Functional characterization of the roles of endocytic recycling regulator EHD1 using in vivo and in vitro analyses

Endocytic recycling is a fundamental cellular process that allows the precise regulation of the membrane components and receptors at the cell surface. Recent studies have established that the C-terminal Eps15 homology domain-containing (EHD) proteins function as key regulators of this process. Four highly-conserved members of the EHD protein family in mammals, EHD1-EHD4, play shared as well as unique roles in endocytic trafficking. Studies presented here demonstrate a critical role of EHD1 in the normal ocular development in mice. Ehd1 knockout mice generated in our laboratory displayed gross ocular phenotypes including the anophthalmia, microphthalmia, and congenital cataracts. Hematoxylin and eosin (H&E) staining revealed defects in the Ehd1 mutant mice that included smaller lens, lack of lens, and persistence of the lens-stalk and hyaloid vasculature. By deleting Ehd1 specifically in the lens (Ehd1 CKO), my studies provide evidence that EHD1 expression in the lens precursor cells within the surface ectoderm is necessary for the normal lens development. Ehd1 CKO mice recapitulated the major ocular phenotypes of the Ehd1-null mice, and exhibited reduced proliferation and increased cell death within the lens epithelium as well as a disorganized corneal endothelium. These data suggest the important roles of EHD1 in the overall process of eye development. EHD proteins are characterized by the presence of a C-terminal Eps15 homology (EH) domain. EH-domains are known to mediate interactions with proteins containing sequences with core Asn-Pro-Phe (NPF), Asp-Pro-Phe (DPF) or Gly-Pro-Phe (GPF) tri-peptide motifs. Such interactions are thought to facilitate EHD protein functions, but only a small number of such partners has been identified. To identify novel EH domain-NPF mediated interaction partners, I carried out a proteome-wide bioinformatics analysis to categorize proteins that possess single or multiple putative EH domain-binding motifs and generated a 9-mer peptide library corresponding to motifs containing the N/D/G-P-F sequence together with 6 C-terminal residues found in known EHD protein interaction partners as well as a subset of the previously uncharacterized proteins identified by the bioinformatics analysis selected based on their potential roles in endocytic traffic and related cellular processes. I developed a quantitative high-throughput fluorescence polarization-based competition assay using the EH domain of EHD1 as a prototype and a fluorescent peptide corresponding to a known EHD1 binding motif on MICAL-L1 protein. Using this assay, and the unlabeled peptides in the library in a competition assay, I determined the interaction affinities of the putative peptide motifs for binding to the EH domain of EHD1. This approach helped identify a large number of new EHD interacting partners of which selected candidates were validated using a GST-EH domain pull-down assay. These studies helped markedly expand the potential EHD protein interactome, and implicate the newly identified candidates in multiple cellular functions of EHD1 and potentially other family members

Facile Synthesis of Manganese Oxyhydroxide (MnOOH) Nanowires for the Capacitors Application

A facile method has been introduced to synthesis manganese oxyhydroxide (MnOOH) nanowires synthesized using hydrothermal reaction. The synthesized nanowires have been characterized using different techniques like X-ray diffractogram, infra-red spectroscopy, field emission scanning electron microscopy (FESEM) and electrochemical methods. FESEM results show that the obtained nanowires have very uniform morphology. The electrochemical test shows to have capacitance of 130 F/gm

A comparative study of intracervical foley’s catheter with oxytocin and vaginal misoprostol for induction of labour

Background: Induction of labour implies stimulation of uterine contraction before spontaneous onset of labour with or without ruptured membranes. Aim of this study was to compare the efficacy and safety of intracervical Foley’s catheter with oxytocin and vaginal misoprostol for labour induction at term.Methods: A total 100 term pregnant women were chosen with bishop score 0.05).Conclusions: Misoprostol was found better for successful induction, decreases induction-to-delivery interval and increases vaginal delivery as compared to Foley’s catheter but it needs constant supervision in view of hyperstimulation and tachysystole

An updated review on Physalis peruviana fruit: Cultivational, nutraceutical and pharmaceutical aspects

Plants have always been rich sources of medicinally active constituents in the quest for curing numerous diseases. Among those, Physalis peruviana has been utilized traditionally as a therapeutic (antispasmodic, diuretic, antiseptic, sedative, and analgesic) and nutraceutical herb. It contains numerous active components like essential minerals, α-linolenic acid, iron, vitamins, carbohydrates, phytosterols etc. Its potential as a multifunctional agent in beverages, foods and nutraceutical industries makes it an important crop for consideration. From the agricultural point of view, this fruit is a profitable commercial crop for arid regions also and does not require much effort and investment for cultivation. It easily grows in wild and arid regions. Despite being a nutraceutical and a medicinally important crop, its utilization is not up to the mark. Thus the objective of the present review was to explore and emphasize the nutraceutical and therapeutic potential of Physalis peruviana. It provides exhaustive insight into the origin, distribution, cultivation, harvesting, active constituents and its prospective utility in food, nutrition and pharmaceutical industries

An updated review on Physalis peruviana fruit: Cultivational, nutraceutical and pharmaceutical aspects

97-110Plants have always been rich sources of medicinally active constituents in the quest for curing numerous diseases. Among those, Physalis peruviana has been utilized traditionally as a therapeutic (antispasmodic, diuretic, antiseptic, sedative, and analgesic) and nutraceutical herb. It contains numerous active components like essential minerals, α-linolenic acid, iron, vitamins, carbohydrates, phytosterols etc. Its potential as a multifunctional agent in beverages, foods and nutraceutical industries makes it an important crop for consideration. From the agricultural point of view, this fruit is a profitable commercial crop for arid regions also and does not require much effort and investment for cultivation. It easily grows in wild and arid regions. Despite being a nutraceutical and a medicinally important crop, its utilization is not up to the mark. Thus the objective of the present review was to explore and emphasize the nutraceutical and therapeutic potential of Physalis peruviana. It provides exhaustive insight into the origin, distribution, cultivation, harvesting, active constituents and its prospective utility in food, nutrition and pharmaceutical industries

GCM2 Silencing in Parathyroid Adenoma is associated with Promoter Hypermethylation and Gain of Methylation on Histone 3

PURPOSE: Glial cells missing 2 (GCM2), a zinc finger-transcription factor, is essentially required for the development of parathyroid glands. We sought to identify if the epigenetic alterations in the GCM2 transcription are involved in the pathogenesis of sporadic parathyroid adenoma. In addition, we examined the association between promoter methylation and histone modifications with disease indices. EXPERIMENTAL DESIGN: mRNA and protein expression of GCM2 were analyzed by RT-qPCR and immunohistochemistry in 33 adenomatous and 10 control parathyroid tissues. DNA methylation and histone methylation/acetylation of GCM2 promoter were measured by bisulfite sequencing and ChIP-qPCR. Additionally, we investigated the role of epigenetic modifications on GCM2 and DNA methyltransferase 1 (DNMT1) expression in PTH-C1 cells by treating with 5-aza 2\u27deoxycytidine (DAC) and BRD4770 and assessed for GCM2 mRNA and DNMT1 protein levels. RESULTS: mRNA and protein expression of GCM2 were lower in sporadic adenomatous than in control parathyroid tissues. This reduction correlated with hypermethylation (P\u3c0.001) and higher H3K9me3 levels in GCM2 promoter (P\u3c0.04) in adenomas. In PTH-C1 cells, DAC treatment resulted in increased GCM2 transcription and decreased DNMT1 protein expression, while cells treated with the BRD4770 showed reduced H3K9me3 levels but a non-significant change in GCM2 transcription. CONCLUSION: These findings suggest the concurrent association of promoter hypermethylation and higher H3K9me3 with the repression of GCM2 expression in parathyroid adenomas. Treatment with DAC restored GCM2 expression in PTH-C1 cells. Our results showed a possible epigenetic landscape in the tumorigenesis of parathyroid adenoma and also that DAC may be promising avenues of research for parathyroid adenoma therapeutics

Loss of Cbl and Cbl-b ubiquitin ligases abrogates hematopoietic stem cell quiescence and sensitizes leukemic disease to chemotherapy.

Cbl and Cbl-b are tyrosine kinase-directed RING finger type ubiquitin ligases (E3s) that negatively regulate cellular activation pathways. E3 activity-disrupting human Cbl mutations are associated with myeloproliferative disorders (MPD) that are reproduced in mice with Cbl RING finger mutant knock-in or hematopoietic Cbl and Cbl-b double knockout. However, the role of Cbl proteins in hematopoietic stem cell (HSC) homeostasis, especially in the context of MPD is unclear. Here we demonstrate that HSC expansion and MPD development upon combined Cbl and Cbl-b deletion are dependent on HSCs. Cell cycle analysis demonstrated that DKO HSCs exhibit reduced quiescence associated with compromised reconstitution ability and propensity to undergo exhaustion. We show that sustained c-Kit and FLT3 signaling in DKO HSCs promotes loss of colony-forming potential, and c-Kit or FLT3 inhibition in vitro protects HSCs from exhaustion. In vivo, treatment with 5-fluorouracil hastens DKO HSC exhaustion and protects mice from death due to MPD. Our data reveal a novel and leukemia therapy-relevant role of Cbl and Cbl-b in the maintenance of HSC quiescence and protection against exhaustion, through negative regulation of tyrosine kinase-coupled receptor signaling