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1.
J Agric Food Chem ; 72(17): 9717-9734, 2024 May 01.
Article En | MEDLINE | ID: mdl-38624258

Plants have a history of being employed in managing breast cancer. However, no scientific evidence supports the idea that these plants can effectively reduce the level of HER2 expression. In this study, extracts from 10 medicinal plants were evaluated for their anticancer properties against HER2-positive breast cancer cells through various methods, including the SRB assay, comet assay, annexin V-FITC dual staining, and immunoblotting. All extracts exerted antiproliferative activity against HER2-positive breast cancer cells. Furthermore, Terminalia chebula (T. chebula), Berberis aristata (B. aristata), and Mucuna pruriens (M. pruriens) reduced HER2 expression in tested cell lines. In addition, an increased Bax/Bcl-2 ratio was observed after the treatment. A comparative proteomics study showed modulation in the proteome profile of breast cancer cells after treatment with T. chebula, B. aristata, Punica granatum, M. pruriens, and Acorus calamus. Metabolic profiling of lead plants revealed the existence of multiple anticancer compounds. Our study demonstrates the considerable potential of the mentioned plants as innovative therapies for HER2-positive breast cancer.


Breast Neoplasms , Cell Proliferation , Down-Regulation , Plant Extracts , Plants, Medicinal , Receptor, ErbB-2 , Humans , Breast Neoplasms/drug therapy , Breast Neoplasms/metabolism , Breast Neoplasms/genetics , Receptor, ErbB-2/metabolism , Receptor, ErbB-2/genetics , Plants, Medicinal/chemistry , Female , Plant Extracts/pharmacology , Plant Extracts/chemistry , Cell Line, Tumor , Down-Regulation/drug effects , Cell Proliferation/drug effects , Antineoplastic Agents, Phytogenic/pharmacology , Antineoplastic Agents, Phytogenic/chemistry , Apoptosis/drug effects , Terminalia/chemistry , Mucuna/chemistry
2.
Biochem Genet ; 2024 Mar 01.
Article En | MEDLINE | ID: mdl-38427123

Salmonella Typhimurium (ST) is a zoonotic pathogen that can cause gastroenteritis in humans when they consume contaminated food or water. When exposed to various stressors, both from living organisms (biotic) and the environment (abiotic), Salmonella Typhimurium produces Universal Stress Proteins (USPs). These proteins are gaining recognition for their crucial role in bacterial stress resistance and the ability to enter a prolonged state of growth arrest. Additionally, USPs exhibit diverse structures due to the fusion of the USP domain with different catalytic motifs, enabling them to participate in various reactions and cellular activities during stressful conditions. In this particular study, researchers cloned and analyzed the uspA gene obtained from poultry-derived strains of Salmonella Typhimurium. The gene comprises 435 base pairs, encoding a USP family protein consisting of 144 amino acids. Phylogenetic analysis demonstrated a close relationship between the uspA genes of Salmonella Typhimurium and those found in other bacterial species. We used molecular dynamics simulations and 3D structure prediction to ensure that the USPA protein was stable. Furthermore, we also carried out motif search and network analysis of protein-protein interactions. The findings from this study offer valuable insights for the development of inhibitors targeted against Salmonella Typhimurium.

3.
Cell Signal ; 117: 111103, 2024 05.
Article En | MEDLINE | ID: mdl-38367792

The in vitro fertilization (IVF) is the first choice of infertile couples worldwide to plan for conception. Besides having a significant advancement in IVF procedure, the success rate is still poor. Although several approaches have been tested to improve IVF protocol, minor changes in culture conditions, physical factors and/or drug treatment generate reactive oxygen species (ROS) in oocytes. Due to large size and huge number of mitochondria, oocyte is more susceptible towards ROS-mediated signalling under in vitro culture conditions. Elevation of ROS levels destabilize maturation promoting factor (MPF) that results in meiotic exit from diplotene as well as metaphase-II (M-II) arrest in vitro. Once meiotic exit occurs, these oocytes get further arrested at metaphase-I (M-I) stage or metaphase-III (M-III)-like stage under in vitro culture conditions. The M-I as well as M-III arrested oocytes are not fit for fertilization and limits IVF outcome. Further, the generation of excess levels of ROS cause oxidative stress (OS) that initiate downstream signalling to initiate various death pathways such as apoptosis, autophagy, necroptosis and deteriorates oocyte quality under in vitro culture conditions. The increase of cellular enzymatic antioxidants and/or supplementation of exogenous antioxidants in culture medium protect ROS-induced deterioration of oocyte quality in vitro. Although a growing body of evidence suggests the ROS and OS-mediated deterioration of oocyte quality in vitro, their downstream signalling and related mechanisms remain poorly understood. Hence, this review article summarizes the existing evidences concerning ROS and OS-mediated downstream signalling during deterioration of oocyte quality in vitro. The use of various antioxidants against ROS and OS-mediated impairment of oocyte quality in vitro has also been explored in order to increase the success rate of IVF during assisted reproductive health management.


Antioxidants , Oocytes , Animals , Reactive Oxygen Species/metabolism , Antioxidants/pharmacology , Oxidative Stress , Mammals/metabolism
4.
Mater Horiz ; 10(11): 5235-5245, 2023 Oct 30.
Article En | MEDLINE | ID: mdl-37740285

Networks and systems which exhibit brain-like behavior can analyze information from intrinsically noisy and unstructured data with very low power consumption. Such characteristics arise due to the critical nature and complex interconnectivity of the brain and its neuronal network. We demonstrate a system comprising of multilayer hexagonal boron nitride (hBN) films contacted with silver (Ag), which can uniquely host two different self-assembled networks, which are self-organized at criticality (SOC). This system shows bipolar resistive switching between the high resistance state (HRS) and the low resistance state (LRS). In the HRS, Ag clusters (nodes) intercalate in the van der Waals gaps of hBN forming a network of tunnel junctions, whereas the LRS contains a network of Ag filaments. The temporal avalanche dynamics in both these states exhibit power-law scaling, long-range temporal correlation, and SOC. These networks can be tuned from one to another with voltage as a control parameter. For the first time, two different neural networks are realized in a single CMOS compatible, 2D material platform.

5.
J Phys Condens Matter ; 35(49)2023 Sep 07.
Article En | MEDLINE | ID: mdl-37586379

Out-of-equilibrium investigation of strongly correlated materials deciphers the hidden equilibrium properties. Herein, we have investigated the out-of-equilibrium magnetic properties of polycrystalline Dy2Ti2O7and Ho2Ti2O7spin ices. Our experimental findings reveal the emergence of magnetic field-induced anomalous hysteresis observed solely in temperature-and magnetic field-dependent AC susceptibility measurements. The observed memory effect (anomalous thermomagnetic hysteresis) exhibits a strong dependence on both thermal and non-thermal driving variables. Owing to the non-collinear spin structure, the applied DC bias magnetic field produces quenched disorder sites in the cooperative Ising spin matrix and suppresses the spin-phonon coupling. These quench disorders create a dynamic spin correlation, having slow spin relaxation and quick decay time, which additionally contribute to AC susceptibility. The initial conditions and measurement protocol decide the magnitude and sign of this dynamical term contributing to AC susceptibility. It is being suggested that such out-of-equilibrium properties arise from the combined influences of geometric frustration, disorder, and the cooperative nature of spin dynamics exhibited by these materials.

6.
Int Immunopharmacol ; 120: 110340, 2023 Jul.
Article En | MEDLINE | ID: mdl-37230033

Interferons play a critical role in the innate immune response against several infections and play a key role in the control of a variety of viral and bacterial infectious diseases such as hepatitis, covid-19, cancer, and multiple sclerosis. Therefore, natural or synthetic IFN production is important and had three common methods, including bacterial fermentation, animal cell culture, and recombinant nucleic acid technology. However, the safety, purity, and accuracy of the most preferred INF production systems have not been extensively studied. This study provides a comprehensive comparative overview of interferon production in various systems that include viral, bacterial, yeast, and mammalian. We aim to determine the most efficient, safe, and accurate interferon production system available in the year 2023. The mechanisms of artificial interferon production were reviewed in various organisms, and the types and subtypes of interferons produced by each system were compared. Our analysis provides a comprehensive overview of the similarities and differences in interferon production and highlights the potential for developing new therapeutic strategies to combat infectious diseases. This review article offers the diverse strategies used by different organisms in producing and utilizing interferons, providing a framework for future research into the evolution and function of this critical immune response pathway.


COVID-19 , Communicable Diseases , Animals , Saccharomyces cerevisiae , Interferons/therapeutic use , Immunity, Innate , Communicable Diseases/drug therapy , Mammals
7.
Microbiol Insights ; 16: 11786361231152438, 2023.
Article En | MEDLINE | ID: mdl-36741475

In MDR-TB, mycobacterium is resistant to battlefront drugs like rifampicin and isoniazid. Now it's an urgent global challenge for treatment & diagnosis because more than 50% of drugs are resistant. Till today's information, 5 reasons are liable for MDR: (1) Errors of physicians/patients in therapy management, (2) Complexity and poor vascularization of granulomatous lesions, which obstruct drug distribution to some sites, leading to resistance development, (3) Intrinsic drug resistance of tubercle bacilli, (4) Formation of non-replicating, drug-tolerant bacilli inside the granulomas, (5) Development of mutations in Mtb genes, which are the foremost important molecular mechanisms of resistance. the most contribution of this work is a brief & clear explanation of things chargeable for resistant development, and recent diagnostic & treatment methods for MDR-TB. This study shall help researchers & scientists to develop replacement rapid diagnostic tools, drugs, and treatment protocols.

8.
Crit Rev Microbiol ; 49(1): 57-81, 2023 Feb.
Article En | MEDLINE | ID: mdl-35220864

Moonlighting proteins (MLPs) are ubiquitous and provide a unique advantage to bacteria performing multiple functions using the same genomic content. Targeting MLPs can be considered as a futuristic approach in fighting drug resistance problem. This review follows the MLP trail from its inception to the present-day state, describing a few bacterial MLPs, viz., glyceraldehyde 3'-phosphate dehydrogenase, phosphoglucose isomerase glutamate racemase (GR), and DNA gyrase. Here, we carve out that targeting MLPs are the beacon of hope in an era of increasing drug resistance in bacteria. Evolutionary stability, structure-functional relationships, protein diversity, possible drug targets, and identification of new drugs against bacterial MLP are given due consideration. Before the final curtain calls, we provide a comprehensive list of small molecules that inhibit the biochemical activity of MLPs, which can aid the development of novel molecules to target MLPs for therapeutic applications.


Bacteria , Bacterial Proteins , Bacteria/genetics , Bacteria/metabolism , Bacterial Proteins/metabolism
9.
Front Public Health ; 10: 920126, 2022.
Article En | MEDLINE | ID: mdl-36052011

Objective: The world continues to face the COVID-19 crisis, and efforts are underway to integrate traditional medicine interventions for its effective management. The study aimed to determine the efficacy of the "AYURAKSHA" kit in terms of post-interventional percentage of COVID-19 IgG positivity, immunity levels, and quality of life (QoL) against COVID-19. Method: This was a non-randomized controlled, prospective intervention trial, done after the distribution of 80,000 AYURAKSHA kits (constituent of Sanshamani Vati, AYUSH Kadha, and Anu Taila) among Delhi police participants in India. Among 47,827 participants, the trial group (n = 101) was evaluated with the positivity percentage of IgG COVID-19 and Immune Status Questionnaire (ISQ) scores as a primary outcome and the WHO Quality of Life Brief Version (QOL BREF) scores along with hematological parameters as a secondary outcome in comparison to the control group (n = 71). Results: The data showed that the percentage of COVID-19 IgG positivity was significantly lower in the trial group (17.5 %) as compared to the control group (39.4 %, p = 0.003), indicating the lower risk (55.6%) of COVID-19 infection in the trial group. The decreased incidence (5.05%) and reduced mortality percentage (0.44%) of COVID-19 among Delhi police officers during peak times of the pandemic also corroborate our findings. The ISQ score and WHO-QOL BREF tool analysis showed the improved scores in the trial group when compared with the controls. Furthermore, no dysregulated blood profile and no increase in inflammation markers like C-reactive protein, erythrocyte sedimentation rate, Interleukin-6 (IL-6) were observed in the trial group. However, significantly enhanced (p = 0.027) IL-6 levels and random blood sugar levels were found in the control group (p = 0.032), compared to a trial group (p = 0.165) post-intervention. Importantly, the control group showed more significant (p = 0.0001) decline in lymphocyte subsets CD3+ (% change = 21.04), CD4+ (% change = 20.34) and CD8+ (% change = 21.54) levels than in trial group, confirming more severity of COVID-19 infection in the control group. Conclusion: The AYURAKSHA kit is associated with reduced COVID-19 positivity and with a better quality of life among the trial group. Hence, the study encourages in-depth research and future integration of traditional medicines for the prevention of the COVID-19 pandemic. Clinical trial registration: http://ctri.nic.in/, identifier: CTRI/2020/05/025171.


COVID-19 , COVID-19/epidemiology , COVID-19/prevention & control , Humans , Immunoglobulin G , Interleukin-6 , Pandemics/prevention & control , Police , Prospective Studies , Quality of Life , SARS-CoV-2
10.
Biochem Biophys Rep ; 32: 101350, 2022 Dec.
Article En | MEDLINE | ID: mdl-36164562

Lung cancer is one of the most frequently diagnosed malignant tumors and the leading cause of cancer-related death worldwide. Mainly, Non-small-cell lung cancer (NSCLC), which accounts for more than eighty-five percent of all lung cancers, consists of two major subtypes: lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC). Novel coronavirus disease (COVID-19) affected millions of people caused by acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) around the globe. Lung cancer patients and COVID-19 present unique and unfortunate lethal combinations because the lungs are the primary target organ of SARS-CoV-2 infection. Clinical studies have demonstrated that an over-activated inflammatory response associated with severe COVID-19 cases is characterized by excessive auto-amplifying cytokine release, which is defined as a "cytokine storm." ACE2 and TMPRSS2 receptors play an essential role in SARS-CoV-2 infection; therefore, using in silico analysis, we did correlation analysis with immune infiltration markers in LUAD and LUSC patient groups. Our study identified a promising correlation between immune-modulators and receptor proteins (ACE-2 and TMPRSS2), creating a domain that requires further laboratory studies for clinical authentication.

11.
Proc Biol Sci ; 289(1973): 20212650, 2022 04 27.
Article En | MEDLINE | ID: mdl-35473372

The collection of caterpillar fungus accounts for 50-70% of the household income of thousands of Himalayan communities and has an estimated market value of $5-11 billion across Asia. However, Himalayan collectors are at multiple economic disadvantages compared with collectors on the Tibetan Plateau because their product is not legally recognized. Using a customized hybrid-enrichment probe set and market-grade caterpillar fungus (with samples up to 30 years old) from 94 production zones across Asia, we uncovered clear geography-based signatures of historical dispersal and significant isolation-by-distance among caterpillar fungus hosts. This high-throughput approach can readily distinguish samples from major production zones with definitive geographical resolution, especially for samples from the Himalayan region that form monophyletic clades in our analysis. Based on these results, we propose a two-step procedure to help local communities authenticate their produce and improve this multi-national trade-route without creating opportunities for illegal exports and other forms of economic exploitation. We argue that policymakers and conservation practitioners must encourage the fair trade of caterpillar fungus in addition to sustainable harvesting to support a trans-boundary conservation effort that is much needed for this natural commodity in the Himalayan region.


Fungi , Asia , Geography
12.
JNMA J Nepal Med Assoc ; 59(235): 267-270, 2021 Mar 31.
Article En | MEDLINE | ID: mdl-34506441

INTRODUCTION: Varicosity is the common problem of various etiology having simple limb aching to worst complications like oedema, ulcer, and skin changes. Minimal invasive endovenous laser therapy is a noble procedure. The aim of the study is to find out the recurrence of the varicose vein after laser therapy in a tertiary care center. METHODS: This descriptive cross-sectional study was done in 38 patients with varicosity of the lower limb in a tertiary care hospital, from January 2019 to June 2019 after taking ethical clearance from Institutional Review Committee. Convenience sampling was done. Data was collected and entry was done in Statistical Package for the Social Science software version 22, point estimate at 90% Confidence Interval was calculated along with frequency and proportion for binary data. RESULTS: We recorded 38 patients with ablated limb out of which none of the ablated veins showed recanalization in six months follow up. Twenty two (58%) patients were male and 16 (42%) patients were female with a mean age of 40.26 years. Major bulk, 23 (60.5%) resumed activity in second postoperative day and only 1 (2.6%) patient waited for 5 days for normal activity with mean of 2.58 days postoperatively. Sixteen (42.1%) patients developed erythema or ecchymosis, 12 (31.6%) patients had induration along the long saphenous vein course, 7 (18.4%) patients had paresthesia, 2 (5.3%) patients had limb swelling and 1 (2.6%) patient had skin burn. CONCLUSIONS: Endovenous laser ablation has very low rate of recurrence of varicosity and has minor complications.


Laser Therapy , Varicose Veins , Adult , Cross-Sectional Studies , Female , Humans , Male , Recurrence , Saphenous Vein , Tertiary Care Centers , Treatment Outcome , Varicose Veins/epidemiology , Varicose Veins/surgery
13.
Proc Natl Acad Sci U S A ; 118(17)2021 04 27.
Article En | MEDLINE | ID: mdl-33893233

Peripheral myelin protein (PMP22) is an integral membrane protein that traffics inefficiently even in wild-type (WT) form, with only 20% of the WT protein reaching its final plasma membrane destination in myelinating Schwann cells. Misfolding of PMP22 has been identified as a key factor in multiple peripheral neuropathies, including Charcot-Marie-Tooth disease and Dejerine-Sottas syndrome. While biophysical analyses of disease-associated PMP22 mutants show altered protein stabilities, leading to reduced surface trafficking and loss of PMP22 function, it remains unclear how destabilization of PMP22 mutations causes mistrafficking. Here, native ion mobility-mass spectrometry (IM-MS) is used to compare the gas phase stabilities and abundances for an array of mutant PM22 complexes. We find key differences in the PMP22 mutant stabilities and propensities to form homodimeric complexes. Of particular note, we observe that severely destabilized forms of PMP22 exhibit a higher propensity to dimerize than WT PMP22. Furthermore, we employ lipid raft-mimicking SCOR bicelles to study PMP22 mutants, and find that the differences in dimer abundances are amplified in this medium when compared to micelle-based data, with disease mutants exhibiting up to 4 times more dimer than WT when liberated from SCOR bicelles. We combine our findings with previous cellular data to propose that the formation of PMP22 dimers from destabilized monomers is a key element of PMP22 mistrafficking.


Myelin Proteins/metabolism , Peripheral Nervous System Diseases/physiopathology , Protein Transport/physiology , Cell Membrane/metabolism , Humans , Ion Mobility Spectrometry/methods , Mass Spectrometry/methods , Membrane Proteins/metabolism , Myelin Proteins/genetics , Myelin Proteins/physiology , Peripheral Nervous System Diseases/diagnostic imaging , Peripheral Nervous System Diseases/metabolism , Protein Folding , Protein Stability , Schwann Cells/metabolism
14.
Stem Cell Rev Rep ; 17(3): 777-784, 2021 06.
Article En | MEDLINE | ID: mdl-33140233

Maintenance of metaphase-II (M-II) arrest in ovum is required to present itself as a right gamete for successful fertilization in mammals. Surprisingly, instability of meiotic cell cycle results in spontaneous exit from M-II arrest, chromosomal scattering and incomplete extrusion of second polar body (PB-II) without forming pronuclei so called abortive spontaneous ovum activation (SOA). It remains unclear what causes meiotic instability in freshly ovulated ovum that results in abortive SOA. We propose the involvement of various signal molecules such as reactive oxygen species (ROS), cyclic 3',5' adenosine monophosphate (cAMP) and calcium (Ca2+) in the induction of meiotic instability and thereby abortive SOA. These signal molecules through their downstream pathways modulate phosphorylation status and activity of cyclin dependent kinase (cdk1) as well as cyclin B1 level. Changes in phosphorylation status of cdk1 and its activity, dissociation and degradation of cyclin B1 destabilize maturation promoting factor (MPF). The premature MPF destabilization and defects in other cell cycle regulators possibly cause meiotic instability in ovum soon after ovulation. The meiotic instability results in a pathological condition of abortive SOA and deteriorates ovum quality. These ova are unfit for fertilization and limit reproductive outcome in several mammalian species including human. Therefore, global attention is required to identify the underlying causes in greater details in order to address the problem of meiotic instability in ova of several mammalian species icluding human. Moreover, these activated ova may be used to create parthenogenetic embryonic stem cell lines in vitro for the use in regenerative medicine.Graphical abstract.


Maturation-Promoting Factor , Oocytes , Animals , Calcium/metabolism , Female , Humans , Mammals/metabolism , Maturation-Promoting Factor/metabolism , Phosphorylation
15.
Cell ; 183(7): 1884-1900.e23, 2020 12 23.
Article En | MEDLINE | ID: mdl-33301709

Eastern equine encephalitis virus (EEEV) is one of the most virulent viruses endemic to North America. No licensed vaccines or antiviral therapeutics are available to combat this infection, which has recently shown an increase in human cases. Here, we characterize human monoclonal antibodies (mAbs) isolated from a survivor of natural EEEV infection with potent (<20 pM) inhibitory activity of EEEV. Cryo-electron microscopy reconstructions of two highly neutralizing mAbs, EEEV-33 and EEEV-143, were solved in complex with chimeric Sindbis/EEEV virions to 7.2 Å and 8.3 Å, respectively. The mAbs recognize two distinct antigenic sites that are critical for inhibiting viral entry into cells. EEEV-33 and EEEV-143 protect against disease following stringent lethal aerosol challenge of mice with highly pathogenic EEEV. These studies provide insight into the molecular basis for the neutralizing human antibody response against EEEV and can facilitate development of vaccines and candidate antibody therapeutics.


Aerosols/administration & dosage , Antibodies, Monoclonal/immunology , Antibodies, Viral/immunology , Encephalitis Virus, Eastern Equine/immunology , Encephalomyelitis, Equine/immunology , Encephalomyelitis, Equine/prevention & control , Adult , Animals , Antibodies, Monoclonal/isolation & purification , Antibodies, Neutralizing/immunology , Antigens, Viral/immunology , Cryoelectron Microscopy , Disease Models, Animal , Encephalitis Virus, Eastern Equine/ultrastructure , Encephalomyelitis, Equine/virology , Epitopes/chemistry , Female , Glycoproteins/immunology , Humans , Mice , Models, Molecular , Mutagenesis/genetics , Neutralization Tests , Protein Binding , Protein Domains , Recombinant Proteins/immunology , Sindbis Virus/immunology , Virion/immunology , Virion/ultrastructure , Virus Internalization
16.
Eur J Pharmacol ; 883: 173293, 2020 Sep 15.
Article En | MEDLINE | ID: mdl-32663542

Cyclic nucleotide phosphodiesterases (PDEs) are group of enzymes responsible for the hydrolysis of cyclic adenosine 3', 5' monophosphate (cAMP) and cyclic guanosine 3', 5' monophosphate (cGMP) levels in wide variety of cell types. These PDEs are detected in encircling granulosa cells or in oocyte with in follicular microenvironment and responsible for the decrease of cAMP and cGMP levels in mammalian oocytes. A transient decrease of cAMP level initiates downstream pathways to cause spontaneous meiotic resumption from diplotene arrest and induces oocyte maturation. The nonspecific PDE inhibitors (caffeine, pentoxifylline, theophylline, IBMX etc.) as well as specific PDE inhibitors (cilostamide, milrinone, org 9935, cilostazol etc.) have been used to elevate cAMP level and inhibit meiotic resumption from diplotene arrest and oocyte maturation, ovulation, fertilization and pregnancy rates both in vivo as well as under in vitro culture conditions. The PDEs inhibitors are used as powerful experimental tools to demonstrate cyclic nucleotide mediated changes in ovarian functions and thereby fertility. Indeed, non-hormonal nature and reversible effects of nonspecific as well as specific PDE inhibitors hold promise for the development of novel therapeutic drugs for female fertility regulation.


Fertility Agents, Female/therapeutic use , Fertility/drug effects , Infertility, Female/drug therapy , Oocytes/drug effects , Ovary/drug effects , Phosphodiesterase Inhibitors/therapeutic use , Animals , Female , Humans , Infertility, Female/enzymology , Infertility, Female/physiopathology , Oocytes/enzymology , Ovary/enzymology , Ovary/physiopathology , Ovulation/drug effects , Pregnancy
17.
J Biol Chem ; 295(19): 6299-6311, 2020 05 08.
Article En | MEDLINE | ID: mdl-32179647

3-Mercaptopyruvate sulfur transferase (MPST) catalyzes the desulfuration of 3-mercaptopyruvate (3-MP) and transfers sulfane sulfur from an enzyme-bound persulfide intermediate to thiophilic acceptors such as thioredoxin and cysteine. Hydrogen sulfide (H2S), a signaling molecule implicated in many physiological processes, can be released from the persulfide product of the MPST reaction. Two splice variants of MPST, differing by 20 amino acids at the N terminus, give rise to the cytosolic MPST1 and mitochondrial MPST2 isoforms. Here, we characterized the poorly-studied MPST1 variant and demonstrated that substitutions in its Ser-His-Asp triad, proposed to serve a general acid-base role, minimally affect catalytic activity. We estimated the 3-MP concentration in murine liver, kidney, and brain tissues, finding that it ranges from 0.4 µmol·kg-1 in brain to 1.4 µmol·kg-1 in kidney. We also show that N-acetylcysteine, a widely-used antioxidant, is a poor substrate for MPST and is unlikely to function as a thiophilic acceptor. Thioredoxin exhibits substrate inhibition, increasing the KM for 3-MP ∼15-fold compared with other sulfur acceptors. Kinetic simulations at physiologically-relevant substrate concentrations predicted that the proportion of sulfur transfer to thioredoxin increases ∼3.5-fold as its concentration decreases from 10 to 1 µm, whereas the total MPST reaction rate increases ∼7-fold. The simulations also predicted that cysteine is a quantitatively-significant sulfane sulfur acceptor, revealing MPST's potential to generate low-molecular-weight persulfides. We conclude that the MPST1 and MPST2 isoforms are kinetically indistinguishable and that thioredoxin modulates the MPST-catalyzed reaction in a physiologically-relevant concentration range.


Sulfurtransferases , Thioredoxins , Animals , Catalysis , HEK293 Cells , Hep G2 Cells , Humans , Isoenzymes/chemistry , Isoenzymes/metabolism , Male , Mice , Mice, Inbred BALB C , Organ Specificity , Sulfurtransferases/chemistry , Sulfurtransferases/metabolism , Thioredoxins/chemistry , Thioredoxins/metabolism
18.
Biochim Biophys Acta Gene Regul Mech ; 1863(3): 194479, 2020 03.
Article En | MEDLINE | ID: mdl-31931179

Cellular prion protein (PrP) misfolds into an aberrant and infectious scrapie form (PrPSc) that lead to fatal transmissible spongiform encephalopathies (TSEs). Association of prions with G-quadruplex (GQ) forming nucleic acid motifs has been reported, but implications of these interactions remain elusive. Herein, we show that the promoter region of the human prion gene (PRNP) contains two putative GQ motifs (Q1 and Q2) that assume stable, hybrid, intra-molecular quadruplex structures and bind with high affinity to PrP. Here, we investigate the ability of PrP to bind to the quadruplexes in its own promoter. We used a battery of techniques including SPR, NMR, CD, MD simulations and cell culture-based reporter assays. Our results show that PrP auto-regulates its expression by binding and resolving the GQs present in its own promoter. Furthermore, we map this resolvase-like activity to the N-terminal region (residues 23-89) of PrP. Our findings highlight a positive transcriptional-translational feedback regulation of the PRNP gene by PrP through dynamic unwinding of GQs in its promoter. Taken together, our results shed light on a yet unknown mechanism of regulation of the PRNP gene. This work provides the necessary framework for a plethora of studies on understanding the regulation of PrP levels and its implications in prion pathogenesis.


G-Quadruplexes , Gene Expression Regulation , Prion Proteins/genetics , Promoter Regions, Genetic , Transcription, Genetic , Cells, Cultured , Feedback, Physiological , Humans , Prion Proteins/biosynthesis , Prion Proteins/chemistry , Prion Proteins/metabolism
19.
Ayu ; 41(2): 72-78, 2020.
Article En | MEDLINE | ID: mdl-34908791

BACKGROUND: Majority of the population relies on traditional medicines for many of their health related problems. Particularly individuals with chronic illness like diabetes mellitus (DM) are more likely to simultaneously use herbal medicines. Many of such users believe that traditional medicines are natural and therefore safe, but this is a dangerous over simplification. Some herbal medicines may be associated with adverse effects, which include interactions with prescribed drugs. Information on such concomitant use of anti-diabetic drugs along with Ayurveda medicines is limited in Indian scenario. AIMS AND OBJECTIVES: To survey the patterns of concomitant use of Ayurveda and conventional anti-diabetic drugs by diabetic patients attending an out-patient department of a tertiary care teaching hospital in New Delhi, India through a validated questionnaire. MATERIALS AND METHODS: This is a questionnaire-based survey, carried out after the approval of the Institutional Ethics Committee, subsequently registered at CTRI. A questionnaire to assess the pattern of concomitant use was developed; content was validated and pre-tested by a pilot study in 40 patients, further refined and used in the survey. The data was analyzed to evaluate the patterns of concomitant use of Ayurvedic and conventional anti-diabetic drugs. RESULTS: About 95.9% of diabetic patients were taking herbo-mineral formulations concomitantly with conventional anti-diabetic drugs. Although 45.3% of diabetics were using Ayurveda interventions under the supervision of qualified AYUSH physicians, remaining involved in procuring the drugs over the counter (OTC) or from the local vendors. In majority of these instances, the use of Ayurveda formulations was not communicated with their physicians. CONCLUSION: The observations reveal that a majority of the diabetics (95.9%) were taking one or the other form of herbal preparations along with their conventional anti-diabetic drugs and about 44% among them were using these concomitantly. Thus, generating awareness on good practices of drug use seems to be essential.

20.
J Comput Biol ; 27(5): 738-754, 2020 05.
Article En | MEDLINE | ID: mdl-31464514

Heat shock protein 70 (Hsp70), a 70-kDa protein, also known as a molecular chaperone, is highly conserved. It plays a major role in cellular functions such as protein folding, regulation of protein degradation, translocation of proteins across membranes, receptor signaling, and protein assembly or disassembly. Vigna radiata is an important legume crop with available whole-genome sequence, but no such study on the HSP70 family is reported. A total of 32 V. radiate HSP70s (Vr-HSP70s) were identified and described. They are phylogenetically clustered into four subgroups. Vr-HSP70s show variations in intron/exon organization. This indicates that introns may play an essential role in gene regulating. The coexpression analysis of Vr-HSP70s revealed that these genes were involved in both abiotic and biotic stresses. Three cytoplasmic hub genes namely Vr-HSP70-C-14, Vr-HSP70-C-29, and Vr-HSP70-C-30 were found common in both stresses. Our findings provide directions for future studies to dissect functional analysis of Vr-HSP70s in response to abiotic and biotic stresses.


Genome-Wide Association Study , HSP70 Heat-Shock Proteins/genetics , Stress, Physiological/genetics , Vigna/genetics , Evolution, Molecular , Gene Expression Regulation, Plant/genetics , Multigene Family/genetics , Phylogeny , Plant Proteins/genetics , Vigna/growth & development
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