bluuebunny/crossref_metadata_2025 huggingface dataset:
Gastric slow waves, gastrointestinal symptoms and peptides in systemic sclerosis patients
Authors: Mcnearney T. A. , Sallam H. S. , Hunnicutt S. E. , Doshi D. , Wollaston D. E. , Mayes M. D. , Chen J. D. Z.
Published: June 2009

Abstract  Impaired gastric slow waves, frequent gastrointestinal (GI) symptoms and altered GI peptides have been reported in Scleroderma (SSc) patients. The aim of this study was to investigate the associations among these three important components in GI dysmotility. Seventeen fasted SSc patients underwent four channel surface electrogastrography, measuring % of normal gastric slow waves or dysrhythmia. Patients completed a questionnaire designed by us to assess demographics, upper and lower GI symptoms (symptom presence, frequency and impact on quality of life, QOL), by YES/NO, Likert Scales and Visual Analogue Scales 1–100 mm (called GI Dysmotility Questionnaire, GIDQ) and health‐related QOL by SF‐36. Fasting plasma vasoactive intestinal peptide (VIP) and motilin levels were measured by peptide immunoassays. There were significant correlations between percentages of gastric dysrhythmias (bradygastria or arrhythmia) and a number of major GI symptoms such as nausea, abdominal bloating and pain. The plasma level of VIP was correlated positively with % dysrhythmia but negatively with % normal slow waves. Motilin was positively correlated with slow wave coupling (coordination). No major differences were noted in the measured peptides or gastric slow waves between limited SSc and diffuse SSc. Correlations were noted between SF‐36 domain scores and our GIDQ scores. In SSc patients, gastric dysrhythmias are correlated with certain GI symptoms. Correlations are also noted between plasma VIP/Motilin levels and gastric slow waves. Thus in SSc, gastric dysrhythmias may be predictive of development of certain dyspeptic symptoms. Plasma VIP may be involved in the development of dysrhythmias.
Major role for CD8+ T cells in the protection against Toxoplasma gondii following dendritic cell vaccination
Authors: GUITON R. , ZAGANI R. , DIMIER‐POISSON I.
Published: June 2009

Toxoplasma gondii is the causative agent of toxoplasmosis, a worldwide zoonosis for which an effective vaccine is needed. Vaccination with pulsed dendritic cells is very efficient but their use in a vaccination protocol is unconceivable. Nevertheless, unravelling the induced effector mechanisms is crucial to design new vaccine strategies. We vaccinated CBA/J mice with parasite extract‐pulsed dendritic cells, challenged them with T. gondii cysts and carried out in vivo depletion of CD4+ or CD8+ T lymphocytes to study the subsequent cellular immune response and protective mechanisms. CD4+ lymphocytes were poorly implicated either in spleen and mesenteric lymph node (MLN) cytokine secretion or in mice protection. By contrast, the increasing number of intracerebral cysts and depletion of CD8+ cells were strongly correlated, revealing a prominent role for CD8+ lymphocytes in the protection of mice. Splenic CD8+ lymphocytes induce a strong Th1 response controlled by a Th2 response whereas CD8+ cells from MLNs inhibit both Th1 and Th2 responses. CD8+ cells are the main effectors following dendritic cell vaccination and Toxoplasma infection while CD4+ T cells only play a minor role. This contrasts with T. gondii infection which elicits the generation of CD4+ and CD8+ T cells that provide protective immunity.
Heterogeneity of Arabidopsis core promoters revealed by high‐density TSS analysis
Authors: Yamamoto Yoshiharu Y. , Yoshitsugu Tomoaki , Sakurai Tetsuya , Seki Motoaki , Shinozaki Kazuo , Obokata Junichi
Published: June 2009

Our limited understanding of plant promoters does not allow us to recognize any core promoter elements for the majority of plant promoters. To understand the promoter architecture of Arabidopsis, we used the combined approach of in silico detection of novel core promoter elements and large‐scale determination of transcription start sites (TSSs). To this end, we developed a novel methodology for TSS identification, using a combination of the cap‐trapper and massively parallel signature sequencing methods. This technique, CT–MPSS, allowed us to identify 158 237 Arabidopsis TSS tags corresponding to 38 311 TSS loci, which provides an opportunity for quantitative analysis of plant promoters. The expression characteristics of these promoters were analyzed with respect to core promoter elements detected by our in silico analyses, revealing that Arabidopsis promoters contain two main types of elements with exclusive characteristics, the TATA type and the GA type. The TATA‐type promoters tend to be associated with the Y Patch and the Inr motif, and cause high expression with sharp‐peak TSS clusters. By contrast, the GA type produces broad‐type TSS clusters. Unlike mammalian promoters, plant promoters are not associated with CpG islands. However, plant‐specific GA‐type promoters share some characteristics with mammalian CpG‐type promoters.
Uracil salvage is necessary for early Arabidopsis development
Authors: Mainguet Samuel E. , Gakière Bertrand , Majira Amel , Pelletier Sandra , Bringel Françoise , Guérard Florence , Caboche Michel , Berthomé Richard , Renou Jean Pierre
Published: June 2009

Uridine nucleotides can be formed by energy‐consuming de novo synthesis or by the energy‐saving recycling of nucleobases resulting from nucleotide catabolism. Uracil phosphoribosyltransferases (UPRTs; EC 2.4.2.9) are involved in the salvage of pyrimidines by catalyzing the formation of uridine monophosphate (UMP) from uracil and phosphoribosylpyrophosphate. To date, UPRTs are described as non‐essential, energy‐saving enzymes. In the present work, the six genes annotated as UPRTs in the Arabidopsis genome are examined through phylogenetic and functional complementation approaches and the available T‐DNA insertion mutants are characterized. We show that a single nuclear gene encoding a protein targeted to plastids, UPP, is responsible for almost all UPRT activity in Arabidopsis. The inability to salvage uracil caused a light‐dependent dramatic pale‐green to albino phenotype, dwarfism and the inability to produce viable progeny in loss‐of‐function mutants. Plastid biogenesis and starch accumulation were affected in all analysed tissues, with the exception of stomata. Therefore we propose that uracil salvage is of major importance for plant development.
Phosphorylation at S384 regulates the activity of the TaALMT1 malate transporter that underlies aluminum resistance in wheat
Authors: Ligaba Ayalew , Kochian Leon , Piñeros Miguel
Published: June 2009

In this study we examined the role of protein phosphorylation/dephosphorylation in the transport properties of the wheat (Triticum aestivum) root malate efflux transporter underlying Al resistance, TaALMT1. Pre‐incubation of Xenopus laevis oocytes expressing TaALMT1 with protein kinase inhibitors (K252a and staurosporine) strongly inhibited both basal and Al3+‐enhanced TaALMT1‐mediated inward currents (malate efflux). Pre‐incubation with phosphatase inhibitors (okadaic acid and cyclosporine A) resulted in a modest inhibition of the TaALMT1‐mediated currents. Exposure to the protein kinase C (PKC) activator, phorbol 12‐myristate 13‐acetate (PMA), enhanced TaALMT1‐mediated inward currents. Since these observations suggest that TaALMT1 transport activity is regulated by PKC‐mediated phosphorylation, we proceeded to modify candidate amino acids in the TaALMT1 protein in an effort to identify structural motifs underlying the process regulating phosphorylation. The transport properties of eight single point mutations (S56A, S183A, S324A, S337A, S351‐352A, S384A, T323A and Y184F) generated in amino acid residues predicted to be phosphorylation sites and examined electrophysiologically. The basic transport properties of mutants S56A, S183A, S324A, S337A, S351‐352A, T323A and Y184F were not altered relative to the wild‐type TaALMT1. Likewise the sensitivity of these mutants to staurosporine resembled that observed for the wild‐type transporter. However, the mutation S384A was noticeable, as in oocytes expressing this mutant protein TaALMT1‐mediated basal and Al‐enhanced currents were significantly inhibited, and the currents were insensitive to staurosporine or PMA. These findings indicate that S384 is an essential residue regulating TaALMT1 activity via direct protein phosphorylation, which precedes Al3+ enhancement of transport activity.
A mutation in the Arabidopsis mTERF‐related plastid protein SOLDAT10 activates retrograde signaling and suppresses 1O2‐induced cell death
Authors: Meskauskiene Rasa , Würsch Marco , Laloi Christophe , Vidi Pierre‐Alexandre , Coll Núria S. , Kessler Felix , Baruah Aiswarya , Kim Chanhong , Apel Klaus
Published: June 2009

The conditional flu mutant of Arabidopsis thaliana generates singlet oxygen (1O2) in plastids during a dark‐to‐light shift. Seedlings of flu bleach and die, whereas mature plants stop growing and develop macroscopic necrotic lesions. Several suppressor mutants, dubbed singlet oxygen‐linked death activator (soldat), were identified that abrogate 1O2‐mediated cell death of flu seedlings. One of the soldat mutations, soldat10, affects a gene encoding a plastid‐localized protein related to the human mitochondrial transcription termination factor mTERF. As a consequence of this mutation, plastid‐specific rRNA levels decrease and protein synthesis in plastids of soldat10 is attenuated. This disruption of chloroplast homeostasis in soldat10 seedlings affects communication between chloroplasts and the nucleus and leads to changes in the steady‐state concentration of nuclear gene transcripts. The soldat10 seedlings suffer from mild photo‐oxidative stress, as indicated by the constitutive up‐regulation of stress‐related genes. Even though soldat10/flu seedlings overaccumulate the photosensitizer protochlorophyllide in the dark and activate the expression of 1O2‐responsive genes after a dark‐to‐light shift they do not show a 1O2‐dependent cell death response. Disturbance of chloroplast homeostasis in emerging soldat10/flu seedlings seems to antagonize a subsequent 1O2‐mediated cell death response without suppressing 1O2‐dependent retrograde signaling. The results of this work reveal the unexpected complexity of what is commonly referred to as ‘plastid signaling’.
Evaluation of cluster‐randomized trials on maternal and child health research in developing countries
Authors: Handlos Line Neerup , Chakraborty Hrishikesh , Sen Pranab Kumar
Published: June 2009

Objective  To summarize and evaluate all publications including cluster‐randomized trials used for maternal and child health research in developing countries during the last 10 years.

Methods  All cluster‐randomized trials published between 1998 and 2008 were reviewed, and those that met our criteria for inclusion were evaluated further. The criteria for inclusion were that the trial should have been conducted in maternal and child health care in a developing country and that the conclusions should have been made on an individual level. Methods of accounting for clustering in design and analysis were evaluated in the eligible trials.

Results  Thirty‐five eligible trials were identified. The majority of them were conducted in Asia, used community as randomization unit, and had less than 10 000 participants. To minimize confounding, 23 of the 35 trials had stratified, blocked, or paired the clusters before they were randomized, while 17 had adjusted for confounding in the analysis. Ten of the 35 trials did not account for clustering in sample size calculations, and seven did not account for the cluster‐randomized design in the analysis. The number of cluster‐randomized trials increased over time, and the trials generally improved in quality.

Conclusions  Shortcomings exist in the sample‐size calculations and in the analysis of cluster‐randomized trials conducted during maternal and child health research in developing countries. Even though there has been improvement over time, further progress in the way that researchers utilize and analyse cluster‐randomized trials in this field is needed.
Predictors for mortality and loss to follow‐up among children receiving anti‐retroviral therapy in Lilongwe, Malawi
Authors: Fetzer Bradley C. , Hosseinipour Mina C. , Kamthuzi Portia , Hyde Lisa , Bramson Brian , Jobarteh Kebba , Torjesen Kristine , Miller William C. , Hoffman Irving , Kazembe Peter , Mwansambo Charles
Published: June 2009

Objectives  To determine predictors of mortality in children on anti‐retroviral therapy (ART) who attended the Paediatric HIV Clinic at Kamuzu Central Hospital in Lilongwe, Malawi.

Methods  Retrospective case cohort study by chart review of children who had started ART between October 2004 and May 2006. Bivariable and multivariable analysis were performed with and without defaulters to evaluate associations according to vital status and to identify independent predictors of mortality.

Results  Forty‐one of 258 children (15.9%) were deceased, 185 (71.7%) were alive, and 32 (12.4%) had defaulted: 51% were female, 7% were under 18 months, 26% were 18 months to 5 years, and 54% were >5 years of age. Most were WHO stage III or IV (56% and 37%, respectively). On multivariate analysis, factors most strongly associated with mortality and defaulting were age <18 months [hazards ratio (HR) 2.11 (95% CI 1.0–4.51)] and WHO stage IV [HR 2.00 (95% CI 1.07–3.76)].

Conclusions  To improve outcomes of HIV‐positive children, they must be identified and treated early, specifically children under 18 months of age. Access to infant diagnostic procedures must be improved to allow effective initiation of ART in infants at higher risk of death.
Spatial distribution and risk factors of dengue and Japanese encephalitis virus infection in urban settings: the case of Vientiane, Lao PDR
Authors: Vallée Julie , Dubot‐Pérès Audrey , Ounaphom Phonepaseuth , Sayavong Chantalay , Bryant Juliet E. , Gonzalez Jean‐Paul
Published: June 2009

Objectives  To evaluate the prevalence of flavivirus infection in Vientiane city (Lao PDR), to describe the spatial distribution of infection within this city, and to explore the link between flavivirus seroprevalence and urbanization levels of residential neighbourhoods.

Methods  A seroprevalence survey was carried out in 2006 including 1990 adults (≥35 years) and 1568 children (≥6months and <6 years) randomly selected.

Results  The prevalence of individuals with previous flavivirus infection (i.e. negative for both DEN and JE IgM but positive for DEN IgG) was 57.7%, with a significantly (P < 0.001) higher prevalence among adults (84.6%; 95% confidence interval (CI) = 82.4–86.8) than children (9.4%; 95% CI = 7.2–11.6). The prevalence of individuals with recent flavivirus infection (i.e. positive for DEN and/or JE IgM) was 6.5% and also significantly (P < 0.001) higher among adults (10.0%; 95% CI = 8.3–11.7) than children (2.5%; 95% CI = 1.5–3.5). In terms of spatial distribution, IgG prevalence was significantly (P < 0.001) higher among individuals living in the central city (60.1%; 95% CI = 56.2–64.1) than among those living in the periphery (44.3%; 95% CI = 41.5–47.2). In contrast, seroprevalence of recent flavivirus infections was significantly (P < 0.001) higher among individuals living in the periphery (8.8%; 95% CI = 6.9–10.7) than in the central city (4.0%; 95% CI = 2.9–5.2). This association was also statistically consistant (P < 0.01) in multivariate logistic regression after controlling for individual risk factors.

Conclusions  Our findings indicate that the level of urbanization of residential neighbourhoods influences the risk of flavivirus infection. The spatial distribution of flavivirus infection varies, even within a small city of less than 300 000 habitants such as Vientiane.
Nasopharyngeal carriage of S. pneumoniae among young children in rural Nepal
Authors: Coles Christian L. , Sherchand Jeevan B. , Khatry Subarna K. , Katz Joanne , LeClerq Steven C. , Mullany Luke C. , Tielsch James M.
Published: June 2009

Objectives  To provide epidemiologic data on Streptococcus pneumoniae (Spn) carriage in Nepal.

Methods  Prospective, population‐based study among children in Sarlahi, Nepal to estimate carriage prevalence, identify risk factors, and determine antibiotic susceptibility patterns and serotype distribution. Between December 2003 and July 2004, NP specimens were collected from 604 children aged 1–36 months with acute lower respiratory infection (ALRI) and 604 healthy, age‐ and season‐matched controls.

Results  Of the 1100 specimens analysed, carriage prevalence was approximately 80% in both groups. In the multivariate analyses, significant risk factors for Spn carriage in controls were Muslim religion [adjusted odds ratio (AOR): 2.93] and no latrine in the household (AOR: 2.41). Those treated for a recent illness had lower carriage rates (AOR: 0.37). Results were similar for ALRI cases with the addition of age ≥12 months (AOR: 1.68), and symptomatic infection (AOR: 3.78) as risk factors. The antibiotics and proportions of isolates resistant to them were as follows: penicillin 4.5%, cotrimoxazole 89.2%, chloramphenicol 1.4%, erythromycin 1.5% and tetracycline 22.7%. The most prevalent serogroups/types were 6, 19, 23, 15, 9 and 10.

Conclusions  Young children in rural Nepal experience high rates of Spn carriage. Most isolates were resistant to cotrimoxazole. Current conjugate Spn vaccines may substantially reduce the risk of a severe pneumonia and other Spn infections.
Clinical outcome and IL‐17, IL‐23, IL‐27 and FOXP3 expression in peripheral blood mononuclear cells of pollen‐allergic children during sublingual immunotherapy
Authors: Nieminen Kaisa , Valovirta Erkka , Savolainen Johannes
Published: June 2009

Nieminen K, Valovirta E, Savolainen J. Clinical outcome and IL‐17, IL‐23, IL‐27 and FOXP3 expression in peripheral blood mononuclear cells of pollen‐allergic children during sublingual immunotherapy. Pediatr Allergy Immunol 2010: 21: e174–e184. © 2009 John Wiley & Sons A/S

Induction of allergen‐specific, tolerogenic, IL‐10 and/or TGF‐β‐producing T‐regulatory (Treg) cells that express transcription factor FOXP3 is considered as one of the key mechanisms of allergen‐specific immunotherapy. However, little is known of the induction of FOXP3 expression in children during sublingual immunotherapy (SLIT). Recently, also, a novel subgroup of T‐helper (Th) cells, the Th17 cells, secreting predominantly IL‐17 (IL‐17A), was identified. The expressions of IL‐17 or the Th17‐regulating cytokines IL‐23 and IL‐27 during SLIT are currently completely unexplored. This randomized, placebo‐controlled dose‐response study was performed to analyze the effects of SLIT on FOXP3, IL‐17, IL‐23, and IL‐27 expressions in peripheral blood mononuclear cells (PBMC) of children with allergic rhinitis and their associations with clinical outcome. Thirty children were included: ten received SLIT with a glycerinated mixture of birch, hazel and alder with a cumulative weekly dose of 24,000 SQ‐U, 10 with dose 200,000 SQ‐U/wk, and ten received placebo. Cytokine and FOXP3 mRNA expressions in allergen‐, purified protein derivative‐stimulated and non‐stimulated PBMC were determined at 0, 1 and 2 yr of SLIT by real‐time RT‐PCR (TaqMan®). Symptoms and medications were recorded using diary cards. Allergen‐induced IL‐17 mRNA expression was significantly increased in the study subjects with elevated combined Symptom Medication Score (SMS) after 2 yr. There was also a significant positive correlation between the allergen‐induced IL‐17 and SMS in whole study group (r = 0.38, p = 0.039) and especially the 200,000 SQ‐U dose‐treated group (r = 0.74, p = 0.027) at 2 yr. Allergen‐induced FOXP3 mRNA expression was significantly increased in the 200,000 SQ‐U dose‐treated children after two study years as compared with baseline (p = 0.016) and placebo‐treated children (p = 0.028). The changes in FOXP3 mRNA expression positively correlated with IL‐10 and TGF‐β mRNAs during SLIT in whole study population. Increased allergen‐induced IL‐17 responses during SLIT are associated with elevated SMS. Increased tolerogenic, allergen‐specific Treg responses are also observed in children during SLIT.
Suicidality among Finnish anaesthesiologists
Authors: LINDFORS P. M. , MERETOJA O. A. , LUUKKONEN R. A. , ELOVAINIO M. J. , LEINO T. J.
Published: June 2009

Background: Suicide rates among physicians have constantly been reported to be higher than in the general population and anaesthesiologists appear to lead the suicide statistics among physicians.

Methods: A cross‐sectional questionnaire study was sent to all working Finnish anaesthesiologists (n=550) investigating their suicidality (ideation and/or planning and/or attempt). The response rate was 60%.

Results: One in four had at some time seriously been thinking about suicide. Respondents with poor health (crude odds ratios 11.2 and 95% confidence interval 3.8–33.0), low social support (10.5, 4.0–27.9), and family problems (6.5, 3.4–12.5) had the highest risk of suicidality. The highest risks at work were conflicts with co‐workers (4.1, 2.3–7.1) and superiors (2.1, 1.2–3.6), on‐call‐related stress symptoms (3.9, 1.9–8.3) and low organizational justice (1.9, 1.1–3.2). If a respondent had several risk factors, the risk of suicidality doubled with each cumulating factor.

Conclusions: The reported level of suicidal ideation among Finnish anaesthesiologists is worth concern. It should be of utmost importance to screen the risk factors and recognize suicidal physicians in order to help them. Interpersonal relationships, decision‐making procedures, and on‐call‐burden should be focused on when aiming to prevent suicidality among physicians.
Low‐dose vasopressin increases glomerular filtration rate, but impairs renal oxygenation in post‐cardiac surgery patients
Authors: BRAGADOTTIR G. , REDFORS B. , NYGREN A. , SELLGREN J. , RICKSTEN S.‐ E.
Published: June 2009

Background: The beneficial effects of vasopressin on diuresis and creatinine clearance have been demonstrated when used as an additional/alternative therapy in catecholamine‐dependent vasodilatory shock. A detailed analysis of the effects of vasopressin on renal perfusion, glomerular filtration, excretory function and oxygenation in man is, however, lacking. The objective of this pharmacodynamic study was to evaluate the effects of low to moderate doses of vasopressin on renal blood flow (RBF), glomerular filtration rate (GFR), renal oxygen consumption (RVO2) and renal oxygen extraction (RO2Ex) in post‐cardiac surgery patients.

Methods: Twelve patients were studied during sedation and mechanical ventilation after cardiac surgery. Vasopressin was sequentially infused at 1.2, 2.4 and 4.8 U/h. At each infusion rate, systemic haemodynamics were evaluated by a pulmonary artery catheter, and RBF and GFR were measured by the renal vein thermodilution technique and by renal extraction of 51chromium–ethylenediaminetetraacetic acid, respectively. RVO2 and RO2Ex were calculated by arterial and renal vein blood samples.

Results: The mean arterial pressure was not affected by vasopressin while cardiac output and heart rate decreased. RBF decreased and GFR, filtration fraction, sodium reabsorption, RVO2, RO2Ex and renal vascular resistance increased dose‐dependently with vasopressin. Vasopressin exerted direct antidiuretic and antinatriuretic effects.

Conclusions: Short‐term infusion of low to moderate, non‐hypertensive doses of vasopressin induced a post‐glomerular renal vasoconstriction with a decrease in RBF and an increase in GFR in post‐cardiac surgery patients. This was accompanied by an increase in RVO2, as a consequence of the increases in the filtered tubular load of sodium. Finally, vasopressin impaired the renal oxygen demand/supply relationship.
Caffeine does not attenuate experimentally induced ischemic pain in healthy subjects
Authors: DELLERMALM J. , SEGERDAHL M. , GRASS S.
Published: June 2009

Background and aims:Caffeine is likely the most widely used psychoactive substance in the world. It is also an analgesic adjuvant and has individual analgesic properties. The latter effect has been attributed to adenosine receptor antagonism, but the site of action is unknown. The aim of this study was to investigate the analgesic properties of caffeine on experimentally induced ischemic pain and to attempt to elucidate whether the site of action is central or peripheral.

Materials and methods:Seventeen healthy subjects received intravenous (i.v.) regional and systemic infusions of caffeine at 10 mg/kg or placebo in a double‐blind, crossover fashion to investigate the site of action for caffeine‐induced analgesia. Subjects underwent a sub‐maximum effort tourniquet test. Pain scores [visual analogue scale (VAS), 0–100] were assessed every minute up to a maximum of 45 min.

Results:The sum of pain scores (SPS, accumulation of VAS scores) was attenuated neither by systemic 2405 (±234) nor by i.v. regional caffeine 2427 (±190) as compared with placebo 2442 (±205),P=0.99 (mean±SEM). Time to maximal VAS score did not differ significantly between treatments,P=0.94. There was no correlation between caffeine concentration in plasma and time to maximal pain score, or between SPS and plasma concentration.

Conclusion:Caffeine does not have an analgesic effect on ischemic pain, either by a peripheral or by a central site of action.
DNA methylation patterns at the IGF2‐H19 locus in sperm of Swiss Landrace and Swiss Large White boars
Authors: Giannini Petra , Braunschweig Martin
Published: June 2009

DNA methylation patterns at the IGF2‐H19 locus were investigated in sperm DNA from Swiss Landrace (SL) and Swiss Large White (LW) boars. The putative IGF2 differentially methylated regions (DMR) 0, 1 and 2, a quantitative trait nucleotide (QTN) region in the intron 3 and a CpG island in the intron 4 of the IGF2 gene as well as three regions around porcine CTCF binding sites within the H19 differentially methylated domain (DMD) were selected for the DNA methylation analysis. In both breeds putative IGF2 DMR0, 1, 2 and H19 DMD were hypermethylated. Significant differences in DNA methylation content were found between the two breeds in the two DMD regions proximal to the H19 gene. The IGF2 QTN region and the CpG island in the IGF2 intron 4 were hypomethylated in sperm DNA of both breeds. The methylation analysis revealed significantly more methylated CpG sites in the intron 4 of sperm from the LW breed than in that from SL. No difference was found in global DNA methylation between the two breeds. These results indicate differences in DNA methylation patterns between breeds and it remains to be established whether variation in DNA methylation patterns impacts on phenotypic traits.
Novel SSR Markers for Polymorphism Detection in Pigeonpea (Cajanus spp.)
Authors: Saxena R. K. , Prathima C. , Saxena K.B. , Hoisington D. A. , Singh N. K. , Varshney R. K.
Published: June 2009

With 1 figure and 4 tables

With an objective to expand the repertoire of molecular markers in pigeonpea (Cajanus cajan), 36 microsatellite or simple sequence repeat (SSR) loci were isolated from a SSR‐enriched genomic library. Primer pairs were designed for 23 SSR loci, of which 16 yielded amplicons of expected size. Thirteen SSR markers were polymorphic amongst 32 cultivated and eight wild pigeonpea genotypes representing six Cajanus species. These markers amplified a total of 72 alleles ranging from two to eight alleles with an average of 5.5 alleles per locus. The polymorphic information content for these markers ranged from 0.05 to 0.55 with an average of 0.32 per marker. Phenetic analysis clearly distinguished all wild species genotypes from each other and from the cultivated pigeonpea genotypes. These markers should be useful for genome mapping, trait mapping, diversity studies and assessment of gene flow between populations in pigeonpea.
Tribble 3, a novel oxidized low‐density lipoprotein‐inducible gene, is induced via the activating transcription factor 4–C/EBP homologous protein pathway
Authors: Shang Yuan‐Yuan , Zhong Ming , Zhang Li‐Ping , Guo Zhong‐Xiu , Wang Zhi‐Hao , Zhang Yun , Deng Jing‐Ti , Zhang Wei
Published: June 2009

1. C/EBP homologueueueous protein (CHOP), an endoplasmic reticulum (ER) stress‐inducible protein, has a critical role in regulation of the cell cycle and apoptosis by forming heterodimers with other C/EBP proteins. However, how CHOP function is regulated remains to be determined. The human homologue of Drosophila tribbles (TRIB3) is associated with CHOP and is upregulated by oxidized low‐density lipoprotein (ox‐LDL). The aim of the present study was to investigate the role of CHOP in ox‐LDL‐induced TRIB3 expression in macrophages.

2. Human monocyte‐derived macrophages were treated with various concentrations of ox‐LDL (0, 2.5, 5, 10, 25 and 50 μg/mL) or 2 μg/mL tunicamycin for 0, 4, 8, 16, 24 and 48 h or were transfected with CHOP or TRIB3 expression plasmid and TRIB3 targeting short interference RNA (siRNA). The expression of CHOP and activating transcription factor 4 (ATF4) mRNA in treated cells was detected by quantitative real‐time polymerase chain reaction (PCR).

3. The expression of CHOP and ATF4 mRNA increased with increasing concentrations of ox‐LDL and duration of time. The ox‐LDL‐induced expression of TRIB3 mRNA was upregulated later than the expression of CHOP and ATF4 mRNA. Overexpression of CHOP increased the mRNA expression of TRIB3, which was further increased in CHOP‐overexpressing macrophages treated with ox‐LDL. Overexpression of TRIB3 suppressed the expression of CHOP, whereas TRIB3 silencing increased CHOP expression following ox‐LDL stimulation by a negative feedback mechanism.

4. In conculsion, the expression of ATF4 and CHOP is upregulated by ox‐LDL in a dose‐ and time‐dependent manner in naturally differentiated human macrophages. Oxidized LDL induces TRIB3 expression via an ATF4/CHOP‐dependent ER stress pathway.
New strategies for the prevention of stroke
Authors: Liu Ai‐Jun , Guo Jin‐Min , Xia Weiliang , Su Ding‐Feng
Published: June 2009

1. Stroke is a major cause of disability and death worldwide. It is preferable to prevent stroke rather than to treat it and, for the prevention of stroke, all risk factors relating to stroke need to be understood. The present paper reviews potential new strategies for the prevention of stroke based on findings of new risk factors, as well as classical risk factors.

2. Recently, new risk factors related to stroke were reported, including dysfunction of the arterial baroreflex, pro‐inflammatory cytokines, vitamins and hormone deficiency. Correspondingly, therapies targeting these risk factors where shown to significantly reduce the incidence and/or severity of stroke.

3. Because the genesis of stroke is multifactorial, the prevention of stroke should not target one risk factor only. Combination therapies with drugs acting on different risk factors may be more effective in the prevention of stroke.
Rosiglitazone‐induced myocardial protection against ischaemia–reperfusion injury is mediated via a phosphatidylinositol 3‐kinase/Akt‐dependent pathway
Authors: Zhang Xue‐Jiao , Xiong Zi‐Bo , Tang An‐Li , Ma Hong , Ma Yue‐Dong , Wu Jing‐Guo , Dong Yu‐Gang
Published: June 2009

1. Rosiglitazone is widely used in the treatment of Type 2 diabetes. However, in recent years it has become evident that the therapeutic effects of peroxisome proliferator‐activated receptor γ ligands reach far beyond their use as insulin sensitizers. Recently, the ability of rosiglitazone pretreatment to induce cardioprotection following ischaemia–reperfusion (I/R) has been well documented; however, the protective mechanisms have not been elucidated. In the present study, examined the role of the phosphatidylinositol 3‐kinase (PI3‐K)/Akt signalling pathway in rosiglitazone cardioprotection following I/R injury.

2. Mice were pretreated with 3 mg/kg per day rosiglitazone for 14 days before hearts were subjected to ischaemia (30 min) and reperfusion (2 h). Wortmannin (1.4 mg/kg, i.p.), an inhibitor of PI3‐K, was administered 10 min prior to myocardial I/R. Then, activation of the PI3‐K/Akt/glycogen synthase kinase (GSK)‐3α signalling pathway was examined. The effects of PI3‐K inhibition on rosiglitazone‐induced cardioprotection were also evaluated.

3. Compared with control rats, the ratio of infarct size to ischaemic area (area at risk) and the occurrence of sustained ventricular fibrillation in rosiglitazone‐pretreated rats was significantly reduced (P < 0.05). Rosiglitazone pretreatment attenuated cardiac apoptosis, as assessed by ELISA to determine cardiomyocyte DNA fragmentation. Rosiglitazone pretreatment significantly increased levels of phosphorylated (p‐) Akt and p‐GSK‐3α in the rat myocardium. Pharmacological inhibition of PI3‐K by wortmannin markedly abolished the cardioprotection induced by rosiglitazone.

4. These results indicate that rosiglitazone‐induced cardioprotection in I/R injury is mediated via a PI3‐K/Akt/GSK‐3α‐dependent pathway. The data also suggest that modulation of PI3‐K/Akt/GSK‐3α‐dependent signalling pathways may be a viable strategy to reduce myocardial I/R injury.
Angiotensin II and neurohumoral control of the renal medullary circulation
Authors: Evans Roger G , Head Geoffrey A , Eppel Gabriela A , Burke Sandra L , Rajapakse Niwanthi W
Published: June 2009

1. Angiotensin (Ang) II has multiple actions in the renal medullary circulation. It can induce vasodilatation and blunt the response of medullary blood flow (MBF) to renal nerve activation through AT1receptor‐mediated release of nitric oxide (NO) and/or vasodilator prostaglandins. These actions require high intravascular and/or intratubular AngII concentrations, so are not apparent under physiological conditions.

2. Nevertheless, these mechanisms blunt the responsiveness of MBF to AT1receptor‐mediated vasoconstriction. When these protective mechanisms fail, as when oxidative stress reduces NO bioavailability in the medullary circulation, AngII reduces MBF. If sustained, reduced MBF leads to the development of hypertension.

3. Chronic activation of the renin–angiotensin system (RAS) induces oxidative stress in the kidney. Therefore, MBF may be reduced in models of hypertension associated with RAS activation both because AngII levelsper seare increased and because of increased responsiveness of MBF to AngII‐induced vasoconstriction.

4. Endogenous AngII enhances the responsiveness of MBF to renal nerve stimulation, whereas NO blunts it. Chronic RAS activation and/or oxidative stress should therefore be expected to enhance MBF responses to renal nerve stimulation. Consistent with this, reductions in MBF induced by renal nerve stimulation are enhanced in rabbits with AngII‐induced hypertension, renovascular hypertension or after 9 weeks of fat feeding.

5. We conclude that the ability of endogenous AngII to reduce MBF and enhance the response of MBF to activation of the renal nerves could contribute to the development of hypertension under conditions of RAS activation, especially if accompanied by increased renal sympathetic nerve activity.
Brain cytokines as neuromodulators in cardiovascular control
Authors: Shi Peng , Raizada Mohan K , Sumners Colin
Published: June 2009

1. The role of cytokines in cardiovascular control, especially in neurogenic hypertension, has received considerable attention during the past few years. Brain cytokines have been shown to exert profound effects on neuronal activity. Recently, a number of studies have shown that administration of pro‐inflammatory cytokines or anti‐inflammatory cytokines into the central nervous system has a significant impact on sympathetic outflow, arterial pressure and cardiac remodelling in experimental models of hypertension and heart failure.

2. Our objective in this review is to present a succinct account of the effect of cytokines on neuronal activity and their role in cardiovascular disease. Furthermore, we propose a hypothesis for a neuromodulatory role of cytokines in the neural control of cardiovascular function.
Ghrelin gene‐related peptides: Multifunctional endocrine / autocrine modulators in health and disease
Authors: Seim Inge , Amorim Laura , Walpole Carina , Carter Shea , Chopin Lisa K , Herington Adrian C
Published: June 2009

1. Ghrelin is a multifunctional peptide hormone that affects various processes, including growth hormone and insulin release, appetite regulation, gut motility, metabolism and cancer cell proliferation. Ghrelin is produced in the stomach and in other normal and pathological cell types. It may act as an endocrine or autocrine/paracrine factor.

2. The present article reviews recent findings in the study of ghrelin and its receptor that suggest that the ghrelin gene locus may give rise to a number of functional molecules (peptides and RNA transcripts) in addition to ghrelin.

3. The ghrelin gene encodes a precursor protein, preproghrelin, from which ghrelin and other potentially active peptides are derived by alternative mRNA splicing and/or proteolytic processing. The metabolic role of the peptide obestatin, derived from the preproghrelin C‐terminal region, is contentious. However, obestatin has direct effects on cell proliferation.

4. The regulation of ghrelin expression and the mechanisms through which the peptide products arise are unclear. We have recently re‐examined the organization of the ghrelin gene and identified several novel exons and transcripts. One transcript, which lacks the ghrelin‐coding region of preproghrelin, contains the coding sequence of obestatin.

5. Furthermore, we have identified an overlapping gene on the antisense strand of ghrelin, namely GHRLOS, which generates transcripts that may function as non‐coding regulatory RNAs or code for novel, short bioactive peptides.

6. The identification of these novel ghrelin‐gene related transcripts and peptides raises critical questions regarding their physiological function and their potential role in obesity, diabetes and cancer.
Multidrug resistance‐associated proteins and implications in drug development
Authors: Liu Ya‐He , Di Yuan‐Ming , Zhou Zhi‐Wei , Mo Sui‐Lin , Zhou Shu‐Feng
Published: June 2009

1. The multidrug resistance‐associated proteins (MRPs) belong to the ATP‐binding cassette superfamily (ABCC family) of transporters that are expressed differentially in the liver, kidney, intestine and blood–brain barrier. There are nine human MRPs that transport a structurally diverse array of endo‐ and xenobiotics as well as their conjugates.

2. Multidrug resistance‐associated protein 1 can be distinguished from MRP2 and MRP3 by its higher affinity for leukotriene C4. Unlike MRP1, MRP2 functions in the extrusion of endogenous organic anions, such as bilirubin glucuronide and certain anticancer agents. In addition to the transport of glutathione and glucuronate conjugates, MRP3 has the additional capability of mediating the transport of monoanionic bile acids.

3. Both MRP4 and MRP5 are able to mediate the transport of cyclic nucleotides and confer resistance to certain antiviral and anticancer nucleotide analogues. Hereditary deficiency of MRP6 results in pseudoxanthoma elasticum. In the body, MRP6 is involved in the transport of glutathione conjugates and the cyclic pentapeptide BQ123.

4. Various MRPs show considerable differences in tissue distribution, substrate specificity and proposed physiological function. These proteins play a role in drug disposition and excretion and thus are implicated in drug toxicity and drug interactions. Increased efflux of natural product anticancer drugs and other anticancer agents mediated by MRPs from cancer cells is associated with tumour resistance.

5. A better understanding of the function and regulating mechanisms of MRPs could help minimize and avoid drug toxicity and unfavourable drug–drug interactions, as well as help overcome drug resistance.
Supraventricular tachycardia as a presenting feature of volvulus in a 15‐month‐old boy
Authors: Jacombs Anita SW , Cross Kate M , Holland Andrew JA , Wilkins Barry H
Published: June 2009

A 15‐month‐old boy presented in shock with a supraventricular tachycardia following a 12‐h history of worsening abdominal pain and vomiting. The supraventricular tachycardia reverted to sinus tachycardia with fluid resuscitation and adenosine. He was noted to have a distended and firm abdomen. A presumptive diagnosis of intestinal ischaemia was subsequently confirmed at laparotomy when an internal hernia with a distal small bowel volvulus and necrosis was found. Intestinal ischaemia presenting with a life‐threatening cardiac dysrhythmia in a child appears not to have been reported previously.
Review article: Coronary artery stenoses: Detection and revascularization in renal disease
Authors: PILMORE HELEN L
Published: June 2009

Cardiovascular events are markedly elevated in those with all degrees of renal impairment compared to the general population. There are well established guidelines in the general population for the management of coronary artery disease, however, similar guidelines have not been established in the renal population. This review examines the current published work on the detection of coronary artery stenoses in addition to summarizing the outcomes of revascularization in patients with kidney disease. Testing for coronary artery disease in the renal population most commonly occurs in dialysis patients as part of their assessment for renal transplantation. While a positive myocardial stress test for the detection of significant coronary artery stenoses is associated with an increased risk of cardiac events, there is no clear information currently showing that cardiovascular testing itself reduces the rate of adverse cardiac events after transplantation. Revascularization of coronary artery stenoses is associated with higher morbidity and mortality in all groups with kidney disease than in the general population, with the exception of renal transplant recipients where the mortality is likely to be similar to that of the general population. There appears to be a benefit in coronary artery bypass surgery compared to percutaneous intervention in those on dialysis and after renal transplant. Currently, there is little data to support coronary artery intervention prior to transplantation in those with asymptomatic coronary artery disease.
Experimental warming causes rapid loss of plant diversity in New England salt marshes
Authors: Gedan Keryn B. , Bertness Mark D.
Published: June 2009

Anthropogenic climate change is predicted to cause widespread biodiversity loss due to shifts in species’ distributions, but these predictions rarely incorporate ecological associations such as zonation. Here, we predict the decline of a diverse assemblage of mid‐latitude salt marsh plants, based on an ecosystem warming experiment. In New England salt marshes, a guild of halophytic forbs occupies stressful, waterlogged pannes. At three sites, experimental warming of < 4 °C led to diversity declines in pannes and rapid takeover by a competitive dominant, Spartina patens. In Rhode Island, near their southern range limit, pannes were more sensitive to warming than farther north, and panne area also declined in control plots over the three‐season experiment. These results suggest that warming will rapidly reduce plant diversity in New England salt marshes by eliminating a high diversity zone. Biodiversity in zoned ecosystems may be more affected by climate‐driven shifts in zonation than by individual species’ distribution shifts.
When can we distinguish between neutral and non‐neutral processes in community dynamics under ecological drift?
Authors: Ruokolainen Lasse , Ranta Esa , Kaitala Veijo , Fowler Mike S.
Published: June 2009

Determining whether the composition of ecological communities (species presence and abundance), can be predicted from species demographic traits, rather than being a result of neutral drift, is a key ecological question. Here we compare the similarity of community composition, from different community assembly models run under identical environmental conditions, where interspecific competition is assumed to be either neutral or niche‐based. In both cases, species colonize a focal patch from a network of neighbouring patches in a metacommunity. We highlight the circumstances (rate and spatial scale of dispersal, and the relative importance of ecological drift) where commonly used community similarity metrics or species rank–abundance relationships are likely to give similar results, regardless of the underlying processes (neutral or non‐neural) driving species’ dynamics. As drift becomes more important in driving species abundances, deterministic niche structure has a smaller influence. Our ability to discriminate between different underlying processes driving community organization depends on the relative importance of different drift processes that operate on different spatial scales.
The ‘Draw‐the‐Family Test’ in the preoperative assessment of patients with anorectal diseases and psychological distress: a prospective controlled study
Authors: Miliacca C. , Gagliardi G. , Pescatori M.
Published: June 2009

Aim  Some benign anorectal diseases may have psychosomatic aetiology, but patients often refuse direct psychological counselling. The Draw‐the‐Family Test (DFT) is a simple indirect investigation. The aim of this study was to evaluate the DFT in patients with psychological problems undergoing surgery for benign anorectal disease and to correlate the results with surgical outcome.

Method  DFT was administered prior to surgery to 62 patients with benign anorectal disease who admitted psychological problems at the time of the outpatient visit. Of these 18 (29%) had functional disease, mainly obstructed defecation (OD) while 44 (71%) had organic disease (haemorrhoids, fissures, pilonidal sinus or fistula). DFT was also administered to 40 healthy control subjects. Each DFT was judged as normal or pathological according to 10 parameters. Patients were followed up for a median of 12 months (range 3–64) and divided into two outcome groups, success (n = 58) and failure (n = 12) according to the results of a questionnaire. The DFT of all patients was then correlated with the outcome.

Results  None of the patients refused DFT. All DFT parameters but one (animal/things instead of human beings) were more frequent in patients compared with controls (P < 0.05). When comparing separately organic or functional disease patients with controls, one parameter (absence of patient in the drawing) was only pathological in the organic disease group (P < 0.05). Another parameter (schematic figures) was more frequently altered in the organic disease group compared with the functional disease group (P = 0.01). Eight out of 10 parameters were more frequently pathological in patients who failed after treatment, but none reached statistical significance.

Conclusion  Results of DFT in patients with anorectal disease admitting to psychological problems are markedly different from healthy controls. Patients with organic disease and those with functional bowel disease have different DFT profiles. In our study group, DFT had an excellent compliance but could not predict the outcome of surgery.
Two‐year‐olds compute syntactic structure on‐line
Authors: Bernal Savita , Dehaene‐Lambertz Ghislaine , Millotte Séverine , Christophe Anne
Published: June 2009

Syntax allows human beings to build an infinite number of new sentences from a finite stock of words. Because toddlers typically utter only one or two words at a time, they have been thought to have no syntax. Using event‐related potentials (ERPs), we demonstrated that 2‐year‐olds do compute syntactic structure when listening to spoken sentences. We observed an early left‐lateralized brain response when an expected verb was incorrectly replaced by a noun (or vice versa). Thus, toddlers build on‐line expectations as to the syntactic category of the next word in a sentence. In addition, the response topography was different for nouns and verbs, suggesting that different neural networks already underlie noun and verb processing in toddlers, as they do in adults.