The fallopian tubes are critical segments of the female reproductive tract and are essential for transporting gametes and embryos. It creates a conducive environment necessary for successful fertilization, early embryo development, and embryo transport. The cellular composition and function of the fallopian tube are tightly regulated by the sex hormones estradiol and progesterone. Therefore, any pathological/ metabolic condition or exposure to exogenous agents with the potential to alter endocrine levels can have a significant impact on fallopian tube function and health. This review summarizes the effects of medications, infections, pathological conditions, lifestyle choices, and environmental factors that can significantly impact the morphology, histology, cellularity, and functionality of the fallopian tube.

Distinct neuronal subtype specification involves a complex network of transcription factors (TF). Previous studies from our laboratory showed Zinc finger transcription factor of cerebellum (ZIC) 3 to regulate the expression of Tyrosine Hydroxylase (TH), a proxy marker for dopaminergic (DA) neurons, in mouse olfactory bulb (OB) and mid brain (MB) neurons by distinct mechanisms. In absence of ZIC3 consensus binding site in proximal region of mouse TH promoter, ZIC3 interacts with ER81 in OB and regulates TH expression whereas in absence of ER81 in midbrain, ZIC3 regulates TH by enhancing the expression of PITX3. To analyze whether this observation is evolutionarily conserved in humans, different human stem cell model systems were utilized to understand the role of ZIC3 in human DA generation. Differentiation of dental pulp stem cells (DPSCs) with proper cues resulted in the expression of TH. Gain and loss of function demonstrated ZIC3 to be essential for the expression of TH. ZIC3 activates the transcription of TH gene by binding to response element within the region of TH promoter. Addition of SHH, a known morphogen that facilitates dopaminergic differentiation, increased the expression of TH, however, this regulation was suppressed in the absence of ZIC3. Mechanistic insight demonstrated SHH influences ZIC3 expression by GLI protein binding to ZIC3 promoter. Similar role of ZIC3 in TH expression was observed in hiPSCs differentiated to DA like neurons. Conclusively, the present study for the first time demonstrates the undetermined importance of ZIC3 in SHH mediated TH specification in cells of human origin.

NT5E, a 5' Ectonucleotidase, is an emerging hotspot for cancer immunotherapies. However, available enzymatic inhibitors have shown limited promise, indicating activity-independent roles. Delineating activity-dependent and independent roles of this protein with precision requisites identification of critical residues impacting activity without hampering surface expression/stability. Conserved Phenylalanine (F) at 417 and 500 positions, which form a π stack with substrate-AMP, was chosen as the region of interest for in silico mutagenesis. F417 and F500 were mutated to Valine (V) individually using Missense3D to obviate broad structural damage. CD73-F417V/F500V dual mutant was then subjected to Molecular Dynamics simulations for prediction of stability and structural variations in the protein over time in comparison to wild-type crystal structure. Gibbs Free energy indicated a significant decrease in ligand binding affinity of CD73-F417V/F500V. RMSD and RMSF analysis of MD trajectories indicated increased mobility, suggesting weaker ligand binding affinity within the catalytic site of the mutant potentially impacting functional dynamics and protein-ligand contacts. Overexpression of CD73-F417V/F500V mutant in CD73 null-HEKs indicated stable surface expression but complete loss of Ecto-nucleotidase activity, thus providing a tool to dissect activity-dependent and independent functions of NT5E.

The fallopian tube microenvironment supports gamete transport, fertilization and early embryo development. Any disturbances in this microenvironment can lead to fertilization failure, infertility or ectopic pregnancy. In this study, we systematically investigated the effects of clomiphene citrate (CC) on human PAX8-positive fallopian tube secretory epithelial cells (hPFTSECs) to assess CC-induced alterations in the tubal microenvironment. Human fallopian tube tissues obtained from women undergoing postpartum tubectomy were enzymatically digested, and the isolated hPFTSECs were cultured with CC. CC exposure reduced hPFTSEC viability, clonogenicity, proliferation and organoid forming efficiency while inducing apoptosis, DNA damage, and senescence in a dose-dependent manner. Further, delayed cell-cycle progression and impaired DNA replication were observed in CC-exposed hPFTSECs. CC when administered to adult female Swiss albino mice, both single-dose (25, 50, and 100 mg/kg; intraperitoneally) and multiple-dose (10 mg/kg; intraperitoneally for four consecutive days) exposure caused several oviductal abnormalities, including epithelial disorganization, loss of ciliation, dysplasia, and hyperplasia. Our findings reveal that CC induces a spectrum of cytotoxic, genotoxic, and structural alterations in the fallopian tube epithelium, with potential implications for tubal function impairment and disruption of the optimal microenvironment essential for fertilization and early embryo development, which might negatively impact overall reproductive outcomes.

Hormonal signals intricately regulate breast growth and development. Any perturbations due to altered endocrine and metabolic conditions, such as in polycystic ovary syndrome (PCOS), may influence breast cancer risk. This review highlights the susceptibility of women with PCOS to breast cancer, with a focus on the potential contribution of its comorbidities, such as hyperandrogenism, obesity, insulin resistance, diabetes, menopausal status, inflammation, and use of ovulation induction agents to this association. This is a narrative review including articles and information collected from PubMed, Scopus, Google Scholar, and the World Health Organization (WHO) websites. Studies were selected based on the availability of an abstract and research publication in English. Reports related to PCOS, its characteristics (hyperandrogenism, hyperinsulinemia, insulin resistance, type 2 diabetes, obesity), fertility treatment, and their association with breast cancer were included. The interrelation between PCOS and breast carcinoma remains ambiguous due to inconsistent and inconclusive epidemiological evidence. The comorbidities associated with the condition are reported to collectively or individually influence this association. Women with PCOS present anovulatory cycles and have a high necessity of undergoing ovulation induction, further altering hormonal dynamics. Evidence in the literature suggests that the number of treatment cycles, age at the onset of treatment, and cumulative doses of agents used for ovulation induction are a few other factors that could affect breast cancer risk, though the results are inconclusive. Existing literature suggests that PCOS and its characteristics may impact the process of breast development and possibly contribute to breast cancer, although the evidence is still conflicting. Moreover, temporary alterations of endocrine profile following fertility medications may have a significant influence on the breast cancer risk, although research has shown inconclusive evidence. Hence, robust experimental studies are required to understand the underlying mechanisms and the long-term health implications for women with PCOS due to the intricate association between these factors. Few studies suggest that an altered metabolic and endocrine profile in PCOS women predisposes them to breast cancer. However, current epidemiological evidence is inconclusive in establishing a strong link between PCOS and the risk of breast cancer. As PCOS women often depend on assisted reproductive technologies for infertility treatment, ovulation induction/ controlled ovarian stimulation protocols may further increase the health risks. The present article is a comprehensive overview highlighting the possible association with breast cancer risk in PCOS women undergoing assisted reproduction.

Tyrosine hydroxylase (TH) is the rate-limiting enzyme involved in the biosynthesis of catecholamines such as dopamine, norepinephrine, and epinephrine expressed in various regions of the brain, including the olfactory bulb (OB) and midbrain (MB). Previous studies demonstrated Zinc Finger transcription factor of the Cerebellum 3 (ZIC3) to regulate forebrain development, and Zic1/Zic3 compound mutant mice displayed reduced OB size. However, the precise role of ZIC3 in TH regulation remains elusive. In this study, we attempted to understand the role of ZIC3 in TH regulation and its underlying mechanism. While loss of function of Zic3 in OB-derived neurons led to down-regulation of TH expression, it could be rescued by over-expression of shRNA-resistant Zic3. Immunohistochemistry of OB of Zic3 null mice showed a similar reduction in expression of TH. Promoter of TH lacks the consensus ZIC3 binding region, and mechanistic insights revealed ZIC3 to regulate TH expression by interacting with ER81, a known TH regulator. ZIC3 interaction with ER81 is indispensable for ER81 binding to the Th promoter, and it fine-tunes ER81-mediated Th regulation in OB. In MB, where TH levels are highest after birth, ZIC3 regulates TH expression both in vitro and in vivo. TH was significantly reduced in P0 Zic3 null mice, as well as in Zic3 shRNA stereotactically delivered in 7-month-old mice. Mechanistically, in the absence of ER81 in MB, ZIC3 chooses an alternative approach of binding to Pitx3 promoter—a Dopaminergic (DA) fate determinant. Under the ectopic expression of ER81 in MB derived neurons, the propensity of ZIC3 binding to Pitx3 promoter is compromised, and its occupancy on Th promoter encompassing ER81 binding site is established, finally culminating in desired TH expression. Together, these findings reveal a unique ZIC3-mediated bimodal regulation of TH in OB and MB derived neurons.

The interaction of amyloid-β (Aβ) peptides with the plasma membrane (PM) is a potential trigger that initiates the formation of higher-order aggregates, membrane alterations/damage, and progressive neurotoxicity in Alzheimer's disease (AD). Recent studies showed neurons initiate PM repair upon damage induced by Aβ aggregates, and dysfunctional repair mechanisms contribute to neurodegeneration. This study uncovers a previously unrecognized molecular coupling between Rab3a-mediated exocytosis and pPAK1-driven endocytosis as a pivotal mechanism of PM repair in neuronal cells and primary neurons exposed to aggregation-prone oligomers of Aβ (oAβ). Unlike earlier reports that broadly associated PM damage and repair with Aβ aggregates, we specifically demonstrate that toxic oAβ1-42, but not oAβ1-40, provokes a highly efficient Rab3a-dependent exocytic repair response, tightly synchronized with pPAK1-mediated endocytosis. Using TIRF microscopy, we dissected the kinetics of vesicle fusion at nanometer-scale resolution and revealed that repair is initiated within minutes of oAβ1-42 exposure, with Rab3a activity dominating the critical first hour of response. Perturbation of this system-via IPA-3-mediated PAK1 inhibition or shRNA knockdown of Rab3a-abolished repair efficiency, establishing a direct causal link between these pathways. Furthermore, the long-term accumulation of oAβ in lysosomes was found to disrupt Rab3a recycling, implicating lipid-microdomain dynamics in the progressive failure of repair machinery in AD model, underscoring their physiological relevance. This study uniquely defines the synchronized action of exocytosis-endocytosis in PM repair via pPAK and Rab3a coordinated machinery as a critical neuronal survival strategy and highlights its specific failure as a mechanistic contributor to AD pathogenesis.

The interaction of amyloid-β (Aβ) peptides with the plasma membrane (PM) is a potential trigger that initiates the formation of higher-order aggregates, membrane alterations/damage, and progressive neurotoxicity in Alzheimers disease (AD). In a previous study, we showed that oligomers of Aβ1-42 (oAβ1-42) induced PM damage, resulting in PM repair cascade via lysosomal exocytosis coupled with endocytosis, and facilitation of tunneling nanotubes (TNTs)-like membrane protrusions to promote direct cell-to-cell transfer of aggregates. In this study, we demonstrated that PM damage induced by oligomers of the aggregation-prone peptide Aβ1-42 significantly facilitates PM repair by enhancing phosphorylated p21-activated kinase 1 (pPAK1)-dependent endocytosis and Rab3a-dependent exocytosis in SH-SY5Y and SK-N-SH neuronal cells compared to control and oAβ1-40 treated cells. We studied the kinetics of pPAK1-dependent endocytosis and the fusion of EGFP-Rab3a vesicles near the PM using total internal reflection fluorescence (TIRF) microscopy. IPA-3, a selective non-ATP competitive inhibitor of PAK1, inhibits endocytosis of oAβ peptides and Rab3a-dependent PM repair. Further, shRNA-mediated knockdown of the Rab3a gene inhibits pPAK1 and disrupts PM repair. Repair of damaged PM is a vital protective mechanism for non-proliferative cells like neurons, as disruption in PM repair leads to gradual neuronal cell death. However, there was no explicit understanding of PM repair in response to Aβ oligomers. This study revealed the interconnected action of Rab3a and pPAK1 in PM repair in response to oAβ -mediated damage, and its potential correlation in AD pathogenesis. ### Competing Interest Statement The authors have declared no competing interest.

Clomiphene citrate (CC) is commonly used for infertility treatments, particularly in intrauterine insemination (IUI) and in vitro fertilization (IVF) programs. Despite its effectiveness in inducing ovulation, it is reported to have a low pregnancy rate compared to letrozole. Further, risks of ectopic pregnancy, miscarriage, and fetal malformations have been reported in CC-induced fertility treatment cycles. The present study aimed at understanding whether these effects are associated with CC-induced changes in the secretome profile of the human PAX8-positive fallopian tube secretory epithelial cells (hPFTSECs). Fallopian tube tissues from healthy women who underwent postpartum tubectomy were enzymatically dissociated and cultured with CC, and their secretomes were analyzed by Liquid Chromatography Tandem Mass Spectrometry (LC–MS/MS) to identify CC-induced alterations. Proteomic analysis identified a total of 1549 proteins in the secretome pool of hPFTSECs cultured in vitro and 258 proteins were found to be differentially abundant in response to CC. KEGG and REACTOME analysis revealed that the differentially abundant proteins (DAPs) were mainly associated with pathways such as estrogen signaling, cell cycle and DNA damage. Further, candidate proteins associated with ovulation, embryo development, embryo implantation, pregnancy complications, ectopic pregnancy, sperm functions, apoptosis, senescence, and ovarian cancers were identified. CC exposure induced changes in secretome profile of hPFTSECs may affect the female reproductive health by potentially altering the tubal microenvironment. The study was registered in the Clinical Trials Registry of India (CTRI) under the registration number CTRI/2020/11/028791 on 02/11/2020.

Tyrosine hydroxylase (TH) is a key enzyme that catalyzes dopamine synthesis in various regions of the brain. Expression of TH is diversely regulated in Olfactory bulb (OB) and Midbrain (MB). Different sets of transcription factors (TFs) control the expression of TH in these two regions and most of them are exclusive to either OB or MB. Zinc finger transcription factor of cerebellum 3 (ZIC3) is known to regulate forebrain development. However, the role of ZIC3 in TH regulation and its underlying mechanism remains unknown. While loss of function of Zic3 in primary neurons led to down-regulation of TH expression, it could be rescued by over-expression of shRNA resistant Zic3. TH promoter lacks the consensus ZIC3 binding region and mechanistic insights revealed ZIC3 to regulate TH expression by interacting with ER81, a known TH regulator. ZIC3 interaction with ER81 is indispensable for ER81 binding to Th promoter and it fine tunes ER81 mediated Th regulation. In midbrain (MB), ZIC3 regulates TH expression both in vitro and in vivo. However, in the absence of ER81 in MB, ZIC3 chooses an alternative approach of binding to Pitx3 promoter- a DA fate determinant. Under the ectopic expression of ER81 in MB derived neurons, propensity of ZIC3 binding to Pitx3 promoter is compromised and its occupancy on Th promoter encompassing ER81 binding site is established, finally culminating in desired TH expression. Together, these findings reveal a unique ZIC3 mediated bimodal regulation of TH in OB and MB derived neurons.

AIMS:The knowledge of the molecular players that regulate the generation of endoderm cells is imperative to obtain homogenous population of pancreatic β-cells from stem cells. The Ubiquitin proteasome system (UPS) has been envisaged as a crucial intracellular protein degradation system, but its role in the generation of β-cells remains elusive. Hence, it would be appropriate to unravel the potential role of UPS in endoderm specification and utilize the understanding to generate β-cells from pluripotent stem cells. MATERIALS AND METHODS:The pluripotent stem cells (mESCs, miPSCs and hIPSCs) were subjected to differentiation towards pancreatic β-cells and assessed the proteasomal activity during endodermal differentiation. Pharmacologic agents MG132 and IU-1 were employed to inhibit and activate proteasomal activity respectively at the definitive endoderm stage to investigate its impact on the generation of β-cells. The expression of stage-specific genes were analyzed at transcript and protein levels. We also explored the role of unfolded protein response and UPS-regulated signalling pathways in endodermal differentiation. KEY FINDINGS:We observed decreased proteasomal activity specifically during endoderm, but not during the generation of other lineages. Extraneous proteasomal inhibition enhanced the expression of endodermal genes while increasing the proteasomal activity hindered definitive endodermal differentiation. Proteasomal inhibition at the definitive endodermal stage culminated in an enriched generation of insulin-positive cells. Elevated endodermal gene expression was consistent in mESCs and hIPSCs upon proteasomal inhibition. Mechanistic insight revealed the proteasome-inhibited enhanced endodermal differentiation to be via modulating the YAP pathway. SIGNIFICANCE:Our study unravels the specific involvement of UPS in endoderm cell generation from pluripotent stem cells and paves the way for obtaining potential definitive endodermal cells for plausible cellular therapy in the future.

Background Every pregnant woman faces the risk of pregnancy related morbidities. It Is better that pregnant women are aware of the pregnancy related symptoms and warning signs. Early intervention or referral to obstetric care facility can be made only if the pregnant woman has adequate knowledge in identifying danger signs. This study aimed to assess the knowledge of obstetric danger signs among pregnant women. Methods Community based cross sectional study was conducted from March 2019 to august 2019. 280 pregnant women were interviewed face to face using semi structured questionnaire. Results Only 86(30.7%) respondents were aware of at least more than four danger signs during antenatal period. Only 7.1% of the study participants were aware of the danger signs during post-partum. Maternal age, socioeconomic scale and parity had significant association with knowledge of obstetric danger signs. Conclusion Mother’s awareness on danger signs of pregnancy was poor during both antenatal and postpartum period. There is probably a need to enhance the health education sessions to all pregnant women irrespective of their demographic characteristics.

Introduction: Immunomodulation is the predominant mechanism via which Mesenchymal stromal cells (MSCs) mediate their therapeutic benefits. However, inconsistent success in numerous clinical trials warrants a better understating of the molecular mechanisms regulating their immunomodulatory properties. CD73, an ecto-5′-nucleotidase is abundantly expressed by MSCs, however its precise role in regulating their immunomodulatory properties is still elusive. The present study explored the role of CD73 in Interferon-gamma (IFNγ) sensing and in turn their ability to suppress “inflammatory” M1 macrophages. Materials and methods: CD73 knockdown MSCs (CD73-KDN) were initially assessed for expression of immunoregulatory molecules and IFNγ sensing ability by analysing expression of IFNγ signalling downstream targets such as pSTAT-1, Interferon-Stimulated Genes (ISG) and Indoleamine 2,3-dioxygnease (IDO), a prototypic IFNγ-induced immunomodulator. Next CD73-KDN MSCs were co-cultured with inflammatory M1 macrophages and evaluated for their ability to suppress them. To delineate the contributory role of CD73 and IFNγ signalling downstream target IDO, they were overexpressed independently in CD73-KDN MSCs and re-evaluated for their ability to suppress M1 macrophages. Results: CD73-KDN MSCs exhibited reduced expression of immunoregulatory molecules and were refractory to IFNγ signalling as indicated by attenuated expression of pSTAT-1, Interferon-Stimulated Genes (ISG) and Indoleamine 2,3-dioxygnease (IDO) upon IFNγ exposure. Since sensing of inflammation is critical for MSC mediated immunomodulation, CD73-KDN MSCs were functionally evaluated for their ability to immune-modulate “inflammatory” M1 macrophages wherein they failed to suppress M1 macrophages. Interestingly, ectopic expression of either CD73 or IFNγ signalling target IDO1 in CD73-KDN MSCs restored their ability to suppress M1 macrophages, establishing the importance of CD73-IFNγ signalling axis in MSC-mediated inflammatory macrophage suppression. Conclusion: The present study uncovers the unexplored role of CD73-IFNγ axis in MSC-mediated M1 macrophage suppression. MSC-educated macrophages are the actual immune-modulators at MSC transplant sites, thus CD73 can serve as a key immune-potency marker for benchmarking therapeutically relevant MSCs.

Pancreatic development is orchestrated by timely synthesis and degradation of stage-specific transcription factors (TFs). The transition from one stage to another stage is dependent on the precise expression of the developmentally relevant TFs. Persistent expression of particular TF would impede the exit from the progenitor stage to the matured cell type. Intracellular protein degradation-mediated protein turnover contributes to a major extent to the turnover of these TFs and thereby dictates the development of different tissues. Since even subtle changes in the crucial cellular pathways would dramatically impact pancreatic β-cell performance, it is generally acknowledged that the biological activity of these pathways is tightly regulated by protein synthesis and degradation process. Intracellular protein degradation is executed majorly by the ubiquitin proteasome system (UPS) and Lysosomal degradation pathway. As more than 90% of the TFs are targeted to proteasomal degradation, this review aims to examine the crucial role of UPS in normal pancreatic β-cell development and how dysfunction of these pathways manifests in metabolic syndromes such as diabetes. Such understanding would facilitate designing a faithful approach to obtain a therapeutic quality of β-cells from stem cells.

Cryopreservation of testicular tissue holds an important role in the field of fertility preservation, particularly for prepubertal boys diagnosed with cancer. However, prepubertal testicular tissue cryopreservation is still considered to be in the experimental stage necessitating the refinement of cryopreservation protocol. Considering the fact that loss of membrane lipids is the primary cause of freeze–thaw-induced loss of testicular cell functions, in this study, we explored the beneficial properties of exogenous supplementation of membrane lipids in the form of liposomes in enhancing the cryosurvival of prepubertal testicular tissue. The freezing medium supplemented with liposomes (prepared from soy lecithin, phosphatidylethanolamine, phosphatidylserine, and cholesterol) was used for the experiments. Prepubertal testicular tissues from Swiss albino mice were cryopreserved in a liposome-containing freezing medium (LFM) composed of 0.25 mg/mL liposomes, 5

The present study aimed to investigate the role of antidiabetic drug metformin on the cytoplasmic organization of oocytes. Germinal vesicle (GV) stage oocytes were collected from adult female Swiss albino mice and subjected to in vitro maturation (IVM) in various experimental groups- control, vehicle control (0.3% ethanol), metformin (50 mu g/mL), high glucose and high lipid (HGHL, 10 mM glucose; 150 mu M palmitic acid; 75 mu M stearic acid and 200 mu M oleic acid in ethanol), and HGHL supplemented with metformin. The metaphase II (MII) oocytes were analyzed for lipid accumulation, mitochondrial and endoplasmic reticulum (ER) distribution pattern, oxidative and ER stress, actin filament organization, cortical granule distribution pattern, spindle organization and chromosome alignment. An early polar body extrusion was observed in the HGHL group. However, the maturation rate at 24 h did not differ significantly among the experimental groups compared to the control. The HGHL conditions exhibited significantly higher levels of oxidative stress, ER stress, poor actin filament organization, increased lipid accumulation, altered mitochondrial distribution, spindle abnormalities, and chromosome misalignment compared to the control. Except for spindle organization, supplementation of metformin to the HGHL conditions improved all the parameters (non-significant for ER and actin distribution pattern). These results show that metformin exposure in the culture media helped to improve the hyperglycemia and hyperlipidemia-induced cytoplasmic anomalies except for spindle organization. Given the crucial role of spindle organization in proper chromosome segregation during oocyte maturation and meiotic resumption, the implications of metformin's limitations in this aspect warrant careful evaluation and further investigation.

The present study explores the advantages of enriching the freezing medium with membrane lipids and antioxidants in improving the outcome of prepubertal testicular tissue cryopreservation. For the study, testicular tissue from Swiss albino mice of prepubertal age group (2 weeks) was cryopreserved by slow freezing method either in control freezing medium (CFM; containing DMSO and FBS in DMEM/F12) or test freezing medium (TFM; containing soy lecithin, phosphatidylserine, phosphatidylethanolamine, cholesterol, vitamin C, sodium selenite, DMSO and FBS in DMEM/F12 medium) and stored in liquid nitrogen for at least one week. The tissues were thawed and enzymatically digested to assess viability, DNA damage, and oxidative stress in the testicular cells. The results indicate that TFM significantly mitigated freeze–thaw-induced cell death, DNA damage, and lipid peroxidation compared to tissue cryopreserved in CFM. Further, a decrease in Cyt C, Caspase-3, and an increase in Gpx4 mRNA transcripts were observed in tissues frozen with TFM. Spermatogonial germ cells (SGCs) collected from tissues frozen with TFM exhibited higher cell survival and superior DNA integrity compared to those frozen in CFM. Proteomic analysis revealed that SGCs experienced a lower degree of freeze–thaw-induced damage when cryopreserved in TFM, as evident from an increase in the level of proteins involved in mitigating the heat stress response, transcriptional and translational machinery. These results emphasize the beneficial role of membrane lipids and antioxidants in enhancing the cryosurvival of prepubertal testicular tissue offering a significant stride towards improving the clinical outcome of prepubertal testicular tissue cryopreservation.

In this paper, we demonstrate a unique FET system composed of vertically aligned MoS2nanosheets for real time, label free monitoring of zeta potential and dimension of mesenchymal stem cells during proliferation. To avoid the interference from non-specifically adsorbed macromolecules, a time varying electric field with an optimized frequency of 10 kHz has been applied at the gate and effective gate bias has been estimated from the real time drain current measurement in an electrical double layer gated cell-coupled FET platform. Special signal patterns corresponding to the alterations in the cell membrane potential have been observed. Extraction of cell parameters from the experimental observations, using closed set of equations reveal that the results correlate well with the gold standards and a high transconductance has enabled tracking of cell count with sufficient accuracy. The MoS2 FET signal has been compared with the pH change of the cell culture medium and the signal follows the cell proliferation related activity more closely than the pH change. Thus, it may be expected that this platform can substitute the existing time consuming and laborious methods.

Loss of insulin-secreting beta-cells in diabetes may be either due to apoptosis or dedifferentiation of beta-cell mass. The ubiquitin-proteasome system comprising E3 ligase and deubiquitinases (DUBs) controls several aspects of beta-cell functions. In this study, screening for key DUBs identified USP1 to be specifically involved in dedifferentiation process. Inhibition of USP1 either by genetic intervention or small-molecule inhibitor ML323 restored epithelial phenotype of beta-cells, but not with inhibition of other DUBs. In absence of dedifferentiation cues, overexpression of USP1 was sufficient to induce dedifferentiation in beta-cells; mechanistic insight showed USP1 to mediate its effect via modulating the expression of inhibitor of differentiation (ID) 2. In an in vivo streptozotocin (STZ)-induced dedifferentiation mouse model system, administering ML323 alleviated hyperglycemic state. Overall, this study identifies USP1 to be involved in dedifferentiation of beta-cells and its inhibition may have a therapeutic application of reducing beta-cell loss during diabetes.

Loss of insulin-secreting β-cells in diabetes may be either due to apoptosis or dedifferentiation of β-cell mass. The ubiquitin-proteasome system comprising E3 ligase and deubiquitinases (DUBs) controls several aspects of β-cell functions. In this study, screening for key DUBs identified USP1 to be specifically involved in dedifferentiation process. Inhibition of USP1 either by genetic intervention or small-molecule inhibitor ML323 restored epithelial phenotype of β-cells, but not with inhibition of other DUBs. In absence of dedifferentiation cues, overexpression of USP1 was sufficient to induce dedifferentiation in β-cells; mechanistic insight showed USP1 to mediate its effect via modulating the expression of inhibitor of differentiation (ID) 2. In an in vivo streptozotocin (STZ)-induced dedifferentiation mouse model system, administering ML323 alleviated hyperglycemic state. Overall, this study identifies USP1 to be involved in dedifferentiation of β-cells and its inhibition may have a therapeutic application of reducing β-cell loss during diabetes.