Analysis of Beta-Cell Gene Expression Reveals Inflammatory Signaling and Evidence of Dedifferentiation following Human Islet Isolation and Culture

The stresses encountered during islet isolation and culture may have deleterious effects on beta-cell physiology. However, the biological response of human islet cells to isolation remains poorly characterized. A better understanding of the network of signaling pathways induced by islet isolation and culturing may lead to strategies aimed at improving islet graft survival and function. Laser capture microdissection (LCM) was used to extract beta-cell RNA from 1) intact pancreatic islets, 2) freshly isolated islets, 3) islets cultured for 3 days, and changes in gene expression were examined by microarray analysis. We identified a strong inflammatory response induced by islet isolation that continues during in-vitro culture manifested by upregulation of several cytokines and cytokine-receptors. The most highly upregulated gene, interleukin-8 (IL-8), was induced by 3.6-fold following islet isolation and 56-fold after 3 days in culture. Immunofluorescence studies showed that the majority of IL-8 was produced by beta-cells themselves. We also observed that several pancreas-specific transcription factors were down-regulated in cultured islets. Concordantly, several pancreatic progenitor cell-specific transcription factors like SOX4, SOX9, and ID2 were upregulated in cultured islets, suggesting progressive transformation of mature beta-cell phenotype toward an immature endocrine cell phenotype. Our findings suggest islet isolation and culture induces an inflammatory response and loss of the mature endocrine cell phenotype. A better understanding of the signals required to maintain a mature beta-cell phenotype may help improve the efficacy of islet transplantation

AN ENHANCED SCHEDULING APPROACH WITH CLOUDLET MIGRATIONS FOR RESOURCE INTENSIVE APPLICATIONS

Cloud computing is one of the most advanced technologies to present computerized generation. Scheduling plays a major role in it. The connectivity of Virtual Machines (VM) to schedule the assigned tasks (cloudlet) is a most attractive field to research. This paper introduces a confined Cloudlet Migration based scheduling algorithm using Enhanced-First Come First Serve (CMeFCFS). The objective of this work is to minimize the makespan, cost and to optimize the resource utilization. The proposed work has been simulated in the CloudSim toolkit package. The results have been compared with pre-existing scheduling algorithms with same experimental configuration. Important parameters like execution time, completion time, cost, makespan and utilization of resources are compared to measure the performance of the proposed algorithm. Extensive simulation results prove that introduced work has better results than existing approaches. 99.8% resource utilization has been achieved by CMeFCFS. Plotted graphs and calculated values show that the proposed algorithm is very effective for cloudlet scheduling

Preliminary Physiochemical Evaluation of Nagaradi Ointment: An Ayurvedic Formulation

Plants serve as a very rich source of bioactive molecules, which are used to treat various acute and chronic diseases in Ayurveda. Medicinal plant materials are formulated into various types of Ayurvedic medicine either by ancient or modern methods where standardization plays a pivotal role for authentication. Standardization confirms the identity, quality and purity of drugs. WHO has set up various parameters to evaluate the crude drugs and their finished products. Now a day’s application of several modern analytical techniques has become inevitable for evaluating quality, safety and efficacy of the polyherbal Ayurvedic formulations. Out of several formulations available in Ayurveda, Anjana is considered as unique in Shalakya Tantra due to its various forms (which are prepared by different ways). This article is about the formation of Nagaradi Anjana as ointment which can be taken as a form of Raskriyanjana. Most of the ingredients of Nagaradi ointment are Chakshushya i.e., good for eyes and also have Kaphashamak properties. That is why they are useful in ocular diseased conditions which have dominance of Kapha dosha like Kaphaja Abhishyanda. Keeping all these points in view, the present study has been undertaken with the aim to make the Ayurvedic formulation in the form of Raskriya Anjana or ointment, from the herbal drugs, mentioned in Astanga Sangrah, an Ayurvedic classical text and to develop the physiochemical profile of it. The eye ointment was prepared by using vaseline as base which is mixed with powdered Ghana Satva of the herbal drugs

A proximal tissue-specific module and a distal negative regulatory module control apolipoprotein(a) gene transcription.

The apo(a) [apolipoprotein(a)] gene is responsible for variations in plasma lipoprotein(a), high levels of which are a risk factor for atherosclerosis and myocardial infarction. The apo(a) promoter stimulates the expression of reporter genes in HepG2 cells, but not in HeLa cells. In the present study, we demonstrate that the 1.4 kb apo(a) promoter comprises two composite regulatory regions: a distal negative regulatory module (positions -1432 to -716) and a proximal tissue-specific module (-716 to -616). The distal negative regulatory module contains two strong negative regulatory regions [polymorphic PNR (pentanucleotide repeat region) and NREbeta (negative regulatory element beta)], which sandwich the postive regulatory region PREbeta (positive regulatory element beta). The PNR was shown to bind to transcription factors in a tissue-specific manner, whereas the ubiquitous transcription factors hepatocyte nuclear factor 3alpha and GATA binding protein 4 bound to NREbeta to repress gene transcription. The proximal tissue-specific module contains two regulatory elements: an activating region (PREalpha) that activates transcription in HepG2 cells, and NREalpha, which is responsible for repressing the apo(a) gene in HeLa cells. NREalpha binds to a HeLa-specific repressor. These multiple regulatory elements might work co-operatively to finely regulate apo(a) gene expression. Although the tissue-specific module is required for apo(a) gene activation and repression in a tissue-specific manner, the combinatorial interplay of the distal and proximal regulators might define the complex pathway(s) of apo(a) gene regulation

Hierarchical clustering of differentially expressed genes in fresh (d0) and cultured islets (d3).

<p>The expression data for intact islets and isolated islets (d0-islets & d3-islets) were analyzed and top 359 genes with p<0.05 and fold change >2.5 were used to create cluster map.</p

NFKB target genes from top 15 upregulated genes and differentially expressed cytokines and chemokines.

<p>(A) Analysis of the top fifteen upregulated genes from both groups (d0 and d3) revealed that 6/15 genes have NFKB binding sites and, (B) among 39 differentially expressed cytokines/chemokines from d3-islets 22 genes have NFKB binding sites.</p