bluuebunny/crossref_metadata_2025 huggingface dataset:
Bos grunniens papillomavirus type 1: a novel deltapapillomavirus associated with fibropapilloma in yak
Authors: Zhu Wei , Dong Jian-Bao , Zhang Jun , Uchida Kazuyuki , Watanabe Ken-ichi , Goto Yoshitaka , Haga Takeshi
Published: October 2012

Papillomaviruses (PVs) have been widely identified among vertebrates, but have not yet been reported to infect yaks. We report, for the first time, a novel deltapapillomavirus that was associated with fibropapilloma in yak herds on the Qinghai–Tibetan Plateau. Six skin papilloma samples were collected and examined using histopathology, immunohistochemistry and PCR assays. The samples were identified as fibropapilloma and were found to contain PV antigens. Sequencing of diagnostic PCR products and the full-length genome revealed that all samples were infected with the same PV type. The whole virus genome was 7946 bp in length and possessed the common PV genomic organization. The virus was identified as a novel PV type and designated Bos grunniens papillomavirus type 1 (BgPV-1) based on the nucleotide sequence alignment of the L1 ORF. It is classified in the Delta-4 species of the genus Deltapapillomavirus based on phylogenetic analysis of the L1 ORF. Identification of this novel PV type provides further information about the pathology, development of diagnostic methods and evolutionary studies of the family Papillomaviridae.
Temporal classification and mapping of non-polyadenylated transcripts of an invertebrate iridovirus
Authors: İnce İkbal Agah , Özcan Kadriye , Vlak Just M. , van Oers Monique M.
Published: October 2012

The temporal expression of the 54 Chilo iridescent virus (CIV) virion protein genes was investigated by combining drug treatments that inhibit protein or DNA synthesis and an RT-PCR strategy particularly suitable for non-polyadenylated mRNAs. This method generates a uniform 3′ terminus by ligation of a 5′-phosphorylated oligonucleotide to the 3′ end of the transcript that is recognized by a complementary primer during RT-PCR. This analysis showed that CIV virion proteins are encoded by genes in all three predetermined temporal classes: 23 immediate-early, 11 delayed-early and seven late virion gene transcripts were identified and assigned to ORFs. Early transcription of many virion protein genes supports the notion that virion proteins may also play essential roles in the initial stages of infection. In addition, some of the early gene products present in the virion may reflect the intracellular path that the virus follows during infection.
Overlapping structure of hepatitis B virus (HBV) genome and immune selection pressure are critical forces modulating HBV evolution
Authors: Cento Valeria , Mirabelli Carmen , Dimonte Salvatore , Salpini Romina , Han Yue , Trimoulet Pascale , Bertoli Ada , Micheli Valeria , Gubertini Guido , Cappiello Giuseppina , Spanò Alberto , Longo Roberta , Bernassola Martina , Mazzotta Francesco , De Sanctis Giuseppe Maria , Zhang Xin Xin , Verheyen Jens , D’Arminio Monforte Antonella , Ceccherini-Silberstein Francesca , Perno Carlo Federico , Svicher Valentina
Published: October 2012

How the overlap between the hepatitis B virus (HBV) reverse transcriptase (RT) and HBV S antigen (HBsAg) genes modulates the extent of HBV genetic variability is still an open question, and was investigated here. The rate of nucleotide conservation (≤1 % variability) followed an atypical pattern in the RT gene, due to an overlap between RT and HBsAg (69.9 % nucleotide conservation in the overlapping region vs 41.2 % in the non-overlapping region;P<0.001), with a consequently lower rate of synonymous substitution within the overlapping region [median(interquartile range)dS = 3.1(1.5–7.4) vs 20.1(10.6–30.0);P = 3.249×10−22]. The most conserved RT regions were located within the YMDD motif and the N-terminal parts of the palm and finger domains, critical for RT functionality. These regions also corresponded to highly conserved HBsAg domains that are critical for HBsAg secretion. Conversely, the genomic region encoding the HBsAg antigenic loop (where immune-escape mutations are localized) showed a sharp decrease in the extent of conservation (40.6 %), which was less pronounced in the setting of human immunodeficiency virus (HIV)-driven immune suppression (48.8 % in HIV–HBV co-infection vs 21.5 % in mono-infected patients;P = 0.020). In conclusion, the overlapping reading frame and the immune system appear to have shaped the patterns of RT and HBsAg genetic variability. Highly conserved regions in RT and HBsAg may deserve further attention as novel therapeutic targets.
ISG56/IFIT1 is primarily responsible for interferon-induced changes to patterns of parainfluenza virus type 5 transcription and protein synthesis
Authors: Andrejeva J. , Norsted H. , Habjan M. , Thiel V. , Goodbourn S. , Randall R. E.
Published: October 2012

Interferon (IFN) induces an antiviral state in cells that results in alterations of the patterns and levels of parainfluenza virus type 5 (PIV5) transcripts and proteins. This study reports that IFN-stimulated gene 56/IFN-induced protein with tetratricopeptide repeats 1 (ISG56/IFIT1) is primarily responsible for these effects of IFN. It was shown that treating cells with IFN after infection resulted in an increase in virus transcription but an overall decrease in virus protein synthesis. As there was no obvious decrease in the overall levels of cellular protein synthesis in infected cells treated with IFN, these results suggested that ISG56/IFIT1 selectively inhibits the translation of viral mRNAs. This conclusion was supported by in vitro translation studies. Previous work has shown that ISG56/IFIT1 can restrict the replication of viruses lacking a 2′-O-methyltransferase activity, an enzyme that methylates the 2′-hydroxyl group of ribose sugars in the 5′-cap structures of mRNA. However, the data in the current study strongly suggested that PIV5 mRNAs are methylated at the 2′-hydroxyl group and thus that ISG56/IFIT1 selectively inhibits the translation of PIV5 mRNA by some as yet unrecognized mechanism. It was also shown that ISG56/IFIT1 is primarily responsible for the IFN-induced inhibition of PIV5.
Correlation between structure, protein composition, morphogenesis and cytopathology of Glossina pallidipes salivary gland hypertrophy virus
Authors: Kariithi Henry M. , van Lent Jan W. M. , Boeren Sjef , Abd-Alla Adly M. M. , İnce İkbal Agah , van Oers Monique M. , Vlak Just M.
Published: October 2012

TheGlossina pallidipessalivary gland hypertrophy virus (GpSGHV) is a dsDNA virus with rod-shaped, enveloped virions. Its 190 kb genome contains 160 putative protein-coding ORFs. Here, the structural components, protein composition and associated aspects of GpSGHV morphogenesis and cytopathology were investigated. Four morphologically distinct structures: the nucleocapsid, tegument, envelope and helical surface projections, were observed in purified GpSGHV virions by electron microscopy. Nucleocapsids were present in virogenic stroma within the nuclei of infected salivary gland cells, whereas enveloped virions were located in the cytoplasm. The cytoplasm of infected cells appeared disordered and the plasma membranes disintegrated. Treatment of virions with 1 % NP-40 efficiently partitioned the virions into envelope and nucleocapsid fractions. The fractions were separated by SDS-PAGE followed by in-gel trypsin digestion and analysis of the tryptic peptides by liquid chromatography coupled to electrospray and tandem mass spectrometry. Using the MaxQuant program with Andromeda as a database search engine, a total of 45 viral proteins were identified. Of these, ten and 15 were associated with the envelope and the nucleocapsid fractions, respectively, whilst 20 were detected in both fractions, most likely representing tegument proteins. In addition, 51 host-derived proteins were identified in the proteome of the virus particle, 13 of which were verified to be incorporated into the mature virion using a proteinase K protection assay. This study provides important information about GpSGHV biology and suggests options for the development of future anti-GpSGHV strategies by interfering with virus–host interactions.
Structures of the ribosome-inactivating protein from barley seeds reveal a unique activation mechanism
Authors: Lee Byung-Gil , Kim Min Kyung , Kim Byeong-Won , Suh Se Won , Song Hyun Kyu
Published: October 2012

Ribosome-inactivating protein (RIP), a defence protein found in various plants, possesses different chain architectures and activation mechanisms. The RIP from barley (bRIP) is a type I RIP and has sequence features that are divergent from those of type I and type II RIPs from dicotyledonous plants and even the type III RIP from maize. This study presents the first crystal structure of an RIP from a cereal crop, barley, in free, AMP-bound and adenine-bound states. For phasing, a codon-optimized syntheticbrip1gene was used and a vector was constructed to overexpress soluble bRIP fusion proteins; such expression has been verified in a number of cases. The overall structure of bRIP shows folding similar to that observed in other RIPs but also shows significant differences in specific regions, particularly in a switch region that undergoes a structural transition between a 310-helix and a loop depending on the liganded state. The switch region is in a position equivalent to that of a proteolytically susceptible and putative ribosome-binding site in type III RIPs. Thus, the bRIP structure confirms the detailed enzymatic mechanism of this N-glycosidase and reveals a novel activation mechanism for type I RIPs from cereal crops.
A monomeric TIM-barrel structure fromPyrococcus furiosusis optimized for extreme temperatures
Authors: Repo Heidi , Oeemig Jesper S. , Djupsjöbacka Janica , Iwaï Hideo , Heikinheimo Pirkko
Published: October 2012

The structure of phosphoribosyl anthranilate isomerase (TrpF) from the hyperthermophilic archaeonPyrococcus furiosus(PfTrpF) has been determined at 1.75 Å resolution. ThePfTrpF structure has a monomeric TIM-barrel fold which differs from the dimeric structures of two other known thermophilic TrpF proteins. A comparison of thePfTrpF structure with the two known bacterial thermophilic TrpF structures and the structure of a related mesophilic protein fromEscherichia coli(EcTrpF) is presented. The thermophilic TrpF structures contain a higher proportion of ion pairs and charged residues compared with the mesophilicEcTrpF. These residues contribute to the closure of the central barrel and the stabilization of the barrel and the surrounding α-helices. In the monomericPfTrpF conserved structural water molecules are mostly absent; instead, the structural waters are replaced by direct side-chain–main-chain interactions. As a consequence of these combined mechanisms, theP. furiosusenzyme is a thermodynamically stable and entropically optimized monomeric TIM-barrel enzyme which defines a good framework for further protein engineering for industrial applications.
The structure ofL-amino-acid ligase fromBacillus licheniformis
Authors: Suzuki Michihiko , Takahashi Yuichi , Noguchi Atsushi , Arai Toshinobu , Yagasaki Makoto , Kino Kuniki , Saito Jun-ichi
Published: October 2012

L-Amino-acid ligases (LALs) are enzymes which catalyze the formation of dipeptides by linking two L-amino acids. Although many dipeptides are known and expected to have medical and nutritional benefits, their practical use has been limited owing to their low availability and high expense. LALs are potentially desirable tools for the efficient production of dipeptides; however, the molecular basis of substrate recognition by LAL has not yet been sufficiently elucidated for the design of ideal LALs for the desired dipeptides. This report presents the crystal structure of the LAL BL00235 derived fromBacillus licheniformisNBRC 12200 determined at 1.9 Å resolution using the multi-wavelength anomalous dispersion method. The overall structure of BL00235 is fairly similar to that of YwfE, the only LAL with a known structure, but the structure around the catalytic site contains some significant differences. Detailed structural comparison of BL00235 with YwfE sheds some light on the molecular basis of the substrate specificities.
Severe adhesive arachnoiditis resulting in progressive paraplegia following obstetric spinal anaesthesia: a case report and review
Authors: Killeen T. , Kamat A. , Walsh D. , Parker A. , Aliashkevich A.
Published: October 2012

A 27‐year‐old woman developed severe adhesive arachnoiditis after an obstetric spinal anaesthetic with bupivacaine and fentanyl, complicated by back pain and headache. No other precipitating cause could be identified. She presented one week postpartum with communicating hydrocephalus and syringomyelia and underwent ventriculoperitoneal shunting and foramen magnum decompression. Two months later, she developed rapid, progressive paraplegia and sphincter dysfunction. Attempted treatments included exploratory laminectomy, external drainage of the syrinx and intravenous steroids, but these were unsuccessful and the patient remains significantly disabled 21 months later. We discuss the pathophysiology of adhesive arachnoiditis following central neuraxial anaesthesia and possible causative factors, including contamination of the injectate, intrathecal blood and local anaesthetic neurotoxicity, with reference to other published cases. In the absence of more conclusive data, practitioners of central neuraxial anaesthesia can only continue to ensure meticulous, aseptic, atraumatic technique and avoid all potential sources of contamination. It seems appropriate to discuss with patients the possibility of delayed, permanent neurological deficit while taking informed consent.
Neurodevelopment in children with and without congenital heart block born to anti‐Ro/SSA‐positive mothers
Authors: Skog Amanda , Tingström Joanna , Salomonsson Stina , Sonesson Sven‐Erik , Wahren‐Herlenius Marie
Published: October 2012

Objective

To define factors influencing neurodevelopment in children with and without complete congenital heart block (CHB) born to mothers with Ro/SSA autoantibodies.

Patients and methods

Medical records of a population‐based cohort of siblings with (n = 60) and without (n = 54) CHB born 1974–2009 to anti‐Ro/SSA‐positive mothers were retrieved from children primary healthcare centres and school health services and used to extract data on neurodevelopment.

Results

Impaired neurodevelopment was reported in 16% of the children (18/114) during the follow‐up time of 13.0 (8.2–17.5) years, median (quartiles). Reported problems included speech (9%), motor (8%) and learning (8%) impairment, attention deficit (5%) and behavioural impairment (4%). Impairment in motor skill development was more common in boys (p < 0.001) if the child was born preterm (p < 0.001). Learning impairment was significantly influenced by maternal SLE (p < 0.005), while attention deficits was influenced by both maternal SLE (p < 0.05) and CHB in the child (p < 0.05).

Conclusions

Our data indicate that in addition to well‐established factors such as male sex and being born preterm, both maternal SLE and CHB may influence neurodevelopment. Follow‐up of neurodevelopment should therefore be considered for children with CHB, especially if the mother is diagnosed with SLE.
Physician specialty is associated with adherence to treatment guidelines for acute otitis media in children
Authors: Grossman Zachi , Silverman Barbara G , Miron Dan
Published: October 2012

Aim

To identify physician and visit characteristics affecting adherence to guidelines recommending delayed antibiotic therapy for acute otitis media (AOM).

Methods

We used physician visit data from an Israeli health maintenance organization to identify children ages 0–15 diagnosed with AOM. We defined early antibiotic treatment as purchase, within 3 days of diagnosis, of antibiotics prescribed by the diagnosing physician. We considered the effect of physician specialty (paediatrician, otolaryngologist, and family physician), day of the week, setting (urgent care/clinic) and clinic location (peripheral/central) on the rate of early treatment.

Results

Proportion of cases receiving early treatment dropped from 2002 to 2009 in cases treated by otolaryngologists (47–36%) and paediatricians (46–42%) and increased in cases treated by family physicians (43–50%). Treatment rate was higher in cases treated on weekends than on weekdays (48% vs. 44%), in urgent care as compared to clinic settings (51% vs. 44%), and in children living in peripheral as compared to central areas (52% vs. 38%).

Conclusions

Successful implementation of delayed treatment of otitis media guidelines requires addressing factors associated with increased early treatment rates.
Review article: the emerging interplay among the gastrointestinal tract, bile acids and incretins in the pathogenesis of diabetes and non‐alcoholic fatty liver disease
Authors: Zarrinpar A. , Loomba R.
Published: October 2012

Background

Recent research has led to an interest in the role of the gut and liver in type 2 diabetes mellitus (T2DM).

Aim

To review the role of the gastrointestinal system in glucose homoeostasis, with particular focus on the effects of incretin hormones, hepatic steatosis and bile acids.

Methods

PubMed and Google Scholar were searched using terms such as incretin, glucose‐dependent insulinotropic polypeptide (GIP), glucagon‐like peptide‐1 (GLP‐1), dipeptidyl peptidase‐4 (DPP‐4), hepatic steatosis, bile acid and gastric bypass. Additional relevant references were identified by reviewing the reference lists of articles.

Results

Perturbations of incretin hormones and bile acid secretion contribute to the pathogenesis ofT2DM, leading to their potential as therapeutic targets. The incretin hormones (GIPandGLP‐1) are deactivated byDPP‐4.GLP‐1 agonists andDPP‐4 inhibitors improve glycaemic control in patients withT2DM. Hepatic steatosis, along with insulin resistance, may precede the development ofT2DM, and may benefit from anti‐diabetes medications. Bile acids play an important role in glucose homoeostasis, with effects mediated via the farnesoidXreceptor (FXR) and the cell surface receptorTGR5. The bile acid sequestrant colesevelam has been shown to be effective in improving glycaemic control in patients withT2DM. Altered gastrointestinal anatomy after gastric bypass surgery may also affect enterohepatic recirculation of bile acids and contribute to improved glycaemic control.

Conclusions

Research in recent years has led to new pathways and processes with a role in glucose homoeostasis, and new therapeutic targets and options for type 2 diabetes mellitus.
The relationship between sub‐clinical obsessive‐compulsive symptoms and social cognition in chronic schizophrenia
Authors: Whitton Alexis E. , Henry Julie D.
Published: October 2012

Objectives

Comorbid obsessive‐compulsive (OC) symptoms in individuals with a primary diagnosis of schizophrenia are related to poorer cognitive performance and functional outcomes, but no study to date has assessed whether this comorbidity might also have implications for social cognition. The aim of the present study was to provide the first test of this possibility.

Design and methods

Individuals with schizophrenia (n = 34) and demographically matched non‐clinical controls (n = 44) were assessed on two of the most important aspects of social cognitive function (1) facial affect recognition and (2) theory of mind, alongside more standard measures of cognitive function.

Results

The presence of OC symptoms was related to poorer performance on some of the cognitive measures, as well as one of the social cognitive measures (facial affect recognition). However, these relationships disappeared after controlling for scores on more general indices of schizophrenia psychopathology.

Conclusions

The presence of OC symptoms in schizophrenia is not only associated with increased cognitive impairment but also increased difficulties with at least some aspects of social cognitive function. However, these relationships appear to reflect the elevated levels of psychopathology seen in this cohort more generally, rather than being uniquely attributable to OC symptomatology.

Practitioner Points

The presence of obsessive‐compulsive (OC) symptoms in chronic schizophrenia is not only associated with greater cognitive impairment but also with increased difficulties in at least some aspects of social cognitive function.

These increased difficulties appear to reflect a more severe presentation of schizophrenia overall rather than the presence of OC symptoms specifically.

A limitation of this research design was the inclusion of only chronic, medicated participants with schizophrenia. Individuals at early or more acute phases of schizophrenia may present with a different profile of impairment.

Further research is needed to assess whether or not different results emerge where there is a comorbid diagnosis of obsessive‐compulsive disorder (as opposed to sub‐clinical OC symptoms).
The Limits on Pay as a Strategic Tool: Obstacles to Alignment in Non‐Union Environments
Authors: Trevor Jonathan , Brown William
Published: October 2012

Strategic human resource management literature emphasizes the potential of pay to secure strategically desirable employee outcomes for the employer. Strategic pay, in contrast with pluralist models of pay determination, assumes an absence of collective bargaining constraints. This article analyses the process of determination of non‐unionized managerial, professional and technical pay in seven leading consumer goods firms that claim to use pay as a strategic tool. It demonstrates that implemented pay practice is often remote from what is aspired to strategically. Despite the absence of collective bargaining constraints, there remain unavoidable obstacles to the ability of management to implement pay systems aligned to strategic goals. These constraints impose fundamental limitations on the use of pay as a strategic tool.
Anxiety and chronic couple relationship stress moderate adrenocortical response to couple interaction in expectant parents
Authors: Feinberg Mark E. , Jones Damon E. , Granger Douglas A. , Bontempo Daniel E.
Published: October 2012

The study examines whether anxiety or chronic relationship stress alter the way that couple conflict affects cortisol levels for women and men during the transition to parenthood. Saliva samples, assayed for cortisol, were collected before and after couple interaction from 128 heterosexual couples expecting their first child. Confirming prior research, expectant mothers had higher cortisol levels than their spouses, and gestational age was linked to women's cortisol level. Negativity during couple interaction was associated with greater cortisol reactivity for men, but not women. Tests of moderation indicated little relation between negativity and cortisol recovery for individuals with a low level of anxiety or little history of chronic arguing with the partner. However, among individuals with elevated levels of either of these two factors, negativity was linked to less cortisol recovery for men, but more cortisol recovery for women. Consistent results were also found for the relation between low warmth in the couple interaction and both reactivity and recovery for men and women high in anxiety. Future research should examine whether pregnancy is responsible for these different gender patterns, or whether the inhibition of negativity is stressful for women with high levels of risk.
A study of subunit selectivity, mechanism and site of action of the delta selective compound 2 (DS2) at human recombinant and rodent native GABAA receptors
Authors: Jensen ML , Wafford KA , Brown AR , Belelli D , Lambert JJ , Mirza NR
Published: October 2012

Background and Purpose

Most GABAA receptor subtypes comprise 2α, 2β and 1γ subunit, although for some isoforms, a δ replaces a γ‐subunit. Extrasynaptic δ‐GABAA receptors are important therapeutic targets, but there are few suitable pharmacological tools. We profiled DS2, the purported positive allosteric modulator (PAM) of δ‐GABAA receptors to better understand subtype selectivity, mechanism/site of action and activity at native δ‐GABAA receptors.

Experimental Approach

Subunit specificity of DS2 was determined using electrophysiological recordings of Xenopus laevis oocytes expressing human recombinant GABAA receptor isoforms. Effects of DS2 on GABA concentration–response curves were assessed to define mechanisms of action. Radioligand binding and electrophysiology utilising mutant receptors and pharmacology were used to define site of action. Using brain‐slice electrophysiology, we assessed the influence of DS2 on thalamic inhibition in wild‐type and δ0/0 mice.

Key Results

Actions of DS2 were primarily determined by the δ‐subunit but were additionally influenced by the α, but not the β, subunit (α4/6βxδ > α1βxδ >> γ2‐GABAA receptors > α4β3). For δ‐GABAA receptors, DS2 enhanced maximum responses to GABA, with minimal influence on GABA potency. (iii) DS2 did not act via the orthosteric, or known modulatory sites on GABAA receptors. (iv) DS2 enhanced tonic currents of thalamocortical neurones from wild‐type but not δ0/0 mice.

Conclusions and Implications

DS2 is the first PAM selective for α4/6βxδ receptors, providing a novel tool to investigate extrasynaptic δ‐GABAA receptors. The effects of DS2 are mediated by an unknown site leading to GABAA receptor isoform selectivity.
Negative automatic thoughts and emotional and behavioural problems in adolescence
Authors: Flouri Eirini , Panourgia Constantina
Published: October 2012

Background

The aim of this study was to test whether maladaptive cognitions or difficulties in emotion regulation mediate the association between change in number of adverse life events (i.e. change in life stress) and emotional and behavioural problems in adolescence.

Method

Our sample consisted of 557 young people, aged 10–19 years from a state secondary school in London. We fitted a multiple mediator model to contrast five mediators: three maladaptive cognitions (negative cognitive errors, negative automatic thoughts, and dysfunctional attitudes) and the two emotion regulatory processes of cognitive reappraisal and expressive suppression. We adjusted for gender, age, ethnicity, special educational needs and family poverty.

Results

Taken as a set, our mediators explained the effect of change in life stress on adolescent emotional and behavioural problems. However, cognitive reappraisal, expressive suppression, dysfunctional attitudes and negative cognitive errors did not contribute to the indirect effect above and beyond negative automatic thoughts.

Conclusions

Only negative automatic thoughts mediated the association between change in life stress and emotional and behavioural problems. This suggests the possibility that negative automatic thoughts is the pathway through which an increase in life stress may lead to emotional and behavioural problems in adolescents.
Genetic factors related to gastric cancer susceptibility identified using a genome‐wide association study
Authors: Saeki Norihisa , Ono Hiroe , Sakamoto Hiromi , Yoshida Teruhiko
Published: October 2012

Gastric cancer (GC) is one of the major malignant diseases worldwide, especially in Asia, where Japan and Korea have the highest incidence in the world. Gastric cancer is classified into intestinal and diffuse types. While the former is almost absolutely caused by Helicobacter pylori infection as the initial insult, the latter seems to include cases in which the role of infection is limited, if any, and a contribution of genetic factors is anticipated. Previously, we performed a genome‐wide association study (GWAS) on diffuse‐type GC by using single nucleotide polymorphisms (SNP) catalogued for Japanese population (JSNP), and identified a prostate stem cell antigen (PSCA) gene encoding a glycosylphosphatidylinositol‐anchored cell surface antigen as a GC susceptibility gene. From the second candidate locus identified using the GWAS, 1q22, we found the Mucin 1 (MUC1) gene encoding a cell membrane‐bound mucin protein as another gene related to diffuse‐type GC. A two‐allele analysis based on risk genotypes of the two genes revealed approximately 95% of Japanese population have at least one of the two risk genotypes, and approximately 56% of the population have both risk genotypes. The two‐SNP genotype might offer ample room to further stratify a high GC risk subpopulation in Japan and Asia by adding another genetic and/or non‐genetic factor. Recently, a GWAS on the Chinese population disclosed an additional three GC susceptibility loci: 3q13.31, 5p13.1 and 10q23. (Cancer Sci 2013; 104: 1–8)
Prevalence and long‐term follow‐up outcomes of testicular adrenal rest tumours in children and adolescent males with congenital adrenal hyperplasia
Authors: Aycan Zehra , Bas Veysel N. , Cetinkaya Semra , Yilmaz Agladioglu Sebahat , Tiryaki Tuğrul
Published: October 2012

Objective

There are a few studies regarding the prevalence of testicular adrenal rest tumours (TARTs) in boys and adolescent males with congenital adrenal hyperplasia (CAH), and there is little information regarding the treatment outcomes in patients with TARTs. The aim of this study was to determine the long‐term treatment outcomes in boys and adolescent males with CAH.

Patients and methods

Sixty boys and adolescent males with CAH, who were between 2 and 18 years of age, were included in the study. Fifty‐five patients had 21‐hydroxylase deficiency (21‐OHD), and five patients had 11‐β hydroxylase deficiency (11β‐OHD). All patients were screened for TARTs by scrotal ultrasonography (US) performed by an experienced radiologist.

Results

TART prevalence was 18·3% in 2–18 years’ of age; eight patients had 21‐OHD, and three had 11β‐OHD. The youngest patient with TART was 4 years old, whereas eight patients with RTs were at puberty. Only two patients had tight metabolic control: eight patients had stage 2, one had stage 4, and two had stage five rest tumours. In four patients with stage 2 TARTs, tumours disappeared after high‐dose steroid treatment and did not recur. Shrinkage of tumour was observed in two patients. Testis‐sparing surgery was performed in one patient with stage five tumour. Gonadal functions were normal in patients with partially regressed tumours. Two patients became fathers of healthy male off‐springs.

Conclusions

Detection and treatment for TARTs in children with CAH at younger ages, earlier stages, may prevent infertility in adulthood. Therefore, we recommend that scrotal US screening should be performed in every 1–2 years starting from early childhood.
Tai Chi in Patients With Heart Failure With Preserved Ejection Fraction
Authors: Yeh Gloria Y. , Wood Malissa J. , Wayne Peter M. , Quilty Mary T. , Stevenson Lynne W. , Davis Roger B. , Phillips Russell S. , Forman Daniel E.
Published: October 2012

Although exercise is an important component of heart failure management, optimal regimens, particularly in heart failure with preserved ejection fraction (HFPEF), are uncertain. Tai chi (TC) is a mind‐body exercise that may have potential benefits but has not been studied in this population. The authors randomized 16 patients with HFPEF to either 12 weeks TC or aerobic exercise. Assessments included peak oxygen uptake, 6‐minute walk, quality of life, echocardiography, mood, and self‐efficacy at baseline and at 12 weeks. Cardiorespiratory measures during exercise were obtained to characterize training intensities. Baseline characteristics were as follows: age 66±12 years, E/A ratio 1.3±0.7, and E/e′ ratio 15.9±4.8. Overall, adherence was excellent (89% attendance). Change in peak oxygen uptake was similar between groups after 12 weeks, but 6‐minute walk distance increased more after TC (69±46 m vs 10±31 m, P=.02). While both groups had improved Minnesota Living With Heart Failure scores and self‐efficacy, Profile of Mood States (POMS)‐Depression scores improved more with TC (−1.7±2.8 vs 1.6±3, P=.05). Cardiorespiratory assessment during TC showed lower oxygen uptake (4.3 mL/kg/min vs 9.4 mL/kg/min, P<.01), respiratory rate, and heart rate. TC is feasible and safe in HFPEF. Therepeutic endpoints appear similar with TC relative to aerobic exercise despite a lower aerobic training workload.
Emergency transanal haemorrhoidal Doppler guided dearterialization for acute and persistent haemorrhoidal bleeding
Authors: Cavazzoni E. , Bugiantella W. , Graziosi L. , Silvia Franceschini M. , Cantarella F. , Rosati E. , Donini A.
Published: October 2012

Aim

The effectiveness of Doppler guided transanal haemorrhoidal dearterialization (THD) for arresting persistent haemorrhoidal bleeding in patients admitted as an emergency was studied.

Method

Eleven patients with severe anal bleeding underwent emergency THD as definitive treatment for haemorrhoids. In the majority of patients antiplatelet or anticoagulant therapy was ongoing and severe anaemia was present in six patients.

Results

The mean operative time was 39.7 min. Six to nine feeding arteries were ligated. Intra‐operative blood loss was nil. Bleeding was well controlled in all patients. No blood transfusion was required. Mean pain score per verbal numeric scale was 3.6 and 1.4 on day 1 and day 3 respectively. The mean time to resumption of normal activities was 8 days. No major complications were experienced. Six months follow‐up demonstrated good control of haemorrhoidal disease.

Conclusion

THD is effective in controlling acute haemorrhoidal bleeding with a low incidence of postoperative complications.
An extended medial to lateral approach to mobilize the splenic flexure during laparoscopic low anterior resection
Authors: Kim H. J. , Kim C. H. , Lim S. W. , Huh J. W. , Kim Y. J. , Kim H. R.
Published: October 2012

Aim

The aim of this retrospective study of laparoscopic low anterior resection was to compare splenic flexure mobilization (SFM) carried out by an extended medial to lateral approach with that by a lateral approach.

Method

Records of patients with rectal cancer on a prospectively maintained database undergoing laparoscopic low anterior resection performed between January 2009 and November 2011 by a single surgeon were analysed. The extended medial to lateral approach involved continuing the medial to lateral approach upwards to enter the lesser sac over the pancreas, thus permitting detachment of the splenic flexure.

Results

Two hundred and thirty‐seven patients, including 164 undergoing a lateral SFM and 73 an extended medial to lateral SFM, were evaluated. Both patient groups had similar characteristics except for operative time (152.7 ± 32.7 min extended medial to lateral; 171.5 ± 40.8 min lateral; P < 0.001), postoperatively the interval to oral intake (3.1 ± 0.8 days extended medial to lateral; 3.7 ± 0.9 lateral; P < 0.001) and duration of hospital stay (8.2 ± 2.8 days extended medial to lateral; 10.3 ± 7.5 days lateral; P = 0.002) favoured the extended medial to lateral group.

Conclusion

An extended medial to lateral approach for SFM during laparoscopic low anterior resection of rectal cancer appears to be an improvement over the previously used lateral approach, because it may provide a shorter operation time and shorter hospital stay.
Colectomy with ileorectal anastomosis has a worse 30‐day outcome when performed for colonic inertia than for a neoplastic indication
Authors: Reshef A. , Gurland B. , Zutshi M. , Kiran R. P. , Hull T.
Published: October 2012

Aim

Whether bowel related dysfunction adversely affects postoperative recovery after total colectomy with ileorectal anastomosis (C + IRA) for colonic inertia (CI) has not been previously well evaluated. This study compared the early postoperative outcome of C + IRA for CI and for other noninflammatory indications.

Method

Patients undergoing elective C + IRA from 1999 to 2010 were identified from a prospectively maintained database. Since inflammation in the rectum or small bowel may influence the outcome, patients with inflammatory bowel disease were excluded. Patients undergoing surgery for CI (group A) were compared with patients having the operation for other benign noninflammatory diseases (group B). Demographics, American Society of Anesthesiologists (ASA) score, body mass index (BMI), surgical procedure and 30‐day complications were assessed.

Results

The study population consisted of 333 patients undergoing elective C + IRA (99 men, mean age 39 ± 16 years). The procedure was laparoscopic in 163 (49%) patients. Groups A (n = 131) and B (n = 202) had similar age and ASA score (39 ± 11 vs 39 ± 19 years, P = 0.4; 2.2 ± 0.5 vs 2.4 ± 0.7). Group A patients had lower BMI (25 ± 5 vs 28 ± 8 kg/m2, P = 0.002), more women (99 vs 51%, P < 0.001) and fewer laparoscopic procedures (43 vs 53%, P = 0.04). Compared with group B, group A had a greater incidence of postoperative ileus (32 vs 19%, P = 0.009), higher overall morbidity (36 vs 15%, P < 0.001) and increased length of stay (8.4 ± 6 vs 7.2 ± 5 days, P < 0.006). These differences persisted when subgroups of patients who underwent laparoscopic or open surgery were compared.

Conclusion

Although CI is considered a ‘benign’ condition, patients undergoing C + IRA for this indication have significant morbidity compared with patients having the operation for other noninflammatory benign conditions.
Use of near‐infrared spectroscopy to identify trends in regional cerebral oxygen saturation in horses
Authors: McConnell E. J. , Rioja E. , Bester L. , Sanz M. G. , Fosgate G. T. , Saulez M. N.
Published: October 2012

Reasons for performing study

Alterations in cerebral haemodynamics may contribute to perianaesthetic complications in horses. Near‐infrared spectroscopy (NIRS) is frequently used intraoperatively in man to provide information regarding cerebral perfusion.

Objectives

To determine whetherNIRScan identify trends in regional cerebral oxygen saturation (rSO2) in horses and whether there is a correlation betweenrSO2and venous oxygen tensions.

Methods

A cerebral oximeter sensor recordedrSO2from the dorsal sagittal sinus of 6 healthy horses. Values forrSO2, arterial and venous oxygen and carbon dioxide tensions (PaO2,PvO2,PaCO2andPvCO2respectively), along with arteriovenous oxygen saturations (SavO2) were recorded in unsedated (recording period [RP] 1), sedated (RP2) and anaesthetised horses (RP3–5) and during recovery (RP6–8). During anaesthesia, horses were ventilated to achieve states of normo‐ (RP3), hyper‐ (RP4) and hypocapnoea (RP5). Data were evaluated descriptively and analysed using linear mixed‐effects models andPearson's correlation coefficient.

Results

Overall mean ± s.d. values forrSO2,PaO2,PvO2,PaCO2,SavO2and mean arterial pressure varied significantly byRP(P<0.001). Significant decreases inrSO2were identified betweenRP1 and the post anaesthetic periods (P<0.001). No significant differences inrSO2values were identified betweenRP1 and the intra‐anaesthesia periods or betweenRP3,RP4 andRP5. Significant correlations were identified betweenrSO2andPaO2(r = 0.448, P<0.001),rSO2andPvO2(r = 0.512, P<0.001) andrSO2andSavO2(r = 0.469, P<0.001).

Conclusions

This is the first study to identify trends inrSO2in horses usingNIRS. A positive correlation was identified betweenrSO2andPvO2, suggesting that alterations in cerebral oxygenation may be reflected inPvO2.

Potential relevance

Near‐infrared spectroscopy may be used to monitor trends inrSO2during equine anaesthesia. DecreasingrSO2values may act as an early warning signal, alerting clinicians to potential cerebral desaturation events and indicating a need for intervention.
Successful isolation of equine mesenchymal stromal cells from cryopreserved umbilical cord blood‐derived mononuclear cell fractions
Authors: De Schauwer C. , van de Walle G. R. , Piepers S. , Hoogewijs M. K. , Govaere J. L. J. , Meyer E. , van Soom A.
Published: October 2012

Reasons for performing study

The therapeutic potential of mesenchymal stromal cells for cellular therapy has generated increasing interest in human as well as veterinary medicine. Considerable research has been performed on the cryopreservation of expanded mesenchymal stromal cells, but little information is available on the cryopreservation of the original mononuclear cell fraction.

Objectives

The present study describes a protocol to expand equine mesenchymal stromal cells after cryopreserving the mononuclear cells of umbilical cord blood.

Methods

To this end, mononuclear cells were isolated from 7 umbilical cord blood samples and cryopreserved at a concentration of 1–2 × 109 cells/l cold freezing solution. Cells were cryopreserved and kept frozen for at least 6 months before thawing. Frozen cryotubes were thawed in a 37°C water bath. Putative equine mesenchymal stromal cells were immunophenotyped using multicolour flow cytometry based on a selected 9 marker panel.

Results

Average cell viability upon thawing was 98.7 ± 0.6%. In 6 out of 7 samples, adherent spindle‐shaped cell colonies were observed within 9.0 ± 2.6 days and attained 80% confluency at 12.3 ± 3.9 days. After 3 passages, putative equine mesenchymal stromal cells were successfully immunophenotyped as CD29, CD44 and CD90 positive, and CD45, CD73, CD79α, CD105, MHC II and monocyte‐marker negative.

Conclusions and potential relevance

Equine mesenchymal stromal cells can be cultured after cryopreservation of the isolated mononuclear cells, a time‐ as well as cost‐efficient approach in equine regenerative medicine.
Structure and mechanism of an inverting alkylsulfatase from Pseudomonas sp. DSM6611 specific for secondary alkyl sulfates
Authors: Knaus Tanja , Schober Markus , Kepplinger Bernhard , Faccinelli Martin , Pitzer Julia , Faber Kurt , Macheroux Peter , Wagner Ulrike
Published: October 2012

A highly enantioselective and stereoselective secondary alkylsulfatase from Pseudomonas sp. DSM6611 (Pisa1) was heterologously expressed in Escherichia coli BL21, and purified to homogeneity for kinetic and structural studies. Structure determination of Pisa1 by X‐ray crystallography showed that the protein belongs to the family of metallo‐β‐lactamases with a conserved binuclear Zn2+ cluster in the active site. In contrast to a closely related alkylsulfatase from Pseudomonas aeruginosa (SdsA1), Pisa1 showed a preference for secondary rather than primary alkyl sulfates, and enantioselectively hydrolyzed the (R)‐enantiomer of rac‐2‐octyl sulfate, yielding (S)‐2‐octanol with inversion of absolute configuration as a result of C–O bond cleavage. In order to elucidate the mechanism of inverting sulfate ester hydrolysis, for which no counterpart in chemical catalysis exists, we designed variants of Pisa1 guided by three‐dimensional structure and docking experiments. In the course of these studies, we identified an invariant histidine (His317) near the sulfate‐binding site as the general acid for crucial protonation of the sulfate leaving group. Additionally, amino acid replacements in the alkyl chain‐binding pocket generated an enzyme variant that lost its stereoselectivity towards rac‐2‐octyl sulfate. These findings are discussed in light of the potential use of this enzyme family for applications in biocatalysis.

Database

The atomic coordinates and structural factors have been deposited in the Protein Data Bank under the accession codes 2YHE (wild type, crystal form I), 4AV7 (double variant Ser233→Tyr/Ph250→Gly) and 4AXH (wild type, crystal form II)

Structured digital abstract

Pisa1 and Pisa1 bind by x-ray crystallography (View interaction)
Corynebacterium glutamicum Zur acts as a zinc‐sensing transcriptional repressor of both zinc‐inducible and zinc‐repressible genes involved in zinc homeostasis
Authors: Teramoto Haruhiko , Inui Masayuki , Yukawa Hideaki
Published: October 2012

Zur is a zinc‐dependent transcriptional repressor of zinc uptake systems in bacteria. In the present study, we examined the role of Corynebacterium glutamicum Zur in the zinc‐inducible expression of two genes: one encoding a cation diffusion facilitator (zrf) and the other a metal‐translocating P‐type ATPase (zra). Both genes were shown to be involved in zinc resistance. Disruption of the zur gene encoding Zur resulted in constitutive expression of zrf and zra mRNAs. An electrophoretic mobility shift assay revealed that the Zur protein binds to the zrf and zra promoters, for which the in vivo activities were up‐regulated in response to excess zinc. Interestingly, the in vitro DNA binding activity of Zur was inhibited by zinc, in contrast to its zinc‐dependent binding to the promoter region of a zinc‐repressible ABC transporter gene znuB2. A 21‐bp motif found in the Zur binding site overlaps the putative –35 region of both the zrf and zra promoters. This new motif is a 10‐1‐10 direct repeat sequence distinct from the 10‐1‐10 inverted repeat sequence of a previously identified Zur box for zinc‐dependent binding. Nevertheless, their 10‐bp elements share some sequence similarities. Overexpression of zur in the zur deletion mutant background, as well as deletion of zur in the zrf and zra double deletion mutant background, resulted in decreased resistance to zinc. These results suggest that the direct negative control of both zinc uptake and export systems by Zur is central to C. glutamicum zinc homeostasis and is effected in distinct ways.
Fish assemblages respond to altered flow regimes via ecological filtering of life history strategies
Authors: MIMS MERYL C. , OLDEN JULIAN D.
Published: October 2012

1. In riverine ecosystems, streamflow determines the physical template upon which the life history strategies of biota are forged. Human freshwater needs and activities have resulted in widespread alteration of the variability, predictability and timing of streamflow, and anticipating the biotic consequences of anthropogenic streamflow alteration is critical for successful environmental flow management. In this study, we examined relationships between dam characteristics, metrics of flow alteration and fish functional community composition according to life history strategies by coupling stream flow records and fish survey data in paired flow‐regulated and free‐flowing rivers across the conterminous United States.

2. Dam operations have generally reduced flow variability and increased flow constancy based on a comparison of pre‐ and post‐dam flow records (respective mean record lengths 26.2 and 43.1 years). In agreement with ecological theory, fish assemblages downstream of dams were characterised by a lower proportion of opportunistic species (a strategy favoured in environmental settings dominated by unpredictable environmental change) and a higher proportion of equilibrium species (a strategy favoured in more stable, predictable environments) compared to free‐flowing, neighbouring locations.

3. Multiple linear regression models provided modest support for links between alteration of specific flow attributes and differential life history representation below dams, and they provided strong support for life history associations with dam attributes (age and release type). We also found support for a relationship of both reduced flow variability and dam age with higher representation of non‐native species below dams.

4. Our study demonstrated that river regulation by large dams has significant hydrological and biological consequences across the United States. We showed that on ecological time scales (i.e. the order of years to decades), dams are effectively changing the functional composition of communities that have established over millennia. Furthermore, the changes are directional and indicate a filtering by dams for some life histories (equilibrium strategists) and against other life histories (opportunists). Finally, our study highlights that dependence upon long‐term flow records and availability of biotic surveys extracted from national survey efforts limit our ability to guide environmental flow standards particularly in data‐poor regions.