bluuebunny/crossref_metadata_2025 huggingface dataset:
Retinopathy Screening in Children and Adolescents with Diabetes
Authors: RAMAN V. , CAMPBELL F. , HOLLAND P. , CHAPMAN T. , DABBS T. , BODANSKY H. J. , O'NEILL D. P.
Published: July 2010

Abstract: Diabetic retinopathy is a complication of diabetes that affects the blood vessels of the retina. The majority of patients with diabetic retinopathy exhibit background retinopathy. In ∼ 10% of patients, background retinopathy will progress to sight‐threatening retinopathy. The aims of our study were to measure the prevalence of retinopathy among our population and to assess the optimal time of screening children and adolescents with type 1 diabetes.
Clinical Association of Autoimmune Diseases with Diabetes Mellitus
Authors: SRIDHAR G. R. , NAGAMANI G.
Published: July 2010

Abstract: The adult form of polyglandular autoimmune syndrome (type 2) is associated with disorders of the thyroid, adrenal, and pancreas (type 1 diabetes mellitus). We profile the clinical association of autoimmune diseases among patients from our center in southern India: autoimmune diseases were diagnosed in 1.68% of persons with diabetes mellitus (147/15,523). Diabetes mellitus was diagnosed in 2.3% of persons with hypothyroidism (33/1435) and in 4.35% with thyrotoxicosis (15/345).
Activation of IL‐1 Signaling Pathway in Schwann Cells during Diabetic Neuropathy
Authors: SKUNDRIC DUSANKA S. , DAI RUJUAN , JAMES JASCINTH , LISAK ROBERT P.
Published: July 2010

Abstract: Our results show activation of the IL‐1 signaling pathway in Schwann cells (SC), prior to and during early stages of diabetic neuropathy, thus suggesting its role in the initiation of SC‐axonal miscommunication.
Class II MHC Alleles in Rheumatoid Arthritis in Tamilnadu, India
Authors: PARTHIBAN M. , MADHAVAN RADHA , PORKODI R. , RAJENDRAN C. PANCHAPAKESA , ZAKE L. , SANJEEVI C. B. , CHANDRASEKARAN A. N.
Published: July 2010

Abstract: Rheumatoid arthritis (RA) was reported to be associated with class II MHC alleles in different ethnic populations. A similar study was undertaken to determine the association of class II MHC with RA patients of Tamilnadu (Tamil‐speaking Hindus), India. Thirty patients with RA and 39 healthy controls were included. Polymorphic second exons of the DRB1, DQA1, and DQB1 genes were amplified and subjected to SSOP typing. No allele was found to be significantly associated with RA. However DRB1*11 (P= 0.01) and DQB1*0302 (P= 0.02) were significantly associated with rheumatoid factor‐positive RA patients. (All the DRB1*11‐positive RA patients had either *04 or *10 allele as their second allele. This study is first of its kind in this population.
Genetic Alterations and Oncogenic Pathways in Hepatocellular Carcinoma
Authors: LÉVY LAURENCE , RENARD CLAIRE ANGÉLIQUE , WEI YU , BUENDIA MARIE ANNICK
Published: July 2010

Abstract: Hepatocellular carcinoma (HCC) is a major type of primary liver cancer and one of the rare human neoplasms etiologically linked to viral factors. Chronic infections with the hepatitis B virus (HBV) and the hepatitis C virus (HCV) have been implicated in about 80% of cases worldwide, and other known environmental risk factors, including alcohol abuse and dietary intake of aflatoxin B1, might synergize with viral infections. Recent insight into the molecular mechanisms leading to HCC development has been provided by the identification of major genetic abnormalities revealed by genomewide allelotype studies and molecular cytogenetic analysis. Moreover, several oncogenic pathways have been implicated in malignant transformation of liver cells. Inactivation of the p53 tumor suppressor gene by mutations and allelic deletions in about 30% of HCC cases has been associated predominantly with exposure to aflatoxin B1 and HBV infection. By contrast, a mutation in the β‐catenin gene in around 22% of HCCs is more rare in HBV‐associated tumors. Activation of cyclin D1 and disruption of the Rb pathway are also commonly involved in liver tumorigenesis. New major challenges include the identification of candidate genes located in frequently altered chromosomal regions and that of oncogenic pathways driven by different risk factors. This search might shed some light on the tumorigenic role of HBV and HCV. It might also permit accurate evaluation of major targets for prognostic and therapeutic intervention.
Circulating IL‐6 and sIL‐6R in Patients with Hepatocellular Carcinoma
Authors: GIANNITRAPANI L. , CERVELLO M. , SORESI M. , NOTARBARTOLO M. , LA ROSA M. , VIRRUSO L. , D'ALESSANDRO N. , MONTALTO G.
Published: July 2010

Abstract: Interleukin‐6 plays a central role in regulating the immune system, hematopoiesis, and acute phase reaction. It interacts with a receptor complex consisting of a specific ligand‐binding protein (IL‐6R, gp80) and a signal transduction protein (gp130). In this report, serum levels of IL‐6 and a soluble form of the interleukin‐6 receptor (sIL‐6R) were evaluated in patients with hepatocellular carcinoma. The correlation between IL‐6 and sIL‐6R values, the stage of hepatocellular carcinoma, and main liver function tests was also studied.
Linkage Analysis in Families with Recurrent Neuroblastoma
Authors: PERRI PATRIZIA , LONGO LUCA , McCONVILLE CARMEL , CUSANO ROBERTO , REES SALLY A. , SERI MARCO , CONTE MASSIMO , ROMEO GIOVANNI , DEVOTO MARCELLA , TONINI GIAN PAOLO
Published: July 2010

Abstract: Neuroblastoma is a neural crest‐derived tumor of childhood with a serious prognosis; only 20% of patients with stage 4 disease survive 5 years from diagnosis. Mechanisms involved in neuroblastoma development are unclear, but the engagement of many neuroblastoma‐related gene(s) is suggested by specific chromosomal alterations. Most prominent among these is the amplification of the MYCN oncogene and the deletion of the 1p36 region. Other genetic aberrations have been discovered over the years such as deletions of 11q and 14q and gain of 17q. Although tumor aggressiveness greatly depends on the most frequent genetic abnormalities, to date no neuroblastoma‐related gene has been discovered. Neuroblastoma usually occurs sporadically, but 1.5% of all diagnosed cases show familial recurrence with an autosomal dominant inheritance and incomplete penetrance. A comparison between hereditary and sporadic neuroblastomas led Knudson and Strong to gather that the two‐hit hypothesis, proposed for retinoblastoma, could be applied to neuroblastoma. To determine if the 1p36 region harbors a predisposition gene for familial neuroblastoma, we carried out linkage analysis at 1p36 loci in two families with recurrent neuroblastoma. Similarly, we analyzed loci of chromosome 16, where a predisposition locus was recently mapped. We also analyzed markers located close to several candidate genes (RET, NF1, GDNF, GFRA1, EDNRB, and EDN3) involved to a different extent in other neurocristopathies. Our findings indicate that the candidate chromosomal regions and genes analyzed are not in linkage with neuroblastoma.
Intercellular Communication and Human Prostate Carcinogenesis
Authors: CARRUBA GIUSEPPE , STEFANO ROSALBA , COCCIADIFERRO LETIZIA , SALADINO FRANCESCA , DI CRISTINA ANTONIETTA , TOKAR ERIK , QUADER SALMAAN T.A. , WEBBER MUKTA M. , CASTAGNETTA LUIGI
Published: July 2010

Abstract: Gap‐junction‐mediated intercellular communication (GJIC) is required for completion of embryonic development, tissue homeostasis, and regulation of cell proliferation and death. Although, as emphasized in several reports, defects or disruption of GJIC may be important in carcinogenesis, the potential role of GJIC in the onset and progression of human prostate cancer remains ill‐defined. The gap junction channel‐forming connexins (Cx) comprise a multigene family of highly conserved proteins that are differentially expressed in a tissue‐ and development‐specific manner; changes in connexin expression are also commonly seen during cellular differentiation. However, when multiple connexins are concurrently expressed, gap junction channels may consist of more than one connexin species. This is important, because only certain pairings give rise to functional channels. In our studies, we investigated GJIC in a panel of both nontumorigenic (RWPE‐1) and malignant (RWPE‐2, LNCaP, DU‐145) human prostate epithelial cells, compared to a normal rat liver epithelial F344 (WB‐1) cell line, as it was found to be junctionally proficient. In addition, expression and regulation of Cx43 and Cx32 were also inspected using western blot analysis. The ability of hormones, antihormones, and the antihypertensive drug forskolin to restore GJIC in nontumorigenic and malignant human prostate epithelial cells was examined by the scrape‐loading/dye transfer (SL/DT) or fluorescence recovery after photobleaching (FRAP) methods using an Ultima laser cytometer. Results from both assays showed that neither nontumorigenic nor malignant prostate cells have functional GJIC. However, both estrone (E1) and forskolin (FK) induced a significant increase (4.4‐ and 2.8‐fold, respectively) in cell‐cell communication only in the RWPE‐1 cells. Interestingly, the use of Matrigel, a solubilized basement membrane, as substrate for cell attachment and growth resulted in the rescue of GJIC activity in RWPE‐1 cells, as revealed by the SL/DT method. Furthermore, E1 induced a twofold increase in connexin 43 (Cx43), whereas forskolin caused a 50% reduction in Cx32 expression in RWPE‐1 cells. These data suggest that agents that increase Cx43:Cx32 ratio may restore GJIC in junctionally deficient cells, providing a basis for the development of new strategies for the prevention and treatment of human prostate cancer.
Src Is an Initial Target of Sex Steroid Hormone Action
Authors: MIGLIACCIO A. , CASTORIA G. , DI DOMENICO M. , DE FALCO A. , BILANCIO A. , AURICCHIO F.
Published: July 2010

Abstract: Recent observations that steroids use pathways universally known to be regulated by growth factors and interleukins highlight the following points: (1) Steroid stimulation of the canonical pathway Src/Ras/Erk signaling from membrane to nuclei or its single members has been observed in different cell types including human cancer‐derived cells, neurons, osteoblasts, osteocytes, and endothelial cells. This stimulation has been reconstituted and analyzed in transiently transfected cells. (2) Cellular context and intracellular localization of receptors are crucial in determining the biological effects evoked by this hormonal stimulation: proliferation, protection from apoptosis, and vasorelaxation. (3) Classical steroid receptors localized in the extranuclear compartment directly and, in some cases, simultaneously interact with Src. They are capable of unexpected cross talks responsible for the observed effects. (4) Other signaling pathways including P13K/AKT are also stimulated by steroids. The aim of future work will be to arrive at an integrated general view of the different signaling pathways activated by steroids and to analyze the concert between these pathways and the hormonal transcriptional action. This general view should be simultaneously verified in different cell contexts, under different physiologic and pathologic conditions. We expect that the new technologies, above all gene and protein microarray, will make this goal feasible.
Methods to Obtain More Clinical and Pathologic Information from Needle Core Biopsy of the Prostate Gland
Authors: GALOSI ANDREA B. , MUZZONIGRO GIOVANNI
Published: July 2010

Abstract: Prostate needle biopsy can disclose important clinical information on tumor extension and grading, useful prognostic parameters for therapeutic choices and prognostic definition. To obtain more histopathologic information on specimens and clinical prognostic parameters, we used a new method to handle and embed tissue fragments of prostate biopsy as well as a new trans‐rectal probe (TRUS) with an end‐fire convex ultrasound transducer to guide ultrasonically precise needle placement in the prostate gland. In our experience these methods combined can be useful in obtaining reliable clinical and prognostic information in the management of patients with prostate cancer.
Multidimensional Flow Cytometry Immunophenotyping of Hematologic Malignancy
Authors: PAGNUCCO GUIDO , VANELLI LAURA , GERVASI FRANCESCO
Published: July 2010

Abstract: Immunophenotyping of hematologic malignancies represents one of the most relevant clinical applications of flow cytometry. Classically, leukemic/lymphomatous cells have been considered to reflect the immunophenotypic characteristics of different precursors and mature healthy cells blocked at certain differentiation stages. Recently, accumulating evidence has shown that neoplastic cells display several aberrant phenotypic patterns. These aberrant phenotypes are believed to reflect genetic abnormalities present in pathologic cells, and recent data have shown that at least in acute leukemias, myelodysplastic syndromes, chronic lymphoproliferative disorders, and plasma cell dyscrasias, they may be present in almost all patients. The aim of this work is to review recent advances in flow cytometry and the role of gating strategies more useful in the identification and characterization of neoplastic cells of different hematologic malignancies.
Digestion Stability as a Criterion for Protein Allergenicity Assessment
Authors: FU TONG‐JEN
Published: July 2010

Abstract: Assessing the allergenic potential of transgenic proteins introduced into genetically engineered food remains a critical part of the overall safety assessment of these foods. Stability to digestion has been proposed as one of the steps in the decision tree approach to assess the allergenic potential of transgenic proteins. The validity of digestion stability as a criterion for protein allergenicity assessment, however, has encountered some criticism in recent years. This chapter gives an overview of the rationale behind the use of digestion stability as a criterion for protein allergenicity assessment and reviews the available data that may or may not support its use. The application of in vitro digestion assays for the assessment of allergenic potential of novel proteins and the factors that affect the assay results are also discussed. There is a need to establish standardized assay conditions so that direct comparison of results from different laboratories can be made. Consensus also needs to be reached on relating the measured digestibility to the allergenic potential of proteins.
Hypothesis: The Protein Body Effect and Other Aspects of Food Matrix Effects
Authors: TEUBER SUZANNE S.
Published: July 2010

Abstract: With regard to the allergenicity of edible seeds and nuts, certain proteins may not be immediately accessible to digestion in the stomach and the upper small intestine because of the nature of the organization of such proteins into protein body organelles. Protein body hydration status, interactions between proteins, phytochemicals, protease inhibitors, and other matrix effects may contribute to the ability of a protein or package of proteins to reach the sites of active immune sampling in the gastrointestinal mucosa and thus be an influence on the potential allergenicity of a protein.
Food Allergy Animal Models
Authors: HELM RICKI M.
Published: July 2010

Abstract:Specific food allergy is characterized by sensitization to innocuous food proteins with production of allergen‐specific IgE that binds to receptors on basophils and mast cells. Upon recurrent exposure to the same allergen, an allergic response is induced by mediator release following cross‐linking of cell‐bound allergen‐specific IgE. The determination of what makes an innocuous food protein an allergen in predisposed individuals is unknown; however, mechanistic and protein allergen predictive models are being actively investigated in a number of animal models. Currently, there is no animal model that will actively profile known food allergens, predict the allergic potential of novel food proteins, or demonstrate clinically the human food allergic sensitization/allergic response. Animal models under investigation include mice, rats, the guinea pig, atopic dog, and neonatal swine. These models are being assessed for production of IgE, clinical responses to re‐exposure, and a ranking of food allergens (based on potency) including a nonfood allergen protein source. A selection of animal models actively being investigated that will contribute to our understanding of what makes a protein an allergen and future predictive models for assessing the allergenicity of novel proteins is presented in this review.
Protein Allergenicity in Mice
Authors: ATHERTON KEITH T. , DEARMAN REBECCA J. , KIMBER IAN
Published: July 2010

Abstract: Food allergy is an important and common health issue, and there is therefore a need to identify and characterize the sensitizing potential of novel food proteins. Approaches currently used include consideration of structural similarity to, or amino acid sequence homology with, known human allergens; immunologic (generally serologic) cross‐reactivity with known allergens; and the measurement of resistance to proteolytic digestion in a simulated gastric fluid. Although these methods provide information that contributes to safety assessment, they do not provide a direct evaluation of the ability of a novel protein to cause allergic sensitization. For this reason considerable interest exists in the design and evaluation of suitable animal models that may provide a more holistic assessment of allergenic potential. The experimental strategy we have adopted is to measure allergenic activity as a function of the ability of proteins to provoke IgG and IgE antibody responses in BALB/c strain mice, a strain that is known to favor immune responses of the quality required for allergic sensitization. To date, emphasis has focused on the characterization of humoral immune responses induced following systemic exposure of mice (by intraperitoneal administration) to the test material. Under these conditions it has been found that proteins known to be associated with food allergy in humans (including peanut proteins and ovalbumin) elicit both specific IgG and IgE antibody responses, measured using enzyme‐linked immunosorbant and homologous passive cutaneous anaphylaxis assays, respectively. In contrast, other proteins derived from foods believed to exhibit little or no allergenic potential (such as those derived from the potato), although immunogenic in BALB/c mice (measured as a function of IgG antibody production), either failed to induce IgE antibody responses at all or were associated with only low‐grade IgE production at relatively high test concentrations. In comparative analyses, exposure of BALB/c mice to proteins by gavage was found to be less sensitive and discriminatory with respect to the induction of IgE. Experience to date indicates that the inherent sensitizing potential of novel food proteins can be evaluated on the basis of antibody responses stimulated by parenteral exposure of BALB/c strain mice.
Ibogaine Signals Addiction Genes and Methamphetamine Alteration of Long‐Term Potentiation
Authors: ONAIVI EMMANUEL S. , ALI SYED F. , CHIRWA SANIKA S. , ZWILLER JEAN , THIRIET NATHALIE , AKINSHOLA B. EMMANUEL , ISHIGURO HIROKI
Published: July 2010

Abstract:The mapping of the human genetic code will enable us to identify potential gene products involved in human addictions and diseases that have hereditary components. Thus, large‐scale, parallel gene‐expression studies, made possible by advances in microarray technologies, have shown insights into the connection between specific genes, or sets of genes, and human diseases. The compulsive use of addictive substances despite adverse consequences continues to affect society, and the science underlying these addictions in general is intensively studied. Pharmacological treatment of drug and alcohol addiction has largely been disappointing, and new therapeutic targets and hypotheses are needed. As the usefulness of the pharmacotherapy of addiction has been limited, an emerging potential, yet controversial, therapeutic agent is the natural alkaloid ibogaine. We have continued to investigate programs of gene expression and the putative signaling molecules used by psychostimulants such as amphetamine inin vivoandin vitromodels. Our work and that of others reveal that complex but defined signal transduction pathways are associated with psychostimulant administration and that there is broad‐spectrum regulation of these signals by ibogaine. We report that the actions of methamphetamine were similar to those of cocaine, including the propensity to alter long‐term potentiation (LTP) in the hippocampus of the rat brain. This action suggests that there may be a “threshold” beyond which the excessive brain stimulation that probably occurs with compulsive psychostimulant use results in the occlusion of LTP. The influence of ibogaine on immediate early genes (IEGs) and other candidate genes possibly regulated by psychostimulants and other abused substances requires further evaluation in compulsive use, reward, relapse, tolerance, craving and withdrawal reactions. It is therefore tempting to suggest that ibogaine signals addiction gene products.
Alcohol Exposure During Adulthood Induces Neuronal and Astroglial Alterations in the Hippocampal CA‐1 Area
Authors: TAGLIAFERRO PATRICIA , VEGA MAITE DUHALDE , EVRARD SERGIO GUSTAVO , RAMOS ALBERTO JAVIER , BRUSCO ALICIA
Published: July 2010

Abstract:Ethanol (ETOH) exposure can result in neuronal damage. Astrocytes are morphologic and functionally related to neurons, and astrocyte‐neuron interactions provide strategic sites for the actions of many chemical compounds. The aim of the present work was to study the morphologic alterations of glial cells and neurons on the hippocampus after long‐term ETOH exposure using GFAP and S‐100β protein, neurofilaments of 200 kDa (Nf200), MAP2, and serotonin transporter (5HT‐T) immunocytochemical staining. Adult Wistar male rats (200‐250 g) were orally exposed to ETOH (6.6% v/v ad libitum) for 6 weeks. Control rats received water ad libitum. Brain sections from control and exposed rats were processed by immunocytochemistry. After ETOH exposure we observed in the CA1 area of the hippocampus: (1) an important astroglial reaction evidenced by the presence of GFAP+reactive astrocytes; (2) an increase in S‐100β immunostaining in astroglial cells; and (3) a decrease in Nf200, 5HT‐T, and MAP2 immunoreactivity. The current study provides evidence that long‐term ETOH exposure induces alterations in the neuronal cytoskeleton and an astroglial reaction, which is a common response to brain injury and may promote functional recovery of the nervous system, as by the release of glial‐derived trophic factors (such as S‐100β) that promote cell survival and neurite growth.
Brain Regions Mediating the Discriminative Stimulus Effects of Nicotine in Rats
Authors: MIYATA HISATSUGU , ANDO KIYOSHI , YANAGITA TOMOJI
Published: July 2010

Abstract: The involvement of cerebral regions in the discriminative stimulus (DS) effects of nicotine was studied using rats. Substitution tests with nicotine administered into the medial prefrontal cortex, nucleus accumbens, and ventral tegmental area, all of which are located on the mesolimbocortical dopaminergic neurons, and into the dorsal hippocampus and medial habenular nucleus, which possess high densities of nicotinic cholinergic receptors, were conducted in rats trained to discriminate nicotine (0.5 mg/kg sc) from saline solution in a two‐lever, food‐reinforced, operant task. Nicotine administered into the medial prefrontal cortex substituted for nicotine (0.5 mg/kg sc), whereas nicotine administered into the nucleus accumbens and ventral tegmental area partially substituted for sc injected nicotine. However, nicotine administered into the dorsal hippocampus and medial habenular nucleus did not substitute for sc injected nicotine. These results suggest that the medial prefrontal cortex is primarily involved in the DS effects of nicotine, whereas the nucleus accumbens and ventral tegmental area are partially involved.
Protein Kinase C and Lipid‐Induced Insulin Resistance in Skeletal Muscle
Authors: SCHMITZ‐PEIFFER CARSTEN
Published: July 2010

Abstract: Insulin resistance of skeletal muscle in humans, animals, and cells is often strongly correlated with increased lipid availability. The elevation of certain intracellular lipid species can lead to the activation of signal transduction pathways that inhibit normal insulin action. Thus, increased diacylglycerol levels in muscle are associated with the activation of one or more isoforms of the protein kinase C family, which is known to attenuate insulin signaling, especially at the level of IRS‐1. In addition, de novo synthesis of ceramide can inhibit more distal sites by the activation of protein phosphatase 2A and hence promote the dephosphorylation and inactivation of protein kinase B. Such mechanisms may account at least in part for the reduced insulin sensitivity occurring in obesity and type 2 diabetes where lipid oversupply is a major factor.
A Custom‐Built Insulin Resistance Gene Chip
Authors: WALDER KEN , SEGAL DAVID , CHEHAB SAM , AUGERT GUY , CAMERON‐SMITH DAVID , HARGREAVES MARK , COLLIER GREG R.
Published: July 2010

Abstract: Objectives/Aim—Microarray (gene chip) technology offers a powerful new tool for analyzing the expression of large numbers of genes in many experimental samples. The aim of this study was to design, construct, and use a gene chip to measure the expression levels of key genes in metabolic pathways related to insulin resistance. Methods—We selected genes that were implicated in the development of insulin resistance, including genes involved in insulin signaling; glucose uptake, oxidation, and storage; fat uptake, oxidation, and storage; cytoskeletal components; and transcription factors. The key regulatory genes in the pathways were identified, along with other recently identified candidate genes such as calpain‐10. A total of 242 selected genes (including 32 internal control elements) were sequence‐verified, purified, and arrayed on aldehyde‐coated slides. Results—Where more than 1 clone containing the gene of interest was available, we chose those containing the genes in the 5′ orientation and an insert size of around 1.5 kb. Of the 262 clones purchased, 56 (21%) were found to contain sequences other than those expected. In addition, 2 (1%) did not grow under standard conditions and were assumed to be nonviable. In these cases, alternate clones containing the gene of interest were chosen as described above. The current version of the Insulin Resistance Gene Chip contains 210 genes of interest, plus 48 control elements. A full list of the genes is available at http://www.hbs.deakin.edu.au/mru/research/gene_chip_tech/genechip_three.htm/. Conclusions—The human Insulin Resistance Gene Chip that we have constructed will be a very useful tool for investigating variation in the expression of genes relevant to insulin resistance under various experimental conditions. Initially, the gene chip will be used in studies such as exercise interventions, fasting, euglycemic‐hyperinsulinemic clamps, and administration of antidiabetic agents.
Neuroimaging and Obesity
Authors: DEL PARIGI ANGELO , GAUTIER JEAN‐FRANCOIS , CHEN KEWEI , SALBE ARLINE D. , RAVUSSIN ERIC , REIMAN ERIC , TATARANNI P. ANTONIO
Published: July 2010

Abstract: The hypothalamus has a major role in the control of food intake. However, neurotracing studies have shown that the hypothalamus receives input from several other regions of the brain that are likely to modulate its activity. Of particular interest to the understanding of human eating behavior is the possible involvement of the cortex. Using positron emission tomography (PET), we generated functional brain maps of the neuroanatomical correlates of hunger (after a 36‐h fast) and satiation (after oral administration of a liquid formula meal) in lean and obese subjects. Results in lean individuals indicate that the neuroanatomical correlates of hunger form a complex network of brain regions including the hypothalamus, thalamus, and several limbic/paralimbic areas such as the insula, hippocampal/parahippocampal formation, and the orbitofrontal cortex. Satiation was associated with preferentially increased neuronal activity in the prefrontal cortex. Our studies also indicate that the brain responses to hunger/satiation in the hypothalamus, limbic/paralimbic areas (commonly associated with the regulation of emotion), and prefrontal cortex (thought to be involved in the inhibition of inappropriate response tendencies) might be different in obese and lean individuals. In conclusion, neuroimaging of the human brain is proving to be an important tool for understanding the complexity of brain involvement in the regulation of eating behavior. PET studies might help to unravel the neuropathophysiology underlying human obesity.
Endocrine Regulation of Subcutaneous Fat Metabolism during Cold Exposure in Humans
Authors: KOSKA JURAJ , KSINANTOVA LUCIA , ŠEBÖKOVÁ ELENA , KVETNANSKY RICHARD , KLIMEŠ IWAR , CHROUSOS GEORGE , PACAK KAREL
Published: July 2010

Abstract: Increased oxidation of carbohydrates and free fatty acids is a well‐known phenomenon during cold stress. Nevertheless, sources of the fuels used have not been fully clarified as yet. Thus, the aim of our study was to evaluate the effect of acute cold exposure on lipid and carbohydrate metabolism in human subcutaneous adipose tissue and to identify the possible regulatory mechanisms involved. Ten volunteers were exposed for 30 min to an ambient temperature of 4°C. Interstitial metabolism was assessed with the aid of the microdialysis technique. Lipolysis intensity was evaluated from changes of glycerol concentration in plasma and in dialysate. Cold exposure induced a significant increase of glycerol concentration both in plasma (by 199 ± 16%, p < 0.01) and in dialysate (by 308 ± 58%, p < 0.001). No changes in glucose concentration were found whether in plasma or in the dialysate. Ethanol concentration in dialysate increased (148 ± 15%, p < 0.01), indicating a slower blood flow in the subcutaneous region. Plasma concentrations of various gluco‐ and/or lipid‐regulatory hormones remained unaffected by the cold exposure, except for norepinephrine, which rose about threefold (309 ± 41%, p < 0.001). The data indicate an important role for subcutaneous adipose tissue in mobilization of free fatty acids during cold exposure. This process seems to be regulated by the sympathetic nervous system, whereas hormones involved in the regulation of lipid metabolism, such as epinephrine, insulin, cortisol, and growth hormone, may play a less significant role—at least under the conditions studied.
Association between a Variant at the GABAAα6 Receptor Subunit Gene, Abdominal Obesity, and Cortisol Secretion
Authors: ROSMOND ROLAND , BOUCHARD CLAUDE , BJÖRNTORP PER
Published: July 2010

Abstract: In the present study, we examined the potential impact of a T‐to‐C substitution at nucleotide 1519 of the GABAAα6 receptor subunit gene (GABRA6) on obesity and obesity‐related phenotypes as well as circulating hormones, including salivary cortisol, in 284 unrelated Swedish men born in 1944. The subjects were genotyped by using PCR amplification followed by digestion with the restriction enzyme AlwNI. The frequency of allele T was 0.54 and that of allele C was 0.46. Carriers for the T allele (n= 211) had higher waist‐to‐hip ratio (p= 0.094) and abdominal sagittal diameter (p= 0.084) compared to homozygotes for the C allele (n= 56). The homozygotes for the T allele had, in comparison to heterozygotes, significantly (p= 0.004‐0.024) higher mean cortisol levels at 11:45 am; at 30, 45, and 60 min after a standardized lunch; and finally at 5:00 pm. In addition, T/T subjects had significantly (p= 0.031) higher diurnal cortisol secretion compared to T/C subjects. Leptin, insulin, and glucose were not different across the genotype groups. In conclusion, these findings suggest a role of the point substitution (T‐to‐C) at nucleotide 1519 of GABRA6 in the predisposition to hypercortisolism and perhaps abdominal obesity. The pathophysiology may involve various environmental factors, particularly stress, that destabilize the GABA‐hypothalamic‐pituitary‐adrenal systems in those with genetic vulnerability.
The Na+/Ca2+ Exchange Molecule
Authors: PHILIPSON KENNETH D. , NICOLL DEBORA A. , OTTOLIA MICHELA , QUEDNAU BEATE D. , REUTER HANNES , JOHN SCOTT , QIU ZHIYONG
Published: July 2010

Abstract: An overview of the molecular physiology of the Na+/Ca2+ exchanger is presented. This includes information on the variety of exchangers that have been described and their regulatory properties. Molecular insight is most detailed for the cardiac Na+/Ca2+ exchanger (NCX1). Parts of the NCS1 molecule involved in regulation and ion transport have been elucidated, and initial information on the topology and structure is available.
Pathways Regulating Na+/Ca2+ Exchanger Expression in the Heart
Authors: MENICK DONALD R. , XU LIN , KAPPLER CHRISTIANA , JIANG WENJING , WITHERS PATRICK R. , SHEPHERD NEAL , CONWAY SIMON J. , MÜLLER JOACHIM G.
Published: July 2010

Abstract: The Na+/Ca2+ exchanger (NCX1) is regulated at the transcriptional level in cardiac hypertrophy, ischemia, and failure. Following pressure overload, activation of MAPKs coincides with the kinetics of NCX1 gene upregulation in adult cardiocytes. Using adenoviral gene delivery, we begin to identify the molecular pathways responsible for upregulation of the exchanger gene. Inhibition of ERK with the MEK inhibitor UO126, the ERK protein phosphatase MKP‐3, inhibited ERK activation, but only inhibited α‐adrenergic‐induced NCX1 upregulation by 30%. Overexpression of DN‐JNK lowered basal NCX1 expression. Overexpression of activated MKK‐3 was sufficient for α‐adrenergic‐stimulated upregulation of the reporter gene. Together, this data indicates that (1) JNK mediates basal cardiac expression of the NCX1 gene, (2) ERK and p38 play a role in α‐adrenergic‐stimulated NCX1 upregulation, and (3) p38 activation alone is sufficient for NCX1 upregulation.
Cyclosporin A Regulates Sodium‐Calcium Exchanger (NCX1) Gene Expression in Vitro and Cardiac Hypertrophy in NCX1 Transgenic Mice
Authors: JORDAN MARIA C. , QUEDNAU BEATE D. , ROOS KENNETH P. , ROSS ROBERT S. , PHILIPSON KENNETH D. , NICHOLAS SUSANNE B.
Published: July 2010

Abstract: The cardiac‐specific sodium‐calcium exchanger (NCX1) is a GATA‐4 dependent gene that is upregulated during cardiac hypertrophy and heart failure. To date, lack of an appropriate inhibitor of NCX1 and embryonic lethality of NCX1 knockout mice have slowed investigation of the relation between NCX1 upregulation and cardiac hypertrophy. Recently, in vitro studies have shown that cyclosporin A (CSA), a calcineurin inhibitor, significantly downregulated expression of the hypertrophic genes atrial natriuretic factor and β‐myosin heavy chain and protected against cardiac hypertrophy and heart failure in calcineurin overexpressing mice. This suggested that CSA might play an important role in the treatment of hypertrophy and heart failure. In an in vitro model of cardiac hypertrophy, we showed that CSA is a potent inhibitor of NCX1 basal expression and NCX1 promoter activity. Female homozygous transgenic mice that overexpress NCX1 develop heart failure and die prematurely after two or more pregnancies. Others have demonstrated that pressure overloaded wild‐type mice treated with CSA do not develop cardiac hypertrophy and downregulate expression of NCX1. We investigated the effect of CSA on NCX1 expression and transverse aortic constriction‐induced cardiac hypertrophy in NCX1 overexpressing mice. We found that CSA blunted these responses.
Regulation of Phosphatidylinositol‐4,5‐Biphosphate Bound to the Bovine Cardiac Na+/Ca2+ Exchanger
Authors: BEAUGÉ LUIS , ASTEGGIANO CARLA , BERBERIÁN GRACIELA
Published: July 2010

Abstract: Western blot and cross immunoprecipitation analysis with specific antibodies demonstrate that in bovine heart sarcolemmal vesicles phosphatidylinositol‐4,5‐biphosphate (PtdIns‐4,5‐P2) binds strongly to the Na+/Ca2+ exchanger (NCX1). This binding is modulated by ATP, Ca2+, vanadate, exchanger inhibitory peptide (XIP), and PLC‐PtdIns specific in a way resembling the ATP regulation of the exchange fluxes. With 1 μM Ca2+, 3 mM Mg2+, and 0.4 mM vanadate, 1 mM ATP increased about twofold the bound PtdIns‐4,5‐P2, reaching a steady state in 3–5 s at 37°C. With 100 μM Ca2+, ATP had no effect on the PtdIns‐4,5‐P2 bound to NCX1 or on the exchange fluxes. Without vanadate the bound PtdIns‐4,5‐P2 was largely reduced; under this condition ATP failed to increase it and did not stimulate the exchanger. XIP inhibits the exchanger, more noticeable in the absence of ATP. With XIP, ATP does not modify the levels of bound PtdIns‐4,5‐P2; however there is a small but distinct ATP stimulation of the exchanger. Vesicles pretreated with PtdIns‐PLC, showed no de novo, [32P]ATP‐induced, production of PtdIns‐4,5‐P2, but some ATP‐stimulated increase in the bound PtdIns‐4,5‐P2 was detected; however, that increase did not exceed the levels found with vanadate and no ATP. These results indicate that in bovine heart sarcolemmal vesicles, ATP upregulation of NCX1 is related to PtdIns‐4,5‐P2 bound to the exchanger, perhaps over a “threshold” or “unspecific” amount. In addition, vanadate could influence the amount of detected PtdIns‐4,5‐P2 either by inhibiting phosphoinositide‐specific phosphatases and/or by inducing a redistribution of PtdIns‐4,5‐P2 molecules associated with the Na+/Ca2+ exchanger.
Na/Ca Exchanger and PMCA Localization in Neurons and Astrocytes
Authors: BLAUSTEIN M. P. , JUHASZOVA M. , GOLOVINA V. A. , CHURCH P. J. , STANLEY E. F.
Published: July 2010

Abstract: Immunocytochemistry reveals that the Na/Ca exchanger (NCX) in neuronal somata and astrocytes is confined to plasma membrane (PM) microdomains that overlie sub‐PM (junctional) endoplasmic reticulum (jER). By contrast, the PM Ca2+ pump (PMCA) is more uniformly distributed in the PM. At presynaptic nerve terminals, the NCX distribution is consistent with that observed in the neuronal somata, but the PMCA is clustered at the active zones. Thus, the PMCA, with high affinity for Ca2+ (Kd≅100 nM), may keep active zone Ca2+ very low and thereby “reprime” the vesicular release mechanism following activity. NCX, with lower affinity for Ca2+ (Kd≅1,000 nM), on the other hand, may extrude Ca2+ that has diffused away from the active zones and been temporarily sequestered in the endoplasmic reticulum. The PL microdomains that contain the NCX also contain Na+ pump high ouabain affinity α2 (astrocytes) or α3 (neurons) subunit isoforms (IC50≅5‐50 nM ouabain). In constrast, the α1 isoform (low ouabain affinity in rodents; IC50 >10,000 nM), like the PMCA, is more uniformly distributed in these cells. The sub‐PM endoplasmic reticulum in neurons (and probably glia and other cell types as well) and the adjacent PM form junctions that resemble cardiac muscle dyads. We suggest that the PM microdomains containing NCX and α2/α3 Na+ pumps, the underlying jER, and the intervening tiny volume of cytosol (<10−18 l) form functional units (PLasmERosomes); diffusion of Na+ and Ca2+ between these cytosolic compartments and “bulk” cytosol may be markedly restricted. The activity of the Na+ pumps with α2/α3 subunits may thus regulate NCX activity and jER Ca2+ content. This view is supported by studies in mice with genetically reduced (by ≅50%) α2 Na+ pumps: evoked Ca2+ transients were augmented in these cells despite normal cytosolic Na+ and resting Ca2+ concentrations ([Na+]CYT and [Ca2+]CYT). We conclude that α2/α3 Na+ pumps control PLasmERosome (local) [Na+]CYT. This, in turn, via NCX, modulates local [Ca2+]CYT, jER Ca2+ storage, Ca2+ signaling, and cell responses.
A Study of the Activity of the Plasma Membrane Na/Ca Exchanger in the Cellular Environment
Authors: BRINI MARISA , MANNI SABRINA , CARAFOLI ERNESTO
Published: July 2010

Abstract: The Na/Ca exchanger is a large‐capacity system that is supposed to be predominant in the ejection of calcium in excitable tissues. Its activity is normally evaluated in electro‐physiological experiments or using Na‐loaded cells or plasma membrane vesicles in experiments in which the activity is estimated by measuring the uptake of isotopic calcium. Here the activity of the exchanger has been evaluated in model cells that coexpress the exchanger together with the (protein) Ca indicator aequorin targeted to the different subcellular compartments. Exchanger isoform 1 (NCX1) has been used for experiments in which the effect of its overexpression has been evaluated on the Ca homeostasis in the cytoplasm at large, in the endoplasmic reticulum, in mitochondria, and in the layer of cytoplasm underneath the plasma membrane. The experiments have shown that, despite its very low calcium affinity, the exchanger was active in cells at rest in which calcium is around 100 nM. They have also shown that expressed NCX1 significantly reduced the cytosolic and mitochondrial calcium transients produced by the emptying of the ER calcium stores with InsP3‐linked agonists. The effect of the expressed NCX1 on the calcium homeostasis in the domain underneath the plasma membrane was more complex: The exchanger apparently catalyzed a reverse operation, leading to increased penetration of calcium.
Theories in Early Embryology
Authors: Van SPEYBROECK LINDA , de WAELE DANI , Van de VIJVER GERTRUDIS
Published: July 2010

Abstract:In current biological and philosophical literature, the use of the termsepigenesisandepigeneticshas increased tremendously. As these terms are often confused, this paper aims at clarifying the distinction between them by drawing their conceptual and historical evolutions. The evolution of the termepigenesisis situated in the context of early embryological studies. Departing from Aristotle's natural philosophy, it is shown thatepigenesisgained alternating attention from the 17th century onwards, as it was introduced into neo‐classical embryology and considered to be the opposite of thepreformationisttradition. Where preformation stated that the germ cells of each organism contain preformed miniature adults that unfold during development, epigenesis held that the embryo forms by successive gradual exchanges in an amorphous zygote. Although both traditions tried to explain developmental organization, religious and metaphysical arguments on the conception of embryonic matter as either active or passive determined the scope of their respective explanations. It is shown that these very arguments still underlie the use of gene‐centric metaphors in the molecular revolution of the 20th century.
From Epigenesis to Epigenetics
Authors: Van Speybroeck Linda
Published: July 2010

Abstract:One continuous thread in this volume is the name of Conrad H. Waddington (1905‐1975), the developmental biologist known as the inventor of the termepigenetics. After some biographical notes on his life, this article explores the meaning of the Waddingtonian equation and the context wherein it was developed. This equation holds thatepigenesis+genetics=epigenetics, and refers in retrospect to the debate onepigenesisversuspreformationismin neoclassical embryology. Whereas Waddington actualized this debate by linking epigenesis to developmental biology and preformation to genetics, thereby stressing the importance of genetic action in causal embryology, today'sepigeneticsmore and more offers the possibility to enfeeble biological thinking in terms of genes only, as it expands the gene‐centric view in biology by introducing a flexible and pragmatically oriented hierarchy of crucial genomic contexts that go beyond the organism.
What Is “Epi” about Epigenetics?
Authors: GRIESEMER JAMES
Published: July 2010

Abstract:What counts as epigenetic depends on what counts as genetic. It is argued that Weismannism, the doctrine of genetic continuity and somatic discontinuity, is the basis for an overly inclusive concept of epigenetics as every inherited resource “beyond the genes.” An alternative theoretical perspective, the “reproducer” concept, is introduced to facilitate analysis of multiple inheritance systems without labeling all nongenetic inheritance “epigenetic.”
Alternative Epigenetic States Understood in Terms of Specific Regulatory Structures
Authors: THIEFFRY DENIS , SÁNCHEZ LUCAS
Published: July 2010

Abstract: Generally speaking, epigenetic states or epigenetic regulation refer to situations in which several states of gene expression may coexist in similar environmental conditions, despite the absence of significant changes in the genomic sequence. In one way or another, the mechanisms behind these phenomena involve vicious circles, so that each epigenetic state tends to sustain itself, even after the disappearance of the inductive signal involved in the selection of that particular state. These vicious circles constitute positive feedback circuits and are found at the core of many developmental regulatory systems. In this paper, we present a qualitative model for the regulatory network formed by maternal and gap gene cross‐regulations. This network controls the initial anterior‐posterior patterning during early Drosophila embryogenesis and encompasses several intertwined feedback circuits. On the basis of our model analysis, we derive interesting insights about how specific expression states of the gap genes are selected along the anterior‐posterior axis, in particular in relation with the activity of one positive feedback circuit, namely that formed by giant and Krüppel cross‐inhibitions. In addition, we are able to qualitatively simulate the patterns of gene expression in the wild‐type, as well as to predict the phenotypes of various loss‐of‐function mutations at the maternal and gap genes, or cis‐regulatory mutations at the gap genes, as well as the effects of ectopic expression of these genes.
Remembering Professor Cesare Maltoni
Authors: MEHLMAN MYRON A.
Published: July 2010

Abstract: Professor Cesare Maltoni, a renowned leader in the research of the hazards of carcinogens in the workplace, died on January 22, 2001 at the age of 70. Born in Faenza (Ravenna), Italy on November 17, 1930, he received his M.D. degree from the University of Bologna in 1954‐1955. He was Director of the Institute of Oncology of Bologna (1964 to 1997), Director of the Bologna Centre for the Prevention and Detection of Tumours and Oncological Research (1966 to 1989), and Scientific Director, European Foundation of Oncology and Environmental Sciences “B. Ramazzini” from 1993 until he died. Maltoni conducted long‐term carcinogenic studies on some 200 agents. He was the first to demonstrate that vinyl chloride is a carcinogen that produces, among other tumors, angiosarcoma of the liver. He was the first to show that benzene is a powerful multipotential carcinogen. Maltoni authored more than 700 original scientific publications, books, and proceedings. He was editor and coeditor of many journals. Among his many awards were the Stokinger Award, American Conference of Governmental Industrial Hygienists (ACGIH), Kansas City, 1995; International Award “B. Ramazzini” of the Collegium Ramazzini, Washington, 1995; International I.J. Selikoff Memorial Award, Washington, 1995; and the Sigillum Magnum of the University of Bologna, 1997. Many fellow Ramazzinians and his coworkers wish to be considered part of the group of, as he once wrote, his “always family friends” and to remember happy moments when we were together. A man of great stature and many contributions, Cesare Maltoni will never be forgotten.