 247SYLTPG252, 634RVYST638, 675QTQTNSPRRARSV687, and 1054QSAPH1058) were selected as potent antigenic determinants. The quantum of information generated by this study will prove beneficial for the development of effective therapeutics, diagnostics, and multi-epitopic vaccines to combat this ongoing menace. © 2021, Indian Virological Society.
Genetic Characterization and Pathology of Porcine Teschovirus (PTV) in India
(Agricultural Research Communication Centre, 2024) Shailesh Kumar Patel; Jigyasa Rana; Mamta Pathak; Alok Singh; Aditya Agrawal; G. Saikumar
Background: The porcine teschovirus (PTV) is common swine pathogen which causes a wide range of illnesses ranging from asymptomatic infection to acute fatal encephalomyelitis, diarrhoea and pneumonia. Despite of its economical importance very limited studies are available on the pathology of PTV. The present study was conducted with the objective to investigate the PTV infection and associated pathology in piglets. Methods: A total of 78 piglets below 3 months of age were necropsied and representative tissue samples along with intestinal content were collected for histopathological examination and molecular investigation during April 2019 to November 2020. The 5' UTR region of PTV was amplified via RT-PCR and further confirmed by sequencing. Thereafter, genetic characterization of the Indian PTV isolate was done on the basis of 5' UTR gene. Result: A total of seven out of 78 cases were found positive for the PTV. On necropsy the intestinal wall was thinned and distended with yellowish coloured diarrhoeic content. Congestion of serosal and mucosal vessels along with severe meningeal congestion was observed. Microscopic lesions included congestion of mucosal vessels and destruction of villous structure of jejunum along with perivascular cuffing, focal gliosis, neuronophagia, congestion of meningeal and cerebral vessels and interstitial pneumonia. The phylogenetic analysis revealed that the isolate PTV/India/IVRI-381/2020 (MW022462) and PTV/India/IVRI-1093/2020 (MW022463) were clustered with PTV-13 strain wild boar/WB2C-TV/2011/HUN (JQ429405) and PTV-2 strain Vir 6711-12/83 (AF296107) of Hungary and Germany respectively. This study reported the genotypic and pathological investigation of PTV from Indian piglets. Further explorative surveillance along with studies in suitable cell lines and animal model will pave the way for better understanding of PTV among Indian pigs. © 2024 Agricultural Research Communication Centre. All rights reserved.
Gold nanoparticle based immunochromatographic biosensor for rapid diagnosis of Mycobacterium avium subspecies paratuberculosis infection using recombinant protein
(Elsevier B.V., 2020) Aditya Agrawal; Rajat Varshney; Anil Gattani; P. Kirthika; Mahvash Hira Khan; Rahul Singh; Shantanu Kodape; Shailesh Kumar Patel; Praveen Singh
Highly infectious and obvious withstand ability of Mycobacterium avium subspecies paratuberculosis (MAP) to environment as well as lack of on-site field diagnostic methods notably hampers the paratuberculosis (PTB) control. The existing intricacy, time-consuming and complicated diagnostic methods of PTB accentuate the development of novel and easy-to-perform on-site test. A gold nanoparticle (GNP) based lateral-flow assay (LFA) using MAP recombinant protein (44 kDa) has been developed for sensitive and specific detection of PTB in field conditions. The diagnostic sensitivity and specificity of the LFA for MAP specific antibodies was found approximately 84.2% and 83.3% in comparison to indirect enzyme-linked immunosorbent assay. Consequently, the newly developed GNP based LFA offers on-site and cost-effective method for the prompt diagnosis of PTB and precludes the time-consuming laboratory screening. © 2020 Elsevier B.V.
Monkeypox: A global threat to domestic and wild animals – Correspondence
(Elsevier Ltd, 2022) Nikhil K. C; Jigyasa Rana; Shailesh Kumar Patel; Aditya Agrawal; Kumar Govil; Alok Singh; Megha Katare Pandey; Dhruv Nitinkumar Desai; Sita Prasad Tiwari
[No abstract available]
Non-infectious outer membrane vesicles derived from Brucella abortus S19Δper as an alternative acellular vaccine protects mice against virulent challenge
(Elsevier B.V., 2021) Khushal Singh Solanki; Rajat Varshney; Salauddin Qureshi; Prasad Thomas; Rahul Singh; Aditya Agrawal; Pallab Chaudhuri
The prime human and animal safety issues accentuate the search of promising newer alternative vaccine candidates to resolve complications associated with the live attenuated Brucella abortus strain19 (S19) vaccine. Outer membrane vesicles (OMVs S19 Δper) extracted from Brucella abortus S19Δper (S19Δper) as an alternative subunit vaccine candidate has been explored in the present study as OMVs are endowed with immunogenic molecules, including LPS and outer membrane proteins (OMPs) and do not cause infection by virtue of being an acellular entity. The LPS defective S19Δper released a higher amount of OMVs than its parent strain S19. Under transmission electron microscopy (TEM), OMVs were seen as nano-sized outward bulge from the surface of Brucella. Dynamic light scattering analysis of OMVs revealed that OMVs S19Δper showed the less polydispersity index (PDI) than OMVs S19 pointing towards relatively more homogenous OMVs populations. Both OMVs S19Δper and OMVs S19 with or without booster dose and S19 vaccine were used for immunization of mice and subsequently challenged with 2 × 105 CFU virulent Brucella abortus strain 544 (S544) to assess protective efficacy of vaccines. The less splenic weight index and less S544 count in OMVs immunized mice in comparison to unimmunized mice after S544 challenge clearly indicated good protective efficacy of OMVs. OMVs S19 Δper induced relatively high titer of IgG than OMVs S19 but conferred nearly equal protection against brucellosis. An ELISA based determination of IgG and its isotype response, Cytometric Bead Array (CBA) based quantitation of serum cytokines and FACS based enumeration of CD4+ and CD8+ T cells revealed high titer of IgG, production of both Th1 (IgG2a) and Th2 (IgG1) related antibodies, stimulation of IL-2, TNF (Th1) and IL-4, IL-6, IL-10 (Th2) cytokines, and induced T cell response suggested that OMVs S19Δper elicited Th1 and Th2 type immune response and ensured protection against S544 challenge in murine model. © 2020 Elsevier B.V.
Pathology and Molecular Characterization of Porcine Sapelovirus in Indian Pigs
(Agricultural Research Communication Centre, 2023) Shailesh Kumar Patel; Mamta Pathak; Alok Singh; Aditya Agrawal; Jigyasa Rana; G. Saikumar
Background: The porcine sapelovirus (PSV) is a small, non-enveloped, single-stranded, positive-sense, RNA virus of the family Picornaviridae. The PSV infections in pigs have been found associated with diarrhoea, polioencephalomyelitis, pneumonia and reproductive disorders with a high morbidity rate. Despite of its economical importance very few studies are available on the pathology of PSV. The present study was conducted with the aim to investigate the PSV infection and associated pathology in Indian pigs. Methods: Tissue samples along with intestinal content were collected from a total of 78 necropsied cases for histopathological examination and molecular investigation during April 2019 to August 2020. The amplification of 5́ UTR region of PSV was carried out via RT-PCR and confirmed by sequencing. The Genetic characterization of Indian isolate of the PSV was done on the basis of viral 5́ UTR gene. Result: A total of eight out of 78 cases were found positive for the PSV. Catarrhal and haemorrhagic enteritis, thickening and clouding of brain meninges along with congestion of brain and pneumonia was observed as common gross lesions. Microscopic lesions included perivascular cuffing, focal gliosis, neuronophagia, congestion of meningeal and cerebral vessels, interstitial pneumonia, inflammatory changes in the intestinal mucosa and sloughing of villi. The genetic characterization revealed maximum identity of 96.89% with PSV-1 strain PSV-46-V (LC508233) and PSV-1 strain PSV-26-B (LC508232) of Zambia. This study reported the pathological and molecular investigation of PSV from Indian pigs. Further explorative surveillance along with experimental studies in suitable animal model and cell lines are highly warranted for better understanding of PSV pathology in Indian pigs. © 2023 Agricultural Research Communication Centre. All rights reserved.
SLAM (CD150) receptor homologous peptides block the peste des petits ruminants virus entry into B95a cells
(John Wiley and Sons Inc, 2024) Aditya Agrawal; Rajat Varshney; Anil Gattani; Perumalraja Kirthika; Rohini Gupta; Deepak Kumar; Rabindra Prasad Singh; Praveen Singh
The fusion of haemagglutinin-neuraminidase (HN) protein of peste des petits ruminant (PPR) virus with signaling lymphocyte activation molecules (SLAM) host cell receptor consequences the virus entry and multiplication inside the host cell. The use of synthetic SLAM homologous peptides (i.e., molecular decoy for HN protein of PPR virus) may check PPR infection at the preliminary stage. Hence, the predicted SLAM homologous peptides using bioinformatics tools were synthesized by solid phase chemistry with standard Merrifield's 9-fluorenylmethoxycarbonyl (Fmoc) chemistry and were purified by reverse phase high performance liquid chromatography. The secondary structures of synthesized peptides were elucidated by circular dichroism spectroscopy. The in vitro interactions of these peptides were studied through indirect Enzyme Linked Immuno Sorbent Assay (ELISA) and visual surface plasmon UV-visible spectroscopy. The SLAM homologous peptides were able to interact with the peste des petits ruminant virus (PPRV) with varying binding efficiency. The interaction of SLAM homologous peptide with the PPR virus was ascertained by the change in the plasmon color from red wine to purple during visual detection and also by bathochromic shift in absorbance spectra under UV-visible spectrophotometry. The cytotoxic and anti-PPRV effect of these peptides were also evaluated in B95a cell line using PPR virus (Sungri/96). The cytotoxic concentration 50 (CC50) value of each peptide was greater than 1000 μg mL−1. The anti-PPRV efficiency of SLAM-22 was relatively high among SLAM homologous peptides, SLAM-22 at 25 μg mL−1 concentration showed a reduction of more than log10 3 virus titer by priming of B95a cell line while the use of SLAM-15 and Muco-17 at the same concentration dropped virus titer from log10 4.8 to log10 2.5 and log10 3.1 respectively. The concentration of SLAM homologous peptide (25 μg mL−1) to exert its anti-PPRV effect was much less than its CC50 level (>1000 μg mL−1). Therefore, the synthetic SLAM homologous peptides may prove to be better agents to target PPRV. © 2023 Wiley Periodicals LLC.