bluuebunny/crossref_metadata_2025 huggingface dataset:
Stathmin reveals dissociable roles of the basolateral amygdala in parental and social behaviors
Authors: Martel Guillaume , Nishi Akinori , Shumyatsky Gleb P.
Published: September 2008

Innate parental behaviors and adult social interactions are essential for survival of the individual along with the species as a whole. Because these behaviors require threat assessment of the environment, it is plausible that they are regulated by the amygdala-associated neural circuitry of fear. However, the amygdala is not a single anatomic and functional unit, and nuclei of the amygdala have multiple inter- and intra-connections. This poses a question as to the exact role of different amygdala nuclei in these behaviors and the mechanisms involved. The basolateral complex of the amygdala nuclei (BLA) is particularly interesting in this regard: although the BLA role in forming memories for learned fear is established, the BLA role in innate behaviors is not well understood. We recently demonstrated that mice without an inhibitor of microtubules,stathmin, a gene enriched in BLA-associated circuitry, have deficiency in innate and learned fear. Here we show that the deficiency in fear processing instathmin−/−females leads to improper threat assessment, which in turn affects innate parental care and adult social interactions. Profound deficiency is observed in maternal behavior ofstathmin−/−females: they lack motivation for retrieving pups and are unable to choose a safe location for nest-building. Remarkably,stathmin−/−females have an enhancement in social interactions. BLA lesions in WT mice produce similar effects in maternal and social behaviors, confirming vital BLA participation. The findings implicatestathminas the critical molecular component linking the BLA-associated neural circuitry with innate parental behaviors and adult social interactions.
Alcohol, oxidative stress and free radical damage
Authors: Albano Emanuele
Published: September 2008

The involvement of free radical mechanisms in the pathogenesis of alcoholic liver disease (ALD) is demonstrated by the detection of lipid peroxidation markers in the liver and the serum of patients with alcoholism, as well as by experiments in alcohol-feed rodents that show a relationship between alcohol-induced oxidative stress and the development of liver pathology. Ethanol-induced oxidative stress is the result of the combined impairment of antioxidant defences and the production of reactive oxygen species by the mitochondrial electron transport chain, the alcohol-inducible cytochrome P450 (CYP) 2E1 and activated phagocytes. Furthermore, hydroxyethyl free radicals (HER) are also generated during ethanol metabolism by CYP2E1. The mechanisms by which oxidative stress contributes to alcohol toxicity are still not completely understood. The available evidence indicates that, by favouring mitochondrial permeability transition, oxidative stress promotes hepatocyte necrosis and/or apoptosis and is implicated in the alcohol-induced sensitization of hepatocytes to the pro-apoptotic action of TNF-α. Moreover, oxidative mechanisms can contribute to liver fibrosis, by triggering the release of pro-fibrotic cytokines and collagen gene expression in hepatic stellate cells. Finally, the reactions of HER and lipid peroxidation products with hepatic proteins stimulate both humoral and cellular immune reactions and favour the breaking of self-tolerance during ALD. Thus, immune responses might represent the mechanism by which alcohol-induced oxidative stress contributes to the perpetuation of chronic hepatic inflammation. Together these observations provide a rationale for the possible clinical application of antioxidants in the therapy for ALD.
Glutamine in critical care: current evidence from systematic reviews
Authors: Avenell Alison
Published: September 2008

Glutamine, the most abundant amino acid in the body, is thought to become conditionally essential in critical illness. Some of the important roles for glutamine are as a carrier for inter-organ N, a preferred fuel for enterocytes and cells of the immune system, a substrate for renal NH3 formation and a precursor for glutathione. Mechanisms by which glutamine could improve recovery include attenuating oxidant damage and inflammatory cytokine production, reducing gut bacterial translocation and improving N balance. The present systematic review has found trends to suggest that parenteral and enteral glutamine supplementation reduce mortality, the development of infection and organ failure in critical illness. Trials of parenteral nutrition containing glutamine with patients after elective surgery also suggest reduction of infection, but it is unlikely that glutamine-containing parenteral nutrition would be used for such patients. The evidence base is limited by the quality of the reported trials and the suggestion that there is publication bias, with trials suggesting reduced infection being more likely to be published.
Hyperglycaemia and pulmonary infection
Authors: Baker Emma H. , Wood David M. , Brennan Amanda L. , Clark Nicholas , Baines Deborah L. , Philips Barbara J.
Published: September 2008

Pathophysiological stress from acute illness causes metabolic disturbance, including altered hepatic glucose metabolism, increased peripheral insulin resistance and hyperglycaemia. Acute hyperglycaemia is associated with increased morbidity and mortality in patients in intensive care units and patients with acute respiratory disease. The present review will consider mechanisms underlying this association. In normal lungs the glucose concentration of airway secretions is approximately 10-fold lower than that of plasma. Low airway glucose concentrations are maintained against a concentration gradient by active glucose transport. Airway glucose concentrations become elevated if normal homeostasis is disrupted by a rise in blood glucose concentrations or inflammation of the airway epithelium. Elevated airway glucose concentrations are associated with and precede increased isolation of respiratory pathogens, particularly methicillin-resistantStaphylococcus aureus, from bronchial aspirates of patients intubated on intensive care. Markers of elevated airway glucose are associated with similar patterns of respiratory infection in patients admitted with acute exacerbations of chronic obstructive pulmonary disease. Glucose at airway concentrations stimulates the growth of respiratory pathogens, over and above the effect of other nutrients. Elevated airway glucose concentrations may also worsen respiratory disease by promoting local inflammation. Hyperglycaemia may thus promote pulmonary infection, at least in part, by an effect on airway glucose concentrations. Therapeutic options, including systemic control of blood glucose and local manipulation of airway glucose homeostasis, will be considered.
Use of fish oil in parenteral nutrition: rationale and reality
Authors: Calder Philip C.
Published: September 2008

Excessive or inappropriate inflammation and immunosuppression are components of the response to surgery, trauma, injury and infection in some individuals and can lead, progressively, to sepsis and septic shock. The hyperinflammation is characterised by the production of inflammatory cytokines, arachidonic acid-derived eicosanoids and other inflammatory mediators, while the immunosuppression is characterised by impairment of antigen presentation and of T-helper lymphocyte type-1 responses. Long-chainn-3 fatty acids from fish oil decrease the production of inflammatory cytokines and eicosanoids. They act both directly (by replacing arachidonic acid as an eicosanoid substrate and by inhibiting arachidonic acid metabolism) and indirectly (by altering the expression of inflammatory genes through effects on transcription factor activation). Thus, long-chainn-3 fatty acids are potentially useful anti-inflammatory agents and may be of benefit in patients at risk of hyperinflammation and sepsis. As a consequence, an emerging application forn-3 fatty acids, in which they may be added to parenteral (or enteral) formulas, is in surgical or critically-ill patients. Parenteral nutrition that includesn-3 fatty acids appears to preserve immune function better than standard formulas and appears to diminish the extent of the inflammatory response. Studies to date are suggestive of clinical benefits from these approaches, especially in patients post surgery, although evidence of clinical benefit in patients with sepsis is emerging.
The biochemical basis of antioxidant therapy in critical illness
Authors: Eaton Simon
Published: September 2008

During critical illness free radical production may increase as a result of, for example, sepsis or tissue trauma. In addition, because of a potential for increased losses, and the possibility of inadequate nutrition, the antioxidant defences of the body may become compromised. Thus, the delicate balance between free radicals and antioxidants may be disturbed. Various nutritional and pharmacological strategies to enhance antioxidant defences have been proposed, which aim either to maintain or enhance endogenous antioxidant stores or to provide alternative antioxidant agents. Trace elements and amino acids are particularly important, and their synergistic role in the maintenance of the body's antioxidant defence network will be discussed.
Moderate alcohol consumption and increased bone mineral density: potential ethanol and non-ethanol mechanisms
Authors: Jugdaohsingh R. , O'Connell M. A. , Sripanyakorn S. , Powell J. J.
Published: September 2008

Mounting epidemiological evidence indicates an association between the moderate ingestion of alcoholic beverages and higher bone mineral density (v. abstainers). More limited findings provide some evidence for translation of this association into reduced fracture risk, but further studies are required. Here, these data are reviewed and caveats in their assimilation, comparison and interpretation as well as in the use and application of bone health indices are discussed. Whilst it is concluded that evidence is now strong for the moderate alcohol–bone health association, at least in relation to bone mineral density, mechanisms are less clear. Both ethanol and non-ethanol components have been implicated as factors that positively affect bone health in the light of moderate consumption of alcoholic beverages, and four particular areas are discussed. First, recent findings suggest that moderate ethanol consumption acutely inhibits bone resorption, in a non-parathyroid hormone- and non-calcitonin-dependent fashion, which can only partly be attributed to an energy effect. Second, critical review of the literature does not support a role for moderate ethanol consumption affecting oestrogen status and leading to a knock-on effect on bone. Third, Si is present at high levels in certain alcoholic beverages, especially beer, and may have a measurable role in promoting bone formation. Fourth, a large body of work indicates that phytochemicals (e.g. polyphenols) from alcoholic beverages could influence bone health, but human data are lacking. With further work it is hoped to be able to model epidemiological observations and provide a clear pathway between the magnitude of association and the relative contribution of these mechanisms for the major classes of alcoholic beverage.
Investigations as a prerequisite for genetic counseling after termination of pregnancy based on sonographic detection of serious central nervous system – or skeletal anomalies
Authors: KAASEN ANNE , PRESCOTT TRINE E. , HEIBERG ARVID , SCOTT HELGE , HAUGEN GUTTORM
Published: September 2008

Objectives. The primary aim was to evaluate which investigation performed after sonographic detection of central nervous system (CNS) or skeletal anomalies that had highest diagnostic yield. The secondary aim was to estimate recurrence risk. Design. Retrospective review of patients’ records. Setting. Tertiary fetal medicine referral center. Sample. Pregnancy terminations (n = 97) because of CNS or skeletal anomalies during a 17‐year period, within 12–24 weeks gestation. Methods. Two medical geneticists and one genetic counselor reviewed charts independently. Main outcome measures. Primary ultrasound diagnosis, change in diagnosis following supplementary examinations in addition to prenatal ultrasound (medical history, autopsy, post‐mortem X‐ray, karyotyping, targeted DNA analysis and investigations for infection), the most useful method to determine diagnosis, and recurrence risk estimate including inter‐rater agreement. Results. Mean gestational age was 19.8 weeks. All three investigators agreed in each case on which investigation constituted the best basis to determine the most precise diagnosis. The examinations performed in addition to prenatal ultrasound provided important diagnostic information in 54 cases (56%) and altered recurrence risk in 22 (23%) cases; in eight of these cases the risk estimate was increased. In nine cases (9%) the investigators disagreed in their estimates of recurrence risk. Kappa for inter‐rater agreement was >0.90. Conclusions. A panel of diagnostic investigations, depending on the organ system involved, allows for a more precise diagnosis and a more reliable estimate of recurrence risk than prenatal ultrasound alone. In some instances, recurrence risk estimation is not straightforward as evidenced by lack of consensus.
Timing of heparin prophylaxis and bleeding complications in hysterectomy a nationwide prospective cohort study of 9,949 Danish women
Authors: HANSEN CHARLOTTE T. , KEHLET HENRIK , MØLLER CHARLOTTE , MØRCH LINA , UTZON JAN , OTTESEN BENT
Published: September 2008

Objective. To examine bleeding complications and thromboembolic events in relation to timing of heparin prophylaxis after hysterectomy. Design. Nationwide prospective cohort study with 30 days post‐operative follow‐up within the Danish Hysterectomy Database (DHD). Setting. All gynecological departments in Denmark (n = 31). Sample. 9,949 women who had an elective hysterectomy for benign indication between October 2003 and May 2006 and were reported to DHD (national response rate: 88–99% throughout 2004–2005). Main outcome measures. Odds ratios (OR) of peri‐operative bleeding complications (≥1,000 ml bleeding during surgery or post‐operative wound/vaginal‐vault/intraabdominal bleeding or hematoma) and number of events of venous thromboembolism. Logistic regression analysis adjusting for: age, body mass index, alcohol, smoking, meno‐/metrorrhagia, uterine weight, department volume, surgeon's experience, route and type of hysterectomy and additional surgery, and stratification on assistant's experience, peri‐operative pain prophylaxis with NSAID and daily use of Acetyl Salicylic Acid (ASA)/NSAID. Results. 9,051 women (92%) received thromboprophylaxis with heparin, initiated pre‐operatively in 48% and post‐operatively in 52%. At least one bleeding complication was noted in 881 women (10%). Post‐operative heparin administration was associated with a reduced risk of bleeding complications; OR=0.85 (95% confidence interval 0.73–0.99) compared to pre‐operative administration. Excluding cases with potential impaired hemostasis at baseline, the OR was 0.78 (0.64–0.94). There was no fatal embolism. Three of seven pulmonary embolisms and one of three symptomatic deep venous thromboses occurred with the post‐operative heparin administration. Conclusion. Post‐operative rather than pre‐operative administration of heparin prophylaxis may reduce the risk of bleeding complications after hysterectomy without apparent risk of increased thromboembolic events.
Partner change among Norwegian teenage girls attending youth health clinics
Authors: SKJELDESTAD FINN EGIL , NILSEN STINE
Published: September 2008

Objective. To examine incidence and risk factors of partner change among Norwegian teenage girls attending youth health clinics. Design. Prospective cohort study. Setting. Study subjects were 691 girls, aged 16 to 19 years, who participated in a study on prescription of oral contraceptives at three youth health clinics in the city of Trondheim, Norway. Methods. Eligible for analysis were participants who were sexually active, and who returned for at least one follow‐up visit more than 120 days after study start. All demographic and behavior data were collected through interview by public health nurses. All analyses were performed with SPSS for Windows (version 15.0) using chi‐square test, survival analyses and logistic regression. Main outcome measure. New sexual male partners. Results. Four out of five girls (80%) did not have any new partner during the study period of 12 months. From survival analyses the 12‐month incidence rate were 29.8 (95% CI: 25.3–34.1) and 10.6 per 100‐women months (95% CI: 7.3–13.9) for one or two or more partners, respectively. Predictors of partner change were partner change during the last six month prior study start, being single and sexual debut within three years after menarche. Conclusion. Being in a regular relationship with a boyfriend reduce considerably the risk of having a new sexual partner. Recent history of partner change is the strongest predictor of future partner change.
Measurement of nitric oxide in the vagina
Authors: SIOUTAS ANGELOS , EHRÉN INGRID , GEMZELL‐DANIELSSON KRISTINA
Published: September 2008

Objective.Local measurement of nitric oxide (NO) gas has been used to detect and monitor inflammatory processes in the airways, the colon and in the urinary bladder, but so far NO has not been studied in the lower female genital tract. The objective of this study is to measure NO gas directly in the vaginal lumen of healthy women and in patients with vaginitis.Setting. The outpatient clinic of a university hospital.Methods.Eighteen non‐pregnant women from 19 to 65 years of age with symptoms of vaginitis, eight healthy women in reproductive age with regular menstrual cycles and nine healthy postmenopausal women were enrolled in the study. NO levels were measured in air incubated for five minutes in a catheter balloon in the vagina.Results.In patients with symptoms of vaginitis, NO concentration was almost 100‐fold increased compared to healthy controls (p<0.001) with no individual overlap. Vaginal NO levels were uniformly low among healthy women, both in reproductive age and in menopause.Conclusions. NO gas can be measured directly in the vagina with a fast, simple and safe method. The levels of NO are increased in patients with vaginitis.
Reference values for α1‐acid glycoprotein, α1‐antitrypsin, albumin, haptoglobin, C‐reactive protein, IgA, IgG and IgM during pregnancy
Authors: LARSSON ANDERS , PALM MARIA , HANSSON LARS‐OLOF , BASU SAMAR , AXELSSON OVE
Published: September 2008

The objective of this study was to establish reference intervals and decision limits for the interpretation of the acute phase proteins α1‐acid glycoprotein (orosomucoid), α1‐antitrypsin, C‐reactive protein (CRP), haptoglobin and albumin, IgA, IgG and IgM during pregnancy by longitudinal sampling from 52 healthy women with normal pregnancies. Each woman was sampled in weeks 7–17; weeks 17–24; weeks 24–28; weeks 28–31; weeks 31–34; weeks 34–38 and predelivery (–14–0 days prior to delivery) and postpartum (>6 weeks after delivery). The 2.5th and 97.5th percentiles were calculated according to the recommendations of the International Federation of Clinical Chemistry on the statistical treatment of reference values. Reference values for α1‐acid glycoprotein, α1‐antitrypsin, albumin, haptoglobin, CRP, IgA, IgG and IgM are reported. Most of these proteins changed during normal pregnancy, as a reflection of the major physiological and biochemical changes that occur in pregnancy. A laboratory test result from a pregnant woman should be compared with pregnancy‐specific reference intervals.
Regulation of caveolin-1 membrane trafficking by the Na/K-ATPase
Authors: Cai Ting , Wang Haojie , Chen Yiliang , Liu Lijun , Gunning William T , Quintas Luis Eduardo M. , Xie Zi-Jian
Published: September 2008

Here, we show that the Na/K-ATPase interacts with caveolin-1 (Cav1) and regulates Cav1 trafficking. Graded knockdown of Na/K-ATPase decreases the plasma membrane pool of Cav1, which results in a significant reduction in the number of caveolae on the cell surface. These effects are independent of the pumping function of Na/K-ATPase, and instead depend on interaction between Na/K-ATPase and Cav1 mediated by an N-terminal caveolin-binding motif within the ATPase α1 subunit. Moreover, knockdown of the Na/K-ATPase increases basal levels of active Src and stimulates endocytosis of Cav1 from the plasma membrane. Microtubule-dependent long-range directional trafficking in Na/K-ATPase–depleted cells results in perinuclear accumulation of Cav1-positive vesicles. Finally, Na/K-ATPase knockdown has no effect on processing or exit of Cav1 from the Golgi. Thus, the Na/K-ATPase regulates Cav1 endocytic trafficking and stabilizes the Cav1 plasma membrane pool.
Effector caspase Dcp-1 and IAP protein Bruce regulate starvation-induced autophagy during Drosophila melanogaster oogenesis
Authors: Hou Ying-Chen Claire , Chittaranjan Suganthi , Barbosa Sharon González , McCall Kimberly , Gorski Sharon M.
Published: September 2008

A complex relationship exists between autophagy and apoptosis, but the regulatory mechanisms underlying their interactions are largely unknown. We conducted a systematic study of Drosophila melanogaster cell death–related genes to determine their requirement in the regulation of starvation-induced autophagy. We discovered that six cell death genes—death caspase-1 (Dcp-1), hid, Bruce, Buffy, debcl, and p53—as well as Ras–Raf–mitogen activated protein kinase signaling pathway components had a role in autophagy regulation in D. melanogaster cultured cells. During D. melanogaster oogenesis, we found that autophagy is induced at two nutrient status checkpoints: germarium and mid-oogenesis. At these two stages, the effector caspase Dcp-1 and the inhibitor of apoptosis protein Bruce function to regulate both autophagy and starvation-induced cell death. Mutations in Atg1 and Atg7 resulted in reduced DNA fragmentation in degenerating midstage egg chambers but did not appear to affect nuclear condensation, which indicates that autophagy contributes in part to cell death in the ovary. Our study provides new insights into the molecular mechanisms that coordinately regulate autophagic and apoptotic events in vivo.
Regulation of Sli15/INCENP, kinetochore, and Cdc14 phosphatase functions by the ribosome biogenesis protein Utp7
Authors: Jwa Miri , Kim Jae-hyun , Chan Clarence S.M.
Published: September 2008

The Sli15–Ipl1–Bir1 chromosomal passenger complex is essential for proper kinetochore–microtubule attachment and spindle stability in the budding yeast Saccharomyces cerevisiae. During early anaphase, release of the Cdc14 protein phosphatase from the nucleolus leads to the dephosphorylation of Sli15 and redistribution of this complex from kinetochores to the spindle. We show here that the predominantly nucleolar ribosome biogenesis protein Utp7 is also present at kinetochores and is required for normal organization of kinetochore proteins and proper chromosome segregation. Utp7 associates with and regulates the localization of Sli15 and Cdc14. Before anaphase onset, it prevents the premature nucleolar release of Cdc14 and the premature concentration of Sli15 on the spindle. Furthermore, Utp7 can regulate the localization and phosphorylation status of Sli15 independent of its effect on Cdc14 function. Thus, Utp7 is a multifunctional protein that plays essential roles in the vital cellular processes of ribosome biogenesis, chromosome segregation, and cell cycle control.
Human squamous cell carcinomas evade the immune response by down-regulation of vascular E-selectin and recruitment of regulatory T cells
Authors: Clark Rachael A. , Huang Susan J. , Murphy George F. , Mollet Ilse G. , Hijnen Dirkjan , Muthukuru Manoj , Schanbacher Carl F. , Edwards Vonetta , Miller Danielle M. , Kim Jenny E. , Lambert Jo , Kupper Thomas S.
Published: September 2008

Squamous cell carcinomas (SCCs) of the skin are sun-induced skin cancers that are particularly numerous in patients on T cell immunosuppression. We found that blood vessels in SCCs did not express E-selectin, and tumors contained few cutaneous lymphocyte antigen (CLA)+ T cells, the cell type thought to provide cutaneous immunosurveillance. Tumors treated with the Toll-like receptor (TLR)7 agonist imiquimod before excision showed induction of E-selectin on tumor vessels, recruitment of CLA+ CD8+ T cells, and histological evidence of tumor regression. SCCs treated in vitro with imiquimod also expressed vascular E-selectin. Approximately 50% of the T cells infiltrating untreated SCCs were FOXP3+ regulatory T (T reg) cells. Imiquimod-treated tumors contained a decreased percentage of T reg cells, and these cells produced less FOXP3, interleukin (IL)-10, and transforming growth factor (TGF)-β. Treatment of T reg cells in vitro with imiquimod inhibited their suppressive activity and reduced FOXP3, CD39, CD73, IL-10, and TGF-β by indirect mechanisms. In vivo and in vitro treatment with imiquimod also induced IL-6 production by effector T cells. In summary, we find that SCCs evade the immune response at least in part by down-regulating vascular E-selectin and recruiting T reg cells. TLR7 agonists neutralized both of these strategies, supporting their use in SCCs and other tumors with similar immune defects.
Activating transcription factor 3 is a negative regulator of allergic pulmonary inflammation
Authors: Gilchrist Mark , Henderson William R. , Clark April E. , Simmons Randi M. , Ye Xin , Smith Kelly D. , Aderem Alan
Published: September 2008

We recently demonstrated the pivotal role of the transcription factor (TF) activating TF 3 (ATF3) in dampening inflammation. We demonstrate that ATF3 also ameliorates allergen-induced airway inflammation and hyperresponsiveness in a mouse model of human asthma. ATF3 expression was increased in the lungs of mice challenged with ovalbumin allergen, and this was associated with its recruitment to the promoters of genes encoding Th2-associated cytokines. ATF3-deficient mice developed significantly increased airway hyperresponsiveness, pulmonary eosinophilia, and enhanced chemokine and Th2 cytokine responses in lung tissue and in lung-derived CD4+ lymphocytes. Although several TFs have been associated with enhanced inflammatory responses in the lung, ATF3 attenuates the inflammatory responses associated with allergic airway disease.
Origin and pathogenesis of nodular lymphocyte–predominant Hodgkin lymphoma as revealed by global gene expression analysis
Authors: Brune Verena , Tiacci Enrico , Pfeil Ines , Döring Claudia , Eckerle Susan , van Noesel Carel J.M. , Klapper Wolfram , Falini Brunangelo , von Heydebreck Anja , Metzler Dirk , Bräuninger Andreas , Hansmann Martin-Leo , Küppers Ralf
Published: September 2008

The pathogenesis of nodular lymphocyte–predominant Hodgkin lymphoma (NLPHL) and its relationship to other lymphomas are largely unknown. This is partly because of the technical challenge of analyzing its rare neoplastic lymphocytic and histiocytic (L&H) cells, which are dispersed in an abundant nonneoplastic cellular microenvironment. We performed a genome-wide expression study of microdissected L&H lymphoma cells in comparison to normal and other malignant B cells that indicated a relationship of L&H cells to and/or that they originate from germinal center B cells at the transition to memory B cells. L&H cells show a surprisingly high similarity to the tumor cells of T cell–rich B cell lymphoma and classical Hodgkin lymphoma, a partial loss of their B cell phenotype, and deregulation of many apoptosis regulators and putative oncogenes. Importantly, L&H cells are characterized by constitutive nuclear factor κB activity and aberrant extracellular signal-regulated kinase signaling. Thus, these findings shed new light on the nature of L&H cells, reveal several novel pathogenetic mechanisms in NLPHL, and may help in differential diagnosis and lead to novel therapeutic strategies.
The tight junction protein CAR regulates cardiac conduction and cell–cell communication
Authors: Lisewski Ulrike , Shi Yu , Wrackmeyer Uta , Fischer Robert , Chen Chen , Schirdewan Alexander , Jüttner Rene , Rathjen Fritz , Poller Wolfgang , Radke Michael H. , Gotthardt Michael
Published: September 2008

The Coxsackievirus-adenovirus receptor (CAR) is known for its role in virus uptake and as a protein of the tight junction. It is predominantly expressed in the developing brain and heart and reinduced upon cardiac remodeling in heart disease. So far, the physiological functions of CAR in the adult heart are largely unknown. We have generated a heart-specific inducible CAR knockout (KO) and found impaired electrical conduction between atrium and ventricle that increased with progressive loss of CAR. The underlying mechanism relates to the cross talk of tight and gap junctions with altered expression and localization of connexins that affect communication between CAR KO cardiomyocytes. Our results indicate that CAR is not only relevant for virus uptake and cardiac remodeling but also has a previously unknown function in the propagation of excitation from the atrium to the ventricle that could explain the association of arrhythmia and Coxsackievirus infection of the heart.
Hypericum triquetrifolium—Derived Factors Downregulate the Production Levels of LPS‐Induced Nitric Oxide and Tumor Necrosis Factor‐αin THP‐1 Cells
Authors: Saad Bashar , AbouAtta Bernadette Soudah , Basha Walid , Hmade Alaa , Kmail Abdalsalam , Khasib Said , Said Omar
Published: September 2008

Based on knowledge from traditional Arab herbal medicine, thisin vitrostudy aims to examine the anti‐inflammatory mechanism ofHypericum triquetrifoliumby measuring the expression and release of pro‐inflammatory cytokines, tumor necrosis factor‐α(TNF‐α) and interleukine‐6 (IL‐6), and inducible nitric oxide synthase (iNOS) in human monocytic cells, THP‐1. The effects were assessed by measuring the levels of secretory proteins and mRNA of TNF‐αand IL‐6, the levels of nitric oxide (NO) secretion and the expression of iNOS in THP‐1 cells. Cells were treated with 5 μg lipopolysaccharide/ml (LPS) in the presence and absence of increasing concentrations of extracts from the aerial parts ofH. triquetrifolium. During the entire experimental period, we used extract concentrations (up to 250 μg mL−1) that had no cytotoxic effects, as measured with MTT and LDH assays.Hypericum triquetrifoliumextracts remarkably suppressed the LPS‐induced NO release, significantly attenuated the LPS‐induced transcription of iNOS and inhibited in a dose‐dependent manner the expression and release of TNF‐α. No significant effects were observed on the release of IL‐6. Taken together, these results suggest thatH. triquetrifoliumprobably exerts anti‐inflammatory effects through the suppression of TNF‐αand iNOS expressions.
Spirulina in Clinical Practice: Evidence‐Based Human Applications
Authors: Karkos P. D. , Leong S. C. , Karkos C. D. , Sivaji N. , Assimakopoulos D. A.
Published: September 2008

Spirulina or Arthrospira is a blue‐green alga that became famous after it was successfully used by NASA as a dietary supplement for astronauts on space missions. It has the ability to modulate immune functions and exhibits anti‐inflammatory properties by inhibiting the release of histamine by mast cells. Multiple studies investigating the efficacy and the potential clinical applications of Spirulina in treating several diseases have been performed and a few randomized controlled trials and systematic reviews suggest that this alga may improve several symptoms and may even have an anticancer, antiviral and antiallergic effects. Current and potential clinical applications, issues of safety, indications, side‐effects and levels of evidence are addressed in this review. Areas of ongoing and future research are also discussed.
Taking and Recording a Genetic Family History
Authors: Farndon Peter
Published: September 2008

During a consultation, John remarked: ‘My grandfather and an aunt and uncle died of bowel cancer’. General practitioners are constantly receiving and documenting such information about family members, but how best can it be recorded and interpreted to decide if an individual is at risk for a genetic disorder?
Antenatal Screening. The UK NHS Antenatal Screening Programmes: Policy and Practice
Authors: Harcombe Joanne , Armstrong Valerie
Published: September 2008

This article outlines the antenatal screening tests available to women as part of NHS maternity care. The tests are part of managed screening programmes which are overseen by the UK National Screening Committee (NSC) and underpinned by standards, education and quality assurance.
Characterization of hepatitis C RNA-containing particles from human liver by density and size
Authors: Nielsen Søren U. , Bassendine Margaret F. , Martin Caroline , Lowther Daniel , Purcell Paul J. , King Barnabas J. , Neely Dermot , Toms Geoffrey L.
Published: September 2008

Hepatitis C virus (HCV) particles foundin vivoare heterogeneous in density and size, but their detailed characterization has been restricted by the low titre of HCV in human serum. Previously, our group has found that HCV circulates in blood in association with very-low-density lipoprotein (VLDL). Our aim in this study was to characterize HCV RNA-containing membranes and particles in human liver by both density and size and to identify the subcellular compartment(s) where the association with VLDL occurs. HCV was purified by density using iodixanol gradients and by size using gel filtration. Both positive-strand HCV RNA (present in virus particles) and negative-strand HCV RNA (an intermediate in virus replication) were found with densities below 1.08 g ml−1. Viral structural and non-structural proteins, host proteins ApoB, ApoE and caveolin-2, as well as cholesterol, triglyceride and phospholipids were also detected in these low density fractions. After fractionation by size with Superose gel filtration, HCV RNA and viral proteins co-fractionated with endoplasmic reticulum proteins and VLDL. Fractionation on Toyopearl, which separates particles with diameters up to 200 nm, showed that 78 % of HCV RNA from liver was >100 nm in size, with a positive-/negative-strand ratio of 6 : 1. Also, 8 % of HCV RNA was found in particles with diameters between 40 nm and 70 nm and a positive-/negative-strand ratio of 45 : 1. This HCV was associated with ApoB, ApoE and viral glycoprotein E2, similar to viral particles circulating in serum. Our results indicate that the association between HCV and VLDL occurs in the liver.
Differences in virulence among porcine circovirus type 2 isolates are unrelated to cluster type 2a or 2b and prior infection provides heterologous protection
Authors: Opriessnig T. , Ramamoorthy S. , Madson D. M. , Patterson A. R. , Pal N. , Carman S. , Meng X. J. , Halbur P. G.
Published: September 2008

Porcine circovirus type 2 (PCV2) is divided into two genetic clusters designated PCV2a and PCV2b. The objectives of this study were to determine whether isolates from different clusters vary in virulence and to determine whether infection with PCV2a isolates induces protective immunity against subsequent infection with a recent PCV2b isolate. One-hundred and thirteen conventional specific-pathogen-free (SPF) pigs were assigned randomly to treatment groups and rooms: pigs inoculated with PCV2a cluster isolates (ISU-40895 or ISU-4838), pigs inoculated with PCV2b cluster isolates (NC-16845 or Can-17639) and uninoculated pigs. Necropsies were performed at 16 or 51 days post-inoculation (p.i.). There were no significant differences in PCV2-associated lymphoid lesions between PCV2a and PCV2b clusters; however, within the same cluster, significant differences were found between isolates: ISU-4838- and Can-17639-inoculated pigs had significantly (P<0.05) less severe lesions compared with ISU-40895- and NC-16845-inoculated pigs. To evaluate cross-protection, six pigs within each group were challenged at 35 days p.i. with an isolate from the heterologous cluster and were necropsied 51 days p.i. The severity of PCV2-associated lesions was reduced in pigs with prior exposure to an isolate from the heterologous cluster in comparison with singly inoculated pigs. Results indicate that the virulence of PCV2a and PCV2b isolates is not different in the conventional SPF pig model; however, the virulence of isolates within the same cluster differs. Increased virulence as reported to be associated with PCV2b isolates in the field was not observed under the conditions of this study. Moreover, cross-protection between PCV2a and PCV2b exists.
Cytokine patterns in a comparative model of arenavirus haemorrhagic fever in guinea pigs
Authors: Scott Erin P. , Aronson Judith F.
Published: September 2008

Arenaviruses such as Lassa virus cause a spectrum of disease in humans ranging from mild febrile illness to lethal haemorrhagic fever. The contributions of innate immunity to protection or pathogenicity are unknown. We compared patterns of expression of cytokines of innate immunity in mild versus severe arenavirus disease using an established guinea pig model based on the macrophage-tropic arenavirus Pichinde virus (PICV). Cytokine transcripts were measured by using real-time RT-PCR in target organs and blood during mild infection (caused by PICV, P2 variant) and lethal haemorrhagic fever (caused by PICV, P18 variant). In the initial peritoneal target cells, virulent P18 infection was associated with significantly increased gamma interferon (IFN-γ) and monocyte chemoattractant protein-1 (MCP-1, CCL2) mRNA levels relative to P2 infection. Peritoneal cells from P18-infected animals had decreased tumour necrosis factor alpha (TNF-α), interleukin (IL)-8 (CXCL-8) and IL-12p40 transcripts relative to mock-infected animals. Late in infection, P18-infected peripheral blood leukocytes (PBL) had decreased TNF-α, IFN-γ, and regulated upon activation, normal T cell expressed and secreted (RANTES, CCL-5) cytokine transcripts relative to P2-infected PBL. We conclude that, in severe arenavirus disease, patterns of cytokine expression in the initially infected cells favour recruitment of additional target monocytes, while inhibiting some of their pro-inflammatory responses. Suppression rather than overexpression of pro-inflammatory cytokines accompanied the terminal shock in this model of arenavirus haemorrhagic fever.
The effects of prion protein proteolysis and disaggregation on the strain properties of hamster scrapie
Authors: Deleault Andrea M. , Deleault Nathan R. , Harris Brent T. , Rees Judy R. , Supattapone Surachai
Published: September 2008

Native mammalian prions exist in self-propagating strains that exhibit distinctive clinical, pathological and biochemical characteristics. Prion strain diversity is associated with variations in PrPScconformation, but it remains unknown precisely which physical properties of the PrPScmolecules are required to encipher mammalian prion strain phenotypes. In this study, we subjected prion-infected brain homogenates derived from three different hamster scrapie strains to either (i) proteinase K digestion or (ii) sonication, and inoculated the modified samples into normal hamsters. The results show that the strain-specific clinical features and neuropathological profiles of inoculated animals were not affected by either treatment. Similarly, the strain-dependent biochemical characteristics of the PrPScmolecules (including electrophoretic mobility, glycoform composition, conformational stability and susceptibility to protease digestion) in infected animals were unaffected by either proteolysis or sonication of the original inocula. These results indicate that the infectious strain properties of native prions do not appear to be altered by PrPScdisaggregation, and that maintenance of such properties does not require the N-domain (approximately residues 23–90) of the protease-resistant PrPScmolecules or protease-sensitive PrPScmolecules.
Isolation and cloning of two variant papillomaviruses from domestic pigs: Sus scrofa papillomaviruses type 1 variants a and b
Authors: Stevens Hans , Rector Annabel , Van Der Kroght Kees , Van Ranst Marc
Published: September 2008

The healthy skin of two female domestic pigs (Sus scrofa domestica) was sampled with cotton-tipped swabs. Total genomic DNA was extracted from the samples and subjected to PCR with degenerate papillomavirus (PV)-specific primers. Similarity searches performed with blastn showed that partial E1 and L1 sequences of two novel PVs were amplified. Subsequently, the complete genomes of these Sus scrofa papillomaviruses (SsPVs) were amplified by long-template PCR, cloned and sequenced using a transposon insertion method. They contained the typical PV open reading frames (ORFs) E1, E2, E4, E6, L1 and L2, but the E7 ORF was absent in both viruses. Pairwise nucleotide sequence alignment of the L1 ORFs of the SsPVs showed 98.5 % similarity, classifying these viruses as SsPV type 1 ‘variants’ (SsPV-1a and -1b). Based on a concatenated alignment of the E1, E2, L1 and L2 ORFs of SsPV-1 variants a and b, and 81 other human and animal PV type species, a neighbour-joining phylogenetic tree was constructed. This phylogenetic analysis showed that the SsPV-1a and -1b variants did not cluster with the other PVs of artiodactyls (cloven-hoofed) host species, but clustered on the edge of the genus Alphapapillomavirus, very near to the root of this genus.
Tolerance to mutations in the foot-and-mouth disease virus integrin-binding RGD region is different in cultured cells and in vivo and depends on the capsid sequence context
Authors: Gutiérrez-Rivas Mónica , Pulido Miguel Rodríguez , Baranowski Eric , Sobrino Francisco , Sáiz Margarita
Published: September 2008

Engineered RNAs carrying substitutions in the integrin receptor-binding Arg-Gly-Asp (RGD) region of foot-and-mouth disease virus (FMDV) were constructed (aa 141–147 of VP1 capsid protein) and their infectivity was assayed in cultured cells and suckling mice. The effect of these changes was studied in the capsid proteins of two FMDVs, C-S8c1, which enters cells through integrins, and 213hs−, a derivative highly adapted to cell culture whose ability to infect cells using the glycosaminoglycan heparan sulfate (HS) as receptor, acquired by multiple passage on BHK-21 cells, has been abolished. The capsid sequence context determined infectivity in cultured cells and directed the selection of additional replacements in structural proteins. Interestingly, a viral population derived from a C-S8c1/L144A mutant, carrying only three substitutions in the capsid, was able to expand tropism to wild-type (wt) and mutant (mt) glycosaminoglycan-deficient CHO cells. In contrast, the 213hs− capsid tolerated all substitutions analysed with no additional mutations, and the viruses recovered maintained the ability of the 213hs− parental virus to infect wt and mt CHO cells. Viruses derived from C-S8c1 with atypical RGD regions were virulent and transmissible for mice with no other changes in the capsid. Substitution of Asp143 for Ala in the C-S8c1 capsid eliminated infectivity in cultured cells and mice. Co-inoculation with a neutralizing monoclonal antibody directed against the type C FMDV RGD region abolished infectivity of C-S8c1 virus on suckling mice, suggesting that FMDV can infect mice using integrins. Sequence requirements imposed for viral entry in vitro and in vivo are discussed.
Genome-wide transcriptional response of primary alveolar macrophages following infection with porcine reproductive and respiratory syndrome virus
Authors: Genini Sem , Delputte Peter L. , Malinverni Roberto , Cecere Maria , Stella Alessandra , Nauwynck Hans J. , Giuffra Elisabetta
Published: September 2008

Porcine reproductive and respiratory syndrome is a major cause of economic loss for the swine industry worldwide. Porcine reproductive and respiratory syndrome virus (PRRSV) triggers weak and atypical innate immune responses, but key genes and mechanisms by which the virus interferes with the host innate immunity have not yet been elucidated. In this study, genes that control the response of the main target of PRRSV, porcine alveolar macrophages (PAMs), were profiledin vitrowith a time-course experiment spanning the first round of virus replication. PAMs were obtained from six piglets and challenged with the Lelystad PRRSV strain, and gene expression was investigated using Affymetrix microarrays and real-time PCR. Of the 1409 differentially expressed transcripts identified by analysis of variance, two, five, 25, 16 and 100 differed from controls by a minimum of 1.5-fold at 1, 3, 6, 9 and 12 h post-infection (p.i.), respectively. A PRRSV infection effect was detectable between 3 and 6 h p.i., and was characterized by a consistent downregulation of gene expression, followed by the start of the host innate immune response at 9 h p.i. The expression of beta interferon 1 (IFN-β), but not ofIFN-α, was strongly upregulated, whilst few genes commonly expressed in response to viral infections and/or induced by interferons were found to be differentially expressed. A predominance of anti-apoptotic transcripts (e.g. interleukin-10), a shift towards a T-helper cell type 2 response and a weak upregulation of tumour necrosis factor-αexpression were observed within 12 h p.i., reinforcing the hypotheses that PRRSV has developed sophisticated mechanisms to escape the host defence.
Analysis of herpes simplex virus type 1 restriction fragment length polymorphism variants associated with herpes gladiatorum and Kaposi's varicelliform eruption in sumo wrestlers
Authors: Ban Fumihiko , Asano Satoe , Ozawa Shigeru , Eda Hiroyuki , Norman James , Stroop William G. , Yanagi Kazuo
Published: September 2008

The geographical distribution of herpes simplex virus type 1 (HSV-1) restriction fragment length polymorphism (RFLP) variants BgKLand BgOLand the high relative frequency (RF) of BgKLin orolabial lesions has led to a dispersion–replacement hypothesis for these variants. The pathogenic properties of HSV-1 variants in mice and professional sumo wrestlers were examined here. The wrestlers herpes gladiatorum (HG) was caused by primary and non-primary HSV-1 infections and recurred in many wrestlers. HSV-1 neutralizing antibody titres in sera from wrestlers who did not develop HG were relatively high. HG was caused by distinct HSV-1 variants and strains from wrestlers living in the same sumo stable. The BgKLRF was significantly higher in HG cases, particularly in those with Kaposi's varicelliform eruption. These data indicated that reactivation and transmission of latent HSV-1 infections, especially BgKL, occurred frequently among wrestlers and was caused by severe skin damage. These results support the BgKLdispersion hypothesis.
Heterogeneous nuclear ribonuclear protein K interacts with the enterovirus 71 5′ untranslated region and participates in virus replication
Authors: Lin Jing-Yi , Li Mei-Ling , Huang Peng-Nien , Chien Kun-Yi , Horng Jim-Tong , Shih Shin-Ru
Published: September 2008

Enterovirus 71 (EV71) is a picornavirus that can cause severe neurological complications in children. Like other picornaviruses, the genomic RNA of EV71 contains a long 5′ untranslated region (UTR). Cellular proteins interact with the EV71 5′ UTR, and these interactions are important for virus replication. Using an RNA pull-down assay and proteomics approaches, this study identified the heterogeneous nuclear ribonucleoprotein K (hnRNP K) as one of the EV71 5′ UTR-associated proteins. The interaction between hnRNP K and the 5′ UTR was further confirmed by mapping the interaction regions to stem–loops I–II and IV in the 5′ UTR. During EV71 infection, hnRNP K was enriched in the cytoplasm where virus replication occurs, whereas hnRNP K was localized in the nucleus in mock-infected cells. Viral yields were found to be significantly lower in hnRNP K knockdown cells and viral RNA synthesis was delayed in hnRNP K knockdown cells in comparison with negative-control cells treated with small interfering RNA. These results suggest that hnRNP K interacts with the EV71 5′ UTR and participates in virus replication.