Microbiology Commons^™

Metabolism Of Herpes Simplex Virus 1 Infected Raw 264.7 Macrophages, Mary K. Jenkins

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Macrophages are immune cells that phagocytize pathogens and act as antigen presenting cells. Macrophages are critical in regulating the adaptive immune response and play a key role in neutralizing infection. Macrophages exhibit a variety of phenotypes which have different energy requirements. In the case of HSV-1 infection the virus has been shown to alter metabolic processes of its host. Utilizing a XF24 Extracellular Flux Analyzer the metabolic activity of M0, M1, and M2 (IL-4, IL-13, and IL-10) RAW 264.7 macrophages was quantified for both uninfected cells and in response to HSV-1 infection. The analysis showed uninfected M0 and M2 (IL-4, …

The Anti-Apoptotic Effect Of Hsv-1 On Murine Macrophages: Raw 246.7 Murine Macrophage Cell Line, Mofeda Abdussalam Alhanghari

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Herpes simplex virus type 1 (HSV-1) is a worldwide pathogen that affects humans and has the ability to establish a latent state of infection in the sensory nerve ganglia after primary infection of epithelial cells (Boutell and Everett, 2003). HSV-1 is a very contagious virus, which can be transmitted from person to person and cause cold sores in the infected person. Rarely, infection can lead to more serious complications, such as encephalitis. Most HSV-1 infections usually occur in childhood with lifelong potential for symptomatic or asymptomatic viral shedding episodes (Looker et al., 2015). HSV- 1 infects 60%-80% of people throughout …

Delineating ΔNp63α'S Function In Epithelial Cells, Suraj Sakaram

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ΔNp63α, the most physiologically relevant isoform of p63, a p53 family member protein, maintains the proliferative capacity of the basal layer of epithelial tissues and is highly expressed in non-melanoma skin cancer. It has been implicated in a variety of different cancers, including breast, lung, and prostate cancers, however the complex role it plays in disease remains poorly understood. Elucidating ΔNp63α’s regulatory network may provide insight into its role in epithelial tissues and in cancer progression. The objective of this thesis is to investigate ΔNp63α’s upstream regulation in the context of non-melanoma skin cancer as well as expand on ΔNp63α’s …

Species Differences In The Effects Of Tcdd On A Transcriptional Regulatory Region Within The Ig Heavy Chain Gene, Zahra J. Alfaheeda

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The immunoglobulin heavy chain gene (Igh) is regulated by numerous regulatory elements including the 3'Igh regulatory region (3'IghRR). Several transcription factors are involved in modulating the 3'IghRR including NF-B, AP-1, and the aryl hydrocarbon receptor (AhR). The AhR is a ligand-activated transcription factor that mediates the transcription of genes involved in the metabolism of environmental toxicants such as TCDD. TCDD binds AhR and regulates immunoglobulin (Ig) expression in B cells of animal models. This modulation appears to be directly mediated by binding of the AhR to dioxin response elements (DRE) within the 3'IghRR. Structural differences have been found between the …

Comparing Antibody-Coated Immune Beads With Flow Cytometry To Measure Ss-2-Microglubulin+ Murine Spleen Cells, Amal Fahad Alajman

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In diseases like AIDS, Multiple Myeloma, Multiple Sclerosis (MS), Type 1 Diabetes, and Systemic Lupus Erythromatosus the detection of major histocompatibility complex type 1 molecules (MHC 1) can be a helpful component of disease diagnosis and prognosis. Most somatic mammalian cells display to varying degrees major histocompatibility complex (MHC) class 1 antigen on the cell surface. MHC1 molecules consist of a polymorphic alpha a chain and a monomorphic beta chain, beta 2 microglubulin (ß2m). The ß2M composition is fairly constant within a species whereas the alpha chain is not. The polymorphic alpha chain is encoded by an MHC complex gene …

The Impact Of Hsv-1 Infection, Socs1 Peptide, And Socs3 Peptide Mimetic On Cell Viability, Morphology, And Cytoskeleton Proteins Of Unpolarized And Cytokine-Polarized M1 Raw 264.7 Murine Macrophages, Mubarak Almutairi

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Mubarak Huraysan Almutairi. M.S. Department of Microbiology and Immunology, Wright State University, 2016. The Impact of HSV-1 Infection, SOCS1 peptide, and SOCS3 peptide mimetic on Cell Viability, Morphology, and Cytoskeleton Proteins of Unpolarized and Cytokine-Polarized M1 RAW 264.7 Murine Macrophages. The immune response against HSV-1 involves macrophages in both innate and adaptive immunity by limiting HSV-1 replication. In this study, the effects of HSV-1 challenge on cell viability, morphology, and cytoskeletal filament in unpolarized and cytokine-polarized murine RAW 264.7 macrophages at 24 and 48 hours were monitored. Because the distribution of cytoskeleton throughout the cell is critical in cell viability …

Correlating Innate Functional Recovery From Stroke Either With Stem Cell Proliferation And/Or Limb Rehabilitation, Devipriyanka Nagarajan

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In the present study 10-12 month female rats were examined for functional recovery from stroke and this recovery was compared with the stem cell/progenitor cell proliferation in the brain (which was measured by Ki67). The cell proliferation indicated by Ki67 showed a 6 fold increase in control animals compared to the rehabilitation animals. The contralateral functional recovery in control animals were 46.6% and in the rehabilitation animals were 24.5%. The physical rehabilitation was carried out to determine if limb rehabilitation can promote greater functional recovery. The results showed that when the animals were made to over use their impaired limb …

Immunotherapy For Human Breast Cancer, Nasrah Ali Al Kamal

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Al Kamal, Nasrah. M.S. Program of Microbiology and Immunology, Wright State University, 2015.Immunotherapy for Human Breast Cancer. The study focuses on the methodologies that are used in breast cancer therapy. The review finds major evolutions of these methods and the influence they have made on the increased survival rates in cancer patients. There is also a projection of the processes that are involved in the therapy practices and the way the protocols have been improved over time. The various stages of cancer treatment project the various aspects of better performance articulations in the therapy sector. There is also reference to …

Application Of Fluorescence In Situ Hybridization For Visualization And Quantification Of Human Gastrointestinal Microbiota, Alexander M. Gordon

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The microbiota of the human gastrointestinal tract is the focus of current research due to their role in human health and disease. Modern methods characterize the communities of gut microbiota through the use of culture independent techniques. Technologies such as microarrays and next generation sequencing (NGS) determine microbial profiles by analyzing the pool of 16S ribosomal small subunit RNA genes in the community. These techniques operate through the measurement of genomic content rather than through direct analysis of cells. This approach predisposes the methods to discrepancies and bias. In contrast, the use of fluorescence in situ hybridization (FISH) to analyze …

Determining The Role Of The Ahr In Immunoglobulin Expression And Class Switch Recombination, Bassam Fawaz Kashgari

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The aryl hydrocarbon receptor (AhR) is a ligand-activated cytosolic transcription factor that regulates xenobiotic-metabolizing enzymes. It mediates the toxicity of various environmental chemicals such as 2,3,7,8-tetracholorodibenzo-p-dioxin (TCDD). TCDD inhibits the differentiation of B cells into antibody-secreting cells and inhibits immunoglobulin (Ig) expression in various animal models. We have previously determined that TCDD-induced inhibition of the mouse Ig heavy chain gene (mo-Igh) is AhR-dependent. This inhibition may be mediated by binding of the AhR to dioxin response elements (DREs) within the 3'Igh regulatory region (3'IghRR) and inhibition of 3'IghRR activity, a significant transcriptional regulator of Ig expression. However, there are structural …

Silver Nanoparticles Inhibit The Binding And Replication Of Dengue Virus, Kelley J. Williams

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Dengue is an emerging hemorrhagic fever virus and widely considered the most important arbovirus in the world. The CDC and the World Health Organization estimates Dengue virus (DENV) infects 50-400 million people annually in the tropical and subtropical regions of the world. More than 500 thousand of these will develop severe infection and approximately 22 thousand will lead to death. Dengue virus (DENV) is a positive-sense RNA virus that exists in 4 antigenic serotypes. An immunological phenomenon called antibody-dependent enhancement (ADE) leaves a DENV victim vulnerable to increased risk of subsequent infections. Secondary infections with DENV are known to increase …

The Impact Of Hsv-1 Infection On Cell Viability, Morphology, And Cd Markers Expression By Unpolarized And Cytokine-Polarized J774a.1 Mouse Macrophages, Sarah Saad Alsharif

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Macrophages play an important role in the immune system, particularly in neutralizing pathogens via phagocytosis and the production of multiple cytokines and chemokines that control infection after exposure to specific stimuli. Macrophages exhibit two different phenotypes, M1 and M2. This study evaluated the role of Suppressors of cytokine signaling (SOCS)1 and SOCS3 on Herpes Simplex Virus (HSV-1) infection of polarized macrophages, cell viability, cell morphology, and the expression of cell surface CD markers. I hypothesized that J774A.1 murine macrophages cells in the naive state (M0), and M1 and M2 phenotypes would display differences in CD markers CD80,CD163, and CD200R, cell …

Inhibition Of Nectin-1 And Herpes Virus Entry Mediator (Hvem) Using Monoclonal Antibodies Decreases Hsv-1 Entry Into Neuro-2a Cells, Erica Marie Rinehart

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This study examines the effects of blocking the nectin-1 and Herpes Virus Entry Mediator (HVEM) receptors on Neuro-2a (N2a) cells in order to prevent HSV-1 infection. These two receptors have been identified as a primary and secondary mode of entry for HSV-1. Since there was some debate about the correct serum level to be used in growth media for healthy N2a cells, three concentrations (2%, 5%, 10%) were used to examine relative growth and neurite development, in addition to examining possible differences in infectivity levels in all other experiments. There was a difference in general morphology, with neurite abundance and …

The Use Of Antibody-Coated Latex Beads To Determine Single Positive And Double Positive Mouse Spleen Cells Expressing Cd5 And/Or Cd19 Glycoproteins, Abdulrazzag Abdulaziz Othman

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Flow cytometry is the standard method used to diagnose, stage, and monitor patients' response to the treatment given by counting the numbers of CD5, CD19 and CD5+ CD19+ B lymphocytes. In this study, a comparison was done between numbers of single CD5+, single CD19+ and dual CD5+ CD19+ mouse spleen B lymphocytes using flow cytometry and antibody-latex beads. The bead method involved antibody-coated latex bead and yielded results similar to those of flow cytometry. For cells exhibiting both markers (CD5+ CD19+), the bead method used antibody-coated beads of two different colors yielded similar results to those of flow cytometry results. …

Morphometric Studies On Subadult Liza Subviridis And Sillago Sihama From Sonmiani Bay (Miani Hor), Balochistan, Pakistan, Muhammad A. Gondal, Naureen A. Qureshi, Noor U. Saher

Journal of Bioresource Management

Length-weight relationship of Liza subviridis and Sillago sihama (Family: Mugilidae; Class: Pisces and Family: Sillaginidae; Class: Pisces) in subadults sampled from Somiani Bay Balochistan, Pakistan during 2002, 2003 and 2006 was analyzed. Liza subviridis showed positive allometry (3.23) in 2002, and negative allometry during 2003 (2.95) and 2006 (1.95). Sub adults of Sillago sihama showed positive allometry during 2002 (3.10) and 2003 (3.13). An isometric condition (3.02) was observed in the samples of Sillago sihama collected from Bhaira in 2006.

On Individual, Sex And Age Differentiation Of Indian House Crow (Corvus Splendens) Call: A Preliminary Study In Potohar, Pakistan, Waqas Hameed, Inayat Ullah, Safia Janjua, Fakhar -I- Abbas, Afsar Mian

Journal of Bioresource Management

Considering importance of acoustics studies in population biology, 500 calls of the Indian House Crow (Corvus splendens) were recorded in morning - mid-afternoon hours (January-February, 2009) from 23 sites of urban areas of Potahar (Punjab, Pakistan). Calls were recorded using Sony CFS 1030 S sound records (sampling rate = 48 KHz) and edited using Sound Analysis Pro (Version 1.02). software using FFT method rate 50%, data window 9.27 ms, advanced window 1.36 ms. Through editing of calls, we selected 60 (37 ♂♂, 17 ♀♀, 6 Juvenile ♂♂) good quality spectrograms for detailed analysis. Spectrograms were characterized by rapid frequency modulations …

Review: Pollination, Pollinated And Pollinators Interaction In Pakistan, Mohammad Irshad, Elizabeth Stephen

Journal of Bioresource Management

The agriculture sector is important in the overall economy and export earnings of Pakistan. Pollination is an essential ecosystem service that depends on symbiosis between species, the pollinated and the pollinator. Animal mediated pollination contributes to the sexual production of over 90% species of modern angiosperms. Effective pollination results in increased crop production, quality improvement and more seed production. Many fruits, vegetables, edible oil crops, stimulant crops and nuts are highly dependent on bee pollination. Worldwide value of pollinators is €153 billion (217 billion US dollars). The production value of pollinated dependent crop in Pakistan is quantified to be 1.59 …

Research Note: Contribution To Mushroom Flora Of Rawalpindi-Islamabad, Pakistan, Kishwar Sultana, Naveeda Riaz, Gulshan Irshad, Ali Nauman Khan

Journal of Bioresource Management

Seven hymenomycetous fungi (mushrooms) were collected from the twin cities area during July 2008. These were identified as the members of the order Agaricales, class Basidiomycetes. They belong to five genera: *Clitocybe fragrans, *Collybia cookie, *Coprinus radians, *Coprinus sterculinus, *Volvariella bingensis, Volvariella parvula, and Termitomyces microcarpus. Five of them, marked with asterisk were reported for the first time from Pakistan.

Diagnostic Accuracy Of C-Reactive Protein In Neonatal Sepsis, Sidra Younis, Muhammad Ali Sheikh, Amjad Ali Raza

Journal of Bioresource Management

Sepsis is the most common cause of neonatal mortality and is responsible for 30-50% of total neonatal deaths each year in developing countries. The objective of the study was to determine the diagnostic accuracy of C-reactive protein (CRP) in neonatal sepsis. Fifty nine consecutive patients with risk factors and clinical features suggestive of CRP sepsis were selected as per operational definition and fulfilling the inclusion and exclusion criteria. Detailed physical examination was carried out. Blood sample for culture and CRP was taken from all the patients. Results of blood culture and CRP were noted down in the performa. Statistical analysis …

Differentiation Of U-937 Monocytes To Macrophage-Like Cells Polarized Into M1 Or M2 Phenotypes According To Their Specific Environment: A Study Of Morphology, Cell Viability, And Cd Markers Of An In Vitro Model Of Human Macrophages, Fatma Husien S. Abdulhadi

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In this study, the human leukemic monocyte lymphoma cell line U-937 was used as an in vitro model for monitoring monocyte/macrophage differentiation. Phorbol 12- myristate (13) (PMA) was used to activate U-937 cells into macrophage-like cells (M0). After 24 hours of PMA treatment, non- adherent U-937 cells became tightly adherent to the culture plates forming M0 cells. M0 cells were then polarized into the M1 macrophage phenotype by treatment with LPS and IFN-γ for another 24 hours. Each of the cytokines IL-4, IL-13, or IL-10 was applied separately to three M0 cultures for 24 hours to induce the M2 macrophage …

Socs1/Socs3 Expression And Virus Replication Of Denv2 And Hsv-1 In Cytokine-Polarized Subsets Of Raw 264.7 Macrophages, Nagarjuna Reddy Cheemarla

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Cytokine activation of macrophages leads to "macrophage polarizatio" The M1 subset is polarized by exposure to lipopolysaccharide (LPS) and interferon-gamma (IFN-γ) and M2 polarized cells develop after exposure to interleukin-4 (IL-4) or IL-13. In this study, the RAW 264.7 murine macrophage cell line was used to study the effect herpes simplex virus-Type I (HSV-1) and Dengue virus serotype-2 (DENV2) infections have on cultures of unpolarized (M0) or polarized subsets. The macrophage subsets were characterized using cluster of differentiation markers CD14 and CD86. Uninfected M1 macrophages showed up regulation in expression of the co-stimulatory factor B7.2 (CD86) compared to expression by …

Development Of A Co-Culture System To Mimic The Transfection Of Hsv-1 From Keratinocytes To Neuronal Cells, David A. Dixon

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To more closely mimic the in vivo progression of HSV-1 a novel in vitro method was created. In this study HEL-30 keratinocytes were infected with a 0.1 MOI of HSV-1 and plated unto a membrane with 8μm diameter pores. The membrane insert was then placed into culture wells that contained Neuro-2a cells (N2A). The neurites from the neuronal cells made cell-to-cell contact with the infected HEL-30 cells and the virus was transmitted into the neuronal cells. To determine if infection occurred in the N2A cells the cells were lysed and the lysate incubated with Vero cells to titrate virus plaque …

White Blood Cell Counts, Parasite Prevalence, And Plasma Cortisol Levels Of Dogs In A County Animal Shelter: Changes Over Days And Impact Of A Program Of Repeated Human Interaction, Emily S. Dudley

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Animal shelter housing is highly stressful for a dog, compromising welfare and leading to undesirable behaviors and unknown health consequences. We documented the changes in circulating numbers of white blood cells, plasma cortisol, and fecal parasite shedding of dogs housed for 10 days at a county animal shelter. White blood cell changes were most prominent on Day 10 after arrival to the shelter. Changes included increased total leukocytes, mature neutrophils, and lymphocytes, with less consistent increases in monocytes and neutrophil to lymphocyte ratio (N:L). Fecal parasite shedding was elevated and not affected by day. Cortisol levels of shelter dogs declined …

Identification Of The Sites Of Action Of Inhibitors Of Mammalian Phospholipase D2 (Pld2) And The Role Of Interacting Protein Partners, Ramya Ganesan

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Phospholipase D (PLD) is a key enzyme for the remodeling of phospholipids in the cell membrane. PLD has been implicated in many physiological functions such as chemotaxis and phagocytosis as well as pathological functions such as ischemia/reperfusion and cancer metastasis. Several small molecule inhibitors of PLD have been recently developed to overcome these pathological effects. We have concentrated on 5-Fluoro-2-Indolyl des-Chlorohalopemide (FIPI) which is an indole derivative of halopemide that inhibits the two mammalian isoforms PLD1 and PLD2. We also concentrate on N-[2-[1-(3-Fluorophenyl)-4-oxo-1,3,-8-triazaspiro[4.5]dec-8-yl]ethyl]-2-naphthalenecarboxamide (NFOT) and N-[2-(4-oxo-1-phenyl-1,3,8-triazaspiro[4,5]dec-8-yl)ethyl]-2-naphthalenecarboxamide (NOPT) that are specific inhibitors of the PLD2 isoform. In spite of knowing their …

Comparison Between Flow Cytometry And Bead Method In Counting Cd4 And Cd8 T Lymphocytes In Mouse Spleen Cells Suspension, Abdulrahman Abdalla Allabidi

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The flow cytometry is gold standard method to count CD4, CD8 human T lymphocytes, and calculate CD8 to CD4 ratio in HIV patients peripheral blood to stage them and monitor their response to treatment. In this study, a comparison was made by counting CD4and CD8 mouse spleen T lymphocytes by flow cytometry, and a method involving antibody coated latex beads. The bead method yielded results comparable to those obtained by flow cytometry. These results indicate that antibody coated beads are suitable to determine CD4 and CD8 + T lymphocytes counts in situation where flow cytometry is not readily available.

Effect Of Herpes Simplex Virus-1 On Macrophage Cd Marker Expression, Hind Obaid Albeshri

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Macrophages are specialized phagocytic cells derived from the peripheral blood mononuclear cells. Macrophages can assume M1 or M2 cellular states upon activation with specific cytokines. Upon activation, macrophages produce either pro- or anti- inflammatory cytokines and thereby perform their functions of antigen engulfment and debris clearance. Macrophages have several cell surface cluster differentiation (CD) markers including CD80, CD163 and CD200R that differentiate M1 macrophages from M2. Downstream signaling and cytokine production varies based on the interaction between the specific CD marker and antigen.

The present study was aimed at analyzing and comparing the effects of HSV-1 infection on un-polarized and …

Elucidating The Effects Of Tcdd On The Polymorphic Human Hs1,2 Enhancer, Abdullah Freiwan

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2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent environmental toxin known to inhibit immunoglobulin (Ig) gene expression in various animal studies. We have identified the mouse 3'Ig heavy chain regulatory region (3'IgH RR) as a sensitive transcriptional target of TCDD, which may mediate the inhibitory effect of TCDD on Ig expression. Interestingly, the human hs1,2 enhancer is polymorphic and has been associated with a number of autoimmune diseases.

Suggesting a species difference, TCDD inhibited mouse hs1,2 enhancer activation and activated basal human hs1,2 (hs-hs1,2) enhancer activity in the mouse B-cell line. The objective of this study was to elucidate the effects of TCDD …

Adeno-Associated Virus (Aav) Transduction Of Primary Human Cd4+T Lymphocytes, Sahar Hussein Kamel

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Development of latent HIV-infected CD4+ T lymphocytes is the major cause of HIV treatment failure. Adeno-associated virus (AAV) is an attractive vector for anti-HIV gene therapy due to its lack of pathogenicity, low immunogenicity, and persistent transgene expression. However, a major limitation for AAV gene transfer is the cell-specific tropism of each serotype. Only AAV serotypes 2 and 5 have been investigated in hematopoietic cells, both of these serotypes have shown low transduction efficiency. Since each of the currently described AAV serotypes demonstrate distinct tissue tropism, I hypothesized that other serotypes such as AAV1, 4, 8, or 9 may demonstrate …

Expression Of The Alpha, Beta, And Gamma Subunits Of The Interleukin-2 Receptor By Human Vascular Smooth Muscle Cells, Sultan Mohammed Alhayyani

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Interleukin 2 (IL-2) is a member of the cytokine family and contributes to the proliferation, survival, and death of lymphocytes [1]. The interleukin-2 receptor (IL-2) is a tripartite receptor commonly expressed on the surfaces of many lymphoid cells and is composed of three non-covalently associated subunits, alpha (CD25), beta (CD122), and gamma (CD132) [2]. Our laboratory has previously described IL-2 receptor Beta expression by vasculature smooth muscle cells (VSMC) in mice and humans [3]. The current work expands our previous observations by assessing the expression of the alpha and gamma subunits of the IL-2R by VSMCs. Analysis of IL-2R expression …

Glut 4 And Insulin Resistance, Salmin Ali

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Five types of glucose transporters stimulate the transport of glucose from the bloodstream into cells such as muscle cells and adipocytes in response to insulin signaling or exercise. Skeletal muscle and adipose tissue are the main sites for glucose uptake and dispose in insulin action dependent pathway. In non-insulin stimulated condition, the GLUT4 storage vesicles stay away from the plasma membrane, so there is no GLUT4 translocation or GLUT4 mediated glucose uptake. However, in insulin stimulated state, insulin initiates insulin signaling cascade which results in rapid GLUT4 translocation and increased glucose uptake at the cell surface. Failure of the translocation …