bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ready, set, internalize: mechanisms and regulation of GLUT4 endocytosis
Authors: Antonescu Costin N. , Foti Michelangelo , Sauvonnet Nathalie , Klip Amira 
Published: December 2008

The facilitative glucose transporter GLUT4, a recycling membrane protein, is required for dietary glucose uptake into muscle and fat cells. GLUT4 is also responsible for the increased glucose uptake by myofibres during muscle contraction. Defects in GLUT4 membrane traffic contribute to loss of insulin-stimulated glucose uptake in insulin resistance and Type 2 diabetes. Numerous studies have analysed the intracellular membrane compartments occupied by GLUT4 and the mechanisms by which insulin regulates GLUT4 exocytosis. However, until recently, GLUT4 internalization was less well understood. In the present paper, we review: (i) evidence supporting the co-existence of clathrin-dependent and independent GLUT4 internalization in adipocytes and muscle cells; (ii) the contrasting regulation of GLUT4 internalization by insulin in these cells; and (iii) evidence suggesting regulation of GLUT4 endocytosis in muscle cells by signals associated with muscle contraction.
Predictors of serious complications due to Clostridium difficile infection
Authors: GUJJA D. , FRIEDENBERG F. K. 
Published: December 2008

SummaryBackground  Identifying individuals with severe Clostridium difficile infection (CDI) at risk for major complications has become an important objective. Presence of clinical variables that predict complications from CDI would have the potential to strongly influence management.Aim  To determine which clinical variables predict complications from CDI.Methods  Cross‐sectional study of all individuals admitted to Temple University Hospital between 12/1/03 and 7/1/08 with the primary discharge diagnosis of CDI were eligible. Only patients experiencing their first episode of CDI were included. Abstracted data included demographic, physiological, laboratory, radiological, endoscopic, pharmacy and outcome data. Response was categorized as none, partial or complete. Complications attributed to CDI were defined as colon resection or death.Results  Overall 32 of 200 patients (16%) experienced a complication due to CDI including death (n = 20) and colectomy (n = 12). White blood cell count above 30,000 cells/mm3 (OR = 4.06; 95% CI, 1.28–12.87) and a rise in the creatinine to over 50% above baseline (OR = 7.13; 95% CI, 3.05–16.68) predicted a complication. AROC for percent rise in serum creatinine was 0.73 (95% CI: 0.64–0.85) and 0.62 (95% CI: 0.58–0.80) for white blood cell count.Conclusions  Severe white blood cell count elevation and a rise in the creatinine to over 50% above baseline are important independent predictors of serious adverse events due to CDI. These patients likely would benefit from more intensive care and early surgical consultation.
Manufacturing of human placenta‐derived mesenchymal stem cells for clinical trials
Authors: Brooke Gary , Rossetti Tony , Pelekanos Rebecca , Ilic Nina , Murray Patricia , Hancock Sonia , Antonenas Vicki , Huang Gillian , Gottlieb David , Bradstock Ken , Atkinson Kerry 
Published: December 2008

SummaryMesenchymal stem cells (MSC) are being used increasingly in clinical trials for a range of regenerative and inflammatory diseases. Bone marrow is the traditional source but is relatively inaccessible in large volume. MSC have now been derived from tissues other than bone marrow including placenta and adipose tissue. We have used placenta obtained after delivery as a source of MSC and have been unable to detect any marked differences from marrow‐derived MSC in terms of cell surface phenotype, chemokine receptor display, mesodermal differentiation capacity or immunosuppressive ability. This report described our manufacturing process for isolating and expanding placenta‐derived human MSC and their safe infusion into the first patient in a clinical trial program of human placenta‐derived MSC.
T regulatory cells distinguish two types of primary hypophysitis
Authors: Mirocha S , Elagin R B , Salamat S , Jaume J C 
Published: December 2008

Summary
Numerous cases of primary hypophysitis have been described over the past 25 years with, however, little insight into the cause(s) of this disease. In order to guide treatment, a better understanding of the pathogenesis is needed. We studied the pathogenesis of primary hypophysitis by analysing systematically the immune response at the pituitary tissue level of consecutive cases of ‘lymphocytic’ hypophysitis who underwent pituitary biopsy. In order to investigate further the pathogenesis of their diseases we characterized two cases at clinical, cellular and molecular levels. We show here, for the first time, that lymphocytic hypophysitis probably encompasses at least two separate entities. One entity, in agreement with the classical description of lymphocytic hypophysitis, demonstrates an autoimmune process with T helper 17 cell dominance and lack of T regulatory cells. The other entity represents a process in which T regulatory cells seem to control the immune response, which may not be self- but foreign-targeted. Our data suggest that it may be necessary to biopsy suspected primary hypophysitis and to analyse pituitary tissue with immune markers to guide treatment. Based on our results, hypophysitis driven by an immune homeostatic process should not be treated with immunosuppression, while autoimmune-defined hypophysitis may benefit from it. We show here for the first time two different pathogenic processes classified under one disease type and how to distinguish them. Because of our findings, changes in current diagnostic and therapeutic approaches may need to be considered.
A critical appraisal of erectile function in animal models of diabetes mellitus
Authors: Gur Serap , Kadowitz Philip J. , Hellstrom Wayne J. G. 
Published: December 2008

SummaryThe study of erectile function in diabetic animal models has revealed physiological alterations in neural, vascular, hormonal and endothelial function. The aims of this review are to further elucidate pathophysiological changes induced by diabetes mellitus and to introduce new concepts in the study of erectile dysfunction (ED) in animal models. The recognized pathophysiological mechanisms causing diabetic ED include oxidative stress and hormonal imbalance. The evolving treatments for ED include advanced glycosylated endproduct (AGE) inhibitors, phosphodiesterase type 5 inhibitors, protein kinase C (PKC) inhibitors, hormone replacement, and gene transfer techniques. Our current understanding of how these multiple pathophysiological mechanisms contribute to ED is discussed. In this review, diabetic animal model studies have documented that oxidative stress is a pre‐eminent pathophysiological mechanism and several anti‐oxidants, such as α‐lipoic acid, vitamin E, sodium selenate, melatonin, and ascorbic acid, reverse both neurogenic and endothelial dysfunction in diabetic models. Further, the peroxynitrite decomposition catalyst – FeTMPyP, PKCβ selective inhibitor – LY333531, IkappaB kinase 2 inhibitor – AS602868, AGE inhibitors – aminoguanidine and ALT‐711 show promise by exploring different cellular mechanisms in treating diabetic problems. A number of vectors have been used to insert genes to increase the expression of nitric oxide synthase, superoxide dismutase, maxi‐K channel (hSlo), neurotrophin‐3, and vasoactive intestinal polypeptide for the treatment of erectile function. Further investigation of the hormonal treatment of diabetes associated with hypogonadism may improve sildenafil responsiveness in diabetic patients. We are optimistic that novel prevention and treatment strategies for diabetic ED are on the horizon.
Retracted: Effects of exposure to a mobile phone on testicular function and structure in adult rabbit
Authors: Salama Nader , Kishimoto Tomoteru , Kanayama Hiro‐omi 
Published: December 2008

SummaryThe accumulating effects of exposure to electromagnetic radiation emitted by a conventional mobile phone (standby position) on the testicular function and structure are not yet fully investigated. To study these effects longitudinally, a total of 24 adult male rabbits were randomly and equally divided into three groups. Rabbits in the first (phone) group were exposed, in specially designed cages, to radio frequency emitted from the mobile phone (800 MHz) in a standby position opposite to that of testes for 8 h daily for 12 weeks. The second group consisted of the stress controls which were kept in the same kind of cages to appreciate any cage‐induced anxiety. The third group included the ordinary controls which were kept in the conventional roomy cages. Semen analysis and sperm function tests (viability, hypo‐osmotic swelling and acridine orange) were conducted weekly. Histological testicular sections and serum total testosterone were also evaluated. A drop in the sperm concentration appeared in the phone group at week 6. This became statistically significant at week 8, compared with the two control (stress and ordinary) groups (133, 339 and 356 × 106/mL, respectively) and to the initial sperm count (341 × 106/mL) of this group. Motile sperm population showed similarity amongst the three study groups until week 10 when it declined significantly, and thereafter in the phone and stress control groups, with more significant decline in the phone animals (50, 61 and 72.4%, respectively). Histological examination showed also a significant decrease in the diameter of seminiferous tubules in the phone group vs. the stress and ordinary controls (191 μm vs. 206 and 226 μm, respectively). The other study points did not show any difference. In conclusion, low intensity pulsed radio frequency emitted by a conventional mobile phone kept in the standby position could affect the testicular function and structure in the adult rabbit.
Biological and clinical significance of DNA damage in the male germ line
Authors: Aitken R. John , De Iuliis Geoffry N. , McLachlan Robert I. 
Published: December 2008

SummaryDNA damage is a common feature of human spermatozoa with purported links to poor rates of conception, impaired embryonic development, an increased incidence of miscarriage and the appearance of various kinds of morbidity in the offspring including childhood cancer. However, difficulties in interpretation arise, because these associations are not consistently observed across all data sets. Such inconsistency reflects the inherent complexity of the reproductive process, large variations in sample size, differences in patient selection, inadequate study design as well as inter‐individual differences in the type of DNA damage being detected and the effectiveness of repair mechanisms in the oocyte. This review considers the type, source and measurement of DNA damage in human spermatozoa. It also addresses the clinical utility of the information generated in such studies, and highlights areas where further research is needed to bridge the gap between an intriguing biological phenomenon and the evidence‐based clinical management of male patients characterized by high levels of DNA damage in their spermatozoa.
The role of light in soybean seed filling metabolism
Authors: Allen Doug K. , Ohlrogge John B. , Shachar‐Hill Yair 
Published: December 2008

SummarySoybean (Glycine max) yields high levels of both protein and oil, making it one of the most versatile and important crops in the world. Light has been implicated in the physiology of developing green seeds including soybeans but its roles are not quantitatively understood. We have determined the light levels reaching growing soybean embryos under field conditions and report detailed redox and energy balance analyses for them. Direct flux measurements and labeling patterns for multiple labeling experiments including [U‐13C6]‐glucose, [U‐13C5]‐glutamine, the combination of [U‐14C12]‐sucrose + [U‐14C6]‐glucose + [U‐14C5]‐glutamine + [U‐14C4]‐asparagine, or 14CO2 labeling were performed at different light levels to give further insight into green embryo metabolism during seed filling and to develop and validate a flux map. Labeling patterns (protein amino acids, triacylglycerol fatty acids, starch, cell wall, protein glycan monomers, organic acids), uptake fluxes (glutamine, asparagine, sucrose, glucose), fluxes to biomass (protein amino acids, oil), and respiratory fluxes (CO2, O2) were established by a combination of gas chromatography‐mass spectrometry, 13C‐ and 1H‐NMR, scintillation counting, HPLC, gas chromatography‐flame ionization detection, C:N and amino acid analyses, and infrared gas analysis, yielding over 750 measurements of metabolism. Our results show: (i) that developing soybeans receive low but significant light levels that influence growth and metabolism; (ii) a role for light in generating ATP but not net reductant during seed filling; (iii) that flux through Rubisco contributes to carbon conversion efficiency through generation of 3‐phosphoglycerate; and (iv) a larger contribution of amino acid carbon to fatty acid synthesis than in other oilseeds analyzed to date.
A BAHD acyltransferase is expressed in the tapetum of Arabidopsis anthers and is involved in the synthesis of hydroxycinnamoyl spermidines
Authors: Grienenberger Etienne , Besseau Sébastien , Geoffroy Pierrette , Debayle Delphine , Heintz Dimitri , Lapierre Catherine , Pollet Brigitte , Heitz Thierry , Legrand Michel 
Published: December 2008

SummaryBAHD acyltransferases catalyze the acylation of many plant secondary metabolites. We characterized the function of At2g19070, a member of the BAHD gene family of Arabidopsis thaliana. The acyltransferase gene was shown to be specifically expressed in anther tapetum cells in the early stages of flower development. The impact of gene repression was studied in RNAi plants and in a knockout (KO) mutant line. Immunoblotting with a specific antiserum raised against the recombinant protein was used to evaluate the accumulation of At2g19070 gene product in flowers of various Arabidopsis genotypes including the KO and RNAi lines, the male sterile mutant ms1 and transformants overexpressing the acyltransferase gene. Metabolic profiling of flower bud tissues from these genetic backgrounds demonstrated a positive correlation between the accumulation of acyltransferase protein and the quantities of metabolites that were putatively identified by tandem mass spectrometry as N1,N5,N10‐trihydroxyferuloyl spermidine and N1,N5‐dihydroxyferuloyl‐N10‐sinapoyl spermidine. These products, deposited in pollen coat, can be readily extracted by pollen wash and were shown to be responsible for pollen autofluorescence. The activity of the recombinant enzyme produced in bacteria was assayed with various hydroxycinnamoyl‐CoA esters and polyamines as donor and acceptor substrates, respectively. Feruloyl‐CoA and spermidine proved the best substrates, and the enzyme has therefore been named spermidine hydroxycinnamoyl transferase (SHT). A methyltransferase gene (At1g67990) which co‐regulated with SHT during flower development, was shown to be involved in the O‐methylation of spermidine conjugates by analyzing the consequences of its repression in RNAi plants and by characterizing the methylation activity of the recombinant enzyme.
The Assessment of Histocompatibility by Mixed Lymphocyte Reaction as Measured by the Macrophage Electrophoretic Mobility (MEM) Test
Authors: SHENTON B. K. , ULDALL P. R. , SWINNEY J. , FIELD E. J. 
Published: December 2008

The mixed lymphocyte reaction (MLR) between donor and recipienTlymphocytes has been measured by the macrophage electrophoretic mobility (MEM) test and the modified (MOD‐MEM) test. Its value as a measure of compatibility has been assessed by comparison with conventional HL‐A serotyping and with the outcome of renal transplantation.Thirty‐six living donor/recipient pairs and 59 cadaver donor/recipient pairs for transplantation have been studied. Whilst uniovular twins gave lymphocyte interactions, measured as macrophage slowings of about 1 %, the slowing produced by paired allogeneic lymphocytes ranged from 2 % to 26 % depending on the number of HL—A matches.The test measurement of lymphocytic interaction was significantly correlated with histocompatibility measured by HL—A serotyping, in both living and cadaver donors. One way MEM‐MLR showed the dominant role of the second HL—A sublocus in mixed lymphocyte reactivity. The long term success of the renal graft correlated with the pre‐transplant initial reaction between donor and recipienTlymphocytes.The test has advantages in the field of human histocompatibility assessment since no particular reference to individual antigens is made and it may be performed in a matter of hours.
Appearance and Evolution of Anti‐Da (B Cell‐Specific) Antibodies after Planned Immunizations
Authors: Belvedere Mariaclara , Richiardi Patricia , Savi Mario , Curtoni Emilio S. , Carbonara Angelo O. , Ceppellini Ruggero 
Published: December 2008

Appearance and evolution of anti‐Da antibodies has been followed in eight volunteers immunized by whole blood transfusions or leukocyte intradermal injections from a single donor incompatible for HLA—A,—B,—C and—D specificities. Several unabsorbed bleedings from each recipient were studied against the specific immunizer with three different complement‐dependent lymphocytotoxicity (CdL) techniques: (1) standard NIH CdL on total peripheral blood lymphocytes (PBL); (2) VII Workshop standard CdL technique on B cell‐enriched suspensions; (3) β2 microglobulin blanketing test (“bb” test) on B cells.Results obtained with the “bb” test were confirmed with platelet‐absorbed sera. The “bb” and the absorbed sera allow discrimination between anti‐Da and anti‐HLA—A,—B,—C antibodies.Stage of appearance and evolution are rather similar for an anti‐HLA—A,—B,—C and anti‐Da. An early appearance of antibodies positive only against B cells is due to weak anti‐HLA—A,—B antibodies which react better with B cells than with total PBL. Immunogenicity of Da antigens seems to be of the same order as HLA—A, and—B. In fact, Da reactivity was present in all eight recipients studied. These reactivities always segregated in families with these HLA haplotypes. On a small panel of unrelated D‐typed donors, three sera showed a significant positive association with D alleles.
Tissue Typing of Cells in Culture. II. Histocompatibility Typing of Diploid Fibroblastic Cultures by the Mixed Hemadsorption (MH) Test
Authors: Espmark Åke , Möller Erna 
Published: December 2008

Skin fibroblasts derived from persons who had been HLA—typed by conventional lymphocyto‐toxocity tests were subjected to HLA—typing by the mixed hemadsorption (MH) test. The agreement between the two methods was good, provided sera for the MH test were carefully selected. Minor discrepancies found in this study can probably be corrected by further serum selection.There was no difficulty in typing late passages of cultured diploid cells (22 doublings) and in fact it was easier to type a culture at the 33rd doubling than at the 11th. Thus no signs of disappearance of HLA—antigens were seen in these tests.
Studies on the Specificity of CML Report from a CML‐Workshop
Authors: Kristensen Tom 
Published: December 2008

In a collaboratory study involving eight different laboratories 30 human, mixed lymphocyte culture educated cytotoxic/ymphocytes (CTLs) were identified yielding reproducible cytolysis on allogenic lymphocyte target cells without detectable HLA—A, B (and C) antigenic sharing between stimulator and target cells.These CTLs were collected in one laboratory (Aarhus) and tested in parallel against a population sample of 100 unrelated, healthy Danes. The testing was only performed once and 11 CTLs did not discriminate in the population, probably due to transportation damage. On the basis of pairwise comparisons between 19 CTLs, three tentative CML‐defined specificities could be recognized. These three groups may have defined monospecific traits of allelic genetic origin as judged by a mutually negative, albeit not significant, correlation and a fit to Hardy‐Weinberg equilibrium.The concept of determinants other than the serologically defined HLA antigens recognized by some CTLs can thus still be maintained as can the approach to CML typing tested in this workshop.
Sources of variance in the double normalized value: an evaluation of its reproducibility as a measure on HLA‐D locus identity
Authors: Ollier W. , Mendell N. , Sachs J. , Jaraquemada D. , Evans S. , Pegrum G. , Festenstein H. 
Published: December 2008

Two different sets of mixed lymphocyte culture (MLC) experiments were performed using HLA‐D homozygous typing cells (HTC). In all stimulator‐responder combinations the median cpm's of four replicate cultures were reduced into double normalized values(DNV).In the first experimental set the responder panel was unselected, whereas the re‐ sponders for the second set were chosen on the basis of either sharing an HLA‐D determinant with the stimulator (ID) or not (non‐ID).The experiments were designed: Set 1: To estimate the technical variability in the DNV's and to observe the distribution of this variability. The standard deviation of an observation on a stimulator‐responder combination was approximately 24 DNV units. Thus, by running 6 experiments on each responder we would have a mean DNV with a standard error of 10 DNV units. Set 2: To determine whether stimulators typing for the same Dw specificity had the same distribution of DNV's and to investigate the variability between responders, between experiments and within experiments. Although the mean DNV is the same for all HTC's, the variability in observations was greater for some HTC's than for others. The variability may be completely technical for some HTC's, whereas for other HTC's there is evidence of responder variability and between experiment variability. Important implications of these results are: (1) that in using a single cut‐off value of 60 for all HTC to define typing responses one will have a very high misclassification rate for a large percentage of ID responders; (2) for some HTC this error rate can be reduced through repeating the experiment 4 times and raising the cut‐off point; (3) this error, together with the technical and/or experimental variance can be further reduced by using 2 or more HTC's of the same specificity in each experiment and by combining their data; (4) misclassification can be reduced in every situation by doing 4 experiments, using 2 HTC per responder and computing a cut‐off which gives a misclassification rate for each Dw type of 10% in ID's and in non‐ID's.Thus the best approach to achieving Dw locus typing with a desirable low rate of misclassification would be to do similar control studies of every HTC, to estimate technical experimental and responder variance and then use this information to determine a cut‐of value and the number of HTC's and experiments per responder required to keep the error rate at a satisfactory level.