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We report here that dynamin 3 in the testis is associated with structures termed tubulobulbar complexes that internalize intact intercellular junctions during sperm release and turnover of the blood–testis barrier. The protein lies adjacent to an actin-Arp2/3 network that cuffs the double plasma membrane tubular invagination at the core of each complex. To explore the possible relationship between dynamin 3 and nectin-based adhesion junctions, we transiently transfected DsRed-tagged dynamin 3 into MDCK cells stably transfected with eGFP-tagged nectin 2, one of the adhesion molecules known to be expressed in Sertoli cells at adhesion junctions. Cells transfected with the dynamin 3 construct had less uniformly distributed nectin 2 at intercellular contacts when compared to control cells expressing only nectin 2 or transfected with the DsRed plasmid alone. Significantly, tubular extensions positive for nectin 2 were visible projecting into the cells from regions of intercellular contact. Our findings are consistent with the conclusion that dynamin 3 is involved with tubulobulbar morphogenesis. Dynamin 3 also occurs in concentrated deposits around the capitulum and striated columns in the connecting piece of sperm tails suggesting that the protein in these cells may function to stabilize the base of the tail or serve as a reservoir for use during or after fertilization. J. Cell. Physiol. 210: 644–654, 2007. © 2006 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Nerve conduction studies were conducted in 67 children to assess the effect of malnutrition on the peripheral nervous system. Significant reduction in nerve conduction velocities was demonstrated in severe protein energy malnutrition and ongoing long-term malnutrition. [ABSTRACT FROM AUTHOR]

Background and Aims: Apoptosis Signal-regulating Kinase 1 (ASK1) is activated by various pathological stimuli and induces cell apoptosis through downstream p38 activation. We studied the effect of pharmacological ASK1 inhibition on cirrhosis and its sequelae using comprehensive preclinical in vivo and in vitro systems., Approach and Results: Short-term (4-6 wk) and long-term (24-44 wk) ASK1 inhibition using small molecule GS-444217 was tested in thioacetamide-induced and BALB/c. Mdr2-/- murine models of cirrhosis and HCC, and in vitro using primary hepatocyte cell death assays. Short-term GS-444217 therapy in both models strongly reduced phosphorylated p38, hepatocyte death, and fibrosis by up to 50%. Profibrogenic release of mitochondrial DAMP mitochondrial deoxyribonucleic acid from dying hepatocytes was blocked by ASK1 or p38 inhibition. Long-term (24 wk) therapy in BALBc.Mdr2 - / - model resulted in a moderate 25% reduction in bridging fibrosis, but not in net collagen deposition. Despite this, the development of cirrhosis was effectively prevented, with strongly reduced p21 + hepatocyte staining (by 72%), serum ammonia levels (by 46%), and portal pressure (average 6.07 vs. 8.53 mm Hg in controls). Extended ASK1 inhibition for 44 wk in aged BALB/c. Mdr2-/- mice resulted in markedly reduced tumor number and size by ~50% compared to the control group., Conclusions: ASK1 inhibition suppresses the profibrogenic release of mitochondrial deoxyribonucleic acid from dying hepatocytes in a p38-dependent manner and protects from liver fibrosis. Long-term ASK1 targeting resulted in diminished net antifibrotic effect, but the progression to liver cirrhosis and cancer in BALBc/ Mdr2- / - mice was effectively inhibited. These data support the clinical evaluation of ASK1 inhibitors in fibrotic liver diseases., (Copyright © 2024 American Association for the Study of Liver Diseases.)

Background & Aims: Cholangiocarcinoma (CCA) is a dreaded complication of primary sclerosing cholangitis (PSC) that is difficult to diagnose and associated with high mortality. Lack of animal models of CCA recapitulating the hepatic microenvironment of sclerosing cholangitis has hindered the development of novel treatments. Herein, we sought to develop a mouse model of PSC-associated CCA., Methods: Ten-week-old Mdr2 -/- mice with congenital PSC-like disease, and healthy wild-type littermates were subjected to either modified retrograde biliary instillation or hydrodynamic tail vein injection of a sleeping beauty transposon-transposase plasmid system with activated AKT (myr-AKT) and Yap (YapS127A) proto-oncogenes (SB AKT/YAP1). The role of TGFβ was interrogated via ALK5 inhibitor (SB-525334) administration. Tumor phenotype, burden and desmoplastic reaction were analyzed histologically and via RNA sequencing., Results: While SB AKT/YAP1 plasmids administered via retrograde biliary injection caused tumors in Mdr2 -/- , only 26.67% (4/15) of these tumors were CCA. Alternatively, hydrodynamic tail vein injection of SB AKT/YAP1 resulted in robust tumorigenesis in all fibrotic Mdr2 -/- mice with high CCA burden compared to healthy mice. Tumors phenotypically resembled human CCA, expressed multiple CCA (but not hepatocellular carcinoma) markers, and exhibited a profound desmoplastic reaction. RNA sequencing analysis revealed profound transcriptional changes in CCA evolving in a PSC-like context, with specific alterations in multiple immune pathways. Pharmacological TGFβ inhibition led to enhanced immune cell tumor infiltration, reduced tumor burden and suppressed desmoplastic collagen accumulation compared to placebo., Conclusion: We established a new high-fidelity cholangiocarcinoma model in mice, termed SB CCA.Mdr2 -/- , which recapitulates the increased susceptibility to CCA in the setting of biliary injury and fibrosis observed in PSC. Through transcriptomics and pharmacological studies, we show dysregulation of multiple immune pathways and TGFβ signaling as potential drivers of CCA in a PSC-like microenvironment., Impact and Implications: Animal models for primary sclerosing cholangitis (PSC)-related cholangiocarcinoma (PSC-CCA) are lacking. Thus, we have developed and characterized a new mouse model of PSC-CCA, termed SB CCA.Mdr2 -/- , which features reliable tumor induction on a PSC-like background of biliary injury and fibrosis. Global gene expression alterations were identified and standardized tools, including automated whole slide image analysis methodology for tumor burden and feature analysis, were established to enable systematic research into PSC-CCA biology and formal preclinical drug testing., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Recently, the exposure of pesticides/herbicides to the living organisms is increased especially due to agricultural malpractices and industrial processes. In particular, the exposure of pesticides/herbicides (e.g., atrazine) can impart several harsh effects on the human health. The development of efficient detection systems can be crucial in monitoring the atrazine in water and food/plant products, which can be decisive in controlling the deadly exposures of atrazine. Herein, we have developed electrochemical immunosensors for atrazine by employing monoclonal anti-atrazine antibody conjugated gold nanoparticles. Two types of gold nanoparticles (i.e., citrate and glutathione (GSH)-capped AuNPs) were used to modify gold working electrode and utilized for the development of atrazine immunosensors. The conjugation of immunoprobe on working electrode was especially designed to obtain stable and efficient sensing signals. The nanosensing immunoprobes fabricated using citrate-AuNPs and GSH-AuNPs exhibited comparable responses for a wide linear working range of 50 ng/L- 30 μg/L with limit of detection (LOD) values of 0.08 and 0.06 ng/L for atrazine, respectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Lysozyme is widely used for the synthesis of nanomaterials (e.g., gold nanoparticle) to fluorescently sense metal ions. However, the effect of metal ions on the fluorescence of lysozyme is not studied yet. Herein, we have explored the interactions of lysozyme with different metal ions to develop a direct sensing platform for Fe(III). It has been observed that the fluorescence of lysozyme was slightly decreased in the presence of Cu(II), Hg(II), As(V), Co(II), Cd(II), Cr(II), Fe(II), Mn(II), Pb(II), and Zn(II), while a significant decrease in the lysozyme fluorescence was observed for Fe(III). The effect of thermal stability on the fluorescence quenching was also studied from 25 to 60 °C. In the present study, the lysozyme sensing probe was able to selectively and accurately detect 0.5-50 ppm of Fe(III) with a LOD of 0.1 ppm (1.8 µM) at 25 °C., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Morphological characteristics of any nanomaterial are critical in defining its properties. In this context, a method to control morphological parameters of polyaniline (PANI) has been investigated by producing its composite with gold nanoparticles (AuNPs). Herein, we report for the first time the successful control on the physical/chemical properties of PANI composites synthesized via interfacial polymerization through functionalization of its AuNP composite component with citrate, ascorbate, glutathione (GSH), and cetyl trimethyl ammonium bromide (CTAB). A significant difference in the polymerization pattern, morphologies, and electrical properties was recognized in these composites according to the functionality of the modified AuNPs. The obtained composites of AuNPs/PANI exhibited highly diverse morphologies (e.g., nodule, hollow hemisphere, flake, and spider-web galaxy type) and electrical characteristics according to functionalization. Hence, this study is expected to offer better insight into control of the polymerization pattern of AuNP/PANI composites and their associated properties.

Herein, we have developed citrate-, glutathione-, and ascorbate-functionalized gold nanoparticles (AuNPs) to examine their interactions with diverse heavy metal ions, such as Cd, Mn, Cr, Fe, Co, Pb, Hg, Zn, and Ti. These interactions are crucial in defining the final outcome of AuNP-based sensing/removal of heavy metals. We have evaluated these interactions by analyzing the variations in the color and spectroscopic signals of functionalized AuNPs. Additionally, the obtained results were also compared and validated with the computational studies. It has been observed that citrate-AuNPs and GSH-AuNPs displayed high selectivity toward Cr and Mn with E force values of -23.4 and -14.0 kJ/mol, respectively. Likewise, the ascorbate-AuNPs displayed sensitivity for multiple ions, for example, Cd, Fe, and Mn, with an E force value of -19.6 kJ/mol. A detailed analysis focusing on the electrostatic charges, ionic sizes, and interaction energy values has been provided to show specific interactions between functionalized AuNPs and heavy metal ions. The respective mechanisms of interaction between heavy metal ions and functionalized AuNPs have been explored with the help of experimental and computational outcomes.

The increasing level of pesticides and herbicides in food and water sources is a growing threat to human health and the environment. The development of portable, sensitive, specific, simple, and cost-effective sensors is hence in high demand to avoid exposure or consumption of these chemicals through efficient monitoring of their levels in food as well as water samples. The use of nanomaterials (NMs) for the construction of an immunosensing system was demonstrated to be an efficient and effective option to realize selective sensing against pesticides/herbicides. The potential of such applications has hence been demonstrated for a variety of NMs including graphene, carbon nanotubes (CNTs), metal nanoparticles, and nano-polymers either in pristine or composite forms based on diverse sensing principles (e.g., electrochemical, optical, and quartz crystal microbalance (QCM)). This article evaluates the development, applicability, and performances of NM-based immunosensors for the measurement of pesticides and herbicides in water, food, and soil samples. The performance of all the surveyed sensors has been evaluated on the basis of key parameters, e.g., detection limit (DL), sensing range, and response time., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Purinergic signaling is important in the activation and differentiation of macrophages, which play divergent roles in the pathophysiology of liver fibrosis. The ectonucleotidase CD39 is known to modulate the immunoregulatory phenotype of macrophages, but whether this specifically impacts cholestatic liver injury is unknown. Here, we investigated the role of macrophage-expressed CD39 on the development of biliary injury and fibrosis in a mouse model of sclerosing cholangitis. Myeloid-specific CD39-deficient mice (LysMCreCd39 fl/fl ) were generated. Global CD39 null (Cd39 -/- ), wild-type (WT), LysMCreCd39 fl/fl , and Cd39 fl/fl control mice were exposed to 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) to induce biliary fibrosis. Hepatic hydroxyproline levels, liver histology, immunohistochemistry, mRNA expression levels, and serum biochemistry were then assessed. Following 3 weeks of DDC-feeding, Cd39 -/- mice exhibited more severe fibrosis, when compared to WT mice as reflected by morphology and increased liver collagen content. Myeloid-specific CD39 deletion in LysMCreCd39 fl/fl mice recapitulated the phenotype of global Cd39 -/- , after exposure to DDC, and resulted in similar worsening of liver fibrosis when compared to Cd39 fl/fl control animals. Further, DDC-treated LysMCreCd39 fl/fl mice exhibited elevated serum levels of transaminases and total bilirubin, as well as increased hepatic expression of the profibrogenic genes Tgf-β1, Tnf-α, and α-Sma. However, no clear differences were observed in the expression of macrophage-elaborated specific cytokines between LysMCreCd39 fl/fl and Cd39 fl/fl animals subjected to biliary injury. Our results in the DDC-induced biliary type liver fibrosis model suggest that loss of CD39 expression on myeloid cells largely accounts for the exacerbated sclerosing cholangitis in global CD39 knockouts. These findings indicate that macrophage expressed CD39 protects from biliary liver injury and fibrosis and support a potential therapeutic target for human hepatobiliary diseases.

Huntington disease (HD) is caused by the expression of mutant huntingtin (mHTT) bearing a polyglutamine expansion. In HD, mHTT accumulation is accompanied by a dysfunction in basal autophagy, which manifests as specific defects in cargo loading during selective autophagy. Here we show that the expression of mHTT resistant to proteolysis at the caspase cleavage site D586 (C6R mHTT) increases autophagy, which may be due to its increased binding to the autophagy adapter p62. This is accompanied by faster degradation of C6R mHTT in vitro and a lack of mHTT accumulation the C6R mouse model with age. These findings may explain the previously observed neuroprotective properties of C6R mHTT. As the C6R mutation cannot be easily translated into a therapeutic approach, we show that a scheduled feeding paradigm is sufficient to lower mHTT levels in YAC128 mice expressing cleavable mHTT. This is consistent with a previous model, where the presence of cleavable mHTT impairs basal autophagy, while fasting-induced autophagy remains functional. In HD, mHTT clearance and autophagy may become increasingly impaired as a function of age and disease stage, because of gradually increased activity of mHTT-processing enzymes. Our findings imply that mHTT clearance could be enhanced by a regulated dietary schedule that promotes autophagy.

Objective: To determine the prevalence and types of intimate partner violence (IPV) during pregnancy, factors linked with IPV, and effects of IPV on maternal-fetal outcomes., Methods: In a prospective observational study at a tertiary care hospital in Delhi, India, 400 women at 20-28 weeks of pregnancy were screened for IPV between December 2013 and April 2015. The women completed a detailed questionnaire and were followed up until delivery., Results: Overall, 49 (12.3%) women experienced IPV during pregnancy. The most prevalent type of IPV was emotional (43/400 [10.7%]), followed by physical (40/400 [10.0%]) and sexual (7/400 [1.8%]). The most prevalent factor triggering IPV was intimate partner's desire for a son (17/49 [34.7%]). Women and their intimate partners were older in the IPV group than in the control group, and duration of marriage was longer (P<0.05 for all). Multigravidity, lower socioeconomic status, low education level of intimate partner, and partners' addiction were more common in the IPV group (P<0.05 for all). Obstetric outcomes were similar in both groups. Depression was diagnosed in 19 (46.3%) women affected by IPV., Conclusion: IPV was documented in approximately 12% of participants. Population-based surveys need to be done to investigate further., (© 2016 International Federation of Gynecology and Obstetrics.)

Caspase-6 (CASP6) has an important role in axonal degeneration during neuronal apoptosis and in the neurodegenerative diseases Alzheimer and Huntington disease. Decreasing CASP6 activity may help to restore neuronal function in these and other diseases such as stroke and ischemia, where increased CASP6 activity has been implicated. The key to finding approaches to decrease CASP6 activity is a deeper understanding of the mechanisms regulating CASP6 activation. We show that CASP6 is posttranslationally palmitoylated by the palmitoyl acyltransferase HIP14 and that the palmitoylation of CASP6 inhibits its activation. Palmitoylation of CASP6 is decreased both in Hip14 -/- mice, where HIP14 is absent, and in YAC128 mice, a model of Huntington disease, where HIP14 is dysfunctional and where CASP6 activity is increased. Molecular modeling suggests that palmitoylation of CASP6 may inhibit its activation via steric blockage of the substrate-binding groove and inhibition of CASP6 dimerization, both essential for CASP6 function. Our studies identify palmitoylation as a novel CASP6 modification and as a key regulator of CASP6 activity.

Huntington disease (HD) is a dominant, genetic neurodegenerative disease characterized by progressive loss of voluntary motor control, psychiatric disturbance, and cognitive decline, for which there is currently no disease-modifying therapy. HD is caused by the expansion of a CAG tract in the huntingtin (HTT) gene. The mutant HTT protein (muHTT) acquires toxic functions, and there is significant evidence that muHTT lowering would be therapeutically efficacious. However, the wild-type HTT protein (wtHTT) serves vital functions, making allele-specific muHTT lowering strategies potentially safer than nonselective strategies. CAG tract expansion is associated with single nucleotide polymorphisms (SNPs) that can be targeted by gene silencing reagents such as antisense oligonucleotides (ASOs) to accomplish allele-specific muHTT lowering. Here we evaluate ASOs targeted to HD-associated SNPs in acute in vivo studies including screening, distribution, duration of action and dosing, using a humanized mouse model of HD, Hu97/18, that is heterozygous for the targeted SNPs. We have identified four well-tolerated lead ASOs that potently and selectively silence muHTT at a broad range of doses throughout the central nervous system for 16 weeks or more after a single intracerebroventricular (ICV) injection. With further validation, these ASOs could provide a therapeutic option for individuals afflicted with HD.

Huntington disease (HD) is an inherited, fatal neurodegenerative disorder caused by a CAG repeat expansion in the huntingtin gene. The mutant protein causes neuronal dysfunction and degeneration resulting in motor dysfunction, cognitive decline, and psychiatric disturbances. Currently, there is no disease altering treatment, and symptomatic therapy has limited benefit. The pathogenesis of HD is complicated and multiple pathways are compromised. Addressing the problem at its genetic root by suppressing mutant huntingtin expression is a promising therapeutic strategy for HD. We have developed and evaluated antisense oligonucleotides (ASOs) targeting single nucleotide polymorphisms that are significantly enriched on HD alleles (HD-SNPs). We describe our structure-activity relationship studies for ASO design and find that adjusting the SNP position within the gap, chemical modifications of the wings, and shortening the unmodified gap are critical for potent, specific, and well tolerated silencing of mutant huntingtin. Finally, we show that using two distinct ASO drugs targeting the two allelic variants of an HD-SNP could provide a therapeutic option for all persons with HD; allele-specifically for roughly half, and non-specifically for the remainder.

HIP14 is the most highly conserved of 23 human palmitoyl acyltransferases (PATs) that catalyze the post-translational addition of palmitate to proteins, including huntingtin (HTT). HIP14 is dysfunctional in the presence of mutant HTT (mHTT), the causative gene for Huntington disease (HD), and we hypothesize that reduced palmitoylation of HTT and other HIP14 substrates contributes to the pathogenesis of the disease. Here we describe the yeast two-hybrid (Y2H) interactors of HIP14 in the first comprehensive study of interactors of a mammalian PAT. Unexpectedly, we discovered a highly significant overlap between HIP14 interactors and 370 published interactors of HTT, 4-fold greater than for control proteins (P = 8 × 10(-5)). Nearly half of the 36 shared interactors are already implicated in HD, supporting a direct link between HIP14 and the disease. The HIP14 Y2H interaction set is significantly enriched for palmitoylated proteins that are candidate substrates. We confirmed that three of them, GPM6A, and the Sprouty domain-containing proteins SPRED1 and SPRED3, are indeed palmitoylated by HIP14; the first enzyme known to palmitoylate these proteins. These novel substrates functions might be affected by reduced palmitoylation in HD. We also show that the vesicular cargo adapter optineurin, an established HTT-binding protein, co-immunoprecipitates with HIP14 but is not palmitoylated. mHTT leads to mislocalization of optineurin and aberrant cargo trafficking. Therefore, it is possible that optineurin regulates trafficking of HIP14 to its substrates. Taken together, our data raise the possibility that defective palmitoylation by HIP14 might be an important mechanism that contributes to the pathogenesis of HD., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Introduction: Tuboperitoneal pathology is responsible for 40-50% cases of infertility. Hysterosalpingography (HSG) & laparoscopy are the two classic methods available for evaluation of tubal pathology and are complementary to each other. Though pelvic sonography and HSG are good enough to exclude gross intrauterine pathology, but subtle changes in the form of small polyps, adhesions and seedling fibroid are better picked up on magnification with hysteroscopy. Combined hysterolaparoscopy may obviate need for HSG, as complete evaluation and treatment is possible in the same sitting., Aim: To assess the utility of Hysterolaparoscopy as one step procedure and compare it with HSG, in the subset of ovulatory infertile women with normal pelvic sonography / seminogram /hormonal assays., Materials and Method: In this analytical prospective study, 193 infertile women aged 19 to 42 years underwent HSG and Hysterolaparoscopy over a period of six months. They were confirmed to have ovulatory cycles and normal seminogram. Patient with active genital infection were excluded. Findings were categorized as normal/abnormal and therapeutic intervention done, if required. Statistical evaluation was carried out using Chi- square test., Result: On comparing HSG and Hysteroscopy, uterine findings matched in 66.3% patients. HSG failed to detect uterine pathology in 32.12% patients (62/193) with a sensitivity of 21.3% and specificity of 97.45%. Ninety three percent of intrauterine adhesions/polyps were missed on HSG. Hysteroscopic intervention was required in 23.83% cases, adhesiolysis being the commonest. On comparing tubal patency on HSG and laparoscopy, the sensitivity of HSG in detecting bilateral tubal block was 80.6% and specificity of 81.5%. With regard to unilateral tubal block, sensitivity was 34.6% and specificity 89.8%. The agreement between the two was 74%. Pathology such as adhesions, fimbrial agglutination and endometriosis were dealt surgically in 65.8% patients. As per HSG, 112/193 women had both tubes patent and 177 revealed normal uterine cavity. When these 112 women (58.03%) with normal HSG report were further subjected to hysterolaparoscopy, only 35/193 (18.13%) of them actually had normal tubes and uterus; rest 77 women (39.89%) were benefited by one step procedure of hysterolaparoscopic evaluation and intervention and further treatment done., Conclusion: Hysterolaparoscopy (Pan Endoscopic) approach is better than HSG and should be encouraged as first and final procedure in selected infertile women.