Browsing by Author "Anima Tripathi"

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A 12-Crown-4 Ether Containing Dipeptide Boc-12-Crown-4-l-DOPA-Gly-OMe Induces Cell Cycle Arrest and Apoptosis in Rat Eggs Cultured in Vitro
(Springer Netherlands, 2016) Garima Tripathi; Anima Tripathi; Gurunath Ramanathan; Shail K. Chaube
The study was designed to investigate whether crown ether containing dipeptide Boc-12-crown-4-l-DOPA-Gly-OMe has potential to induce meiotic cell cycle arrest and apoptosis in rat eggs cultured in vitro. The immature female rats were subjected to superovulation induction protocol and ovulated eggs were collected from ampulla of the fallopian tube. Ovulated eggs arrested at metaphase-II (M-II) stage of meiotic cell cycle were cultured in media-199 with or without various concentrations (0.0, 0.025, 0.050, 0.10, and 0.20 mM) of dipeptide for 3 h in vitro. Morphological apoptotic changes, hydrogen peroxide (H2O2) concentration, cytochrome c level, caspase-3 level as well as activity and DNA fragmentation were analysed in eggs cultured in vitro. Culture of M-II arrested eggs in plain medium for 3 h in vitro induced meiotic exit from M-II arrest in majority of eggs as evidenced by initiation of extrusion of second polar body (II PB). The dipeptide induced maintenance of M-II arrest and morphological apoptotic features in a concentration-dependent manner prior to degeneration. The dipeptide-induced morphological features were associated with increased H2O2 and cytochrome c levels in treated eggs. The increased cytochrome c induced caspase-3 level and activity and thereby DNA fragmentation as evidenced by DAB positive staining in treated eggs. Our results suggest that dipeptide Boc-12-C-4-l-DOPA-Gly-OMe induces cell cycle arrest at M-II stage and apoptosis in rat eggs cultured in vitro. © 2015 Springer Science+Business Media New York.
A comparative analysis of phytochemicals versus synthetic drugs/nanomedicines in the treatment of uterine fibroid: a systematic review
(Korean Society of Environmental Risk Assessment and Health Science, 2024) Sonal Upadhyay; Vivek Pandey; Anima Tripathi; Alakh N. Sahu; Anjali Rani; Amita Diwakar; Lavina Chaubey; Rashmi Gupta; Pawan K. Dubey
Most women experience uterine fibroids (UFs), a common benign gynecological tumor, at some point in their reproductive age. There are several pharmacological treatments available to shrink fibroids and lessen the UF symptoms. These medications cost a lot of money, though, and frequently have serious side effects. Therefore, due to their low cost, comparable and powerful therapeutic efficiency and lower side effects, phytochemical-based medications are gaining popularity in these days. This review's goal is to provide a summary of the knowledge that is currently unavailable regarding the mechanisms of the action of various phytochemical-based medications with anti-uterine fibroid efficacy. The present results showed that dietary phytocompounds (dehydroxyelephantopin, butein, capsaicin, fisetin, kaempferol, resveratrol, silibinin and curcumin) could probably be effective as therapeutic compounds for uterine leiomyoma. These phytochemicals indicated their capability to regulate main fibroid promoting and initiating events for instance, proliferation, inflammation, angiogenesis and fibrosis in various experimental setups through modulating various signaling pathways, such as Smad 2/3, PI3K/AKT/mTOR, ERK 1/2 and β-catenin indicating that they could serve as targets for prevention and/or treatment of UFs. This review provides key molecular insights for the development of phytochemical-based novel personalized therapy for non-surgical management of UFs which may help to avoid hysterectomy. © The Author(s), under exclusive licence to Korean Society of Environmental Risk Assessment and Health Science 2023.
Apoptosis in mammalian oocytes: A review
(Springer New York LLC, 2015) Meenakshi Tiwari; Shilpa Prasad; Anima Tripathi; Ashutosh N. Pandey; Irfan Ali; Arvind K. Singh; Tulsidas G. Shrivastav; Shail K. Chaube
Apoptosis causes elimination of more than 99 % of germ cells from cohort of ovary through follicular atresia. Less than 1 % of germ cells, which are culminated in oocytes further undergo apoptosis during last phases of oogenesis and depletes ovarian reserve in most of the mammalian species including human. There are several players that induce apoptosis directly or indirectly in oocytes at various stages of meiotic cell cycle. Premature removal of encircling granulosa cells from immature oocytes, reduced levels of adenosine 3′,5′-cyclic monophosphate and guanosine 3′,5′-cyclic monophosphate, increased levels of calcium (Ca2+) and oxidants, sustained reduced level of maturation promoting factor, depletion of survival factors, nutrients and cell cycle proteins, reduced meiotic competency, increased levels of proapoptotic as well as apoptotic factors lead to oocyte apoptosis. The BH3-only proteins also act as key regulators of apoptosis in oocyte within the ovary. Both intrinsic (mitochondria-mediated) as well as extrinsic (cell surface death receptor-mediated) pathways are involved in oocyte apoptosis. BID, a BH3-only protein act as a bridge between both apoptotic pathways and its cleavage activates cell death machinery of both the pathways inside the follicular microenvironment. Oocyte apoptosis leads to the depletion of ovarian reserve that directly affects reproductive outcome of various mammals including human. In this review article, we highlight some of the important players and describe the pathways involved during oocyte apoptosis in mammals. © 2015 Springer Science+Business Media.
Aqueous extract of Azadirachta indica (neem) leaf induces generation of reactive oxygen species and mitochondria-mediated apoptosis in rat oocytes
(2012) Anima Tripathi; Tulsidas G. Shrivastav; Shail K. Chaube
Objective: Present study was aimed to determine whether aqueous neem leaf extract (NLE) induces generation of reactive oxygen species (ROS) and apoptosis through mitochondria-mediated pathway in rat oocytes. Design: A controlled prospective study. Setting: Laboratory research setting at Department of Zoology of Banaras Hindu University. Animal(s): Forty eight sexually immature female rats that were 20-30 days of age. Intervention(s): Sexually immature female rats were fed palatable dose of NLE (10 mg/g dry feed palate) for 10 days and then subjected to superovulation induction protocol. Thereafter, rats were euthanized, ovulated cumulus oocyte complexes were collected from oviduct and oocytes were denuded. Main outcome measure(s): Rate of morphological apoptotic changes, measurement of hydrogen peroxide, nitric oxide and cytochrome c concentrations, caspase-9, caspases-3 activities and DNA fragmentation in oocytes. Results: In vivo NLE treatment induced morphological apoptotic changes were associated with increased hydrogen peroxide, nitric oxide and cytochrome c concentrations, caspase-9, caspase-3 activities and DNA fragmentation in oocyte. Conclusion: NLE induces generation of ROS that leads to oocytes apoptosis through mitochondria-mediated pathway. © 2011 Springer Science+Business Media, LLC.
Assisted reproductive technologies in infertility treatment: Opportunities and challenges
(Springer Singapore, 2017) Pawan K. Dubey; Anima Tripathi; Akhtar Ali
A tremendous rise in the fertility clinics providing ART services is seen worldwide with the birth of first IVF baby (Louise Joy Brown) in 1978. ART comprises various types of medical treatments designed to assist in achieving pregnancy. IVF and other ART-associated technologies of fertilization (ICSI, IUI, PZD, SUZI, MESA, and PESA) offer an opportunity to become parent even in severe cases of infertility. These technologies have allowed millions of individuals to fulfill their parenting wish. A positive attitude combined with an appropriate treatment can help most of the infertile couples experience the joy of parenthood. This chapter provides a thorough overview of the assisted reproductive technologies with opportunities for patients and challenges for clinical professionals or researchers. © Springer Nature Singapore Pte Ltd. 2017. All rights reserved.
Changes in signal molecules and maturation promoting factor levels associate with spontaneous resumption of meiosis in rat oocytes
(Wiley-Blackwell Publishing Ltd, 2015) Shilpa Prasad; Meenakshi Tiwari; Anima Tripathi; Ashutosh N. Pandey; Shail K. Chaube
The present study was aimed to find out changes in signal molecules and maturation promoting factor (MPF) levels during meiotic cell cycle progression from diplotene and metaphase-II (M-II) arrest, a period during which oocyte achieves meiotic competency. Data suggest that high levels of adenosine 3'-5'-cyclic monophosphate (cAMP), guanosine 3'-5'-cyclic monophosphate (cGMP), and nitric oxide (NO) are associated with diplotene arrest, while reduction in their levels correlates with reduced MPF level and meiotic resumption from diplotene arrest. On the other hand, increased intracellular NO, calcium (Ca2+) as well as hydrogen peroxide (H2O2) levels correlate with decreased cAMP, Thr-161 phosphorylated cyclin-dependent kinase-1 (Cdk1) as well as cyclin B1 levels. The decreased Thr-161 phosphorylated Cdk1 and cyclin B1 level reduce MPF level leading to exit from M-II arrest in oocytes cultured in vitro. These data suggest that the decrease of cAMP level and increase of cytosolic free Ca2+ as well as H2O2 levels associate with the reduced MPF level and meiotic resumption from diplotene arrest. On the other hand, increase of NO, cGMP, Ca2+ as well as H2O2 levels are associated with reduced MPF and spontaneous exit from M-II arrest in rat oocytes cultured in vitro. © 2015 International Federation for Cell Biology.
Correction: Intervention of Phytochemicals During Endometriosis and Their Conceivable Mechanisms (Revista Brasileira de Farmacognosia, (2023), 33, 6, (1126-1140), 10.1007/s43450-023-00426-2)
(Springer Science and Business Media Deutschland GmbH, 2023) Safiya Ayesha; Alka Sharma; Jayhind Kumar Chauhan; Vivek Pandey; Garima Tripathi; Pawan K. Dubey; Anima Tripathi
The graphical abstract in the original online version of this article was incorrect. It has been corrected. © 2023, The Author(s) under exclusive licence to Sociedade Brasileira de Farmacognosia.
Deoxyelephantopin, a novel naturally occurring phytochemical impairs growth, induces G2/M arrest, ROS-mediated apoptosis and modulates lncRNA expression against uterine leiomyoma
(Elsevier Masson s.r.l., 2020) Vivek Pandey; Anima Tripathi; Anjali Rani; Pawan K. Dubey
Deoxyelephantopin (DOE), a phytochemical, extracted and purified from Elephantopus scaber, has been shown to exhibit antitumor activities. Objective of the present study was to investigate anti-tumor and apoptosis-inducing properties of DOE against uterine leiomyoma (UL) and to explore their molecular mechanisms. Primary cell cultures from fresh UL tissue were established and maintained up to 12 passages. The cells exhibited continuous proliferation with 24 -h doubling time until 12 passages and was then subjected to molecular characterization. The growth inhibitory effect of DOE on UL cells was confirmed by colony formation, cellular senescence, AO/PI and DAPI staining. Fluorescent-activated cell sorting (FACS) assay for apoptosis and cell cycle arrest analysis revealed that DOE significantly (p < 0.05) inhibited the UL cell proliferation via cell cycle arrest at G2/M phase and induced apoptosis via ROS production by lowering mitochondrial membrane potential. Subsequently, the DOE induced ROS was markedly attenuated by co-treatment of N-Acetyl-Cysteine (NAC). Our quantitative RT-PCR and western blot results showed up-regulation of Bax, Caspase-3 and down-regulation of Bcl2, P53, αSMA, COL4A2, VEGF, PCNA, Cyclin B1 and oncogenic lncRNAs (H19, HOTAIR, BANCR and ROR) in DOE treated UL cells which further strengthen our findings. In conclusion, DOE inhibits growth of UL cells via cell cycle arrest at G2/M phase, induces ROS-dependent caspase-3-mediated mitochondrial intrinsic apoptotic pathway and down-regulation of oncogenic lncRNA in UL cells. Our findings suggest that DOE deserves for further systematic investigation in the uterine leiomyoma animal model as a novel apoptosis inducer for potential applications in the prevention or treatment of uterine leiomyoma. © 2020 The Authors
Di-(2-ethylhexyl) phthalate (DEHP) inhibits steroidogenesis and induces mitochondria-ROS mediated apoptosis in rat ovarian granulosa cells
(Royal Society of Chemistry, 2019) Anima Tripathi; Vivek Pandey; Alakh N. Sahu; Alok Singh; Pawan K. Dubey
Increased oxidative stress (OS) due to ubiquitous exposure to di-(2-ethylhexyl) phthalate (DEHP) can affect the quality of oocytes by inducing apoptosis and hampering granulosa cell mediated steroidogenesis. This study was carried out to investigate whether DEHP induced OS affects steroidogenesis and induces apoptosis in rat ovarian granulosa cells. OS was induced by exposing granulosa cells to various concentrations of DEHP (0.0, 100, 200, 400 and 800 μM) for 72 h in vitro. Intracellular reactive oxygen species (ROS), oxidative stress (OS), mitochondrial membrane potential, cellular senescence, apoptosis, steroid hormones (estradiol & progesterone) and gene expression were analyzed. The results showed that an effective dose of DEHP (400 μg) significantly increased OS by elevating the ROS level, mitochondrial membrane potential, and β-galactosidase activity with higher mRNA expression levels of apoptotic genes (Bax, cytochrome-c and caspase3) and a lower level of an anti-apoptotic gene (Bcl2) as compared to the control. Further, DEHP significantly (P > 0.05) decreased the level of steroid hormones (estradiol and progesterone) in a conditioned medium and this effect was reciprocated with a lower expression (P > 0.05) of steroidogenic responsive genes (Cyp11a1, Cyp19A1, Star, ERβ1) in treated granulosa cells. Furthermore, co-treatment with N-Acetyl-Cysteine (NAC) rescues the effects of DEHP on OS, ROS, β-galactosidase levels and gene expression activities. Altogether, these results suggest that DEHP induces oxidative stress via ROS generation and inhibits steroid synthesis via modulating steroidogenic responsive genes, which leads to the induction of apoptosis through the activation of Bax/Bcl-2-cytochrome-c and the caspase-3-mediated mitochondrial apoptotic pathway in rat granulosa cells. © 2019 The Royal Society of Chemistry and the Centre National de la Recherche Scientifique.
Differential gene expression profile evaluation between uterine leiomyoma and leiomyosarcoma using a machine learning approach
(KeAi Communications Co., 2023) Sonal Upadhyay; Ravi Bhushan; Anima Tripathi; Lavina Chaubey; Amita Diwakar; Pawan K. Dubey
Objective: The objective of this study is to differentiate between uterine leiomyomas (ULM) and uterine leiomyosarcomas (ULMS) by conducting molecular differential analysis and identifying potential prognostic biomarkers for diagnosis. Methods: The microarray datasets (GSEID: GSE64763 and GSE185543) were retrieved from the Gene Expression Omnibus database. Data preprocessing and differential gene expressions (DEGs) analysis were performed. The DEGs were further intersected to find the common DEGs in ULM and ULMS and further validation of selected DEGs were performed. Further, a machine learning classifier was also applied in the selection of biomarkers. Protein-protein interaction network based upon STRING v 10.5, was constructed. Additionally, Gene Ontology (GO) and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analyses were also performed to dissect possible functions and pathways. Results: A total of 50 significant DEGs for ULM while 321 DEGs for ULMS have been identified with their official gene symbol. Between ULM and ULMS, a total of 14 common DEGs were identified of which 8 were up-regulated while 6 were down-regulated. The DEGs of (GSE185543) were also analyzed and the significant genes were retrieved common in both datasets for further analysis. Using a machine learning approach, 10 feature genes were identified. Using the expression profiles of these genes, a sequential minimal optimization (SMO) prediction model was built on the training set, and it accurately and reliably classified features expression in ULM and ULMS in the independent test set. Furthermore, Co- Enrichment analysis was also performed. Conclusion: The study identified several DEGs, including ZNF365, EPYC, COL11A1, SHOX2, MMP13, TNN, GPM6A, and GATA2, through cross-validation, machine learning classifier, and Co- Enrichment analysis. These candidate disease genes may provide valuable insight into the underlying mechanisms and could be used as potential diagnostic biomarkers for ULM and ULMS. However, further validation of these genes is necessary to better understand their roles in the pathogenesis of ULM and ULMS. © 2023 The Authors
Encircling granulosa cells protects against di-(2-ethylhexyl)phthalate-induced apoptosis in rat oocytes cultured in vitro
(Cambridge University Press, 2019) Anima Tripathi; Vivek Pandey; A.N. Sahu; Alok K. Singh; Pawan K. Dubey
The present study investigated if the presence of encircling granulosa cells protected against di(2-ethylhexyl)phthalate (DEHP)-induced oxidative stress in rat oocytes cultured in vitro. Denuded oocytes and cumulus-oocyte complexes (COCs) were treated with or without various doses of DEHP (0.0, 25.0, 50.0, 100, 200, 400 and 800 μM) in vitro. Morphological apoptotic changes, levels of oxidative stress and reactive oxygen species (ROS), mitochondrial membrane potential, and expression levels of apoptotic markers (Bcl2, Bax, cytochrome c) were analyzed. Our results showed that DEHP induced morphological apoptotic changes in a dose-dependent manner in denuded oocytes cultured in vitro. The effective dose of DEHP (400 Âg) significantly (P>0.05) increased oxidative stress by elevating ROS levels and the mitochondrial membrane potential with higher mRNA expression and protein levels of apoptotic markers (Bax, cytochrome c). Encircling granulosa cells protected oocytes from DEHP-induced morphological changes, increased oxidative stress and ROS levels, as well as increased expression of apoptotic markers. Taken together our data suggested that encircling granulosa cells protected oocytes against DEHP-induced apoptosis and that the presence of granulosa cells could act positively towards the survival of oocytes under in vitro culture conditions and may be helpful during assisted reproductive technique programmes. © Cambridge University Press 2019.
Expression and intracellular localization of Nanos2-homologue protein in primordial germ cells and spermatogonial stem cells
(Cambridge University Press, 2019) Vivek Pandey; Anima Tripathi; Pawan K. Dubey
The decision by germ cells to differentiate and undergo either oogenesis or spermatogenesis takes place during embryonic development and Nanos plays an important role in this process. The present study was designed to investigate the expression patterns in rat of Nanos2-homologue protein in primordial germ cells (PGCs) over different embryonic developmental days as well as in spermatogonial stem cells (SSCs). Embryos from three different embryonic days (E8.5, E10.5, E11.5) and SSCs were isolated and used to detect Nanos2-homologue protein using immunocytochemistry, western blotting, reverse transcription polymerase chain reaction (RT-PCR) and flow cytometry. Interestingly, Nanos2 expression was detected in PGCs at day E11.5 onwards and up to colonization of PGCs in the genital ridge of fetal gonads. No Nanos2 expression was found in PGCs during early embryonic days (E8.5 and 10.5). Furthermore, immunohistochemical and immunofluorescence data revealed that Nanos2 expression was restricted within a subpopulation of undifferentiated spermatogonia (As, single type A SSCs and Apr, paired type A SSCs). The same results were confirmed by our western blot and RT-PCR data, as Nanos2 protein and transcripts were detected only in PGCs from day E11.5 and in undifferentiated spermatogonia (As and Apr). Furthermore, Nanos2-positive cells were also immunodetected and sorted using flow cytometry from the THY1-positive SSCs population, and this strengthened the idea that these cells are stem cells. Our findings suggested that stage-specific expression of Nanos2 occurred on different embryonic developmental days, while during the postnatal period Nanos2 expression is restricted to As and Apr SSCs. © 2019 Cambridge University Press.
Extracellular calcium protects against verapamil-induced metaphase-II arrest and initiation of apoptosis in aged rat eggs
(2009) S.K. Chaube; Anima Tripathi; Sabana Khatun; S.K. Mishra; P.V. Prasad; T.G. Shrivastav
Non-specific L-type calcium channel blockers, such as verapamil (≥50 μM), induce metaphase-II (M-II) arrest and apoptosis in aged rat eggs cultured in Ca2+-deficient medium. However, the effects of extracellular Ca2+ on verapamil-induced M-II arrest and apoptosis have not yet been reported. We have demonstrated that postovulatory aging induced exit from M-II arrest by extruding a second polar body, a morphological sign of spontaneous egg activation (SEA). Verapamil inhibited SEA and induced egg apoptosis in a dose-dependent manner in Ca2+-deficient medium. The initiation of apoptotic features was observed at 50 μM of verapamil. Extracellular Ca2+ (1.80 mM) reduced intracellular H2O2 level, bax protein expression, caspase-3 activity, DNA fragmentation and protected against 50 μM, but not higher concentrations of ≥100 μM in verapamil-induced egg apoptosis. These results suggest that extracellular Ca2+ ions have a role during SEA and protect against verapamil-induced apoptosis in aged rat eggs. © 2009 International Federation for Cell Biology.
Genetic variants related to insulin metabolism are associated with gestational diabetes mellitus
(Elsevier B.V., 2024) Ravi Bhushan; Shafiul Haque; Rakesh Kumar Gupta; Anjali Rani; Amita Diwakar; Sakshi Agarwal; Anima Tripathi; Pawan K. Dubey
The current study sought to investigate the associations of common genetic risk variants with gestational diabetes mellitus (GDM) risk in the north Indian population and to evaluate their utility in identifying GDM cases. A case-control study, including 300 pregnant women, was included, and clinical and pathological information was collected. The amplification-refractory mutation system (ARMS) was used for genotyping four single nucleotide polymorphisms (SNPs), namely FTO (rs9939609), PPARG2 (rs1801282), SLC30A8 (rs13266634), and TCF7L2 (rs12255372). The odds ratio and confidence interval were determined for each SNP in different genetic models. Further, attributable risk, population penetrance, and relative risk were also calculated. The risk allele A of FTO (rs9939609) poses a two times higher risk of GDM (p = 0.02, OR = 2.5). The CG and GG genotypes of PPARG2 (rs1801282) have half a lower risk of GDM. In SLC30A8 (rs13266634), the recessive model analysis showed a two times higher risk of having GDM, while the recessive model (TT vs. GG + GT) analysis in TCF7L2 (rs12255372) indicates a lower risk of GDM. Finally, the relative risk, population penetrance, and attributable risk for risk allele in all four variants was higher in GDM mothers. All four polymorphisms were found to be significantly associated with BMI, HbA1c, and insulin. Our study first time confirmed a significant association with GDM for four variants, FTO, PPARG2, SLC30A8, and TCF7L2, in the North Indian population. © 2024 Elsevier B.V.
High cytosolic free calcium level signals apoptosis through mitochondria-caspase mediated pathway in rat eggs cultured in vitro
(2012) Anima Tripathi; Shail K. Chaube
The present study was aimed to find out whether an increase of cytosolic free calcium level induces egg apoptosis through mitochondria-caspase mediated pathway. To increase cytosolic free calcium level and morphological apoptotic changes, ovulated eggs were cultured in Ca 2+/Mg 2+ free media-199 with or without various concentrations of calcium ionophore (0.5, 1, 2, 3, 4 μM) for 3 h in vitro. The morphological apoptotic changes, cytosolic free calcium level, hydrogen peroxide (H 2O 2) concentration, catalase activity, cytochrome c concentration, caspase-9 and caspase-3 activities and DNA fragmentation were analyzed. Calcium ionophore induced morphological apoptotic features in a concentration-dependent manner followed by degeneration at higher concentrations (3 and 4 μM). Calcium ionophore increased cytosolic free calcium level, induced generation of hydrogen peroxide (H 2O 2) and inhibited catalase activity in treated eggs. The increased H 2O 2 concentration was associated with increased cytochrome c concentration, caspase- 9 and caspase-3 activities that resulted in the induction of morphological features characteristic of egg apoptosis. The increased caspase-3 activity finally induced DNA fragmentation as evidenced by TUNEL positive staining in calcium ionophore-treated eggs. These findings suggest that high cytosolic free calcium level induces generation of H 2O 2 that leads to egg apoptosis through mitochondria-caspase mediated pathway. © Springer Science+Business Media, LLC 2012.
Identification and molecular characterization of genes modulating progression of an oocyte from M-I to M-II in rat ovary
(John Wiley and Sons Inc, 2024) Alka Sharma; Pawan K. Dubey; Pradeep Kumar; Kavindra Nath Tiwari; Anima Tripathi
Background: To achieve oocyte competence for successful fertilization, bidirectional communication between oocyte and granulosa cells is crucial. The acquisition of meiotic competency in oocyte is facilitated by various regulatory genes however, expression pattern of these genes is not well documented during meiotic transition from Metaphase-I to Metaphase-II stage. Therefore, the present research analyzed the expression pattern of regulatory genes that are involved in the transition from M-I to M-II stages in rat oocyte. Methods: The analysis of the data was conducted by applying an array of bioinformatic tools. The investigation of gene group interactions was carried out by employing the STRING database, which relies on co-expression information. The gene ontology (GO) analysis was performed utilizing the comparative GO database. Functional annotation for GO and pathway enrichment analysis were performed for genes involved in networking. The GO obtained through computational simulations was subsequently validated using quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. Results: The findings of our study suggest that there is a distinct gene expression pattern in both the oocyte and granulosa cells. This pattern indicates that oocyte-secreted factors, such as BMP15 and GDF9, play a crucial role in regulating the progression of the meiotic cell cycle from the M-I to M-II stages. We have also examined the level of mRNA expression of genes including CYP11A1, CYP19A1, and STAR, which are crucial for the steroidogenesis. Conclusions: It is fascinating to observe that the oscillatory pattern of specific key genes may hold significance in the process of in vitro oocyte maturation, specifically during the transition from the M-I to M-II stage. It might be useful for determining biomarker genes and potential pathways that play a role in attaining oocyte competency, thereby aiding in the assessment of oocyte quality for the purpose of achieving successful fertilization. © 2024 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Induction and characterization of a rat model of endometriosis
(Nature Research, 2024) Jayhind Kumar Chauhan; Pawan K. Dubey; Sangeeta Rai; Anima Tripathi
Endometriosis is a common condition that affects 5% to 10% of women during their reproductive years, although the aetiology and pathophysiology are still unknown. This study aimed to create an endometriosis model in rats to investigate the efficacy of natural and synthetic medications in treating endometriosis. An in vivo endometriotic model was established using a surgical induction method and the endocrine-disrupting drug diethylstilbestrol (DES). In brief, the experiment is categorised into three different groups. Each group contains five rats. The first group had no surgery, while in the in the second group of rats (n = 5), two small tissue grafts were fixed at the right and left walls of the abdomen. But in the in the third group of rats (n = 5), two small pieces of tissue have been grafted on the right and left abdomen walls by surgically along with DES treatments. Noninvasive photoacoustic imaging (PAI) was employed in the study to measure factors such as haemoglobin levels, oxygen saturation, and the size of endometriotic lesions. Histopathological analysis was carried out utilising staining techniques such as Hematoxylin and Eosin, Masson's Trichrome, and Periodic Acid Schiff, as well as immunohistochemistry with marker antibodies. Molecular markers in uterine tissue were examined using Western blots and real-time PCR. The developed endometriosis rat model showed a significant increase in the expression of anti-apoptotic Bcl-2, angiogenic marker VEGF and pro-inflammatory (COX-2 and IL-6) protein markers. In contrast to the control group, the treatment group had considerably lower Caspase-3 expression levels. Photoacoustic imaging (PAI) data demonstrated a constant increase in lesion size, as well as a decrease in oxygen saturation levels. The findings suggest that the in vivo endometriosis rat model may accurately assess the efficacy of natural or synthetic endometriosis treatments. This model may help in the improvement of disease understanding and the development of targeted therapeutic drugs. © The Author(s) 2024.
Integration of transcriptomics and metabolomics data revealed role of insulin resistant SNW1 gene in the pathophysiology of gestational diabetes
(Nature Research, 2025) Ravi Bhushan; Rimjhim Trivedi; Ritu Raj; Anjali Rani; Sangeeta Rai; Anima Tripathi; Shiv Govind Rawat; Ajay Kumar; Dinesh Kumar; Pawan Kumar Dubey
Gestational Diabetes Mellitus (GDM) is an emerging maternal health problem with increasing incidences. The lack of complete understanding of its pathophysiological mechanisms and novel regulatory biomarkers makes early diagnosis difficult. High-throughput RNA sequencing and computational bioinformatics analyses were conducted to identify novel hub genes, and their regulatory mechanisms were validated through qRT-PCR, western blot, and siRNA-mediated knockdown studies. Intermediate metabolites and circulatory levels of amino acids in the serum of GDM patients and healthy controls were measured. Transcriptomic studies identified SNW1 as the most sensitive and specific biomarker, significantly up-regulated in GDM (fold change = 1.09; p < 0.001). Metabolomic studies indicated significantly elevated gluconeogenesis in GDM, evidenced by decreased levels of alanine and increased levels of pyruvate and glucose compared to controls. siRNA-mediated knockdown of SNW1 in PANC1 cells resulted in significant down-regulation of alanine aminotransferase (ALT/GPT) and insulin receptor substrate (IRS1), while glucose transporters (GLUT2/GLUT4) and insulin (INS) were significantly up-regulated at both mRNA and protein levels. This study identified SNW1 as a novel insulin-resistant gene that induces hyperglycemia by elevating gluconeogenesis and decreasing glucose uptake. SNW1 may be considered a potential therapeutic target with clinical utility for the management of GDM. © The Author(s) 2025.
Intervention of Phytochemicals During Endometriosis and Their Conceivable Mechanisms
(Springer Science and Business Media Deutschland GmbH, 2023) Safiya Ayesha; Alka Sharma; Jayhind Kumar Chauhan; Vivek Pandey; Garima Tripathi; Pawan K. Dubey; Anima Tripathi
Endometriosis is a chronic and complex endocrine disorder that affects women of reproductive age. This syndrome is benign and is characterized by a combination of ovarian dysfunction and estrogen dependency symptoms, as well as pain and infertility. The most commonly stated symptoms include dysmenorrhea, significant dyspareunia, dyschezia, and dysuria. It is a severe public health issue. The prevalence rate is quite high and continues to rise on a daily basis. Starting with its diagnosis, pathophysiology, repercussions, and treatment options, there are numerous areas of disagreement. This study aims to provide an overview of the development of endometriosis diagnosis, symptoms, risk factors, etiology, medicinal plants, phytochemicals, and treatment, with a focus on research and the creation of novel therapeutic strategies. We conclude by making predictions and recommendations for the future of endometriosis-related research and prospective therapy approaches. Graphical Abstract: [Figure not available: see fulltext.] © 2023, The Author(s) under exclusive licence to Sociedade Brasileira de Farmacognosia.
Intracellular levels of hydrogen peroxide and nitric oxide in oocytes at various stages of meiotic cell cycle and apoptosis
(2009) Anima Tripathi; Sabana Khatun; A.N. Pandey; S.K. Mishra; Radha Chaube; T.G. Shrivastav; S.K. Chaube
The objective was to find out the functional roles of hydrogen peroxide (H2O2) and nitric oxide (NO) during various stages of meiotic cell cycle and apoptosis in rat oocytes. For this purpose, 30 oocytes from each stage such as diplotene, metaphase-I (M-I), metaphase-II (M-II) and apoptosis were collected and intracellular H2O2, total nitrite level and inducible nitric oxide synthase (iNOS) expression were analysed. This study demonstrated that generation of a tonic level of H2O2 induces meiotic resumption in diplotene-arrested oocytes and further increase may lead to apoptosis. Conversely, reduction in iNOS expression and total nitrite level are associated with meiotic resumption in diplotene-arrested oocytes, but induce apoptosis in aged oocytes. These results suggest that generation of a tonic level of H2 O2, reduced iNOS expression and total nitrite level are associated with meiotic resumption, while more generation of H2O2 and sustained reduced total nitrite level are linked with oocyte apoptosis in rat.