E-Governance Implementation: Challenges of Effective Service Delivery in Civil Service of Nepal

Civil servants are often charged not providing effective services though they have sufficient resources and technological knowhow. The author is trying to explore the challenges of effective service delivery in civil service as implementing e-governance in Nepal. Survey method was used to generate data and adopted quantitative research technique. The study has claimed that Nepal has been facing numerous challenges during service delivery while implementing e-governance. Infrastructure development, human resource development and management, digital divide, are identified as the major challenges. Unnecessary influence of middleman and syndicate created by some hidden groups make government service holdup. Needless expansion of government agencies and its employees; too many layers in decision-making process; more process oriented service delivery rather than result oriented, failure to make individual officials responsible; and a lack of decentralization of necessary authority to the officials are the other challenges

Modulation of mitochondrial dynamics, bioenergetics and the innate antiviral immune response in Zika Virus infected Retinal cells

Modulation of mitochondrial dynamics and bioenergetics in Zika Virus infected retinal cells Karim Dirani1, Sneha Singh1, Shailendra Giri2, Ashok Kumar1 1 Department of Ophthalmology, Visual and Anatomical Sciences, and Kresge Eye Institute, Wayne State University School of Medicine, Detroit, MI 2 Department of Neurology, Henry Ford Health Systems, Detroit, MI Purpose: In addition to the powerhouse of the cells and regulation of apoptosis, mitochondria are being increasingly recognized in playing crucial role in modulating innate immune response in viral infections. In this study, we sought to determine the impact of mitochondrial dynamic (fission, fusion, and mitophagy) and bioenergetics status in regulating antiviral innate response to Zika virus (ZIKV) infection. Methods: Human ARPE-19 cells were infected with ZIKV (strain PRVABC59) at MOI of 1. The mitochondria were visualized for fission and fusion process using TOM20 and mitotracker dyes while the mitochondrial membrane potential was measured using JC-1 staining. The temporal expression of key genes/molecules involved in mitochondrial dynamic was assessed by RT-qPCR and Western blot. ARPE-19 cells stably expressing individual ZIKV genes were used to assess the functional role of viral proteins in altering mitochondrial dynamics. The bioenergetics activity of the cells was assessed using Seahorse XF analyzer for monitoring oxygen consumption rate (OCR) and Extracellular acidification rate (ECAR) upon ZIKV infection and with individual viral proteins. Results: All cells tested were permissive to ZIKV infection, as evidenced by a time dependent increase in virion production and immunostaining of viral antigen in the cells. Infected cells exhibited an increase in the expression of antiviral (e.g., IFNs, ISG15, MX1, OAS2) and inflammatory mediators (e.g., CCL5, TNFa). Amongst the molecules regulating the mitochondrial fission and fusion, levels of p-Drp1, mitofusins (MFN1, MFN2), and Opa1 were altered at both mRNA and protein levels. Similarly, molecules regulating mitophagy, PINK1/Parkin -dependent and independent pathways were modulated in ZIKV infected cells. Among individual viral proteins, the non-structural proteins exerted profound effects on mitochondrial dynamic. This correlated with a decrease in OCR response upon ZIKV infection. Interestingly, the pharmacological inhibition of Drp1 reduced ZIKV replication. Conclusion: Collectively, our data indicates that ZIKV virus modulates mitochondrial dynamics and individual ZIKV proteins exert differential effects on bioenergetics and antiviral innate immune response of infected cells. Targeting mitochondrial dynamic may provide new therapeutic avenues for antiviral therapy against ZIKV infection

Inhibition of Angiogenesis Associated with Ovarian Cancer By Nanoparticles of Cerium Oxide (CIP)

The invention provides a method of treating ovarian cancer in a mammal, the method inhibits angiogenesis associated with the cancer and comprises the parenteral administration to the mammal of an effective amount of cerium oxide nanoparticles having a predominance of Ce+3

15-deoxy-delta12,14-prostaglandin J2 attenuates endothelial-monocyte interaction: implication for inflammatory diseases

<p>Abstract</p> <p>Background</p> <p>The Infiltration of leukocytes across the brain endothelium is a hallmark of various neuroinflammatory disorders. Under inflammatory conditions, there is increased expression of specific cell adhesion molecules (CAMs) on activated vascular endothelial cells which increases the adhesion and infiltration of leukocytes. TNFα is one of the major proinflammatory cytokines that causes endothelial dysfunction by various mechanisms including activation of transcription factor NF-κB, a key transcription factor that regulates expression of CAMs. Peroxisome proliferator-activated receptor gamma (PPARγ) is a member of the nuclear hormone superfamily of ligand-activated transcriptional factors. 15-deoxy-δ 12, 14-prostaglandin J2 (15d-PGJ2) is a well recognized natural ligand of PPARγ and possesses anti-inflammatory properties both <it>in vitro </it>and <it>in vivo</it>. This study aims to elucidate the mechanism of 15-PGJ2 on the adhesion of mononuclear cells to activated endothelial cells.</p> <p>Methods</p> <p>To delineate the signaling pathway of 15d-PGJ2 mediated effects, we employed an <it>in vitro </it>adhesion assay model of endothelial-monocyte interaction. Expression of CAMs was examined using flow cytometry and real time PCR techniques. To define the mechanism of 15d-PGJ2, we explored the role of NF-κB by EMSA (Electrophoretic Mobility Shift Assay) gels, NF-κB reporter and p65-transcriptional activities by transient transfection in the brain-derived endothelial cell line (bEND.3).</p> <p>Results</p> <p>Using an <it>in vitro </it>adhesion assay model, we demonstrate that 15d-PGJ2 inhibits TNFα induced monocyte adhesion to endothelial cells, which is mediated by downregulation of endothelial cell adhesion molecules in a PPARγ independent manner. 15d-PGJ2 modulated the adhesion process by inhibiting the TNFα induced IKK-NF-κB pathway as evident from EMSA, NF-κB reporter and p65 mediated transcriptional activity results in bEND.3 cells.</p> <p>Conclusion</p> <p>These findings suggest that 15d-PGJ2 inhibits inflammation at multiple steps and thus is a potential therapeutic target for various inflammatory diseases.</p

5-Aminoimidazole-4-Carboxamide-1-β-4-Ribofuranoside Inhibits Proinflammatory Response in Glial Cells: A Possible Role of AMP-Activated Protein Kinase

AMP-activated protein kinase (AMPK) is tightly regulated by the cellular AMP:ATP ratio and plays a central role in the regulation of energy homeostasis and metabolic stress. A pharmacological activator of AMPK, 5-amino-4-imidazole carboxamide riboside (AICAR) inhibited lipopolysaccharide (LPS)-induced expression of proinflammatory cytokines (tumor necrosis factor α, interleukin-1β, and interleukin-6) and inducible nitric oxide synthase in primary rat astrocytes, microglia, and peritoneal macrophages. AICAR attenuates the LPS-induced activation of nuclear factor κB via downregulation of IκB kinase α/β activity. It also inhibits nuclear translocation of CCAAT/enhancer-binding protein (C/EBP) transcription factor by inhibiting the expression of C/EBP-δ in brain glial cells. The dominant negative form of AMPKα2 (D157A) and its antisense documents a possible role of AMPK in the regulation of the cellular proinflammatory process. AICAR also inhibited the production of inflammatory mediators in serum and their expression in CNS of rats injected with a sublethal dose of LPS by intraperitoneal injection. These observations in cultured cells as well as in the animal model suggest that AICAR may be of therapeutic value in treating inflammatory diseases

An extract of Artemisia dracunculus L. stimulates insulin secretion from β cells, activates AMPK and suppresses inflammation

© 2015 Elsevier Ireland Ltd. All rights reserved. Ethnopharmacological relevance Artemisia dracunculus L. (Russian tarragon) is a perennial herb belonging to the family Compositae and has a history of medicinal use in humans, particularly for treatment of diabetes. Aim of the study: In this study a defined plant extract from A. dracunculus L. (termed PMI-5011) is used to improve beta(β) cells function and maintain β cell number in pancreatic islets as an alternative drug approach for successful treatment of diabetes. Materials and methods Mouse and human pancreatic beta cells were treated with defined plant extract of A. dracunculus L. (PMI-5011) to understand the mechanism(s) that influence beta cell function and β cell number. Results We found that the PMI-5011 enhances insulin release from primary β cells, isolated mouse and human islets and it maintains β cell number. Insulin released by PMI-5011 is associated with the activation of AMP-activated protein kinase (AMPK), and protein kinase B (PKB). Furthermore, PMI-5011 suppresses LPS/INFγ-induced inflammation and inflammatory mediator(s) in macrophages. PMI-5011 inhibited Nitric oxide (NO) production and expression of inducible nitric oxide synthase (iNOS) at the protein level and also attenuated pro-inflammatory cytokine (IL-6) production in macrophages. Conclusion PMI-5011 has potential therapeutic value for diabetes treatment via increasing insulin release from β cells and decreases capacity of macrophages to combat inflammation

Inhibition of NF-κB activation by 5-lipoxygenase inhibitors protects brain against injury in a rat model of focal cerebral ischemia

BACKGROUND: Stroke is one of the leading causes of death worldwide and a major cause of morbidity and mortality in the United States of America. Brain ischemia-reperfusion (IR) triggers a complex series of biochemical events including inflammation. Leukotrienes derived from 5-lipoxygenase (5-LOX) cause inflammation and are thus involved in the pathobiology of stroke injury. METHODS: To test the neuroprotective efficacy of 5-LOX inhibition in a rat model of focal cerebral IR, ischemic animals were either pre- or post-treated with a potent selective 5-LOX inhibitor, (N- [3-[3-(-fluorophenoxy) phenyl]-1-methyl-2-propenyl]-N-hydroxyurea (BW-B 70C). They were evaluated at 24 h after reperfusion for brain infarction, neurological deficit score, and the expression of 5-LOX. Furthermore, the mechanism and the anti-inflammatory potential of BW-B 70C in the regulation of nuclear factor kappa B (NF-κB) and inflammatory inducible nitric oxide synthase (iNOS) were investigated both in vivo and in vitro. RESULTS AND DISCUSSION: Both pre- and post-treatment with BW-B 70C reduced infarctions and improved neurological deficit scores. Immunohistochemical study of brain sections showed IR-mediated increased expression of 5-LOX in the neurons and microglia. BW-B 70C down-regulated 5-LOX and inhibited iNOS expression by preventing NF-κB activation. Two other structurally different 5-LOX inhibitors were also administered post IR: caffeic acid and 2, 3, 5-trimethyl-6- [12-hydroxy-5, 10-dodecadiynyl]-1, 4-benzoquinone (AA-861). As with BW-B 70C, they provided remarkable neuroprotection. Furthermore, in vitro, BW-B 70C inhibited lipopolysaccharide (LPS) mediated nitric oxide production, iNOS induction and NF-κB activation in the BV2 microglial cell line. Treating rat primary microglia with BW-B70C confirmed blockage of LPS-mediated translocation of the p65 subunit of NF-κB from cytosol to nucleus. CONCLUSION: The study demonstrates the neuroprotective potential of 5-LOX inhibition through down-regulation of NF-κB in a rat model of experimental stroke

AICAR inhibits adipocyte differentiation in 3T3L1 and restores metabolic alterations in diet-induced obesity mice model

BACKGROUND: Obesity is one of the principal causative factors involved in the development of metabolic syndrome. AMP-activated protein kinase (AMPK) is an energy sensor that regulates cellular metabolism. The role of AMP-activated protein kinase in adipocyte differentiation is not completely understood, therefore, we examined the effect of 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside (AICAR), a pharmacological activator of AMP-activated protein kinase (AMPK) on adipocyte differentiation in 3T3L1 cells and in a mouse Diet induced obesity (DIO) model. METHODS: To examine the effect of AICAR on adipocyte differentiation in 3T3L1 cells and in a mouse Diet induced obesity (DIO) model, 3T3L1 cells were differentiatied in the presence or absence of different concentration of AICAR and neutral lipid content and expression of various adipocyte-specific transcription factors were examined. In vivo study, treated and untreated mice with AICAR (0.1–0.5 mg/g body weight) were fed high-fat diet (60% kcal% fat) to induce DIO and several parameters were studied. RESULTS: AICAR blocked adipogenic conversion in 3T3L1 cells along with significant decrease in the neutral lipid content by downregulating several adipocyte-specific transcription factors including peroxisome proliferators-activated receptor γ (PPARγ), C/EBPα and ADD1/SREBP1, which are critical for adipogenesis in vitro. Moreover, intraperitoneal administration of AICAR (0.5 mg g/body weight) to mice fed with high-fat diet (60% kcal% fat) to induce DIO, significantly blocked the body weight gain and total content of epididymal fat in these mice over a period of 6 weeks. AICAR treatment also restored normal adipokine levels and resulted in significant improvement in glucose tolerance and insulin sensitivity. The reduction in adipose tissue content in AICAR treated DIO mice was due to reduction in lipid accumulation in the pre-existing adipocytes. However, no change was observed in the expression of PPARγ, C/EBPα and ADD1/SREBP1 transcription factors in vivo though PGC1α expression was significantly induced. CONCLUSION: This study suggests that AICAR inhibits adipocyte differentiation via downregulation of expression of adipogenic factors in vitro and reduces adipose tissue content in DIO mice by activating expression of PGC1α without inhibiting adipocyte-specific transcription factors in DIO mice

Dietary Isoflavones Alter Gut Microbiota and Lipopolysaccharide Biosynthesis to Reduce Inflammation

The etiopathogenesis of multiple sclerosis (MS) is strongly affected by environmental factors such as diet and the gut microbiota. An isoflavone-rich (ISO) diet was previously shown to reduce the severity of MS in the animal model experimental autoimmune encephalomyelitis (EAE). Translation of this concept to clinical trial where dietary isoflavones may be recommended for MS patients will require preliminary evidence that providing the isoflavone-rich diet to people with MS (PwMS) who lack phytoestrogen-metabolizing bacteria has beneficial effects. We have previously shown that the gut microbiota of PwMS resembles the gut microbiota of mice raised under a phytoestrogen-free (phyto-free) diet in that it lacks phytoestrogen-metabolizing bacteria. To investigate the effects of phytoestrogens on the microbiota inflammatory response and EAE disease severity we switched the diet of mice raised under a phyto-free (PF) diet to an isoflavone-rich diet. Microbiota analysis showed that the change in diet from one that is ISO to one that is PF reduces beneficial bacteria such as Bifidobacterium species. In addition we observed functional differences in lipopolysaccharide (LPS) biosynthesis pathways. Moreover LPS extracted from feces of mice fed an ISO diet induced increased production of anti-inflammatory cytokines from bone marrow-derived macrophages relative to fecal-LPS isolated from mice fed a PF diet. Eventually mice whose diet was switched from a PF diet to an ISO diet trended toward reduced EAE severity and mortality. Overall we show that an isoflavone-rich diet specifically modulates LPS biosynthesis of the gut microbiota imparts an anti-inflammatory response and decreases disease severity