Browsing by Author "G. Saikumar"

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Comparative study on characterization and wound healing potential of goat (Capra hircus) mesenchymal stem cells derived from fetal origin amniotic fluid and adult bone marrow
(Elsevier B.V., 2017) M.D. Pratheesh; Pawan K. Dubey; Nitin E. Gade; Amar Nath; T.B. Sivanarayanan; D.N. Madhu; Anjali Somal; Indu Baiju; T.R. Sreekumar; V.L. Gleeja; Irfan A. Bhatt; Vikash Chandra; Amarpal; Bhaskar Sharma; G. Saikumar; G. Taru Sharma
Caprine amniotic fluid (cAF) and bone marrow cells (cBM) were isolated, expanded and phenotypically characterized by mesenchymal stem cells (MSCs) specific cell surface markers. Both cell types were compared for multilineage differentiation potential by flow cytometry using specific antibodies against lineage specific markers. Furthermore, in vitro expanded cAF-MSCs showed higher expression of trophic factors viz. VEGF and TGF-β1 as compared to cBM-MSCs. Full-skin thickness excisional wounds created on either side of the dorsal midline (thoracolumbar) of New Zealand White rabbits were randomly assigned to subcutaneous injection of either fetal origin cAF-MSCs (n = 4) or adult cBM-MSCs (n = 4) or sterile PBS (control, n = 4). The rate of wound closure was found faster (p < 0.05) in cAF-MSCs treated wounds as compared with cBM-MSCs and PBS treated wounds especially on 21st day post-skin excision. Histomorphological examination of the healing tissue showed that wound healing was improved (p < 0.05) by greater epithelialization, neovascularization and collagen development in cAF-MSCs as compared to cBM-MSCs and PBS treated wounds. © 2017 Elsevier Ltd
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.
Evaluation of persistence and distribution of intra-dermally administered PKH26 labelled goat bone marrow derived mesenchymal stem cells in cutaneous wound healing model
(Springer Netherlands, 2017) M.D. Pratheesh; Nitin E. Gade; Amar Nath; Pawan K. Dubey; T.B. Sivanarayanan; D.N. Madhu; T.R. Sreekumar; Amarpal; G. Saikumar; G. Taru Sharma
The current study was designed to study the persistence and distribution of caprine bone marrow derived mesenchymal stem cells (cBM-MSCs) when administered intra-dermally in experimentally induced cutaneous wounds in rabbits. MSC’s from goat bone marrow were isolated and their differentiation potential towards adipogenic and osteogenic lineages were assayed in vitro. The isolated cells were phenotypically analysed using flow cytometry for the expression of MSC specific matrix receptors (CD73, CD105 and Stro-1) and absence of hematopoietic lineage markers. Further, these in vitro expanded MSCs were stained with PKH26 lipophilic cell membrane red fluorescent dye and prepared for transplantation into cutaneous wounds created on rabbits. Five, 2 cm linear full thickness skin incisions were created on either side of dorsal midline of New Zealand white rabbits (n = 4). Four wounds in each animal were implanted intra-dermally with PKH26 labelled cBM-MSCs suspended in 500 µl of Phosphate Buffer Saline (PBS). Fifth wound was injected with PBS alone and treated as negative control. The skin samples were collected from respective wounds on 3, 7, 10 and 14 days after the wound creation, and cryosections of 6 µM were made from it. Fluorescent microscopy of these cryosections showed that the PKH26 labelled transplanted cells and their daughter cells demonstrated a diffuse pattern of distribution initially and were later concentrated towards the wound edges and finally appeared to be engrafted with the newly developed skin tissues. The labelled cells were found retained in the wound bed throughout the period of 14 days of experimental study with a gradual decline in their intensity of red fluorescence probably due to the dye dilution as a result of multiple cell division. The retention of transplanted MSCs within the wound bed even after the complete wound healing suggests that in addition to their paracrine actions as already been reported, they may have direct involvement in various stages of intricate wound healing process which needs to be explored further. © 2017, Springer Science+Business Media Dordrecht.
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.
Isolation and propagation of classical swine fever virus in porcine Wharton’s Jelly mesenchymal stem cells
(Taylor and Francis Ltd., 2022) Neelam R. Tomar; Irfan A. Bhat; Mukesh K. Bharti; Jeny K. John; Veena Sharma; Vikash Chandra; G. Taru Sharma; G. Saikumar
Classical Swine Fever (CSF) is an extremely infectious and deadly disease of pigs and wild boars caused by the CSF virus (CSFV) which is a member of the Pestivirus genus and the family Flaviviridae. This study was designed to detect the permissibility and replication of CSFV in mesenchymal stem cells (MSCs) monolayer derived from Porcine Wharton’s jelly. Porcine Wharton’s jelly MSCs (pWJ-MSCs) were ex vivo expanded and propagated for more than 81 generations and third passage pWJ-MSCs were characterized as per standard criteria i.e., growth characteristics, trilineage differentiation potential and molecular characterization for pluripotency and stem cell surface markers. Porcine WJ tissue samples found negative for CSFV by RT-PCR test were processed further for the isolation of pWJ-MSCs and CSFV was propagated over the characterized pWJ-MSCs monolayer. No cytopathic effect was observed, which was consistent with non-cytopathic nature of CSFV. The replication of CSFV in pWJ-MSCs was affirmed by RT-PCR and demonstration of viral antigen in the cytoplasm of virus infected cells by immuno-staining technique. In total, three different CSFV isolates were propagated in pWJ-MSCs. Primary pWJ-MSCs permitted CSFV replication to good titer. To the best of our information, this is the first ever report of isolation of CSFV in pWJ-MSCs. © 2020 Taylor & Francis Group, LLC.
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.