• Narcan (naloxone nasal spray) availability in Georgia retail pharmacies

      Johnson, J. Aaron; Chung, Yunmi; Covington, Katherine; Augusta University (2018)
    • Neural population-level memory traces in the mouse hippocampus.

      Chen, Guifen; Wang, Lei Phillip; Tsien, Joe Z.; Brain & Behavior Discovery Institute; Department of Neurology (2009-12-17)
      One of the fundamental goals in neurosciences is to elucidate the formation and retrieval of brain's associative memory traces in real-time. Here, we describe real-time neural ensemble transient dynamics in the mouse hippocampal CA1 region and demonstrate their relationships with behavioral performances during both learning and recall. We employed the classic trace fear conditioning paradigm involving a neutral tone followed by a mild foot-shock 20 seconds later. Our large-scale recording and decoding methods revealed that conditioned tone responses and tone-shock association patterns were not present in CA1 during the first pairing, but emerged quickly after multiple pairings. These encoding patterns showed increased immediate-replay, correlating tightly with increased immediate-freezing during learning. Moreover, during contextual recall, these patterns reappeared in tandem six-to-fourteen times per minute, again correlating tightly with behavioral recall. Upon traced tone recall, while various fear memories were retrieved, the shock traces exhibited a unique recall-peak around the 20-second trace interval, further signifying the memory of time for the expected shock. Therefore, our study has revealed various real-time associative memory traces during learning and recall in CA1, and demonstrates that real-time memory traces can be decoded on a moment-to-moment basis over any single trial.
    • Neuropathology of Anxiety Disorders Comorbid with Alcohol Dependence

      O'Connor, Tara; Keough, Kelsey; Layton, James; Carpenter, Timothy; Crethers, Danielle; Patton, Tadd; Vazdarjanova, Almira; College of Science and Mathematics (2015-08-11)
      Anxiety disorders commonly occur along with other mental or physical illnesses, including alcohol use disorders (AUDs). Like most coexisting disorders, treatment for individuals who suffer from an anxiety disorder and an AUD is particularly challenging, contributing to an increased risk for suicide attempts, more intense withdrawal symptoms, and a higher probability of alcoholism relapse. Previous research has shown that a dysregulation of certain neuronal plasticity-related events in the prefrontal cortex (PFC) is implicated in anxiety disorders and alcohol dependence. However, the exact role this dysregulation plays in the comorbidity of these disorders is not well understood. The experiments conducted here were part of a larger study aimed at understanding the neuropathological characteristics present when anxiety disorders and AUDs coexist. We examined anxiety-like behavior and plasticity-related activity in the brains of rats bred to consume large or small quantities ethanol. After obtaining a prestress baseline of alcohol drinking behavior, rats were exposed to a standard shock procedure in which a mild footshock (paired with a tone) was administered. Drinking activity and anxiety-like behavior were assessed on multiple days following footshock. The rats were then euthanized so that the brains could be examined for activation of plasticity-related activity in the PFC. Significant differences in alcohol consumption and anxiety-like behavior were observed between alcohol-preferring and alcohol-non-preferring rats and are discussed in relation to drinking alcohol as a means to reduce anxiety. These findings will be included in our larger study and will ultimately lead to a better understanding of the neuropathological substrates associated with comorbid anxiety disorders and AUDs.
    • Neurotransmitter Gases as Modulators of GnRH and the Preovulatory LH Surge

      Lamar, Charisee; Institute of Molecular Medicine and Genetics (1998-04)
      Until recently nitric oxide (NO) and carbon monoxide (CO) were viewed only as toxic substances. However, there has been a substantial amount of evidence in the past decade that has redefined these gaseous molecules as physiological messengers. Along these lines, NO and CO are now recognized as modulators of immunological defense, vasodilation, endocrine signaling, and neurotransmission (1-8). As neurotransmitters, NO and CO are unique when compared to classical neurotransmitters. For instance, unlike all other classical neurotransmitters NO and CO are, 1) lipophilic gases with short half lives, 2) not stored in synaptic vesicles, 3) their effects are not mediated through classic receptor proteins - rather their effects result from NO and CO binding to the heme moiety of heme-proteins such as guanylate cyclase and cyclooxygenase, and 4) their effects are terminated by diffusion from target tissues ( 1,2,5,7,9). Production of NO and CO relies on the activity of the enzymes nitric oxide synthase (NOS) and heme oxygenase (HO), respectively. NOS uses the substrate Larginine to generate NO (2, 10), while HO uses the substrate heme to generate CO (9,11-16). NOS exists as three isoforms, macrophage NOS (mNOS), endothelial NOS (eNOS), and neuronal NOS (nNOS) (17-21). HO also exists as three isoforms, heme oxygenase-1 (HO-1), heme oxygenase-2 (HO-2), and heme oxygenase-3 (HO-3) HO-1 is inducible, while HO-2 and HO-3 are constitutive enzymes; however, HO-3 is currently viewed as a poor heme catalyst (12,13,22). There is a growing body of evidence that suggests that NO and CO regulate hypothalamic function. For example, recent studies have shown that the hypothalamus produces significant quantities of NO, primarily through the actions of nNOS (23,24). Likewise, the hypothalamus possesses one of the largest production rates of CO in the brain (25,26). That NO and CO can regulate neurohormone release from the hypothalamus is evidenced by findings demonstrating that NO and CO regulate corticotropin-releasing hormone (CRH) (27-33), vasopressin (29-31,34,35), and oxytocin secretion (29,36). With respect to reproductive function, numerous studies (23,37-45) have provided evidence for a significant role of NO in the control of the hypothalamic releasing factor, gonadotropin-releasing hormone (GnRH). For instance, it has been demonstrated (37,46,47) that NO neurons are located in close proximity to GnRH generating cells bodies in the hypothalamus and thus strategically located to exert regulatory effects over GnRH neurons. That NO can exert regulatory effects on GnRH neurons was demonstrated in studies where exogenously applied NO markedly stimulated GnRH release from male hypothalamic fragments (1,48,49) and immortalized GnRH (GT1-7) cells in vitro (1,48). A physiological role for NO in the steroid-induced luteinizing hormone (LH) surge has been suggested based on studies in which the LH surge was attenuated by the administration of NOS inhibitors (43,44) or NOS antisense oligonucleotides (38) No studies, however, have reported nNOS mRNA, protein, or NOS activity levels in the hypothalamus during the LH surge so as to verify that an increased NO tone actually occurs during this critical time. To address this deficit, Aim 1 of this study was designed to examine NO tone on proestrus in the cycling female rat-the day the natural preovulatory LH surge occurs. Since heme oxygenase, the enzyme that produces CO from heme molecule metabolism, is located in the hypothalamus (2,11,12,14,15), and CO production in the hypothalamus is one of the highest in the brain (25,26), it is conceivable that CO could play a role in regulating GnRH secretion. This possibility has not been investigated and thus studies on this issue appear warranted. Therefore, Aims 2-4 of this study were designed to assess the possible role of CO in the control of GnRH and LH secretion.
    • Neurovascular Injury After Retinal Ischemia Reperfusion Insult: Contrasting Roles Of Arginase Enzyme Isoforms

      Shosha, Esraa; Vascular Biology Center (2016-03-08)
      Purpose: We have previously shown the involvement of arginase enzyme in retinal neurovascular injury. The present study was undertaken to determine the distinct roles of arginase 1 (A1) and arginase 2 (A2) in neurovascular damage following ischemia/reperfusion (I/R) injury. Methods: We used wild type (WT) mice, A2 knock out mice (A2-/-) and mice lacking one copy of A1 (A1+/-). Western blotting, RT-PCR, vascular digests, immunofluorescence, Propidium Iodide (PI) labeling and electroretinography (ERG) were used to evaluate retinal injury and function. Results: I/R injury caused significant increases in A2 expression along with thinning of the neural retina, decreases in NeuN+ GCL neurons and formation of acellular capillaries. Increases in PI labeling and RIP-3 expression showed that cell death occurred by necroptosis. Neurovascular injury was accompanied by microglial activation along with increased expression of GFAP and impairment of the ERG. Neuronal cell loss, capillary degeneration, necroptosis, gliosis and ERG impairment were all significantly reduced by deletion of A2. On the other hand, A1 deletion exacerbated I/R-induced neuronal and vascular injury and further increased necroptosis and gliosis as compared with WT retinas. Conclusions: This study shows for the first time the different roles of arginase isoforms after I/R insult. I/R-induced necrotic cell death and gliosis are mediated by A2, whereas upregulation of A1 may play a role in limiting the pathology.
    • A New Antifibrotic Target of Ac-SDKP: Inhibition of Myofibroblast Differentiation in Rat Lung with Silicosis

      Xu, Hong; Yang, Fang; Sun, Ying; Yuan, Yuan; Cheng, Hua; Wei, Zhongqiu; Li, Shuyu; Cheng, Tan; Brann, Darrell W; Wang, Ruimin; et al. (2012-07-3)
      Background: Myofibroblast differentiation, characterized by a-smooth muscle actin (a-SMA) expression, is a key process in organ fibrosis, and is induced by TGF-b. Here we examined whether an anti-fibrotic agent, N-acetyl-seryl-aspartyllysylproline (Ac-SDKP), can regulate induction of TGF-b signaling and myofibroblast differentiation as a potential key component of its anti-fibrotic mechanism in vivo and in vitro.
    • New Orientation Curriculum with Simulation and Contest Improves Background Knowledge and Clinical Training Experience

      Boyer, Tanna; Braun, Kelli; Odo, Nadine; Arthur, Mary E.; Anesthesiology and Perioperative Medicine & Obstetrics and Gynecology (2016-03)
      After completing their intern year, all residents in anesthesiology must complete an intensive one month training program. The goal of this curriculum is to ensure our residents have the appropriate background knowledge and clinical skills to begin practicing anesthesia independently (in supervised settings). Our goal was to develop a new one month curriculum that was engaging, interactive, low cost, and utilized existing validated resources.
    • NF-kB2 mutation targets survival, proliferation and differentiation pathways in the pathogenesis of plasma cell tumors

      McCarthy, Brian A.; Yang, Liqun; Ding, Jane; Ren, MingQiang; King, William; ElSalanty, Mohammed; Zakhary, Ibrahim; Sharawy, Mohamed; Cui, Hongjuan; Ding, Han-Fei; et al. (2012-05-29)
      Background: Abnormal NF-κB2 activation has been implicated in the pathogenesis of multiple myeloma, a cancer of plasma cells. However, a causal role for aberrant NF-κB2 signaling in the development of plasma cell tumors has not been established. Also unclear is the molecular mechanism that drives the tumorigenic process. We investigated these questions by using a transgenic mouse model with lymphocyte-targeted expression of p80HT, a lymphoma-associated NF-κB2 mutant, and human multiple myeloma cell lines.
    • NMDA Receptors Are Not Required for Pattern Completion During Associative Memory Recall

      Mei, Bing; Li, Fei; Gu, Yiran; Cui, Zhenzhong; Tsien, Joe Z.; Brain & Behavior Discovery Institute; Department of Neurology (2011-04-29)
      Pattern completion, the ability to retrieve complete memories initiated by subsets of external cues, has been a major focus of many computation models. A previously study reports that such pattern completion requires NMDA receptors in the hippocampus. However, such a claim was derived from a non-inducible gene knockout experiment in which the NMDA receptors were absent throughout all stages of memory processes as well as animal's adult life. This raises the critical question regarding whether the previously described results were truly resulting from the requirement of the NMDA receptors in retrieval. Here, we have examined the role of the NMDA receptors in pattern completion via inducible knockout of NMDA receptors limited to the memory retrieval stage. By using two independent mouse lines, we found that inducible knockout mice, lacking NMDA receptor in either forebrain or hippocampus CA1 region at the time of memory retrieval, exhibited normal recall of associative spatial reference memory regardless of whether retrievals took place under full-cue or partial-cue conditions. Moreover, systemic antagonism of NMDA receptor during retention tests also had no effect on full-cue or partial-cue recall of spatial water maze memories. Thus, both genetic and pharmacological experiments collectively demonstrate that pattern completion during spatial associative memory recall does not require the NMDA receptor in the hippocampus or forebrain.
    • Normal colon epithelium: a dataset for the analysis of gene expression and alternative splicing events in colon disease.

      Mojica, Wilfrido; Hawthorn, Lesleyann; GHSU Cancer Center (2010-02-18)
      BACKGROUND: Studies using microarray analysis of colorectal cancer have been generally beleaguered by the lack of a normal cell population of the same lineage as the tumor cell. One of the main objectives of this study was to generate a reference gene expression data set for normal colonic epithelium which can be used in comparisons with diseased tissues, as well as to provide a dataset that could be used as a baseline for studies in alternative splicing. RESULTS: We present a dependable expression reference data set for non-neoplastic colonic epithelial cells. An enriched population of fresh colon epithelial cells were obtained from non-neoplastic, colectomy specimens and analyzed using Affymetrix GeneChip EXON 1.0 ST arrays. For demonstration purposes, we have compared the data derived from these cells to a publically available set of tumor and matched normal colon data. This analysis allowed an assessment of global gene expression alterations and demonstrated that adjacent normal tissues, with a high degree of cellular heterogeneity, are not always representative of normal cells for comparison to tumors which arise from the colon epithelium. We also examined alternative splicing events in tumors compared to normal colon epithelial cells. CONCLUSIONS: The findings from this study represent the first comprehensive expression profile for non-neoplastic colonic epithelial cells reported. Our analysis of splice variants illustrate that this is a very labor intensive procedure, requiring vigilant examination of the data. It is projected that the contribution of this set of data derived from pure colonic epithelial cells will enhance studies in colon-related disease and offer a vital baseline for studies aimed at elucidating the mechanisms of alternative splicing.
    • Notch2 and Notch3 Function Together to Regulate Vascular Smooth Muscle Development

      Wang, Qingqing; Zhao, Ning; Kennard, Simone; Lilly, Brenda; Vascular Biology Center (2012-05-17)
      Notch signaling has been implicated in the regulation of smooth muscle differentiation, but the precise role of Notch receptors is ill defined. Although Notch3 receptor expression is high in smooth muscle, Notch3 mutant mice are viable and display only mild defects in vascular patterning and smooth muscle differentiation. Notch2 is also expressed in smooth muscle and Notch2 mutant mice show cardiovascular abnormalities indicative of smooth muscle defects. Together, these findings infer that Notch2 and Notch3 act together to govern vascular development and smooth muscle differentiation. To address this hypothesis, we characterized the phenotype of mice with a combined deficiency in Notch2 and Notch3. Our results show that when Notch2 and Notch3 genes are simultaneously disrupted, mice die in utero at mid-gestation due to severe vascular abnormalities. Assembly of the vascular network occurs normally as assessed by Pecam1 expression, however smooth muscle cells surrounding the vessels are grossly deficient leading to vascular collapse. In vitro analysis show that both Notch2 and Notch3 robustly activate smooth muscle differentiation genes, and Notch3, but not Notch2 is a target of Notch signaling. These data highlight the combined actions of the Notch receptors in the regulation of vascular development, and suggest that while these receptors exhibit compensatory roles in smooth muscle, their functions are not entirely overlapping.
    • Notch3 Signaling Mediates Heterotypic Cell Interactions During Blood Vessel Formation

      Liu, Hua; Vascular Biology Center (2010-11)
      Blood vessel formation is essential for embryogenesis, wound healing, menstruation, and pregnancy [1, 2]. While much emphasis has been placed on understanding the initial event of endothelial-tube formation, relatively little attention has been paid to the interactions of endothelial cells and the surrounding mural cells (pericytes, smooth muscle cells and fibroblasts). Increasing evidence suggests that the communication of endothelial cells and mural cells is crucial for the assembly, subsequent maturation, and stabilization of blood vessels [3-5]. Abnormal interactions between these two cell types have been implicated in many pathological conditions, including tumor angiogenesis, diabetic microangiopathy, tissue calcification and stroke. However, the molecules mediating the heterotypic interaction are still largely unknown. Our previous studies have shown that in a three-dimensional (3-D) angiogenesis assay, mural cells enhance blood vessel formation and directly interact with endothelial cells [6]. During this process, Notch3 is one gene that is strongly induced in mural cells upon coculture with endothelial cells [6]. Notch3, the causative gene of the neurovascular disorder CADASIL [7], belongs to an evolutionarily conserved family of transmembrane receptors that are known to govern cell fate determination in diverse cell types [8]. Given that Notch receptors and ligands are expressed on both endothelial and mural cells and - 2 - Notch3 is upregulated in mural cells by coculturing with endothelial cells, it is reasonable to assume that the Notch3 receptor might regulate the association of endothelial and mural cells through receptor-ligand interaction during blood vessel formation. The goal of my thesis is to investigate how Notch3 gene expression is regulated in mural cells by endothelial cells and whether the Notch3 receptor is involved in the communication between endothelial and mural cells during blood vessel formation. To achieve these goals, three aims were proposed: Specific Aim 1: To define how Notch3 expression in mural cells is upregulated by endothelial cells. Specific Aim 2: To determine if endothelial cell-induced Notch3 expression is critical for mural cell differentiation. Specific Aim 3: To determine whether Notch3 expression in mural cells modulates blood vessel formation under both physiological and pathological conditions.
    • Novel mechanisms of endothelial dysfunction in diabetes

      Yang, Guang; Lucas, Rudolf; Caldwell, Ruth B.; Yao, Lin; Romero, Maritza J.; Caldwell, Robert William; Vascular Biology Center; Department of Pharmacology and Toxicology (2010-04)
      Diabetes mellitus is a major risk factor for cardiovascular morbidity and mortality. This condition increases the risk of developing coronary, cerebrovascular, and peripheral arterial disease fourfold. Endothelial dysfunction is a major contributor to the pathogenesis of vascular disease in diabetes mellitus patients and has recently received increased attention. In this review article, some recent developments that could improve the knowledge of diabetes-induced endothelial dysfunction are discussed.
    • Novel Role of Heat Shock Protein (HSP) 90 in Regulating ATR-CHK1 DNA Damage Response Pathway in Cancer Cells

      Ha, Kyungsoo; Institute of Molecular Medicine and Genetics (2011-10)
      DNA damage caused by environmental mutagens or reactive metabolic byproducts induces DNA damage response (DDR), which regulates cell cycle transit, DNA repair and apoptosis. DDR involves the phosphorylation and activation of Ataxia Telangiectasia Mutated (ATM) and ATM and RAD3-related (ATR) proteins. ATR regulates the firing of the replication forks during S phase, and the repair of damaged replication forks to prevent premature onset of mitosis. ATR phosphorylates and activates CHK1 which phosphorylates and inactivates CDC25, thereby inhibiting CDK1 activation and cell cycle progression. In the present studies, we determined that treatment with an hsp90 inhibitor AUY922, without affecting the mRNA levels, dose-dependently depletes the protein levels of p-ATR (Ser 428), ATR and CHK1 in human breast and cervical cancer cells. Additionally, treatment with the pan-histone deacetylase inhibitor panobinostat (PS), which is known to induce hyperacetylation and inhibition of hsp90 function, also depleted ATR and CHK1 levels in cancer cells. Co-treatment with the proteasome inhibitor bortezomib (BZ) partially reversed AUY922- or PS-mediated depletion of ATR and CHK1 expression, indicating proteasome-mediated degradation of ATR and CHK1. Treatment with either AUY922 or PS markedly inhibited the binding of ATR with hsp90, induced polyubiquitylation of ATR, and decreased the half-life of both ATR and CHK1 proteins. Treatment with AUY922 also abrogated ionizing radiation (IR)-induced cell cycle arrest and increased the amount of DNA damage in the cancer cells following IR. Treatment with AUY922 also inhibited the recruitment of p-ATR, ATR and 53BP1 to the site of DNA damage. In addition, HDAC3 binds to and deacetylates hsp90 in the nucleus. Depletion of HDAC3 by either short hairpin RNA or genetic knockout induced hyperacetylation of nuclear hsp90, resulting in the inhibition of chaperone association of ATR with hsp90 and depletion of ATR. These findings demonstrate that 1) ATR is chaperoned by hsp90, 2) Inhibition of chaperone function of hsp90 results in proteasomal degradation of ATR and inhibition of DDR, 3) pan-HDAC inhibitors abrogate ATRCHK1 cell cycle checkpoint pathway by modulating chaperone activity of hsp90 and 4) HD AC3 plays a critical role in the regulation of DNA damage response by stabilizing the chaperone activity of nuclear hsp90.
    • Novel Somatic Mutations to PI3K Pathway Genes in Metastatic Melanoma

      Shull, Austin Y.; Latham-Schwark, Alicia; Ramasamy, Poornema; Leskoske, Kristin; Oroian, Dora; Birtwistle, Marc R.; Buckhaults, Phillip J.; GHSU Cancer Center (2012-08-17)
      Background: BRAFV600 inhibitors have offered a new gateway for better treatment of metastatic melanoma. However, the overall efficacy of BRAFV600 inhibitors has been lower than expected in clinical trials, and many patients have shown resistance to the drugâ s effect. We hypothesized that somatic mutations in the Phosphoinositide 3-Kinase (PI3K) pathway, which promotes proliferation and survival, may coincide with BRAFV600 mutations and contribute to chemotherapeutic resistance.
    • A novel subnetwork based analysis reveals shared pathways in T-cell mediated autoimmunity

      Pabla, Simarjot Singh; Center for Biotechnology and Genomic Medicine (2016-03)
      Thymocyte auto-reactivity is an underlying theme of several autoimmune disorders. The precise role of auto-reactive T cells in the initiation and subsequent progression of autoimmune disorders has been studied extensively. However, these disease specific studies ignore pathways that may be in common to several T cell mediated autoimmune pathologies. This can be attributed in part to the shortcomings of traditional gene list based gene expression studies. Here we report a novel method to identify unifying gene expression changes in several autoimmune diseases. In order to uncover pathologically important pathways common to T-cell mediated autoimmune disorders, we used human gene expression data from Multiple Sclerosis, Rheumatoid Arthritis, Juvenile Idiopathic Arthritis and Sjögren’s syndrome. Unlike traditional gene expression analysis, we used jointly active connected subnetwork enrichment to identify subnetworks for each disorder, followed by topological network alignment, which led to identification of shared pathways. We report four pathways shared in these disorders, which include DNA damage response, gonadotropin, innate and adaptive immunity pathways. Importantly, our method did not reveal any common pathways in unrelated diseases. In order to experimentally validate our findings, RNA sequencing of mRNA isolated from salivary glands excised from a murine model of Sjögren’s syndrome was performed. High similarities were observed between Human T-cell mediated autoimmune disorders and Sjögren’s murine model. Collectively, these studies have identified a shared landscape of pathologically significant pathways, including DNA damage response, gonadotropin, innate and adaptive immunity in autoimmune disorders and provide a new methodology to identify common alterations in diseases with similar underlying etiologies.
    • De novo transcriptome sequencing in a songbird, the dark-eyed junco (Junco hyemalis): genomic tools for an ecological model system

      Peterson, Mark P; Whittaker, Danielle J; Ambreth, Shruthi; Sureshchandra, Suhas; Buechlein, Aaron; Podicheti, Ram; Choi, Jeong-Hyeon; Lai, Zhao; Mockatis, Keithanne; Colbourne, John K.; et al. (2012-07-9)
      Background: Though genomic-level data are becoming widely available, many of the metazoan species sequenced are laboratory systems whose natural history is not well documented. In contrast, the wide array of species with very well-characterized natural history have, until recently, lacked genomics tools. It is now possible to address significant evolutionary genomics questions by applying high-throughput sequencing to discover the majority of genes for ecologically tractable species, and by subsequently developing microarray platforms from which to investigate gene regulatory networks that function in natural systems. We used GS-FLX Titanium Sequencing (Roche/454-Sequencing) of two normalized libraries of pooled RNA samples to characterize a transcriptome of the dark-eyed junco (Junco hyemalis), a North American sparrow that is a classically studied species in the fields of photoperiodism, speciation, and hormone-mediated behavior.
    • NUB1, an interferon-inducible protein, mediates anti-proliferative actions and apoptosis in renal cell carcinoma cells through cell-cycle regulation.

      Hosono, T; Tanaka, Tomoaki; Tanji, K; Nakatani, T; Kamitani, Tetsu; Center for Molecular Chaperone/Radiobiology & Cancer Virology (2010-03-03)
      BACKGROUND: NEDD8 ultimate buster 1 (NUB1) is an interferon (IFN)-inducible protein that downregulates NEDD8 expression and its conjugation system. Although overexpression of NUB1 induces a growth-inhibitory effect in cells, the mechanisms underlying the anti-mitogenic actions of NUB1 in cancer cells remain uncertain. We investigated the anti-cancer effects of NUB1 in human renal cell carcinoma (RCC) cells. METHODS: Nine human RCC cells were used for this study. The proliferation of RCC cells exposed to IFN-alpha was measured by water-soluble tetrazolium salt assay. The expression level of NUB1 in cells was measured by quantitative reverse transcriptase PCR or western blot analysis. Apoptosis and cell-cycle analysis were performed by flow cytometry. Silencing of NUB1 was performed using a small interfering RNA. RESULTS: Both NUB1 messenger RNA and protein were significantly induced by IFN-alpha in seven out of nine selected RCC cell lines, and the NUB1 expressions induced by IFN-alpha correlated positively with cell growth inhibition. Overexpression of NUB1 remarkably induced S-phase transition during cell cycle and apoptosis in IFN-alpha-resistant A498 cells, in which NUB1 is not induced by IFN-alpha. The expression levels of two cell-cycle regulator proteins, cyclin E and p27, were increased under the aforementioned conditions. The knockdown of NUB1 enhanced cell proliferation of IFN-alpha-resistant A498 cells and suppressed IFN-alpha-induced growth inhibition in IFN-alpha-sensitive 4TUHR cells. CONCLUSION: NUB1 may be a key factor involved not only in cell growth inhibition by IFN-alpha in RCC cells but also in the anti-cancer effect against IFN-alpha-resistant RCC cells.
    • Obesity related methylation changes in DNA of peripheral blood leukocytes

      Wang, Xiaoling; Zhu, Haidong; Snieder, Harold; Su, Shaoyong; Munn, David H.; Harshfield, Gregory; Maria, Bernard L.; Dong, Yanbin; Treiber, Frank A.; Gutin, Bernard; et al. (2010-12-21)
      Background: Despite evidence linking obesity to impaired immune function, little is known about the specific mechanisms. Because of emerging evidence that immune responses are epigenetically regulated, we hypothesized that DNA methylation changes are involved in obesity induced immune dysfunction and aimed to identify these changes.
    • Oxygen Glucose Deprivation in Rat Hippocampal Slice Cultures Results in Alterations in Carnitine Homeostasis and Mitochondrial Dysfunction

      Rau, Thomas F.; Lu, Qing; Sharma, Shruti; Sun, Xutong; Leary, Gregory; Beckman, Matthew L.; Hou, Yali; Wainwright, Mark S.; Kavanaugh, Michael; Poulsen, David J.; et al. (2012-09-11)
      Mitochondrial dysfunction characterized by depolarization of mitochondrial membranes and the initiation of mitochondrial-mediated apoptosis are pathological responses to hypoxia-ischemia (HI) in the neonatal brain. Carnitine metabolism directly supports mitochondrial metabolism by shuttling long chain fatty acids across the inner mitochondrial membrane for beta-oxidation. Our previous studies have shown that HI disrupts carnitine homeostasis in neonatal rats and that L-carnitine can be neuroprotective. Thus, this study was undertaken to elucidate the molecular mechanisms by which HI alters carnitine metabolism and to begin to elucidate the mechanism underlying the neuroprotective effect of L-carnitine (LCAR) supplementation. Utilizing neonatal rat hippocampal slice cultures we found that oxygen glucose deprivation (OGD) decreased the levels of free carnitines (FC) and increased the acylcarnitine (AC): FC ratio. These changes in carnitine homeostasis correlated with decreases in the protein levels of carnitine palmitoyl transferase (CPT) 1 and 2. LCAR supplementation prevented the decrease in CPT1 and CPT2, enhanced both FC and the ACâ ¶FC ratio and increased slice culture metabolic viability, the mitochondrial membrane potential prior to OGD and prevented the subsequent loss of neurons during later stages of reperfusion through a reduction in apoptotic cell death. Finally, we found that LCAR supplementation preserved the structural integrity and synaptic transmission within the hippocampus after OGD. Thus, we conclude that LCAR supplementation preserves the key enzymes responsible for maintaining carnitine homeostasis and preserves both cell viability and synaptic transmission after OGD.