bluuebunny/crossref_metadata_2025_split huggingface dataset:
Deleting two C-terminal α-helices is effective to crystallize the bacterial ABC transporterEscherichia coliMsbA complexed with AMP-PNP
Authors: Terakado Kanako , Kodan Atsushi , Nakano Hiroaki , Kimura Yasuhisa , Ueda Kazumitsu , Nakatsu Toru , Kato Hiroaki 
Published: February 2010

An MsbA deletion mutant ΔC21 that lacks the two C-terminal α-helices was expressed inEscherichia colistrain C41 and purified by metal-affinity and gel-filtration chromatography. Purified ΔC21 retained 26% of the activity of the wild-type ATPase and had a similar binding affinity to fluorescent nucleotide derivatives. Although crystals of wild-type MsbA complexed with adenosine 5′-(β,γ-imido)triphosphate could not be obtained, crystals of ΔC21 that diffracted to 4.5 Å resolution were obtained. The preliminary ΔC21 structure had the outward-facing conformation, in contrast to the previously reportedE. coliMsbA structure. This result suggests that deletion of the C-terminal α-helices may play a role in facilitating the outward-facing nucleotide-bound crystal structure of EcMsbA.
DJ‐1: A novel independent prognostic marker for survival in glottic squamous cell carcinoma
Authors: Zhu Xiao‐Lin , Wang Zhang‐Feng , Lei Wen‐Bin , Zhuang Hui‐Wen , Jiang Hong‐Yan , Wen Wei‐Ping 
Published: February 2010

DJ‐1 is frequently overexpressed in a large variety of solid tumors, but the DJ‐1 expression in laryngeal squamous cell cancer and its clinical/prognostic significance is unclear. We aimed to evaluate DJ‐1 protein expression in glottic squamous cell carcinoma (GSCC) and to correlate this with clinicopathological data including patient survival. The expression of DJ‐1 in GSCCs (60) and adjacent normal tissue (44) was assessed by immunohistochemistry and western blot analysis. In addition, the role of DJ‐1 was investigated in tumorigenesis by transfecting DJ1‐specific siRNA into laryngeal squamous cell carcinoma (LSCC) Hep‐2 cells. Our data showed that positive expression of DJ‐1 was found in 85% of GSCCs. In univariate survival analysis of the GSCC cohorts, a highly significant association between DJ‐1 expression with shortened patient overall survival (5‐year survival rate 92.9%vs 66.6%; P = 0.001; log rank test) was demonstrated. In multivariate analyses, DJ‐1, tumor grading, and pT status were significant prognostic parameters for shortened patient overall survival. Furthermore, siRNA targeting DJ‐1 can effectively inhibit DJ‐1 expression, resulting in enhanced apoptosis and less proliferation of Hep‐2 cells. We concluded that DJ‐1 overexpression might be a novel independent molecular marker for poor prognosis (shortened overall survival) of patients with GSCC.(Cancer Sci 2010; 101: 1320–1325)
Association between nitrous oxide and the incidence of postoperative nausea and vomiting in adults: a systematic review and meta‐analysis
Authors: Fernández‐Guisasola J. , Gómez‐Arnau J. I. , Cabrera Y. , del Valle S. García 
Published: February 2010

SummarySome, but not all studies have suggested intra‐operative use of nitrous oxide is correlated with postoperative nausea and vomiting. We performed a meta‐analysis of randomised controlled trials to compare the incidence of nausea and vomiting in adults following general anaesthesia with or without nitrous oxide. We retrieved 30 studies (incorporating 33 separate trials) that investigated a ‘nitrous oxide group’ (total 2297 patients) vs a ‘no‐nitrous oxide group’ (2301 patients). Omitting nitrous oxide significantly reduced postoperative nausea and vomiting (pooled relative risk 0.80, 95% CI 0.71–0.90, p = 0.0003). However, the absolute incidence of nausea and vomiting was high in both the nitrous oxide and no‐nitrous oxide groups (33% vs 27%, respectively). In subgroup analysis, the maximal risk reduction was obtained in female patients (pooled relative risk 0.76, 95% CI 0.60–0.96). When nitrous oxide was used in combination with propofol, the antiemetic effect of the latter appeared to compensate the emetogenic effect of nitrous oxide (pooled relative risk 0.94, 95% CI 0.77–1.15). We conclude that avoiding nitrous oxide does reduce the risk of postoperative nausea and vomiting, especially in women, but the overall impact is modest.
Developing a scale for teacher integration of information and communication technology in grades 1–9
Authors: Hsu S. 
Published: February 2010

AbstractThere is no unified view about how teachers' integration of information and communication technology (ICT) should be measured. While many instruments have focused on the technological aspects, recent studies have suggested teachers' pedagogical considerations, professional development, and emerging ethical and safety issues should be included when assessing teachers' ICT literacy. Using the performance standards created by the International Society for Technology in Education, a group of Taiwanese teachers developed equivalent items. After consulting expert panels and interviewing teachers, a scale was constructed. Using this instrument, 3729 teachers from grades 1 through 9 in Taiwan were sampled. Half of the data was analysed by exploratory factor analysis to find the underpinning structure, and the second half was analysed by confirmatory factor analysis to verify the subscales. The results confirmed six subscales for teachers' ICT integration: (1) information collection and preparation; (2) material production and troubleshooting; (3) communication and sharing; (4) planning, teaching and evaluation; (5) professional development and self‐study; and (6) ethical, health and safety issues. Using two teacher attributes, school levels and course or degree obtained, the scale was further verified for its feasibility. The established scale examines the existing concerns for technology, pedagogy and professional development at once with a new addition of ethical and safety issues, which demand growing attention in teachers of future generation.
Viral haemorrhagic septicaemia virus IVb experimental infection of rainbow trout, Oncorhynchus mykiss (Walbaum), and fathead minnow, Pimphales promelas (Rafinesque)
Authors: Al‐Hussinee L , Huber P , Russell S , LePage V , Reid A , Young K M , Nagy E , Stevenson R M W , Lumsden J S 
Published: February 2010

AbstractViral haemorrhagic septicaemia virus (VHSV) in the Great Lakes has had a dramatic impact on fish husbandry because of the implications of the presence of a reportable disease. Experimental infections with VHSV IVb were conducted in rainbow trout, Oncorhynchus mykiss (Walbaum), and fathead minnows, Pimphales promelas (Rafinesque), to examine their susceptibility and the clinical impact of infection. Triplicate groups of rainbow trout (n = 40) were injected intraperitoneally (i.p.) with 100 μL 106.550% tissue culture infective doses (TCID50) or waterborne exposed to graded doses (104.5, 106.5, and 108.5 TCID50 mL−1) of VHSV IVb. Duplicate groups of fathead minnows (n = 15) were i.p. injected with (106.5 TCID50 100 μL) or waterborne exposed (106.5 TCID50 mL−1). All experiments were performed with single‐pass well water maintained at 12 °C. Following either i.p. or waterborne exposure, VHSV RNA was detectable in both rainbow trout and fathead minnows by nested reverse transcription polymerase chain reaction (nRT‐PCR) as early as 4–7 days post‐infection (p.i.). Infected fathead minnow and rainbow trout exhibited lesions characteristic of VHS at 9 and 15 days p.i., respectively. Route of exposure had little effect on the onset of clinical signs. Cumulative mean mortality in rainbow trout was 4.4%, 2.6%, 2.6% and less than 1% in the i.p., high, medium and low dose waterborne exposures, respectively. Cumulative average mortality of 50% and 13% occurred in i.p. and waterborne‐exposed fathead minnows, respectively. VHSV was detected from pooled rainbow trout tissue by RT‐PCR and virus isolation at 38 days p.i., but not at 74 days p.i., regardless of the exposure route. Immunohistochemistry (IHC) with a rabbit antibody to VHSV IVb revealed the viral tissue tropisms following infection, with the identification of viral antigen in myocardium and necrotic branchial epithelium of both species and in gonadal tissue of fathead minnows. Rainbow trout, but not fathead minnows, are relatively refractory to experimental infection with VHSV IVb.
Impact of yellow head virus outbreaks in the whiteleg shrimp, Penaeus vannamei (Boone), in Thailand
Authors: Senapin S , Thaowbut Y , Gangnonngiw W , Chuchird N , Sriurairatana S , Flegel T W 
Published: February 2010

AbstractYellow head virus (YHV) is known as a major pathogen in the black tiger shrimp, Penaeus (Penaeus) monodon. It can also cause serious mortality in farmed whiteleg shrimp, Penaeus (Litopenaeus) vannamei. However, there is no published information on the economic and/or production impact of the disease in P. vannamei. Shrimp with gross signs of YHV disease (faded body colour and 60–70% mortality) were observed in 20 study farms rearing P. vannamei in the central part of Thailand from the end of 2007 through early 2008. The estimated economic loss for these farms according to the Thai Animal Aquaculture Association was approximately US$3 million. Detailed sequence analysis of RT‐PCR amplicons from shrimp in all the study ponds revealed the presence of YHV Type 1b (YHV‐1b) alone (characterized by a 162‐bp deletion in the ORF3 region encoding the structural gene for gp116) and the absence of YHV Type 1a (YHV‐1a), the original YHV type reported from Thailand. Despite the large 162‐bp deletion (= 54 deduced amino acids) in the gp116 structural gene, histopathology of YHV‐1b infections was identical to that of YHV‐1a infections, and electron microscopy revealed that YHV‐1b virions were morphologically indistinguishable from those previously reported for YHV‐1a. In addition, an existing commercial RT‐PCR detection kit and an immunochromatographic test strip for the detection of YHV were proven to have been valid tests for both YHV‐1b and YHV‐1a. The source of the virus for these outbreaks was unlikely to have been the post‐larvae used to stock the ponds, as they were derived from domesticated specific pathogen‐free stocks free of YHV. Thus, it is possible that they originated from an unknown, natural reservoir.
Phenotype determining alleles in GM1 gangliosidosis patients bearing novel GLB1 mutations
Authors: Hofer D , Paul K , Fantur K , Beck M , Roubergue A , Vellodi A , Poorthuis BJ , Michelakakis H , Plecko B , Paschke E 
Published: February 2010

Hofer D, Paul K, Fantur K, Beck M, Roubergue A, Vellodi A, Poorthuis BJ, Michelakakis H, Plecko B, Paschke E. Phenotype determining alleles in GM1 gangliosidosis patients bearing novel GLB1 mutations.GM1 gangliosidosis manifests with progressive psychomotor deterioration and dysostosis of infantile, juvenile, or adult onset, caused by alterations in the structural gene coding for lysosomal acid ß‐galactosidase (GLB1). In addition, allelic variants of this gene can result in Morquio B disease (MBD), a phenotype with dysostosis multiplex and entire lack of neurologic involvement. More than 100 sequence alterations in the GLB1 gene have been identified so far, but only few could be proven to be predictive for one of the GM1 gangliosidosis subtypes or MBD. We performed genotype analyses in 16 GM1 gangliosidosis patients of all phenotypes and detected 28 different genetic lesions. Among these, p.I55FfsX16, p.W65X, p.F107L, p.H112P, p.C127Y, p.W161X, p.I181K, p.C230R, p.W273X, p.R299VfsX5, p.A301V, p.F357L, p.K359KfsX23, p.L389P, p.D448V, p.D448GfsX8, and the intronic mutation IVS6‐8A>G have not been published so far. Due to their occurrence in homozygous patients, four mutations could be correlated to a distinct GM1 gangliosidosis phenotype. Furthermore, the missense mutations from heteroallelic patients and three artificial nonsense mutations were characterized by overexpression in COS‐1 cells, and the subcellular localization of the mutant proteins in fibroblasts was assessed. The phenotype specificity of 10 alleles can be proposed on the basis of our results and previous data.
Two percent of patients suspected of having Angelman syndrome have TCF4 mutations
Authors: Takano K , Lyons M , Moyes C , Jones J , Schwartz CE 
Published: February 2010

Takano K, Lyons M, Moyes C, Jones J, Schwartz CE. Two percent of patients suspected of having Angelman syndrome have TCF4 mutations.The TCF4 gene encodes a basic helix–loop–helix (bHLH) transcription factor which belongs to the family of E‐proteins. E‐proteins form homo‐ and heterodimers with other members of the HLH family and bind to the common DNA sequence called E‐box. Haploinsufficiency of the TCF4 gene has been found to be associated with the Pitt–Hopkins syndrome (PTHS). PTHS is characterized by severe mental retardation, a wide mouth plus other distinctive facial features (fleshy lips, beaked nose, broad nasal bridge) and breathing abnormalities. Because of some phenotypical overlap with Angelman syndrome (AS), it has been suggested that PTHS be considered in its differential diagnosis. To explore this possibility, we screened 86 patients who were suspected of having AS. All the patients were negative for UBE3A testing, and 53 were known to be negative for methylation analysis. We identified two TCF4 mutations in this cohort. The p.S384Tfsx7 mutation lacks the bHLH domain. The p.R582P mutation lies within the bHLH domain in which seven other missense mutations have been reported. Both mutations most likely affect the critical function of the bHLH domain of the TCF4 protein. In summary, we found two TCF4 mutations in 86 patients (2%) suspected to have AS. Screening for mutations in this gene should be considered in patients who present with findings of AS but who have been negative for methylation and UBE3A testing.
Congenital central hypoventilation syndrome: genotype–phenotype correlation in parents of affected children carrying a PHOX2B expansion mutation
Authors: Parodi S , Vollono C , Baglietto MP , Balestri M , Di Duca M , Landri PA , Ceccherini I , Ottonello G , Cilio MR 
Published: February 2010

Parodi S, Vollono C, Baglietto MP, Balestri M, Di Duca M, Landri PA, Ceccherini I, Ottonello G, Cilio MR. Congenital central hypoventilation syndrome: genotype–phenotype correlation in parents of affected children carrying a PHOX2B expansion mutation.Congenital Central Hypoventilation Syndrome (CCHS) is a rare genetic disorder. Although most CCHS associated PHOX2B mutations occur de novo, about 10% of the cases are inherited from apparently asymptomatic parents, thus confirming variable expressivity and incomplete penetrance of PHOX2B mutations. Three asymptomatic parents of children affected with CCHS, and found to carry the same PHOX2B expansion mutations as their siblings, were studied by overnight polysomnography and somatic mosaicism analysis. In one case, significant sleep breathing control anomalies were detected, while the other two resulted in normal. In tissue‐specific allele studies, mosaicism with a comparatively low mutant allele proportion was showed in the two unaffected adult carriers. Accurate polysomnography and assessment of the degree of somatic mosaicism should be conducted in asymptomatic carriers of PHOX2B mutations, as they may unmask subclinical but significant anomalies
A homozygous FKRP start codon mutation is associated with Walker–Warburg syndrome, the severe end of the clinical spectrum
Authors: Van Reeuwijk J , Olderode‐Berends MJW , Van Den Elzen C , Brouwer OF , Roscioli T , Van Pampus MG , Scheffer H , Brunner HG , Van Bokhoven H , Hol FA 
Published: February 2010

van Reeuwijk J, Olderode‐Berends MJW, van den Elzen C, Brouwer OF, Roscioli T, van Pampus MG, Scheffer H, Brunner HG, van Bokhoven H, Hol FA. A homozygous FKRP start codon mutation is associated with Walker–Warburg syndrome, the severe end of the clinical spectrum.Dystroglycanopathies are a heterogeneous group of disorders caused by defects in the glycosylation pathway of α‐dystroglycan. The clinical spectrum ranges from severe congenital muscular dystrophy with structural brain and eye involvement to a relatively mild adult onset limb‐girdle muscular dystrophy without brain abnormalities and normal intelligence. Mutations have been identified in one of six putative or demonstrated glycosyltransferases. Many different FKRP mutations have been identified, which cover the complete clinical spectrum of dystroglycanopathies. In contrast to the other known genes involved in these disorders, genotype–phenotype correlations are not obvious for FKRP mutations. To date, no homozygous or compound heterozygous null mutations have been identified in FKRP, suggesting that null mutations in FKRP could result in embryonic lethality. We report a family with two siblings carrying a homozygous mutation in the start codon of FKRP that is likely to result in a loss of functional FKRP protein. The clinical phenotype of the patients was consistent with Walker–Warburg syndrome, the most severe disorder in the disease spectrum of dystroglycanopathies.
Molecular characterization of five patients with homocystinuria due to severe methylenetetrahydrofolate reductase deficiency
Authors: Urreizti R , Moya‐García AA , Pino‐Ángeles A , Cozar M , Langkilde A , Fanhoe U , Esteves C , Arribas J , Vilaseca MA , Pérez‐Dueñas B , Pineda M , González V , Artuch R , Baldellou A , Vilarinho L , Fowler B , Ribes A , Sánchez‐Jiménez F , Grinberg D , Balcells S 
Published: February 2010

Urreizti R, Moya‐García AA, Pino‐ Ángeles A, Cozar M, Langkilde A, Fanhoe U, Esteves C, Arribas J, Vilaseca MA, Pérez‐Dueñas B, Pineda M, González V, Artuch R, Baldellou, A, Vilarinho L, Fowler B, Ribes A, Sánchez‐Jiménez F, Grinberg D, Balcells S. Molecular characterization of five patients with homocystinuria due to severe MTHFR deficiency.Methylenetetrahydrofolate reductase (MTHFR) plays a major role in folate metabolism. Disturbed function of the enzyme results in hyperhomocysteinemia and causes severe vascular and neurological disorders and developmental delay. Five patients suspected of having non‐classical homocystinuria due to MTHFR deficiency were examined with respect to their symptoms, MTHFR enzyme activity and genotypes of the MTHFR gene. All patients presented symptoms of severe central nervous system disease. Two patients died, at the ages of 15 months and 14 years. One patient is currently 32 years old, and is being treated with betaine and folinic acid. The other two patients, with an early diagnosis and a severe course of the disease, are currently improving under treatment. MTHFR enzyme activity in the fibroblasts of four of the patients was practically undetectable. We found four novel mutations, three of which were missense changes c.664G> T (p.V218L), c.1316T> C (p.F435S) and c.1733T> G (p.V574G), and the fourth was the 1‐bp deletion c.1780delC (p.L590CfsX72). We also found the previously reported nonsense mutation c.1420G> T (p.E470X). All the patients were homozygous. Molecular modelling of the double mutant allele (p.V218L; p.A222V) revealed that affinity for FAD was not affected in this mutant. For the p.E470X mutation, the evidence pointed to nonsense‐mediated mRNA decay. In general, genotype–phenotype analysis predicts milder outcomes for patients with missense changes than for those in which mutations led to severe alterations of the MTHFR protein.
Multi‐trait BLUP in half‐sib selection of annual crops
Authors: Viana J. M. S. , Sobreira F. M. , De Resende M. D. V. , Faria V. R. 
Published: February 2010

With 2 tables AbstractThe Best Linear Unbiased Prediction (BLUP) methodology, which is widely used in animal and forestry genetic evaluation, can also be applied to annual crop breeding. The objective of this study was to compare the accuracy and efficiency of among‐ and within‐half‐sib family selection through the use of single‐ and multi‐trait BLUP, and BLUE/anova methods. Expansion volume and yield data from two recurrent selection cycles of a popcorn population were analyzed. Progeny tests were designed as a lattice. To maximize accuracy of the prediction of breeding values, the BLUP analyses included phenotypic values of the two cycles. All BLUP analyses were performed using the ASReml software. The multi‐trait BLUP method demonstrated greater accuracy and efficiency in family selection. In the case of within‐family selection, both accuracy and efficiency of multi‐ and single‐trait BLUP methods were equivalent. The selection efficiency of the multi‐trait BLUP was dependent on the estimated genetic parameters, particularly the difference between the genetic and environmental correlations of the traits.
Neutrophil gelatinase‐associated lipocalin levels in chronic haemodialysis patients
Authors: BOLIGNANO DAVIDE , COPPOLINO GIUSEPPE , ROMEO ADOLFO , LACQUANITI ANTONIO , BUEMI MICHELE 
Published: February 2010

ABSTRACT:Neutrophil gelatinase‐associated lipocalin (NGAL), a small 25 kDa protein strongly induced in injured renal tubular cells, represents an interesting emerging biomarker in the field of clinical nephrology. The aim of the present pilot study was to analyze circulating NGAL levels in a small cohort of 30 patients on chronic haemodialysis (HD), in order to assess any relationships with different laboratory and clinical parameters. Pre‐ and post‐HD levels were higher in patients than in healthy subjects (485.2 ± 49.7 vs 51.2 ± 4.6 ng/mL; P < 0.001; and 167.4 ± 48.0 vs 51.2 ± 4.6 ng/mL; P = 0.01). Furthermore, a single HD session decreased NGAL levels by approximately fourfold (485.2 ± 49.7 vs 167.4 ± 48.0 ng/mL; p:0.01), with a reduction ratio of 73 ± 14%. At baseline, direct and independent correlations were found between NGAL and, respectively, high‐sensitivity C‐reactive protein (β = 0.34; P = 0.03) and spKt/V (β = 0.35; P = 0.02). The findings showed that HD patients have chronically increased levels of circulating NGAL. However, with a single HD session, a marked reduction was achieved in circulating NGAL values, probably as a result of an important dialytic removal, similar to that observed for other cytokines. Finally, the direct independent correlation found between NGAL and spKt/V raises the question of whether, in the future, NGAL may also become a useful tool in predicting the adequacy of dialysis and in guiding the management of dialysis prescriptions.
Neuronal circuits of fear extinction
Authors: Herry Cyril , Ferraguti Francesco , Singewald Nicolas , Letzkus Johannes J. , Ehrlich Ingrid , Lüthi Andreas 
Published: February 2010

AbstractFear extinction is a form of inhibitory learning that allows for the adaptive control of conditioned fear responses. Although fear extinction is an active learning process that eventually leads to the formation of a consolidated extinction memory, it is a fragile behavioural state. Fear responses can recover spontaneously or subsequent to environmental influences, such as context changes or stress. Understanding the neuronal substrates of fear extinction is of tremendous clinical relevance, as extinction is the cornerstone of psychological therapy of several anxiety disorders and because the relapse of maladaptative fear and anxiety is a major clinical problem. Recent research has begun to shed light on the molecular and cellular processes underlying fear extinction. In particular, the acquisition, consolidation and expression of extinction memories are thought to be mediated by highly specific neuronal circuits embedded in a large‐scale brain network including the amygdala, prefrontal cortex, hippocampus and brain stem. Moreover, recent findings indicate that the neuronal circuitry of extinction is developmentally regulated. Here, we review emerging concepts of the neuronal circuitry of fear extinction, and highlight novel findings suggesting that the fragile phenomenon of extinction can be converted into a permanent erasure of fear memories. Finally, we discuss how research on genetic animal models of impaired extinction can further our understanding of the molecular and genetic bases of human anxiety disorders.