61 results on '"Shyam Sundar Nandi"'
Search Results
2. A rapid point of care CC16 kit for screening of occupational silica dust exposed workers for early detection of silicosis/silico-tuberculosis
- Author
-
Shyam Sundar Nandi, Upendra P. Lambe, Kamalesh Sarkar, Sonali Sawant, and Jagadish Deshpande
- Subjects
Medicine ,Science - Abstract
Abstract Silicosis is an irreversible, incurable and progressive occupational disease caused by prolonged exposure to crystalline-silica dust while working in the relevant industries. Conventionally diagnosis is done by chest radiology, often in an advanced stage as early symptoms often go unnoticed. Early detection and necessary intervention (secondary prevention) could be a realistic possible control strategy for controlling silicosis as no effective treatment is available to stop and/or reverse the pathological process. Additionally, these patients are also vulnerable to pulmonary tuberculosis, which often becomes difficult to treat and with uncertain treatment outcome. Considering India has a huge burden of silicosis and silico-tuberculosis, a rapid and inexpensive screening method was realized to be an urgent need for early detection of silicosis among silica dust exposed workers. Serum club cell protein 16 (CC16) is evidenced to be a useful proxy screening marker for early detection of silicosis as evidenced from the recent research work of ICMR-National Institute of Occupational Health (ICMR-NIOH), India. In this study a lateral-flow assay for semi-quantitative estimation of serum CC16 level was developed. The detection was performed using gold nanoparticles conjugated anti-CC16 monoclonal antibodies. A sum of 106 serum samples was tested to do the performance evaluation of the assay. A concentration of 6 ng/ml or less produced one band, 6.1–9 ng/ml produced two bands, while more than 9 ng/ml produced all the three bands at the test zone. The sensitivity of the assay was 100% while the specificity was 95%. This assay may be used as a sensitive tool for periodic screening of silica dust exposed vulnerable workers for early detection of silicosis in them.
- Published
- 2021
- Full Text
- View/download PDF
3. Potential of Synthetic and Natural Compounds as Novel Histone Deacetylase Inhibitors for the Treatment of Hematological Malignancies
- Author
-
Dilipkumar Pal, Khushboo Raj, Shyam Sundar Nandi, Surajit Sinha, Abhishek Mishra, Arijit Mondal, Ricardo Lagoa, Jack T. Burcher, and Anupam Bishayee
- Subjects
leukemia ,multiple myeloma ,DNA damage ,HDAC inhibitors ,oxidative stress ,chromatin remodeling ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Histone deacetylases (HDACs) and histone acetyltransferases (HATs) are enzymes that remove or add acetyl groups to lysine residues of histones, respectively. Histone deacetylation causes DNA to more snugly encircle histones and decreases gene expression, whereas acetylation has the opposite effect. Through these small alterations in chemical structure, HATs and HDACs regulate DNA expression. Recent research indicates histone deacetylase inhibitors (HDACis) may be used to treat malignancies, including leukemia, B-cell lymphoma, virus-associated tumors, and multiple myeloma. These data suggest that HDACis may boost the production of immune-related molecules, resulting in the growth of CD8-positive T-cells and the recognition of nonreactive tumor cells by the immune system, thereby diminishing tumor immunity. The argument for employing epigenetic drugs in the treatment of acute myeloid leukemia (AML) patients is supported by evidence that both epigenetic changes and mutations in the epigenetic machinery contribute to AML etiology. Although hypomethylating drugs have been licensed for use in AML, additional epigenetic inhibitors, such as HDACis, are now being tested in humans. Preclinical studies evaluating the efficacy of HDACis against AML have shown the ability of specific agents, such as anobinostat, vorinostat, and tricostatin A, to induce growth arrest, apoptosis, autophagy and cell death. However, these inhibitors do not seem to be successful as monotherapies, but instead achieve results when used in conjunction with other medications. In this article, we discuss the mounting evidence that HDACis promote extensive histone acetylation, as well as substantial increases in reactive oxygen species and DNA damage in hematological malignant cells. We also evaluate the potential of various natural product-based HDACis as therapeutic agents to combat hematological malignancies.
- Published
- 2023
- Full Text
- View/download PDF
4. Immune Response to SARS-CoV-2 Vaccines
- Author
-
Navya Bellamkonda, Upendra Pradeep Lambe, Sonali Sawant, Shyam Sundar Nandi, Chiranjib Chakraborty, and Deepak Shukla
- Subjects
COVID-19 ,SARS-CoV-2 ,mRNA vaccine ,adenoviral vectored vaccine ,inactivated vaccine ,subunit vaccine ,Biology (General) ,QH301-705.5 - Abstract
COVID-19 vaccines have been developed to confer immunity against the SARS-CoV-2 infection. Prior to the pandemic of COVID-19 which started in March 2020, there was a well-established understanding about the structure and pathogenesis of previously known Coronaviruses from the SARS and MERS outbreaks. In addition to this, vaccines for various Coronaviruses were available for veterinary use. This knowledge supported the creation of various vaccine platforms for SARS-CoV-2. Before COVID-19 there are no reports of a vaccine being developed in under a year and no vaccine for preventing coronavirus infection in humans had ever been developed. Approximately nine different technologies are being researched and developed at various levels in order to design an effective COVID-19 vaccine. As the spike protein of SARS-CoV-2 is responsible for generating substantial adaptive immune response, mostly all the vaccine candidates have been targeting the whole spike protein or epitopes of spike protein as a vaccine candidate. In this review, we have compiled the immune response to SARS-CoV-2 infection and followed by the mechanism of action of various vaccine platforms such as mRNA vaccines, Adenoviral vectored vaccine, inactivated virus vaccines and subunit vaccines in the market. In the end we have also summarized the various adjuvants used in the COVID-19 vaccine formulation.
- Published
- 2022
- Full Text
- View/download PDF
5. H2S and homocysteine control a novel feedback regulation of cystathionine beta synthase and cystathionine gamma lyase in cardiomyocytes
- Author
-
Shyam Sundar Nandi and Paras Kumar Mishra
- Subjects
Medicine ,Science - Abstract
Abstract Hydrogen sulfide (H2S), a cardioprotective gas, is endogenously produced from homocysteine by cystathionine beta synthase (CBS) and cystathionine gamma lyase (CSE) enzymes. However, effect of H2S or homocysteine on CBS and CSE expression, and cross-talk between CBS and CSE are unclear. We hypothesize that homocysteine and H2S regulate CBS and CSE expressions in a dose dependent manner in cardiomyocytes, and CBS deficiency induces cardiac CSE expression. To test the hypothesis, we treated murine atrial HL1 cardiomyocytes with increasing doses of homocysteine or Na2S/GYY4137, a H2S donor, and measured the levels of CBS and CSE. We found that homocysteine upregulates CSE but downregulates CBS whereas Na2S/GYY4137 downregulates CSE but upregulates CBS in a dose-dependent manner. Moreover, the Na2S-treatment downregulates specificity protein-1 (SP1), an inducer for CSE, and upregulates miR-133a that targets SP1 and inhibits cardiomyocytes hypertrophy. Conversely, in the homocysteine-treated cardiomyocytes, CBS and miR-133a were downregulated and hypertrophy was induced. In vivo studies using CBS+/−, a model for hyperhomocysteinemia, and sibling CBS+/+ control mice revealed that deficiency of CBS upregulates cardiac CSE, plausibly by inducing SP1. In conclusion, we revealed a novel mechanism for H2S-mediated regulation of homocysteine metabolism in cardiomyocytes, and a negative feedback regulation between CBS and CSE in the heart.
- Published
- 2017
- Full Text
- View/download PDF
6. MiR-133a Mimic Alleviates T1DM-Induced Systolic Dysfunction in Akita: An MRI-Based Study
- Author
-
Shyam Sundar Nandi, Hamid Reza Shahshahan, Quanliang Shang, Shelby Kutty, Michael Boska, and Paras Kumar Mishra
- Subjects
Ins2+/- Akita ,miR-133a ,cardiac dysfunction ,MRI ,fibrosis ,hypertrophy ,Physiology ,QP1-981 - Abstract
Diabetic cardiomyopathy is a leading cause of heart failure. Developing a novel therapeutic strategy for diabetic cardiomyopathy and characterizing animal models used for diabetes mellitus (DM) are important. Insulin 2 mutant (Ins2+/-) Akita is a spontaneous, genetic, mouse model for T1DM, which is relevant to humans. There are contrasting reports on systolic dysfunction and pathological remodeling (hypertrophy and fibrosis) in Akita heart. Here, we used magnetic resonance imaging (MRI) approach, a gold standard reference for evaluating cardiac function, to measure ejection fraction (indicator of systolic dysfunction) in Akita. Moreover, we performed Wheat Germ Agglutinin (WGA) and hematoxylin and Eosin stainings to determine cardiac hypertrophy, and Masson’s Trichrome and picrosirius red stainings to determine cardiac fibrosis in Akita. MiR-133a, an anti-hypertrophy and anti-fibrosis miRNA, is downregulated in Akita heart. We determined if miR-133a mimic treatment could mitigate systolic dysfunction and remodeling in Akita heart. Our MRI results revealed decreased ejection fraction in Akita as compared to WT and increased ejection fraction in miR-133a mimic-treated Akita. We also found that miR-133a mimic treatment mitigates T1DM-induced cardiac hypertrophy and fibrosis in Akita. We conclude that Akita shows cardiac hypertrophy, fibrosis and systolic dysfunction and miR-133a mimic treatment to Akita could ameliorate them.
- Published
- 2018
- Full Text
- View/download PDF
7. Diabetic Cardiomyopathy: An Immunometabolic Perspective
- Author
-
Paras K. Mishra, Sushil K. Mahata, Wei Ying, Shyam Sundar Nandi, Gautam K. Bandyopadhyay, and Kaushik K. Patel
- Subjects
obesity ,insulin resistance ,inflammation ,cardiomyopathy ,innate and adaptive immunity ,glucose metabolism ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
The heart possesses a remarkable inherent capability to adapt itself to a wide array of genetic and extrinsic factors to maintain contractile function. Failure to sustain its compensatory responses results in cardiac dysfunction, leading to cardiomyopathy. Diabetic cardiomyopathy (DCM) is characterized by left ventricular hypertrophy and reduced diastolic function, with or without concurrent systolic dysfunction in the absence of hypertension and coronary artery disease. Changes in substrate metabolism, oxidative stress, endoplasmic reticulum stress, formation of extracellular matrix proteins, and advanced glycation end products constitute the early stage in DCM. These early events are followed by steatosis (accumulation of lipid droplets) in cardiomyocytes, which is followed by apoptosis, changes in immune responses with a consequent increase in fibrosis, remodeling of cardiomyocytes, and the resultant decrease in cardiac function. The heart is an omnivore, metabolically flexible, and consumes the highest amount of ATP in the body. Altered myocardial substrate and energy metabolism initiate the development of DCM. Diabetic hearts shift away from the utilization of glucose, rely almost completely on fatty acids (FAs) as the energy source, and become metabolically inflexible. Oxidation of FAs is metabolically inefficient as it consumes more energy. In addition to metabolic inflexibility and energy inefficiency, the diabetic heart suffers from impaired calcium handling with consequent alteration of relaxation–contraction dynamics leading to diastolic and systolic dysfunction. Sarcoplasmic reticulum (SR) plays a key role in excitation–contraction coupling as Ca2+ is transported into the SR by the SERCA2a (sarcoplasmic/endoplasmic reticulum calcium-ATPase 2a) during cardiac relaxation. Diabetic cardiomyocytes display decreased SERCA2a activity and leaky Ca2+ release channel resulting in reduced SR calcium load. The diabetic heart also suffers from marked downregulation of novel cardioprotective microRNAs (miRNAs) discovered recently. Since immune responses and substrate energy metabolism are critically altered in diabetes, the present review will focus on immunometabolism and miRNAs.
- Published
- 2017
- Full Text
- View/download PDF
8. Ablation of matrix metalloproteinase-9 prevents cardiomyocytes contractile dysfunction in diabetics
- Author
-
Priyanka ePrathipati, Naira eMetreveli, Shyam Sundar Nandi, Suresh Chand Tyagi, and Paras Kumar eMishra
- Subjects
Diabetes Mellitus ,Heart Failure ,Calcium transient ,Akita ,Serca-2a ,Physiology ,QP1-981 - Abstract
Elevated expression and activity of matrix metalloproteinase-9 (MMP9) and decreased contractility of cardiomyocytes are documented in diabetic hearts. However, it is unclear whether MMP is involved in the regulation of contractility of cardiomyocytes in diabetic hearts. In the present study, we tested the hypothesis that MMP9 regulates contractility of cardiomyocytes in diabetic hearts, and ablation of matrix metalloproteinase-9 (MMP9) prevents impaired contractility of cardiomyocytes in diabetic hearts. To determine the specific role of MMP9 in cardiomyocyte contractility, we used twelve - fourteen week male WT (normoglycemic sibling of Akita), Akita, and Ins2+/- /MMP9-/- (DKO) mice. DKO mice were generated by cross-breeding male Ins2+/- Akita (T1D) with female MMP9 knockout (MMP9-/-) mice. We isolated cardiomyocytes from the heart of the above three groups of mice and measured their contractility and calcium transients. Moreover, we determined mRNA and protein levels of sarco-endoplasmic reticulum calcium ATPase-2a (SERCA-2a), which is involved in calcium handling during contractility of cardiomyocytes, in WT, Akita, and DKO hearts using qPCR, Western blotting and immunoprecipitation, respectively. Our results revealed that in Akita hearts where increased expression and activity of MMP9 is reported, the rates of shortening and re-lengthening (± dL/dt) of cardiomyocytes were decreased, time to 90% peak height and baseline during contractility was increased, rate of calcium decay was increased, and calcium transient was decreased as compared to WT cardiomyocytes. However, these changes in Akita were blunted in DKO cardiomyocytes. The molecular analyses of SERCA-2a in the hearts showed that it was downregulated in Akita as compared to WT but was comparatively upregulated in DKO. These results suggest that abrogation of MMP9 gene prevents contractility of cardiomyocytes, possibly by increasing SERCA-2a and calcium transients. We conclude that MMP9 plays crucial role in the regulation of contractility of cardiomyocytes in diabetic hearts.
- Published
- 2016
- Full Text
- View/download PDF
9. Aspirin and low-molecular weight heparin combination therapy effectively prevents recurrent miscarriage in hyperhomocysteinemic women.
- Author
-
Pratip Chakraborty, Sayani Banerjee, Piyali Saha, Shyam Sundar Nandi, Sunita Sharma, Sourendra K Goswami, Baidyanath Chakravarty, and Syed N Kabir
- Subjects
Medicine ,Science - Abstract
The management of recurrent pregnancy loss (RPL) still remains a great challenge, and women with polycystic ovarian syndrome (PCOS) are at a greater risk for spontaneous abortion. Treatment with low-molecular-weight heparin (LMWH) has become an accepted treatment option for women with RPL; however, the subgroup of women, who are likely to respond to LMWH, has not been precisely identified. The present study evaluated the efficacy of LMWH with reference to PCOS and associated metabolic phenotypes including hyperhomocysteinemia (HHcy), insulin resistance (IR) and obesity. This prospective observational study was conducted at Institute of Reproductive Medicine, Kolkata, India. A total of 967 women with history of 2 or more consecutive first trimester abortions were screened and 336 were selected for the study. The selected patients were initially divided on the basis of presence or absence of PCOS, while subsequent stratification was based on HHcy, IR and/or obesity. The subjects had treatment with aspirin during one conception cycle and aspirin-LMWH combined anticoagulant therapy for the immediate next conception cycle, if the first treated cycle was unsuccessful. Pregnancy salvage was the sole outcome measure. The overall rate of pregnancy salvage following aspirin therapy was 43.15%, which was mostly represented by normohomocysteinemic women, while the salvage rate was lower in the HHcy populations irrespective of the presence or absence of PCOS, IR, or obesity. By contrast, aspirin-LMWH combined therapy could rescue 66.84% pregnancies in the aspirin-failed cases. Logistic regression analyses showed that HHcy remained a significant factor in predicting salvage rates in the PCOS, IR, and obese subpopulations controlled for other confounding factors. With regard to pregnancy salvage, combined anticoagulant therapy with aspirin and LMWH conferred added benefit to those with HHcy phenotype.
- Published
- 2013
- Full Text
- View/download PDF
10. CD155: A Key Receptor Playing Diversified Roles
- Author
-
Snehasis Jana, Trupti Gohil, Upendra P. Lambe, Shyam Sundar Nandi, Sonali Ankush Sawant, and Sudip Ghosh
- Subjects
Cell signaling ,biology ,Chemistry ,General Medicine ,Ligands ,Biochemistry ,Transmembrane protein ,Hedgehog signaling pathway ,Cell biology ,TIGIT ,Viral Receptor ,Neoplasms ,biology.protein ,Humans ,Receptors, Virus ,Molecular Medicine ,Hedgehog Proteins ,CD155 ,Receptors, Immunologic ,Receptor ,Molecular Biology ,Poliovirus Receptor - Abstract
Cluster of differentiation (CD155), formerly identified as poliovirus receptor (PVR) and later as immunoglobulin molecule, is involved in cell adhesion, proliferation, invasion and migration. It is a surface protein expressed mostly on normal and transformed malignant cells. The expression of the receptor varies based on the origin of tissue. The expression of the protein is determined by factors involved in the sonic hedgehog pathway, Ras-MEK-ERK pathway and during stressful conditions like DNA damage response. The protein uses an alternate splicing mechanism, producing four isoforms, two being soluble (CD155β and CD155γ) and two being transmembrane protein (CD155α and CD155δ). Apart from being a viral receptor, researchers have identified CD155 to play important roles in cancer research and the cell signaling field. The receptor is recognized as a biomarker for identifying cancerous tissue. The receptor interacts with molecules involved in the cells’ defense mechanism. The immunesurveillance role of CD155 is being deciphered to understand the mechanistic approach it utilizes as an onco-immunologic molecule. CD155 is a non-MHC-I ligand which helps in identifying non-self to NK cells via an inhibitory TIGIT ligand. The TIGIT–CD155 pathway is a novel MHC-I-independent education mechanism for cell tolerance and activation of NK cells. The receptor also has a role in metastasis of cancer and transendothelial mechanism. In this review, the authors discuss the virus-host interaction that occurs via a single transmembrane receptor, the poliovirus infection pathway, which is being exploited as a therapeutic pathway. The oncolytic virotherapy is now a promising modality for curing cancer.
- Published
- 2022
11. Medicinal Plants Against Herpes Simplex Virus (HSV) Type 1 Infections: Ethnopharmacology, Chemistry, and Clinical and Preclinical Studies
- Author
-
Shyam Sundar Nandi, Sakshi Phalke, Sonali Ankush Sawant, Priyanka Paul, Upendra Lambe, Navya Bellamkonda, and Deepak Shukla
- Published
- 2023
12. Central Ang II (Angiotensin II)-Mediated Sympathoexcitation
- Author
-
Andréa S. Haibara, Neeru M. Sharma, Hong Zheng, Xuefei Liu, Kenichi Katsurada, Shyam Sundar Nandi, and Kaushik P. Patel
- Subjects
medicine.medical_specialty ,business.industry ,Neurogenic hypertension ,030204 cardiovascular system & hematology ,Hypoxia (medical) ,Angiotensin II ,03 medical and health sciences ,Glutamatergic ,0302 clinical medicine ,Endocrinology ,medicine.anatomical_structure ,Hypoxia-inducible factors ,Hypothalamus ,Internal medicine ,Internal Medicine ,Medicine ,Sympathetic outflow ,medicine.symptom ,business ,Nucleus ,hormones, hormone substitutes, and hormone antagonists ,030217 neurology & neurosurgery - Abstract
Central infusion of Ang II (angiotensin II) has been associated with increased sympathetic outflow resulting in neurogenic hypertension. In the present study, we appraised whether the chronic increase in central Ang II activates the paraventricular nucleus of the hypothalamus (PVN) resulting in elevated sympathetic tone and altered baro- and chemoreflexes. Further, we evaluated the contribution of HIF-1α (hypoxia-inducible factor-1α), a transcription factor involved in enhancing the expression of N-methyl-D-aspartate receptors and thus glutamatergic-mediated sympathetic tone from the PVN. Ang II infusion (20 ng/minute, intracerebroventricular, 14 days) increased mean arterial pressure (126±9 versus 84±4 mm Hg), cardiac sympathetic tone (96±7 versus 75±6 bpm), and decreased cardiac parasympathetic tone (16±2 versus 36±3 versus bpm) compared with saline-infused controls in conscious rats. The Ang II-infused group also showed an impaired baroreflex control of heart rate (−1.50±0.1 versus −2.50±0.3 bpm/mm Hg), potentiation of the chemoreflex pressor response (53±7 versus 30±7 mm Hg) and increased number of FosB-labeled cells (53±3 versus 19±4) in the PVN. Concomitant with the activation of the PVN, there was an increased expression of HIF-1α and N-Methyl-D-Aspartate-type1 receptors in the PVN. Further, Ang II-infusion showed increased renal sympathetic nerve activity (20.5±2.3% versus 6.4±1.9% of Max) and 3-fold enhanced renal sympathetic nerve activity responses to microinjection of N-methyl-D-aspartate (200 pmol) into the PVN of anesthetized rats. Further, silencing of HIF-1α in NG108 cells abrogated the expression of N-methyl-D-aspartate-N-methyl-D-aspartate-type1 induced by Ang II. Taken together, our studies suggest a novel Ang II-HIF-1α-N-methyl-D-aspartate receptor-mediated activation of preautonomic neurons in the PVN, resulting in increased sympathetic outflow and alterations in baro- and chemoreflexes.
- Published
- 2021
13. MMP9 inhibition increases autophagic flux in chronic heart failure
- Author
-
Kenichi Katsurada, Sushil K. Mahata, Kaushik P. Patel, Shyam Sundar Nandi, Neeru M. Sharma, and Daniel R. Anderson
- Subjects
Male ,0301 basic medicine ,Physiology ,Cardiac fibrosis ,Matrix Metalloproteinase Inhibitors ,030204 cardiovascular system & hematology ,Pharmacology ,Biochemistry ,Ventricular Function, Left ,Cell Line ,Rats, Sprague-Dawley ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Fibrosis ,Physiology (medical) ,Autophagy ,Genetics ,medicine ,Animals ,Myocytes, Cardiac ,cardiovascular diseases ,Myocardial infarction ,Molecular Biology ,Mechanistic target of rapamycin ,PI3K/AKT/mTOR pathway ,Benzofurans ,Heart Failure ,Ventricular Remodeling ,biology ,Chemistry ,Fibroblasts ,medicine.disease ,body regions ,Disease Models, Animal ,030104 developmental biology ,Matrix Metalloproteinase 9 ,Heart failure ,cardiovascular system ,biology.protein ,Cardiology and Cardiovascular Medicine ,Flux (metabolism) ,Research Article ,Signal Transduction ,Biotechnology - Abstract
Increased matrix metalloprotease 9 (MMP9) after myocardial infarction (MI) exacerbates ischemia-induced chronic heart failure (CHF). Autophagy is cardioprotective during CHF; however, whether increased MMP9 suppresses autophagic activity in CHF is unknown. This study aimed to determine whether increased MMP9 suppressed autophagic flux and MMP9 inhibition increased autophagic flux in the heart of rats with post-MI CHF. Sprague-Dawley rats underwent either sham surgery or coronary artery ligation 6–8 wk before being treated with MMP9 inhibitor for 7 days, followed by cardiac autophagic flux measurement with lysosomal inhibitor bafilomycin A1. Furthermore, autophagic flux was measured in vitro by treating H9c2 cardiomyocytes with two independent pharmacological MMP9 inhibitors, salvianolic acid B (SalB) and MMP9 inhibitor-I, and CRISPR/cas9-mediated MMP9 genetic ablation. CHF rats showed cardiac infarct, significantly increased left ventricular end-diastolic pressure (LVEDP), and increased MMP9 activity and fibrosis in the peri-infarct areas of left ventricular myocardium. Measurement of the autophagic markers LC3B-II and p62 with lysosomal inhibition showed decreased autophagic flux in the peri-infarct myocardium. Treatment with SalB for 7 days in CHF rats decreased MMP9 activity and cardiac fibrosis but increased autophagic flux in the peri-infarct myocardium. As an in vitro corollary study, measurement of autophagic flux in H9c2 cardiomyocytes and fibroblasts showed that pharmacological inhibition or genetic ablation of MMP9 upregulates autophagic flux. These data are consistent with our observations that MMP9 inhibition upregulates autophagic flux in the heart of rats with CHF. In conclusion, the results in this study suggest that the beneficial outcome of MMP9 inhibition in pathological cardiac remodeling is in part mediated by improved autophagic flux. NEW & NOTEWORTHY This study elucidates that the improved cardiac extracellular matrix (ECM) remodeling and cardioprotective effect of matrix metalloprotease 9 (MMP9) inhibition in chronic heart failure (CHF) are via increased autophagic flux. Autophagy is cardioprotective; however, the mechanism of autophagy suppression in CHF is unknown. We for the first time demonstrated here that increased MMP9 suppressed cardiac autophagy and ablation of MMP9 increased cardiac autophagic flux in CHF rats. Restoring the physiological level of autophagy in the failing heart is a challenge, and our study addressed this challenge. The novelty and highlights of this report are as follows: 1) MMP9 regulates cardiomyocyte and fibroblast autophagy, 2) MMP9 inhibition protects CHF after myocardial infarction (MI) via increased cardiac autophagic flux, 3) MMP9 inhibition increased cardiac autophagy via activation of AMP-activated protein kinase (AMPK)α, Beclin-1, Atg7 pathway and suppressed mechanistic target of rapamycin (mTOR) pathway.
- Published
- 2020
14. Does glucagon-like peptide-1 induce diuresis and natriuresis by modulating afferent renal nerve activity?
- Author
-
Neeru M. Sharma, Kaushik P. Patel, Xuefei Liu, Hong Zheng, Kenichi Katsurada, and Shyam Sundar Nandi
- Subjects
Male ,0301 basic medicine ,endocrine system ,medicine.medical_specialty ,Sympathetic Nervous System ,Physiology ,Calcitonin Gene-Related Peptide ,medicine.medical_treatment ,Natriuresis ,Hemodynamics ,Diuresis ,030204 cardiovascular system & hematology ,Kidney ,Glucagon-Like Peptide-1 Receptor ,Renal Circulation ,Rats, Sprague-Dawley ,Renal nerve ,03 medical and health sciences ,0302 clinical medicine ,Glucagon-Like Peptide 1 ,Internal medicine ,Afferent ,medicine ,Animals ,Humans ,Kidney Pelvis ,Afferent Pathways ,business.industry ,Sodium ,digestive, oral, and skin physiology ,Denervation ,Glucagon-like peptide-1 ,Natriuretic Effects ,Rats ,Urodynamics ,HEK293 Cells ,030104 developmental biology ,Endocrinology ,Diuretic ,business ,hormones, hormone substitutes, and hormone antagonists ,Research Article - Abstract
Glucagon-like peptide-1 (GLP-1), an incretin hormone, has diuretic and natriuretic effects. The present study was designed to explore the possible underlying mechanisms for the diuretic and natriuretic effects of GLP-1 via renal nerves in rats. Immunohistochemistry revealed that GLP-1 receptors were avidly expressed in the pelvic wall, the wall being adjacent to afferent renal nerves immunoreactive to calcitonin gene-related peptide, which is the dominant neurotransmitter for renal afferents. GLP-1 (3 μM) infused into the left renal pelvis increased ipsilateral afferent renal nerve activity (110.0 ± 15.6% of basal value). Intravenous infusion of GLP-1 (1 µg·kg−1·min−1) for 30 min increased renal sympathetic nerve activity (RSNA). After the distal end of the renal nerve was cut to eliminate the afferent signal, the increase in efferent renal nerve activity during intravenous infusion of GLP-1 was diminished compared with the increase in total RSNA (17.0 ± 9.0% vs. 68.1 ± 20.0% of the basal value). Diuretic and natriuretic responses to intravenous infusion of GLP-1 were enhanced by total renal denervation (T-RDN) with acute surgical cutting of the renal nerves. Selective afferent renal nerve denervation (A-RDN) was performed by bilateral perivascular application of capsaicin on the renal nerves. Similar to T-RDN, A-RDN enhanced diuretic and natriuretic responses to GLP-1. Urine flow and Na+ excretion responses to GLP-1 were not significantly different between T-RDN and A-RDN groups. These results indicate that the diuretic and natriuretic effects of GLP-1 are partly governed via activation of afferent renal nerves by GLP-1 acting on sensory nerve fibers within the pelvis of the kidney.
- Published
- 2019
15. Enhanced expression and function of renal SGLT2 (sodium-glucose cotransporter 2) in heart failure: Role of renal nerves
- Author
-
Neeru M. Sharma, Shyam Sundar Nandi, Kenichi Katsurada, and Kaushik P. Patel
- Subjects
Male ,medicine.medical_specialty ,Sympathetic Nervous System ,Sodium ,chemistry.chemical_element ,Kidney ,Article ,Norepinephrine (medication) ,Rats, Sprague-Dawley ,Sodium-Glucose Transporter 2 ,Internal medicine ,Medicine ,Animals ,Sodium-Glucose Transporter 2 Inhibitors ,Denervation ,Heart Failure ,business.industry ,medicine.disease ,medicine.anatomical_structure ,Endocrinology ,Glucose ,chemistry ,Sodium/Glucose Cotransporter 2 ,Heart failure ,Cardiology and Cardiovascular Medicine ,business ,Cotransporter ,Function (biology) ,medicine.drug - Abstract
Background: Recent clinical studies demonstrate that SGLT2 (sodium-glucose cotransporter 2) inhibitors ameliorate heart failure (HF). The present study was conducted to assess the expression and function of renal SGLT2 and the influence of enhanced renal sympathetic tone in HF. Methods: Four weeks after coronary artery ligation surgery to induce HF, surgical bilateral renal denervation (RDN) was performed in rats. Four groups of rats (Sham-operated control [Sham], Sham+RDN, HF and HF+RDN; n=6/group) were used. Immunohistochemistry and Western blot analysis were performed to evaluate the renal SGLT2 expression. One week after RDN (5 weeks after induction of HF), intravenous injection of SGLT2 inhibitor dapagliflozin were performed to assess renal excretory responses. In vitro, human embryonic kidney cells were used to investigate the fractionation of SGLT2 after norepinephrine treatment. Results: In rats with HF, (1) SGLT2 expression in the proximal tubule of the kidney was increased; (2) the response of increases in urine flow, sodium excretion, and glucose excretion to dapagliflozin were greater; and (3) RDN attenuated renal SGLT2 expression and normalized renal functional responses to dapagliflozin. In vitro, norepinephrine promoted translocation of SGLT2 to the cell membrane. Conclusions: These results indicate that the enhanced tonic renal sympathetic nerve activation in HF increases the expression and functional activity of renal SGLT2. Potentiated trafficking of SGLT2 to cell surface in renal proximal tubules mediated by norepinephrine may contribute to this functional activation of SGLT2 in HF. These findings provide critical insight into the underlying mechanisms for the beneficial effects of SGLT2 inhibitors on HF reported in the clinical studies.
- Published
- 2021
16. Neurogenic Hypertension Mediated Mitochondrial Abnormality Leads to Cardiomyopathy: Contribution of UPRmt and Norepinephrine-miR- 18a-5p-HIF-1α Axis
- Author
-
Kaushik P. Patel, Sushil K. Mahata, Shyam Sundar Nandi, and Kenichi Katsurada
- Subjects
Mitochondrial ROS ,medicine.medical_specialty ,hypertension ,Cardiac fibrosis ,business.industry ,mitochondrial damage ,miR-18a-5p ,Physiology ,cardiac hypertrophy ,cardiac fibrosis ,Hypertrophic cardiomyopathy ,Cardiomyopathy ,HIF-1α ,Neurogenic hypertension ,medicine.disease ,Endocrinology ,Mitochondrial biogenesis ,Physiology (medical) ,Internal medicine ,Mitochondrial unfolded protein response ,medicine ,QP1-981 ,Mitochondrial fission ,business - Abstract
Aims: Hypertension increases the risk of heart disease. Hallmark features of hypertensive heart disease is sympathoexcitation and cardiac mitochondrial abnormality. However, the molecular mechanisms for specifically neurally mediated mitochondrial abnormality and subsequent cardiac dysfunction are unclear. We hypothesized that enhanced sympatho-excitation to the heart elicits cardiac miR-18a-5p/HIF-1α and mitochondrial unfolded protein response (UPRmt) signaling that lead to mitochondrial abnormalities and consequent pathological cardiac remodeling.Methods and Results: Using a model of neurogenic hypertension (NG-HTN), induced by intracerebroventricular (ICV) infusion of Ang II (NG-HTN; 20 ng/min, 14 days, 0.5 μl/h, or Saline; Control, 0.9%) through osmotic mini-pumps in Sprague-Dawley rats (250–300 g), we attempted to identify a link between sympathoexcitation (norepinephrine; NE), miRNA and HIF-1α signaling and UPRmt to produce mitochondrial abnormalities resulting in cardiomyopathy. Cardiac remodeling, mitochondrial abnormality, and miRNA/HIF-1α signaling were assessed using histology, immunocytochemistry, electron microscopy, Western blotting or RT-qPCR. NG-HTN demonstrated increased sympatho-excitation with concomitant reduction in UPRmt, miRNA-18a-5p and increased level of HIF-1α in the heart. Our in silico analysis indicated that miR-18a-5p targets HIF-1α. Direct effects of NE on miRNA/HIF-1α signaling and mitochondrial abnormality examined using H9c2 rat cardiomyocytes showed NE reduces miR-18a-5p but increases HIF-1α. Electron microscopy revealed cardiac mitochondrial abnormality in NG-HTN, linked with hypertrophic cardiomyopathy and fibrosis. Mitochondrial unfolded protein response was decreased in NG-HTN indicating mitochondrial proteinopathy and proteotoxic stress, associated with increased mito-ROS and decreased mitochondrial membrane potential (ΔΨm), and oxidative phosphorylation. Further, there was reduced cardiac mitochondrial biogenesis and fusion, but increased mitochondrial fission, coupled with mitochondrial impaired TIM-TOM transport and UPRmt. Direct effects of NE on H9c2 rat cardiomyocytes also showed cardiomyocyte hypertrophy, increased mitochondrial ROS generation, and UPRmt corroborating the in vivo data.Conclusion: In conclusion, enhanced sympatho-excitation suppress miR-18a-5p/HIF-1α signaling and increased mitochondrial stress proteotoxicity, decreased UPRmt leading to decreased mitochondrial dynamics/OXPHOS/ΔΨm and ROS generation. Taken together, these results suggest that ROS induced mitochondrial transition pore opening activates pro-hypertrophy/fibrosis/inflammatory factors that induce pathological cardiac hypertrophy and fibrosis commonly observed in NG-HTN.
- Published
- 2021
17. Deep learning research should be encouraged for diagnosis and treatment of antibiotic resistance of microbial infections in treatment associated emergencies in hospitals
- Author
-
Chiranjib Chakraborty, Manojit Bhattacharya, Ashish Ranjan Sharma, Sanjiban Sekhar Roy, Md Aminul Islam, Sandip Chakraborty, Shyam Sundar Nandi, and Kuldeep Dhama
- Subjects
Drug Resistance, Microbial ,Microbial Sensitivity Tests ,DNA ,General Medicine ,Hospitals ,Article ,Anti-Bacterial Agents ,Computational biophysics ,Deep Learning ,Machine learning ,Next-generation sequencing ,Humans ,Surgery ,Emergencies - Abstract
Targeted high-throughput DNA sequencing is a primary approach for genomics and molecular diagnostics, and more recently as a readout for DNA information storage. Oligonucleotide probes used to enrich gene loci of interest have different hybridization kinetics, resulting in non-uniform coverage that increases sequencing costs and decreases sequencing sensitivities. Here, we present a deep learning model (DLM) for predicting Next-Generation Sequencing (NGS) depth from DNA probe sequences. Our DLM includes a bidirectional recurrent neural network that takes as input both DNA nucleotide identities as well as the calculated probability of the nucleotide being unpaired. We apply our DLM to three different NGS panels: a 39,145-plex panel for human single nucleotide polymorphisms (SNP), a 2000-plex panel for human long non-coding RNA (lncRNA), and a 7373-plex panel targeting non-human sequences for DNA information storage. In cross-validation, our DLM predicts sequencing depth to within a factor of 3 with 93% accuracy for the SNP panel, and 99% accuracy for the non-human panel. In independent testing, the DLM predicts the lncRNA panel with 89% accuracy when trained on the SNP panel. The same model is also effective at predicting the measured single-plex kinetic rate constants of DNA hybridization and strand displacement., DNA probes used in next generation sequencing (NGS) have variable hybridisation kinetics, resulting in non-uniform coverage. Here, the authors develop a deep learning model to predict NGS depth using DNA probe sequences and apply to human and non-human sequencing panels.
- Published
- 2022
18. Comparative genomics, evolutionary epidemiology, and RBD-hACE2 receptor binding pattern in B.1.1.7 (Alpha) and B.1.617.2 (Delta) related to their pandemic response in UK and India
- Author
-
Chiranjib, Chakraborty, Ashish Ranjan, Sharma, Manojit, Bhattacharya, Bidyut, Mallik, Shyam Sundar, Nandi, and Sang-Soo, Lee
- Subjects
Microbiology (medical) ,SARS-CoV-2 ,COVID-19 ,Genomics ,Microbiology ,United Kingdom ,Infectious Diseases ,Mutation ,Spike Glycoprotein, Coronavirus ,Genetics ,Humans ,Angiotensin-Converting Enzyme 2 ,Pandemics ,Molecular Biology ,Ecology, Evolution, Behavior and Systematics - Abstract
The massive increase in COVID-19 infection had generated a second wave in India during May-June 2021 with a critical pandemic situation. The Delta variant (B.1.617.2) was a significant factor during the second wave. Conversely, the UK had passed through the crucial phase of the pandemic from November to December 2020 due to B.1.1.7. The study tried to comprehend the pandemic response in the UK and India to the spread of the B.1.1.7 (Alpha, UK) variant and B.1.617.2 (Delta, India) variant.This study was performed in three directions to understand the pandemic response of the two emerging variants. First, we served comparative genomics, such as genome sequence submission patterns, mutational landscapes, and structural landscapes of significant mutations (N501Y, D614G, L452R, E484Q, and P681R). Second, we performed evolutionary epidemiology using molecular phylogenetics, scatter plots of the cluster evaluation, country-wise transmission pattern, and frequency pattern. Third, the receptor binding pattern was analyzed using the Wuhan reference strain and the other two variants.The study analyzed the country-wise and region-wise genome sequences and their submission pattern, molecular phylogenetics, scatter plot of the cluster evaluation, country-wise geographical distribution and transmission pattern, frequency pattern, entropy diversity, and mutational landscape of the two variants. The structural pattern was analyzed in the N501Y, D614G L452R, E484Q, and P681R mutations. The study found increased molecular interactivity between hACE2-RBD binding of B.1.1.7 and B.1.617.2 compared to the Wuhan reference strain. Our receptor binding analysis showed a similar indication pattern for hACE2-RBD of these two variants. However, B.1.617.2 offers slightly better stability in the hACE2-RBD binding pattern through MD simulation than B.1.1.7.The increased hACE2-RBD binding pattern of B.1.1.7 and B.1.617.2 might help to increase the infectivity compared to the Wuhan reference strain.
- Published
- 2022
19. A Rapid Point of Care CC16 Kit for Screening of Occupational Silica Dust Exposed Workers for Early Detection of Silicosis/Silico-Tuberculosis
- Author
-
Upendra P. Lambe, Kamalesh Sarkar, Sonali Sawant, Shyam Sundar Nandi, Jagadish M. Deshpande, and Trupti Gohil
- Subjects
Silica dust ,Tuberculosis ,Silicosis ,business.industry ,Environmental health ,medicine ,Early detection ,medicine.disease ,business ,Point of care - Abstract
Silicosis is an irreversible, incurable and progressive occupational disease caused by prolonged exposure to crystalline-silica while working in related industries. Conventionally diagnosis is done by chest radiology in advanced stage as early symptoms often go unnoticed. Early detection and secondary prevention could be the only realistic possible control strategy for controlling silicosis as no other method of treatment is available. Additionally, these patients are also vulnerable to drug resistant tuberculosis. Developing countries like India has a huge burden of silico-tuberculosis. Hence, a rapid and inexpensive screening method is a need for early detection of silicosis among silica dust exposed workers. Serum CC16 is a useful proxy screening marker for early detection of silicosis. In this study a lateral-flow assay for semi-quantitative estimation of serum CC16 level was developed. The detection was performed using gold nanoparticles conjugated anti-CC16 monoclonal antibodies. A sum of 47 serum samples was tested to do performance evaluation of the assay. A concentration of 6ng/ml or less produced one band, 6.1 – 9 ng/ml produced two bands, while more than 9 ng/ml produced all the three bands at the test zone. This assay can be used as a proxy marker for periodic screening and early detection of silicosis in vulnerable workers.
- Published
- 2021
20. Central Glucagon-like Peptide-1 Receptor Signaling via Brainstem Catecholamine Neurons Counteracts Hypertension in Spontaneously Hypertensive Rats
- Author
-
Toshihiko Yada, Yuko Maejima, Neeru M. Sharma, Boyang Zhang, Masanori Nakata, Kazuomi Kario, Kenichi Katsurada, Toshinobu Saito, Kaushik K. Patel, and Shyam Sundar Nandi
- Subjects
Agonist ,Male ,medicine.medical_specialty ,Saporin ,medicine.drug_class ,lcsh:Medicine ,Dopamine beta-Hydroxylase ,030204 cardiovascular system & hematology ,Rats, Inbred WKY ,Glucagon-Like Peptide-1 Receptor ,Article ,03 medical and health sciences ,Norepinephrine ,0302 clinical medicine ,Dopamine ,Internal medicine ,Rats, Inbred SHR ,medicine ,Animals ,lcsh:Science ,Multidisciplinary ,biology ,Liraglutide ,business.industry ,Dopaminergic Neurons ,Area postrema ,lcsh:R ,Glucagon-like peptide-1 ,Metabolic syndrome ,Rats ,Endocrinology ,nervous system ,Hypertension ,Catecholamine ,biology.protein ,lcsh:Q ,business ,030217 neurology & neurosurgery ,medicine.drug ,Brain Stem ,Signal Transduction ,Neuroscience ,circulatory and respiratory physiology - Abstract
Glucagon-like peptide-1 receptor (GLP-1R) agonists, widely used to treat type 2 diabetes, reduce blood pressure (BP) in hypertensive patients. Whether this action involves central mechanisms is unknown. We here report that repeated lateral ventricular (LV) injection of GLP-1R agonist, liraglutide, once daily for 15 days counteracted the development of hypertension in spontaneously hypertensive rats (SHR). In parallel, it suppressed urinary norepinephrine excretion, and induced c-Fos expressions in the area postrema (AP) and nucleus tractus solitarius (NTS) of brainstem including the NTS neurons immunoreactive to dopamine beta-hydroxylase (DBH). Acute administration of liraglutide into fourth ventricle, the area with easy access to the AP and NTS, transiently decreased BP in SHR and this effect was attenuated after lesion of NTS DBH neurons with anti-DBH conjugated to saporin (anti-DBH-SAP). In anti-DBH-SAP injected SHR, the antihypertensive effect of repeated LV injection of liraglutide for 14 days was also attenuated. These findings demonstrate that the central GLP-1R signaling via NTS DBH neurons counteracts the development of hypertension in SHR, accompanied by attenuated sympathetic nerve activity.
- Published
- 2019
21. Abstract 16639: A Novel Approach to Model Heart Failure With Preserved Ejection Fraction in Rats
- Author
-
Neeru M. Sharma, Shyam Sundar Nandi, Kaushik P Patel, and Kenichi Katsurada
- Subjects
medicine.medical_specialty ,Ideal (set theory) ,business.industry ,Diastole ,medicine.disease ,Muscle hypertrophy ,Animal model ,Fibrosis ,Physiology (medical) ,Heart failure ,Internal medicine ,medicine ,Cardiology ,Diastolic function ,Cardiology and Cardiovascular Medicine ,Heart failure with preserved ejection fraction ,business - Abstract
Introduction: Diastolic dysfunction and cardiac hypertrophic remodeling are hallmark features of heart failure with preserved ejection fraction (HFpEF) in humans. However, an ideal animal model that mimics all these basic clinical features of HFpEF remains to be developed. Methods: Male Sprague-Dawley rats (250-300 g) were subjected to central infusion of either Angiotensin II (Ang II at 20 ng/min, 0.5 μl/h, i.c.v) or isotonic saline (0.5 μl/h, i.c.v) through osmotic mini-pumps for 14 days. Echocardiography was used to measure systolic (ejection fraction) and diastolic functions (E/A and E/E’ ratio) at day 14. Mean arterial pressure (MAP), heart rate (HR), left ventricular pressure (LVP), changes in ±dP/dt, and cardiac responses to isoproterenol (ISO) were also measured in anesthetized rats. Results: Central infusion of Ang II in rats resulted in increased sympathoexcitation and mean arterial pressure. Concomitantly, there was decreased E/A ratio (AngII: 1.2 ± 0.1 vs. Cont: 1.5 ± 0.1) and increased E/E’ ratio (AngII: 14.3 ± 0.8 vs. Cont: 12.9 ± 0.6), indicating diastolic dysfunction. Ejection fractions were comparable between the groups (AngII: 67 ± 2% vs. Cont: 69 ± 5%). Ang II infused rats demonstrated increased heart to body weight ratio (AngII: 4.5 ± 0.5 vs. Cont: 2.9 ± 0.1), cachexia (AngII: 235 ± 18 g vs. Cont: 331 ± 11 g), hypertrophy (AngII: 46 ± 5 μm/unit area vs. Cont: 36 ± 7 μm/unit area), and fibrosis (AngII: 5.2 ± 0.5 intensity/unit area vs. Cont: 2.6 ± 0.4 intensity/unit area). At the same time cardiac contractile responsiveness to ISO (0.5 μg/kg) was significantly lower for negative dP/dt in Ang II rats (AngII: -6998 mmHg/s vs. Cont: -11675 mmHg/s), which is indicative of abnormal diastole or relaxation. However, positive dP/dt response to ISO was similar in the two groups, indicating preserved systolic function in the Ang II group. Conclusions: Central Ang II infusion in rats shows sympathoexcitation, diastolic dysfunction, and pathological cardiac hypertrophy and fibrosis mimicking abnormalities seen in patients with HFpEF. This model of cardiac dysfunction in rats may represent a suitable platform to investigate the underlying mechanisms of HFpEF.
- Published
- 2020
22. Abstract MP49: Central Angiotensin II Mediates Neurogenic Hypertension Through Hif1-Alpha/NMDAR Axis In The Paraventricular Nucleus Of The Hypothalamus (PVN)
- Author
-
Andrea Siqueira Haibara, Shyam Sundar Nandi, Kaushik P Patel, Kenichi Katsurada, and Neeru M. Sharma
- Subjects
medicine.medical_specialty ,business.industry ,Alpha (ethology) ,Neurogenic hypertension ,Angiotensin II ,Pathogenesis ,Endocrinology ,Blood pressure ,medicine.anatomical_structure ,Hypothalamus ,Internal medicine ,Internal Medicine ,medicine ,NMDA receptor ,business ,Nucleus - Abstract
Central activation of the renin-angiotensin system and subsequent sympathoexcitation plays a pivotal role in the pathogenesis of hypertension. The specific neural substrates and the possible molecular mechanisms for the sympathoexcitation remain unclear. The glutamatergic activation via N-methyl-d-aspartate receptors (NMDAR) in the PVN enhances sympathetic outflow in hypertension. Previously, using Chromatin immunoprecipitation assay (ChiP) we have shown that Hypoxia-Inducible Factor-1 (heterodimer of HIF-1 α and HIF-1 β subunits ) binds to HREs elements in the NMDA-NR1(structural subunit of NMDAR) promoter in nucleus and increases transcription. In this study, central infusion of Ang II (20 ng/min, 14days, 0.5μl/h, i.c.v) through osmotic mini-pumps in Sprague-Dawley rats increased renal sympathetic nerve activity (20.5 ± 2.3 vs. 6.4 ± 1.9 % of Max) and mean arterial pressure (126 ± 9 vs. 84 ± 4 mmHg). At the same time, there was an increased expression of HIF-1α mRNA (2.79 fold) and HIF-1α protein (1.54 fold), as well as the expression of NMDA-NR1 mRNA (3.17 fold) and NMDA-NR1 protein levels (1.52 fold) in the PVN of Ang II infused group. Direct application of Ang II (1μM) induced an increase in the expression of HIF-1α protein (0.74 ± 0.03* vs. 0.50 ± 0.04) as well as decreased expression of prolyl hydroxylase domain protein 2 (an enzyme hydroxylating the proline residues of HIF-α, inducing subsequent ubiquitination and proteasomal degradation) (1.47 ± 0.14* vs. 0.50 ± 0.03) in NG108-15 neuronal cells. Tandem Ubiquitin-Binding Entities assay showed decreased HIF-α -Ub conjugates in Ang II-treated cells (0.65 ± 0.09* vs. 1.31 ± 0.10) suggesting a role for Ang II in post-translational stabilization of HIF-α. Further, silencing of HIF-1α (~60%) with siRNA in NG108-15 cells leads to a decrease in the expression of NMDA-NR1 induced by Ang II, compared with scrambled siRNA (0.47 ± 0.05* vs 0.80 ± 0.04). Taken together, these studies suggest that elevated central Ang II upregulates the expression of HIF-1α at transcriptional as well as post-translational level, which potentiates glutamatergic tone by enhanced expression of NMDA-NR1 in the PVN leading to heightened glutamatergic tone with a consequent increase in sympathetic outflow resulting in hypertension.
- Published
- 2020
23. Vaccines and Therapies in Development for SARS-CoV-2 Infections
- Author
-
Raghuram Koganti, Shyam Sundar Nandi, Deepak Shukla, David Wu, Tejabhiram Yadavalli, and Upendra P. Lambe
- Subjects
Drug ,medicine.medical_specialty ,2019-20 coronavirus outbreak ,Coronavirus disease 2019 (COVID-19) ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,media_common.quotation_subject ,viruses ,therapies ,coronavirus ,lcsh:Medicine ,Review ,medicine.disease_cause ,Virus ,03 medical and health sciences ,0302 clinical medicine ,Pandemic ,medicine ,030212 general & internal medicine ,Intensive care medicine ,030304 developmental biology ,Coronavirus ,media_common ,0303 health sciences ,business.industry ,SARS-CoV-2 ,lcsh:R ,Outbreak ,COVID-19 ,General Medicine ,vaccines ,business - Abstract
The current COVID-19 pandemic is caused by the novel coronavirus SARS-CoV-2. The virus causes severe respiratory symptoms which manifest disproportionately in the elderly. Currently, there are over 6.5 million cases and 380,000 deaths reported. Given the current severity of the outbreak, there is a great need for antiviral therapies and vaccines to treat and prevent COVID-19. In this review, we provide an overview of SARS-CoV-2 and discuss the emerging therapies and vaccines that show promise in combating COVID-19. We also highlight potential viral targets that could be exploited by researchers and drug manufacturers.
- Published
- 2020
24. GLP-1 mediated diuresis and natriuresis are blunted in heart failure and restored by selective afferent renal denervation
- Author
-
Xuefei Liu, Neeru M. Sharma, Hong Zheng, Kenichi Katsurada, Kaushik K. Patel, and Shyam Sundar Nandi
- Subjects
Male ,Glucagon-like peptide-1 ,endocrine system ,lcsh:Diseases of the circulatory (Cardiovascular) system ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Receptor expression ,medicine.medical_treatment ,Natriuresis ,Sympathetic nerve activity ,Diuresis ,Heart failure ,030204 cardiovascular system & hematology ,Kidney ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Glucagon-Like Peptide 1 ,Renal afferent ,Internal medicine ,medicine ,Animals ,Diuretics ,Infusions, Intravenous ,Original Investigation ,030304 developmental biology ,Denervation ,0303 health sciences ,business.industry ,Sympathectomy, Chemical ,Sodium and water homeostasis ,medicine.disease ,Disease Models, Animal ,medicine.anatomical_structure ,Endocrinology ,lcsh:RC666-701 ,Capsaicin ,Diuretic ,Cardiology and Cardiovascular Medicine ,business ,Renal pelvis - Abstract
BackgroundGlucagon-like peptide-1 (GLP-1) induces diuresis and natriuresis. Previously we have shown that GLP-1 activates afferent renal nerve to increase efferent renal sympathetic nerve activity that negates the diuresis and natriuresis as a negative feedback mechanism in normal rats. However, renal effects of GLP-1 in heart failure (HF) has not been elucidated. The present study was designed to assess GLP-1-induced diuresis and natriuresis in rats with HF and its interactions with renal nerve activity.MethodsHF was induced in rats by coronary artery ligation. The direct recording of afferent renal nerve activity (ARNA) with intrapelvic injection of GLP-1 and total renal sympathetic nerve activity (RSNA) with intravenous infusion of GLP-1 were performed. GLP-1 receptor expression in renal pelvis, densely innervated by afferent renal nerve, was assessed by real-time PCR and western blot analysis. In separate group of rats after coronary artery ligation selective afferent renal denervation (A-RDN) was performed by periaxonal application of capsaicin, then intravenous infusion of GLP-1-induced diuresis and natriuresis were evaluated.ResultsIn HF, compared to sham-operated control; (1) response of increase in ARNA to intrapelvic injection of GLP-1 was enhanced (3.7 ± 0.4 vs. 2.0 ± 0.4 µV s), (2) GLP-1 receptor expression was increased in renal pelvis, (3) response of increase in RSNA to intravenous infusion of GLP-1 was enhanced (132 ± 30% vs. 70 ± 16% of the baseline level), and (4) diuretic and natriuretic responses to intravenous infusion of GLP-1 were blunted (urine flow 53.4 ± 4.3 vs. 78.6 ± 4.4 µl/min/gkw, sodium excretion 7.4 ± 0.8 vs. 10.9 ± 1.0 µEq/min/gkw). A-RDN induced significant increases in diuretic and natriuretic responses to GLP-1 in HF (urine flow 96.0 ± 1.9 vs. 53.4 ± 4.3 µl/min/gkw, sodium excretion 13.6 ± 1.4 vs. 7.4 ± 0.8 µEq/min/gkw).ConclusionsThe excessive activation of neural circuitry involving afferent and efferent renal nerves suppresses diuretic and natriuretic responses to GLP-1 in HF. These pathophysiological responses to GLP-1 might be involved in the interaction between incretin-based medicines and established HF condition. RDN restores diuretic and natriuretic effects of GLP-1 and thus has potential beneficial therapeutic implication for diabetic HF patients.
- Published
- 2020
25. Poliovirus Non-Permissive CD155 Knockout Cells Derived from RD Cell Line Using CRISPR Technology for Handling Poliovirus Potentially Infectious Materials in Virology Laboratories
- Author
-
Shyam Sundar Nandi, Sonali Sawant, Ujjala Ghoshal, Jagadish Deshpande, Lucky Sangal, Pauline Harvey, Trupti Gohil, Kaveri Krishnasamy, and Disha Patel
- Subjects
biology ,viruses ,Poliovirus ,Clone (cell biology) ,medicine.disease_cause ,complex mixtures ,Virology ,Cell culture ,biology.protein ,medicine ,Enterovirus ,CRISPR ,CD155 ,Gene ,Poliovirus Receptor - Abstract
Background: Destruction of all poliovirus containing materials, and safe and secure handling of retained polioviruses for vaccine production and research will be obligatory to eliminate facility-associated risks. As defined in WHO-GAPIII, poliovirus potentially infectious materials (PIM) include fecal or respiratory samples collected for any purpose in time and geographic area of wild poliovirus or during OPV use and products of such materials from poliovirus permissive cells or animals. Non-polio laboratories culturing viruses from PIM are most affected as cell cultures of human and monkey origin are also poliovirus permissive. Methods: CRISPR gene editing technology was used to knockout poliovirus receptor (PVR/CD155) gene in RD cell line. Poliovirus non-permissive cells were identified by resistance to poliovirus infection, cell surface immunofluorescence and DNA sequencing. PVR knockout RD cell susceptibility was tested using known non-polio enterovirus types. A selected clone (RD-SJ40) was field evaluated for virus isolation from 626 stool samples of AFP cases. Findings: Poliovirus non-permissive cells derived from RD cell line did not show CD155-specific cell surface immunofluorescence. CD155 gene sequencing confirmed nucleotide base pair deletions within exon2 and exon3. The CD155 knockout RD-SJ40 cells did not support growth of poliovirus from positive stool samples. All NPEV types isolated in parental RD cells were also isolated in RD-SJ40. Interpretation: CRISPR correctly edited CD155 gene of RD cells to render them poliovirus non-permissive while susceptibility to NPEV remained unchanged. RD-SJ40 cells are safe for NPEV isolation from poliovirus PIM without derogating GAPIII containment requirements. Funding: Department of Health Research, Govt. of India. Project Grant No.R.12020/08/2018-HR Declaration of Interests: SSN, SS and JD have IPR interests. Other authors have no conflict of interests.
- Published
- 2020
26. Epitope Mapping of SERCA2a Identifies an Antigenic Determinant That Induces Mainly Atrial Myocarditis in A/J Mice
- Author
-
Muhammad Afzal, Chandirasegaran Massilamany, Rajkumar A. Rajasekaran, Arunakumar Gangaplara, Paras Kumar Mishra, Raymond A. Sobel, Rakesh H. Basavalingappa, David J. Steffen, Vahid Khalilzad-Sharghi, Shyam Sundar Nandi, Jennifer L. Strande, Bharathi Krishnan, N R Jayagopala Reddy, You Zhou, and Jean-Jack M Riethoven
- Subjects
0301 basic medicine ,SERCA ,Myocarditis ,Heart Ventricles ,T cell ,Immunology ,Fluorescent Antibody Technique ,Gene Expression ,Mice, Inbred Strains ,Inflammation ,Article ,Epitope ,Sarcoplasmic Reticulum Calcium-Transporting ATPases ,Epitopes ,Mice ,03 medical and health sciences ,Bacterial Proteins ,T-Lymphocyte Subsets ,medicine ,Animals ,Immunology and Allergy ,Myocytes, Cardiac ,Heart Atria ,Alleles ,MHC class II ,biology ,Immunodominant Epitopes ,Endoplasmic reticulum ,Histocompatibility Antigens Class II ,medicine.disease ,Immunohistochemistry ,Disease Models, Animal ,030104 developmental biology ,medicine.anatomical_structure ,Epitope mapping ,cardiovascular system ,biology.protein ,Cytokines ,Epitopes, B-Lymphocyte ,medicine.symptom ,Peptides ,Epitope Mapping ,circulatory and respiratory physiology - Abstract
Sarcoplasmic/endoplasmic reticulum Ca2+ adenosine triphosphatase (SERCA)2a, a critical regulator of calcium homeostasis, is known to be decreased in heart failure. Patients with myocarditis or dilated cardiomyopathy develop autoantibodies to SERCA2a suggesting that they may have pathogenetic significance. In this report, we describe epitope mapping analysis of SERCA2a in A/J mice that leads us to make five observations: 1) SERCA2a contains multiple T cell epitopes that induce varying degrees of myocarditis. One epitope, SERCA2a 971–990, induces widespread atrial inflammation without affecting noncardiac tissues; the cardiac abnormalities could be noninvasively captured by echocardiography, electrocardiography, and magnetic resonance microscopy imaging. 2) SERCA2a 971–990-induced disease was associated with the induction of CD4 T cell responses and the epitope preferentially binds MHC class II/IAk rather than IEk. By creating IAk/and IEk/SERCA2a 971–990 dextramers, the T cell responses were determined by flow cytometry to be Ag specific. 3) SERCA2a 971–990-sensitized T cells produce both Th1 and Th17 cytokines. 4) Animals immunized with SERCA2a 971–990 showed Ag-specific Abs with enhanced production of IgG2a and IgG2b isotypes, suggesting that SERCA2a 971–990 can potentially act as a common epitope for both T cells and B cells. 5) Finally, SERCA2a 971–990-sensitized T cells were able to transfer disease to naive recipients. Together, these data indicate that SERCA2a is a critical autoantigen in the mediation of atrial inflammation in mice and that our model may be helpful to study the inflammatory events that underlie the development of conditions such as atrial fibrillation in humans.
- Published
- 2018
27. Stem Cell-Derived Exosomes, Autophagy, Extracellular Matrix Turnover, and miRNAs in Cardiac Regeneration during Stem Cell Therapy
- Author
-
Paras Kumar Mishra, Priyanka Prathipati, and Shyam Sundar Nandi
- Subjects
0301 basic medicine ,Cancer Research ,medicine.medical_treatment ,Cellular differentiation ,Cell- and Tissue-Based Therapy ,Cellular homeostasis ,Biology ,Exosomes ,Article ,Extracellular matrix ,03 medical and health sciences ,Paracrine signalling ,Autophagy ,medicine ,Humans ,Regeneration ,Myocardium ,Stem Cells ,Regeneration (biology) ,Heart ,Cell Biology ,Stem-cell therapy ,Microvesicles ,Extracellular Matrix ,Cell biology ,MicroRNAs ,030104 developmental biology ,Immunology ,Stem cell ,Stem Cell Transplantation - Abstract
Stem cell therapy (SCT) raises the hope for cardiac regeneration after ischemic heart disease. However, the molecular mechanisms underlying repair of dead myocardium in the ischemic heart is poorly understood. Growing evidences suggest that cardiac matrix stiffness and differential expressions of miRNAs play a crucial role in stem cell survival and differentiation. However, their roles on transplanted stem cells, for the myocardial repair of the ischemic heart, remain unclear. Transplanted stem cells may act in an autocrine and/or paracrine manner to regenerate the dead myocardium. Paracrine mediators such as stem cell-derived exosomes are emerging as a novel therapeutic strategy to overcome some of the limitations of SCT. These exosomes carry microRNAs (miRNAs) that may regulate stem cell differentiation into a specific lineage. MicroRNAs may also contribute to stiffness of surrounding matrix by regulating extracellular matrix (ECM) turnover. The survival of transplanted stem cell depends on its autophagic process that maintains cellular homeostasis. Therefore, exosomes, miRNAs, extracellular matrix turnover, and autophagy may have an integral role in improving the efficacy of SCT. This review elaborates the specific roles of these regulatory components on cardiac regeneration in ischemic hearts during SCT.
- Published
- 2016
28. Abstract 088: Overexpression of Neuronal Nitric Oxide Synthase Associated Protein in the Paraventricular Nucleus Contributes to Increased Sympathoexcitation in Male Mice by Modulating Subcellular Localization of Neuronal Nitric Oxide Synthase
- Author
-
Neeru M. Sharma, Shyam Sundar Nandi, Kaushik P. Patel, Kenichi Katsurada, and Lie Gao
- Subjects
medicine.anatomical_structure ,Chemistry ,Internal Medicine ,medicine ,Male mice ,Subcellular localization ,Neuronal Nitric Oxide Synthase ,Nucleus ,Cell biology - Abstract
Nitric oxide (NO•) bioavailability within the paraventricular nucleus (PVN) plays a pivotal role in the pathogenesis of many cardiovascular diseases including hypertension. Aligning neuronal nitric oxide synthase (NOS1) with N -methyl-D-aspartate (NMDA) receptor by tethering to PSD95 (postsynaptic density 95 protein) allows the production of NO• initiated by an influx of Ca 2+ in neurons within the PVN. NOS1 associated protein (Nos1ap) competes with PSD95 for NOS1 binding and thus influences the production of NO•. We investigated the effect of specifically overexpressing Nos1ap in the PVN on sympathetic outflow using cre recombinase-conditional Nos1ap over-expression transgenic(Tg) mice injected with lentivirus expressing Cre-GFP or GFP into the PVN. Two weeks of post-viral administration, the sympathetic outflow was evaluated in both male and female mice. Male-Cre-Nos1ap mice displayed a significantly elevated urinary excretion of norepinephrine, an index of overall sympathetic outflow compared to GFP-Nos1ap (0.64±0.11* vs. 0.17±0.03 ug/day) while female littermates showed no difference (0.44±0.06 vs. 0.36±0.03 ug/day). Basal renal sympathetic nerve activity assessed as a percent of maximal activity was elevated 1.5-fold in Cre-Nos1ap male mice (46.1 ± 3.6* vs.31.0 ± 3.2) while there was no significant difference in Cre-Nos1ap female mice(22.6± 1.2 vs. 24.8 ± 2.6) compared to respective GFP controls. Interestingly, female mice had higher levels of basal NOS1 in the PVN compared to male littermates (0.79 ±0.01* vs. 0.37±0.05). Further, a robust overexpression of Nos1ap in the PVN of Cre-Nos1ap (0.49±0.04* vs. 0.21±0.03 male; 0.53±0.08* vs. 0.28±0.04 female) compared to GFP-Nos1ap mice does not lead to any change in NOS1, NMDA receptor(NR)2A, NR2B, while NR1 expression tended to increase in both sexes. Overexpression of Nos1ap in NG108 neurons, in vitro, led to relocalization of 46% of membrane-associated NOS1 to the cytosolic compartment, suggesting that the decrease in NMDA receptor-mediated NOS1 activity leads to reduced NO• production. Taken together, our data indicate that overexpression of Nos1ap within the PVN reduces the levels of NO• generated by NOS1 leading to increased sympathetic outflow in male mice but not in female mice.
- Published
- 2019
29. Role of the Neurogenic Signaling on Cardiac miR‐18‐5p/HIF‐1α Axis to Enhance Mitochondrial Abnormality in Neurogenic Hypertension
- Author
-
Neeru M. Sharma, Shyam Sundar Nandi, Kenichi Katsurada, Andrea Siqueira Haibara, Sushil K. Mahata, and Kaushik P. Patel
- Subjects
business.industry ,Genetics ,Cancer research ,Medicine ,Neurogenic hypertension ,Abnormality ,business ,Molecular Biology ,Biochemistry ,Biotechnology - Published
- 2019
30. Role of the Renal Nerves in Regulating SGLT2 inhibitor‐induced Diuresis and Natriuresis in rats with Heart Failure
- Author
-
Shyam Sundar Nandi, Kenichi Katsurada, Kaushik P. Patel, and Neeru M. Sharma
- Subjects
medicine.medical_specialty ,business.industry ,medicine.disease ,Biochemistry ,Natriuresis ,Endocrinology ,Internal medicine ,Heart failure ,Induced diuresis ,Genetics ,medicine ,SGLT2 Inhibitor ,business ,Molecular Biology ,Biotechnology - Published
- 2020
31. Neuronal Nitric Oxide Synthase Associated Protein: Nos1ap mediates Sympathoexcitation through Paraventricular Nucleus of the Hypothalamus
- Author
-
Lie Gao, Neeru M. Sharma, Shyam Sundar Nandi, Kenichi Katsurada, and Kaushik P. Patel
- Subjects
medicine.anatomical_structure ,Hypothalamus ,Chemistry ,NOS1AP ,Genetics ,medicine ,Molecular Biology ,Biochemistry ,Nucleus ,Neuronal Nitric Oxide Synthase ,Biotechnology ,Cell biology - Published
- 2020
32. Cardiac‐specific Overexpression of MiR‐133a in the Diabetic Heart Mitigates Mitochondrial Abnormality by Targeting TIM17A
- Author
-
Shyam Sundar Nandi and Paras K Mishra
- Subjects
business.industry ,Genetics ,Cancer research ,Medicine ,Mir 133a ,Abnormality ,Diabetic heart ,business ,Molecular Biology ,Biochemistry ,Biotechnology - Published
- 2018
33. Targeting miRNA for Therapy of Juvenile and Adult Diabetic Cardiomyopathy
- Author
-
Paras Kumar Mishra and Shyam Sundar Nandi
- Subjects
Adult ,0301 basic medicine ,medicine.medical_specialty ,endocrine system diseases ,Diabetic Cardiomyopathies ,medicine.medical_treatment ,Cardiomyopathy ,Exosomes ,Exosome ,Article ,03 medical and health sciences ,Insulin resistance ,Internal medicine ,Diabetic cardiomyopathy ,Diabetes mellitus ,Humans ,Medicine ,Myocytes, Cardiac ,Molecular Targeted Therapy ,Child ,Inflammation ,biology ,business.industry ,Myocardium ,Insulin ,medicine.disease ,Fibrosis ,MicroRNAs ,Oxidative Stress ,Insulin receptor ,Diabetes Mellitus, Type 1 ,030104 developmental biology ,Endocrinology ,Diabetes Mellitus, Type 2 ,Gene Expression Regulation ,biology.protein ,RNA Interference ,Insulin Resistance ,Beta cell ,business - Abstract
Prevalence of diabetes mellitus (DM), a multifactorial disease often diagnosed with high blood glucose levels, is rapidly increasing in the world. Association of DM with multi-organ dysfunction including cardiomyopathy makes it a leading cause of morbidity and mortality. There are two major types of DM: type 1 DM (T1D) and type 2 DM (T2D). T1D is diagnosed by reduced levels of insulin and high levels of glucose in the blood. It is caused due to pancreatic beta cell destruction/loss, and mostly found in juveniles (juvenile DM). T2D is diagnosed by increased levels of insulin and glucose in the blood. It is caused due to insulin receptor dysfunction, and mostly found in the adults (adult DM). Both T1D and T2D impair cardiac muscle function, which is referred to as diabetic cardiomyopathy. We and others have reported that miRNAs, a novel class of tiny non-coding regulatory RNAs, are differentially expressed in the diabetic heart and they contribute to diabetic cardiomyopathy. Here, we elaborated the biogenesis of miRNA, how miRNA regulates a gene, cardioprotective roles of different miRNAs including miRNAs present in exosomes, underlying molecular mechanisms by which miRNA ameliorates diabetic cardiomyopathy, and the role of miRNA as a potential therapeutic target for juvenile and adult diabetic cardiomyopathy.
- Published
- 2018
34. Hydrogen sulfide mitigates homocysteine-mediated pathological remodeling by inducing miR-133a in cardiomyocytes
- Author
-
Paras Kumar Mishra, Hamid R. Shahshahan, Surender Kumar Sharawat, Shyam Sundar Nandi, and Varun Kesherwani
- Subjects
Hyperhomocysteinemia ,medicine.medical_specialty ,Homocysteine ,Clinical Biochemistry ,Biology ,Article ,Muscle hypertrophy ,Mice ,chemistry.chemical_compound ,Atrial natriuretic peptide ,Internal medicine ,medicine ,Animals ,Humans ,Myocyte ,Myocytes, Cardiac ,MEF2C ,Hydrogen Sulfide ,Ventricular remodeling ,Molecular Biology ,Ventricular Remodeling ,MEF2 Transcription Factors ,Cell Biology ,General Medicine ,equipment and supplies ,medicine.disease ,HDAC1 ,MicroRNAs ,Endocrinology ,Gene Expression Regulation ,chemistry - Abstract
An elevated level of homocysteine called hyperhomocysteinemia (HHcy) is associated with pathological cardiac remodeling. Hydrogen sulfide (H2S) acts as a cardioprotective gas; however, the mechanism by which H2S mitigates homocysteine-mediated pathological remodeling in cardiomyocytes is unclear. We hypothesized that H2S ameliorates HHcy-mediated hypertrophy by inducing cardioprotective miR-133a in cardiomyocytes. To test the hypothesis, HL1 cardiomyocytes were treated with (1) plain medium (control, CT), (2) 100 µM of homocysteine (Hcy), (3) Hcy with 30 µM of H2S (Hcy + H2S), and (4) H2S for 24 h. The levels of hypertrophy markers: c-fos, atrial natriuretic peptide (ANP), and beta-myosin heavy chain (β-MHC), miR-133a, and its transcriptional inducer myosin enhancer factor-2C (MEF2C) were determined by Western blotting, RT-qPCR, and immunofluorescence. The activity of MEF2C was assessed by co-immunoprecipitation of MEF2C with histone deacetylase-1(HDAC1). Our results show that H2S ameliorates homocysteine-mediated up-regulation of c-fos, ANP, and β-MHC, and down-regulation of MEF2C and miR-133a. HHcy induces the binding of MEF2C with HDAC1, whereas H2S releases MEF2C from MEF2C-HDAC1 complex causing activation of MEF2C. These findings elicit that HHcy induces cardiac hypertrophy by promoting MEF2C-HDAC1 complex formation that inactivates MEF2C causing suppression of anti-hypertrophy miR-133a in cardiomyocytes. H2S mitigates hypertrophy by inducing miR-133a through activation of MEF2C in HHcy cardiomyocytes. To our knowledge, this is a novel mechanism of H2S-mediated activation of MEF2C and induction of miR-133a and inhibition of hypertrophy in HHcy cardiomyocytes.
- Published
- 2015
35. H2S and homocysteine control a novel feedback regulation of cystathionine beta synthase and cystathionine gamma lyase in cardiomyocytes
- Author
-
Paras Kumar Mishra and Shyam Sundar Nandi
- Subjects
Male ,0301 basic medicine ,Homocysteine ,030204 cardiovascular system & hematology ,Mice ,chemistry.chemical_compound ,0302 clinical medicine ,Myocyte ,Myocytes, Cardiac ,Inducer ,Hydrogen Sulfide ,3' Untranslated Regions ,Feedback, Physiological ,Mice, Knockout ,chemistry.chemical_classification ,Regulation of gene expression ,Multidisciplinary ,biology ,Cystathionine gamma-lyase ,3. Good health ,Medicine ,Female ,RNA Interference ,inorganic chemicals ,congenital, hereditary, and neonatal diseases and abnormalities ,Hyperhomocysteinemia ,medicine.medical_specialty ,Science ,Cystathionine beta-Synthase ,Models, Biological ,Article ,03 medical and health sciences ,Cystathionine ,Internal medicine ,parasitic diseases ,medicine ,Animals ,Dose-Response Relationship, Drug ,organic chemicals ,Cystathionine gamma-Lyase ,nutritional and metabolic diseases ,medicine.disease ,Cystathionine beta synthase ,MicroRNAs ,030104 developmental biology ,Enzyme ,Endocrinology ,Gene Expression Regulation ,chemistry ,biology.protein - Abstract
Hydrogen sulfide (H2S), a cardioprotective gas, is endogenously produced from homocysteine by cystathionine beta synthase (CBS) and cystathionine gamma lyase (CSE) enzymes. However, effect of H2S or homocysteine on CBS and CSE expression, and cross-talk between CBS and CSE are unclear. We hypothesize that homocysteine and H2S regulate CBS and CSE expressions in a dose dependent manner in cardiomyocytes, and CBS deficiency induces cardiac CSE expression. To test the hypothesis, we treated murine atrial HL1 cardiomyocytes with increasing doses of homocysteine or Na2S/GYY4137, a H2S donor, and measured the levels of CBS and CSE. We found that homocysteine upregulates CSE but downregulates CBS whereas Na2S/GYY4137 downregulates CSE but upregulates CBS in a dose-dependent manner. Moreover, the Na2S-treatment downregulates specificity protein-1 (SP1), an inducer for CSE, and upregulates miR-133a that targets SP1 and inhibits cardiomyocytes hypertrophy. Conversely, in the homocysteine-treated cardiomyocytes, CBS and miR-133a were downregulated and hypertrophy was induced. In vivo studies using CBS+/−, a model for hyperhomocysteinemia, and sibling CBS+/+ control mice revealed that deficiency of CBS upregulates cardiac CSE, plausibly by inducing SP1. In conclusion, we revealed a novel mechanism for H2S-mediated regulation of homocysteine metabolism in cardiomyocytes, and a negative feedback regulation between CBS and CSE in the heart.
- Published
- 2017
36. A novel role for miR-133a in centrally mediated activation of the renin-angiotensin system in congestive heart failure
- Author
-
Hong Zheng, Paras Kumar Mishra, Kaushik P. Patel, Neeru M. Sharma, and Shyam Sundar Nandi
- Subjects
0301 basic medicine ,Male ,medicine.medical_specialty ,Physiology ,Angiotensinogen ,Kidney ,Losartan ,Receptor, Angiotensin, Type 1 ,Rats, Sprague-Dawley ,Renin-Angiotensin System ,03 medical and health sciences ,Downregulation and upregulation ,Physiology (medical) ,Internal medicine ,Cell Line, Tumor ,Renin–angiotensin system ,microRNA ,parasitic diseases ,medicine ,Animals ,3' Untranslated Regions ,Heart Failure ,business.industry ,medicine.disease ,Angiotensin II ,Rats ,MicroRNAs ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,Hypothalamus ,Heart failure ,Mir 133a ,Cardiology and Cardiovascular Medicine ,business ,Nucleus ,Angiotensin II Type 1 Receptor Blockers ,hormones, hormone substitutes, and hormone antagonists ,Paraventricular Hypothalamic Nucleus ,Research Article - Abstract
An activated renin-angiotensin system (RAS) within the central nervous system has been implicated in sympathoexcitation during various disease conditions including congestive heart failure (CHF). In particular, activation of the RAS in the paraventricular nucleus (PVN) of the hypothalamus has been recognized to augment sympathoexcitation in CHF. We observed a 2.6-fold increase in angiotensinogen (AGT) in the PVN of CHF. To elucidate the molecular mechanism for increased expression of AGT, we performed in silico analysis of the 3′-untranslated region (3′-UTR) of AGT and found a potential binding site for microRNA (miR)-133a. We hypothesized that decreased miR-133a might contribute to increased AGT in the PVN of CHF rats. Overexpression of miR-133a in NG108 cells resulted in 1.4- and 1.5-fold decreases in AGT and angiotensin type II (ANG II) type 1 receptor (AT1R) mRNA levels, respectively. A luciferase reporter assay performed on NG108 cells confirmed miR-133a binding to the 3′-UTR of AGT. Consistent with these in vitro data, we observed a 1.9-fold decrease in miR-133a expression with a concomitant increase in AGT and AT1R expression within the PVN of CHF rats. Furthermore, restoring the levels of miR-133a within the PVN of CHF rats with viral transduction resulted in a significant reduction of AGT (1.4-fold) and AT1R (1.5-fold) levels with a concomitant decrease in basal renal sympathetic nerve activity (RSNA). Restoration of miR-133a also abrogated the enhanced RSNA responses to microinjected ANG II within the PVN of CHF rats. These results reveal a novel and potentially unique role for miR-133a in the regulation of ANG II within the PVN of CHF rats, which may potentially contribute to the commonly observed sympathoexcitation in CHF.NEW & NOTEWORTHY Angiotensinogen (AGT) expression is upregulated in the paraventricular nucleus of the hypothalamus through posttranscriptional mechanism interceded by microRNA-133a in heart failure. Understanding the mechanism of increased expression of AGT in pathological conditions leading to increased sympathoexcitation may provide the basis for the possible development of new therapeutic agents with enhanced specificity.
- Published
- 2016
37. Lack of miR-133a Decreases Contractility of Diabetic Hearts: A Role for Novel Cross Talk Between Tyrosine Aminotransferase and Tyrosine Hydroxylase
- Author
-
Neeru M. Sharma, Shyam Sundar Nandi, Paras Kumar Mishra, Hong Zheng, Kaushik P. Patel, and Hamid R. Shahshahan
- Subjects
0301 basic medicine ,Male ,medicine.medical_specialty ,Tyrosine 3-Monooxygenase ,Endocrinology, Diabetes and Metabolism ,Blotting, Western ,Mice, Transgenic ,Biology ,Pathophysiology ,Diabetes Mellitus, Experimental ,Contractility ,Rats, Sprague-Dawley ,03 medical and health sciences ,Norepinephrine ,Tyrosine aminotransferase ,Downregulation and upregulation ,Internal medicine ,Receptors, Adrenergic, beta ,Internal Medicine ,medicine ,Animals ,Humans ,Myocytes, Cardiac ,Tyrosine ,Receptor ,Tyrosine Transaminase ,Tyrosine hydroxylase ,Reverse Transcriptase Polymerase Chain Reaction ,Myocardium ,Hemodynamics ,Streptozotocin ,Immunohistochemistry ,Myocardial Contraction ,Rats ,MicroRNAs ,030104 developmental biology ,Endocrinology ,HEK293 Cells ,Catecholamine ,medicine.drug - Abstract
MicroRNAs (miRNAs) have a fundamental role in diabetic heart failure. The cardioprotective miRNA-133a (miR-133a) is downregulated, and contractility is decreased in diabetic hearts. Norepinephrine (NE) is a key catecholamine that stimulates contractility by activating β-adrenergic receptors (β-AR). NE is synthesized from tyrosine by the rate-limiting enzyme, tyrosine hydroxylase (TH), and tyrosine is catabolized by tyrosine aminotransferase (TAT). However, the cross talk/link between TAT and TH in the heart is unclear. To determine whether miR-133a plays a role in the cross talk between TH and TAT and regulates contractility by influencing NE biosynthesis and/or β-AR levels in diabetic hearts, Sprague-Dawley rats and miR-133a transgenic (miR-133aTg) mice were injected with streptozotocin to induce diabetes. The diabetic rats were then treated with miR-133a mimic or scrambled miRNA. Our results revealed that miR-133a mimic treatment improved the contractility of the diabetic rat’s heart concomitant with upregulation of TH, cardiac NE, β-AR, and downregulation of TAT and plasma levels of NE. In miR-133aTg mice, cardiac-specific miR-133a overexpression prevented upregulation of TAT and suppression of TH in the heart after streptozotocin was administered. Moreover, miR-133a overexpression in CATH.a neuronal cells suppressed TAT with concomitant upregulation of TH, whereas knockdown and overexpression of TAT demonstrated that TAT inhibited TH. Luciferase reporter assay confirmed that miR-133a targets TAT. In conclusion, miR-133a controls the contractility of diabetic hearts by targeting TAT, regulating NE biosynthesis, and consequently, β-AR and cardiac function.
- Published
- 2016
38. Involvement of Pitx2, a Homeodomain Transcription Factor, in Hypothyroidism Associated Reproductive Disorders
- Author
-
Sib Sankar Roy, Samir Kumar Saha, Pamela Ghosh, Shyam Sundar Nandi, and Samir Bhattacharya
- Subjects
Homeodomain Proteins ,Genetics ,Granulosa Cells ,PITX2 ,Physiology ,Granulosa cell ,Gonadal Disorders ,Estrous Cycle ,Biology ,Models, Biological ,Rats ,Protein Transport ,Gene Expression Regulation ,Hypothyroidism ,PITX2 Gene ,Immunology ,Molecular mechanism ,Animals ,Paired Box Transcription Factors ,Triiodothyronine ,Ovarian dysfunction ,Homeobox ,Female ,Transcription factor ,Transcription Factors - Abstract
Hypothyroid-associated reproductive disorders have now become a striking phenomenon worldwide but the molecular mechanism behind these disorders is not fully known. Pitx2 gene encodes homeodomain transcription factor, which regulates Plod2 gene in brain tissue, transactivates gonadotropin genes in pituitary and plays a substantial role in cell growth and proliferation in different tissues. Pitx2 binds to Plod2 promoter and activates this gene in rat ovary. In this report, we show that Pitx2's expression is markedly reduced in hypothyroid ovary as well as in ovarian granulosa cells, which is recovered with T(3)-supplementation both in vitro and in vivo conditions. Reduced Pitx2 expression could decrease Plod2 expression and hence facilitate ovarian ECM degradation. We have also observed similar pattern of expression of Pitx1 and -3 in hypothyroid and T(3)-supplemented ovaries. The temporal expression of Pitx2 across the estrous cycle shows that it is expressed through all the 4 phases of the cycle and reaches its maximum in the proestrus phase suggesting its possible role in ovulation followed by luteinization. The present study reveals that the reduced Pitx2 expression in hypothyroid ovary could lead to ovarian dysfunction by modulating the Pitx2-Plod2 interaction, further study will be necessary to unravel the complete regulatory mechanism of Pitx2 in ovarian function.
- Published
- 2007
39. MiR‐133a Mitigates Mitophagy in Ins2 +/‐ Diabetic Heart
- Author
-
Shyam Sundar Nandi and Paras Kumar Mishra
- Subjects
endocrine system ,Chemistry ,Depolarization ,Mitochondrion ,Biochemistry ,Molecular biology ,Parkin ,Blot ,Downregulation and upregulation ,In vivo ,Mitophagy ,Genetics ,Mitochondrial fission ,Molecular Biology ,Biotechnology - Abstract
To test the hypothesis that inhibition of miR-133a induces mitophagy in diabetic hearts and it is ameliorated by overexpression of miR-133a, we used four in vivo groups: C57BL/6J, Akita (T1D), Akita+miR-133a, and Akita+scramble; and four in vitro groups of HL1 cardiomyocytes: low (LG-5mM), high (HG-25mM) doses of D-glucose, HG+miR-133a, and HG+scramble; and determined mitoSOX generation, depolarization of damaged mitochondria (ΔΨ, by JC-1), mitochondrial fission (by DRP1 and Parkin), autophagosome formation (by Beclin1, ATG14, ATG3, LC3B) and its lysosomal degradation (by RFP-GFP-LC3B) using qPCR, Western blotting, immunofluorescence, and transmission electron microscopy (TEM). Our results demonstrate that in HG cardiomyocytes, mitoSOX generation, depolarization of mt-ΔΨ and DRP1 protein are elevated by 2.5±0.3, 1.44±0.08 and 1.94±0.29 folds, respectively, but ameliorated (mitoSOX:4.9±0.04, mt-ΔΨ:2.11±0.08, and DRP1:0.85±0.17 fold down regulation) by overexpression of miR-133a. The levels of Beclin1, ATG1...
- Published
- 2015
40. Reduced miR‐133a Results in Upregulation of Angiotensinogen in the Paraventricular Nucleus of Rats with Chronic Heart Failure
- Author
-
Shyam Sundar Nandi, Neeru M. Sharma, Paras Kumar Mishra, Hong Zheng, and Kaushik P. Patel
- Subjects
Untranslated region ,medicine.medical_specialty ,Chemistry ,medicine.disease ,Biochemistry ,Endocrinology ,Downregulation and upregulation ,Cell culture ,Hypothalamus ,Neuroblastoma ,Internal medicine ,Glioma ,Heart failure ,parasitic diseases ,microRNA ,Genetics ,medicine ,Molecular Biology ,hormones, hormone substitutes, and hormone antagonists ,Biotechnology - Abstract
Activation of renin-angiotensin system (RAS) within the paraventricular nucleus (PVN) of the hypothalamus has been suggested to enhance sympathetic activation in chronic heart failure (CHF) condition. The contribution due to changes in angiotensinogen (AGT) per se remains to be examined. In this study, we observed a 12-fold increase in levels of AGT gene in the PVN of CHF rats compared to sham-operated controls. In order to elucidate the molecular mechanism for increased expression of AGT, we performed in silico analysis of 3'-UTR of AGT gene and found potential binding site for microRNA (miR)-133a. We found a 1.9-fold decrease in mir-133a expression in the PVN of rats with CHF. We hypothesized that decreased expression of mir-133a may be responsible for the increased expression of AGT in the PVN. To test this hypothesis we used NG108 (neuroblastoma x glioma) cell line, which endogenously expresses RAS, as our in vitro model. Overexpression of miR133a (pEZX-MR03-miR133a) in NG108 cells resulted in 1.4 and...
- Published
- 2015
41. Induction of autophagy markers is associated with attenuation of miR-133a in diabetic heart failure patients undergoing mechanical unloading
- Author
-
Shyam Sundar, Nandi, Michael J, Duryee, Hamid R, Shahshahan, Geoffrey M, Thiele, Daniel R, Anderson, and Paras K, Mishra
- Subjects
Original Article - Abstract
Autophagy is ubiquitous in all forms of heart failure and cardioprotective miR-133a is attenuated in human heart failure. Previous reports from heart failure patients undergoing left ventricular assist device (LVAD) implantation demonstrated that autophagy is upregulated in the LV of the failing human heart. Studies in the murine model show that diabetes downregulates miR-133a. However, the role of miR-133a in the regulation of autophagy in diabetic hearts is unclear. We tested the hypothesis that diabetes exacerbates cardiac autophagy by inhibiting miR-133a in heart failure patients undergoing LVAD implantation. The miRNA assay was performed on the LV of 15 diabetic (D) and 6 non-diabetic (ND) heart failure patients undergoing LVAD implantation. Four ND with highly upregulated and 5 D with highly downregulated miR-133a were analyzed for autophagy markers (Beclin1, LC3B, ATG3) and their upstream regulators (mTOR and AMPK), and hypertrophy marker (beta-myosin heavy chain) by RT-qPCR, Western blotting and immunofluorescence. Our results demonstrate that attenuation of miR-133a in diabetic hearts is associated with the induction of autophagy and hypertrophy, and suppression of mTOR without appreciable difference in AMPK activity. In conclusion, attenuation of miR-133a contributes to the exacerbation of diabetes mediated cardiac autophagy and hypertrophy in heart failure patients undergoing LVAD implantation.
- Published
- 2015
42. Prion Disease: A Deadly Disease for Protein Misfolding
- Author
-
Snehasis Jana, Shyam Sundar Nandi, and Chiranjib Chakraborty
- Subjects
Fatal familial insomnia ,Protein Folding ,Transmissible mink encephalopathy ,Prions ,animal diseases ,Bovine spongiform encephalopathy ,Brain ,Pharmaceutical Science ,Scrapie ,Disease ,Chronic wasting disease ,Biology ,medicine.disease ,Virology ,Prion Diseases ,nervous system diseases ,Structure-Activity Relationship ,Amyloid disease ,Kuru ,medicine ,Animals ,Humans ,Biotechnology - Abstract
An infectious particle, termed prion, composed largely and perhaps solely of a single protein, is the likely causative agent of prion disease. It produces lethal decline of cognitive and motor function. The responsible protein arrives at a pathogenic state by misfolding from a normal form that has ubiquitous tissue distribution. Prion diseases are often called spongiform encephalopathies. Probably most mammalian species develop these diseases. Specific examples in various animals are -Scrapie, Transmissible Mink Encephalopathy (TME ), Chronic Wasting Disease(CWD) and bovine spongiform encephalopathy (BSE). Humans are also susceptible to several prion diseases: Creutzfeld-Jacob Disease (CJD), Gerstmann-Straussler-Scheinker Syndrome (GSS), Fatal Familial Insomnia (FFI), Kuru and Alpers Syndrome. This paper reviews transmission of this diseases, protein involvement, nature of protein, the conversion process from PrP(c) to PrP(Sc), conversion of prion protein in vitro, the different proposed models for the conversion of PrP(c) to PrP(Sc), prion and other amyloid diseases, prion strains, structure of PrP(c) the particular process that may induce prion disease, and immunization against these diseases.
- Published
- 2005
43. Overexpression and Purification of Recombinant Eel Calcitonin and Its Phylogenetic Analysis
- Author
-
Shyam Sundar Nandi, Surajit Sinha, Biplab Sarkar, and Chiranjib Chakraborty
- Subjects
Calcitonin ,medicine.medical_specialty ,Calcitonin Gene-Related Peptide ,Molecular Sequence Data ,Osteoporosis ,Biology ,Peptide hormone ,Biochemistry ,law.invention ,Structural Biology ,law ,Internal medicine ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Phylogeny ,chemistry.chemical_classification ,Eels ,Sequence Homology, Amino Acid ,Thyroid ,Biological activity ,General Medicine ,biology.organism_classification ,medicine.disease ,Recombinant Proteins ,Streptomyces ,Amino acid ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Recombinant DNA ,Sequence Alignment ,Streptomyces avermitilis - Abstract
Calcitonin (CT), a peptide hormone that is widely used for the treatment of osteoporosis, Paget's disease, hypercalcemic shock and chronic pain in terminal cancer patients, is produced by the para-follicular cells of the thyroid gland in mammals and by the ultimobranchial gland of birds and fish. Fish calcitonin, like eel calcitonin (eCT), is more potent and longer lasting than human CT and is one of the many bioactive peptides that require C-terminal amidation for full biological activity. In this study we describe the over-expression and over-production of C-terminal amidated eCT in recombinant Streptomyces avermitilis. A phylogenetic analysis was performed with all the known CT amino acid sequences.
- Published
- 2005
44. Overexpression, Purification and Characterization of Recombinant Salmon Calcitonin, A Therapeutic Protein, in Streptomyces Avermitilis
- Author
-
Surajit Sinha, Chiranjib Chakraborty, and Shyam Sundar Nandi
- Subjects
Calcitonin ,medicine.medical_specialty ,DNA, Complementary ,Molecular Sequence Data ,Osteoporosis ,Thyroid Gland ,Pain ,Biology ,Pharmacology ,Peptide hormone ,Biochemistry ,law.invention ,chemistry.chemical_compound ,Structural Biology ,law ,Internal medicine ,medicine ,Peptide synthesis ,Animals ,Amino Acid Sequence ,Analgesics ,Thyroid ,Biological activity ,General Medicine ,medicine.disease ,biology.organism_classification ,Recombinant Proteins ,Streptomyces ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Recombinant DNA ,Protein Processing, Post-Translational ,Sequence Alignment ,Streptomyces avermitilis ,Plasmids - Abstract
Calcitonin (CT) is a peptide hormone produced by the parafollicular cells of the thyroid gland in mammals and by the ultimobranchial gland of birds and fish. Salmon calcitonin (sCT), which is more potent and longer lasting than human CT, has been used widely for the treatment of osteoporosis, paget's disease, hypercalcemic shock and chronic pain in terminal cancer patients. sCT is one of the many bioactive peptides that require C-terminal amidation for full biological activity. In this study we describe the over-expression and over-production of C-terminal amidated sCT in recombinant Streptomyces avermitilis. With this approach the utilization of expensive peptide synthesis can be circumvented.
- Published
- 2004
45. Mdivi‐1 mitigates cardiac dysfunction by attenuating mitophagy in diabetes (1155.3)
- Author
-
Vishalakshi Chavali, Shyam Sundar Nandi, and Paras K Mishra
- Subjects
medicine.medical_specialty ,Mechanism (biology) ,business.industry ,Autophagy ,medicine.disease ,Biochemistry ,Cardiac dysfunction ,Internal medicine ,Diabetes mellitus ,Heart failure ,Mitophagy ,Genetics ,medicine ,Cardiology ,business ,Molecular Biology ,Biotechnology - Abstract
Although Mdivi-1 ameliorates heart failure by inhibiting autophagy, the mechanism of Mdivi-1 mediated mitigation of cardiac dysfunction in diabetes is unclear. We hypothesize that Mdivi-1 improves ...
- Published
- 2014
46. miR‐133a alleviates cardiac autophagy by targeting AMPK in Ins2+/‐ diabetic mice (868.3)
- Author
-
Paras Kumar Mishra and Shyam Sundar Nandi
- Subjects
microRNA ,Autophagy ,Genetics ,AMPK ,Mir 133a ,Diabetic mouse ,Biology ,Molecular Biology ,Biochemistry ,Function (biology) ,Biotechnology ,Cell biology - Abstract
Optimum level of autophagy is indispensable for survival and cellular function, and miRNA tightly regulates biological functions. In diabetic hearts, miR-133a (anti-hypertrophy and anti-fibrosis) i...
- Published
- 2014
47. miR‐133a ameliorates cardiac dysfunction in diabetes: possibly by restoring β‐adrenergic receptor function and expression (1078.6)
- Author
-
Kaushik P. Patel, Shyam Sundar Nandi, Paras Kumar Mishra, Hong Zheng, Vishalakshi Chavali, Xuefei Liu, and Neeru M. Sharma
- Subjects
medicine.medical_specialty ,business.industry ,medicine.disease ,Biochemistry ,Cardiac dysfunction ,Endocrinology ,Fibrosis ,Internal medicine ,Diabetes mellitus ,Cardiac hypertrophy ,microRNA ,cardiovascular system ,Genetics ,Medicine ,Mir 133a ,β adrenergic receptor ,business ,Molecular Biology ,Function (biology) ,Biotechnology - Abstract
Inhibition of microRNA (miR)-133a causes cardiac hypertrophy and fibrosis. However, the beneficial effects of miR-133a in cardiac dysfunction remain to be examined. Furthermore, impaired contractil...
- Published
- 2014
48. Generating double knockout mice to model genetic intervention for diabetic cardiomyopathy in Humans
- Author
-
Vishalakshi Chavali, Shree Ram Singh, Shyam Sundar Nandi, and Paras Kumar Mishra
- Subjects
Male ,medicine.medical_specialty ,endocrine system ,endocrine system diseases ,Genotyping Techniques ,Cardiac fibrosis ,Diabetic Cardiomyopathies ,Wheat Germ Agglutinins ,Transgene ,Blotting, Western ,MMP9 ,Polymerase Chain Reaction ,Article ,Muscle hypertrophy ,Gene Knockout Techniques ,Mice ,Internal medicine ,Diabetic cardiomyopathy ,Diabetes mellitus ,Genetic model ,medicine ,Animals ,Humans ,Insulin ,Electrophoresis, Agar Gel ,Mice, Knockout ,business.industry ,Cell Membrane ,DNA ,Genetic Therapy ,Hypertrophy ,medicine.disease ,body regions ,Endocrinology ,Matrix Metalloproteinase 9 ,Heart failure ,Mutation ,Hybridization, Genetic ,Female ,business - Abstract
Diabetes is a rapidly increasing disease that enhances the chances of heart failure twofold to fourfold (as compared to age and sex matched nondiabetics) and becomes a leading cause of morbidity and mortality. There are two broad classifications of diabetes: type1 diabetes (T1D) and type2 diabetes (T2D). Several mice models mimic both T1D and T2D in humans. However, the genetic intervention to ameliorate diabetic cardiomyopathy in these mice often requires creating double knockout (DKO). In order to assess the therapeutic potential of a gene, that specific gene is either overexpressed (transgenic expression) or abrogated (knockout) in the diabetic mice. If the genetic mice model for diabetes is used, it is necessary to create DKO with transgenic/knockout of the target gene to investigate the specific role of that gene in pathological cardiac remodeling in diabetics. One of the important genes involved in extracellular matrix (ECM) remodeling in diabetes is matrix metalloproteinase-9 (Mmp9). Mmp9 is a collagenase that remains latent in healthy hearts but induced in diabetic hearts. Activated Mmp9 degrades extracellular matrix (ECM) and increases matrix turnover causing cardiac fibrosis that leads to heart failure. Insulin2 mutant (Ins2+/-) Akita is a genetic model for T1D that becomes diabetic spontaneously at the age of 3-4 weeks and show robust hyperglycemia at the age of 10-12 weeks. It is a chronic model of T1D. In Ins2+/- Akita, Mmp9 is induced. To investigate the specific role of Mmp9 in diabetic hearts, it is necessary to create diabetic mice where Mmp9 gene is deleted. Here, we describe the method to generate Ins2+/-/Mmp9-/- (DKO) mice to determine whether the abrogation of Mmp9 ameliorates diabetic cardiomyopathy.
- Published
- 2014
49. Assay for identification of heterozygous single-nucleotide polymorphism (Ala67Thr) in human poliovirus receptor gene
- Author
-
Jagadish M. Deshpande, Shyam Sundar Nandi, and Deepa Kailash Sharma
- Subjects
0301 basic medicine ,Heterozygote ,Virus genetics ,Genotype ,lcsh:Medicine ,poliovirus receptor ,SNP ,Single-nucleotide polymorphism ,Biology ,Molecular Inversion Probe ,Polymorphism, Single Nucleotide ,General Biochemistry, Genetics and Molecular Biology ,DNA sequencing ,03 medical and health sciences ,Humans ,Genetics ,Genetic polymorphism ,lcsh:R ,Sequence Analysis, DNA ,General Medicine ,030112 virology ,Molecular biology ,Genetic polymorphism - poliomyelitis - poliovirus receptor - PVR gene - SNP ,Poliovirus ,PVR gene ,Receptors, Virus ,Original Article ,Poliovirus Receptor ,Poliomyelitis ,SNP array - Abstract
Background & objectives : It is important to understand the role of cell surface receptors in susceptibility to infectious diseases. CD155 a member of the immunoglobulin super family, serves as the poliovirus receptor (PVR). Heterozygous (Ala67Thr) polymorphism in CD155 has been suggested as a risk factor for paralytic outcome of poliovirus infection. The present study pertains to the development of a screening test to detect the single nucleotide (SNP) polymorphism in the CD155 gene. Methods: New primers were designed for PCR, sequencing and SNP analysis of Exon2 of CD155 gene. DNAs extracted from either whole blood (n=75) or cells from oral cavity (n=75) were used for standardization and validation of the SNP assay. DNA sequencing was used as the gold standard method. Results: A new SNP assay for detection of heterozygous Ala67Thr genotype was developed and validated by testing 150 DNA samples. Heterozygous CD155 was detected in 27.33 per cent (41/150) of DNA samples tested by both SNP detection assay and sequencing. Interpretation & conclusions: The SNP detection assay was successfully developed for identification of Ala67Thr polymorphism in human PVR / CD155 gene. The SNP assay will be useful for large scale screening of DNA samples.
- Published
- 2016
50. Expression of PITX2 homeodomain transcription factor during rat gonadal development in a sexually dimorphic manner
- Author
-
Pamela Ghosh, Sib Sankar Roy, and Shyam Sundar Nandi
- Subjects
Male ,congenital, hereditary, and neonatal diseases and abnormalities ,endocrine system ,medicine.medical_specialty ,Gonad ,Physiology ,Somatic cell ,Molecular Sequence Data ,Embryonic Development ,Biology ,Rats, Sprague-Dawley ,Epitopes ,stomatognathic system ,Pregnancy ,Internal medicine ,Testis ,medicine ,Animals ,Protein Isoforms ,Amino Acid Sequence ,Gonads ,Homeodomain Proteins ,Sex Characteristics ,PITX2 ,Embryogenesis ,Ovary ,Embryo, Mammalian ,Embryonic stem cell ,Cell biology ,Rats ,stomatognathic diseases ,Endocrinology ,medicine.anatomical_structure ,Homeobox ,Female ,sense organs ,Development of the gonads ,Stem cell ,Transcription Factors - Abstract
PITX2, a multifunctional Paired-like homeodomain transcription factor, plays obligatory role during development of organs like heart, brain and pituitary. It regulates differentiation of vascular smooth muscle cells and hematopoietic stem cells. Although we earlier reported the Pitx2/PITX2 expression in gonad, but the expression pattern of its different isoforms in mammalian gonads especially during development is still not known. As PITX2 participates in the development of multiple organs and different homeobox genes have been shown to control gonadal functions, we wanted to investigate the role of PITX2 in gonadal development and its function. The objective of our study was to know the expression profile of different Pitx2/PITX2 isoforms and its localization throughout the development of gonads. Here we show the temporal and spatial expression pattern of Pitx2/PITX2 and its localization throughout the embryonic and postnatal stages of rat gonads. Pitx2/PITX2 expression profile reveals the differential and dimorphic expression pattern of its two isoforms PITX2B2 and-Cβ throughout the embryonic development stages and also in the postnatal stages, where it becomes more prominent. This is the first report where PITX2 homeodomain transcription factor shows isoform-specific sexually dimorphic expression. In addition, PITX2 localization was found in the embryonic ovarian primordial germ cell clusters and germ cells inside the testicular cords and also in somatic cells. In adults, ovarian granulosa and theca cells as well as germ cells inside the seminiferous tubules in testis express PITX2. All the evidences suggest that the differential expression of PITX2 might be associated with sex-specific embryonic and postnatal gonadal development and the physiological processes.
- Published
- 2011
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.