bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic Diversity of Thanatephorus cucumeris Infecting Tomato in Iran
Authors: Pourmahdi Atena , Taheri Parissa 
Published: May 2014

AbstractThe necrotrophic fungus Thanatephorus cucumeris (anamorph Rhizoctonia solani) is among the most important soil‐borne pathogens which causes tomato foot and root rot worldwide. We investigated virulence and genetic relationships among and within different taxonomic groups of R. solani from the tomato‐growing regions in the north‐east of Iran. Characterization of R. solani taxonomic groups revealed that, of 56 isolates, four were AG‐2‐1, 16 were AG‐3 PT, 21 were AG‐4 HG‐I and 15 were AG‐4 HG‐II. Because interprimer binding site (iPBS), which is based on amplification of retrotransposons, is known as novel and powerful DNA fingerprinting technology, we selected four iPBS primers, which can detect polymorphisms of tomato foot root and root rot pathogen, for investigating genotypic variability of the isolates. The iPBS analyses separated various taxonomic groups of R. solani and showed great diversity among the isolates, demonstrating that the R. solani isolates obtained from tomato were not a clonal population. Crop rotation strategies and geographic location seem to be important factors affecting genetic structure of the isolates. Pathogenicity tests on tomato cultivar ‘Mobil’ showed significant differences in the virulence of various isolates. The overall results indicated that isolates of AG‐3 and AG‐4 were more virulent than AG‐2‐1. There was no significant correlation between genetic diversity and virulence of the isolates. This is the first report of R. solani AG‐4 HG‐II, causing tomato foot and root rot. Also, our research is the first in assessment of genetic diversity in fungal populations using iPBS molecular markers.
Development of an enzyme-linked immunosorbent assay for Bartonella henselae infection detection
Authors: Ferrara F. , Di Niro R. , D'Angelo S. , Busetti M. , Marzari R. , Not T. , Sblattero D. 
Published: May 2014

Abstract
Several serological diagnostics rely on enzyme-linked immunosorbent assay (ELISA) to detect bacterial infections. However, for some pathogens, including Bartonella henselae, diagnosis still depends on manually intensive, time-consuming assays including micro-immunofluorescence, Western blotting or indirect immunofluorescence. For such pathogens, there is obviously still a need to identify antigens to establish a reliable, fast and high-throughput assay (Dupon et al. 8). We evaluated two B. henselae proteins to develop a novel serological ELISA: a well-known antigen, the 17-kDa protein, and GroEL, identified during this study by a proteomic approach. When serum IgG were tested, the specificity and sensitivity were 76 and 65·7% for 17-kDa, respectively, and 82 and 42·9% for GroEL, respectively. IgM were found to be more sensitive and specific for both proteins: 17-kDa protein, specificity 86·2% and sensitivity 75%; GroEL, specificity 97·7% and sensitivity 45·3%. IgM antibodies were also measured in lymphoma patients and patients with Mycobacterium tuberculosis infection to assess the usefulness of our ELISA to distinguish them from B. henselae infected patients. The resulting specificities were 89·1 and 93·5% for 17-kDa protein and GroEL, respectively. Combining the results from the two tests, we obtained a sensitivity of 82·8% and a specificity of 83·9%. Our work described and validated a proteomic approach suitable to identify immunogenic proteins useful for developing a serological test of B. henselae infection.

Significance and Impact of the Study
A reliable serological assay for the diagnosis of Cat Scratch Disease (CSD) – a pathological condition caused by Bartonella henselae infection – has not yet been developed. Such an assay would be extremely useful to discriminate between CSD and other pathologies with similar symptoms but different aetiologies, for example lymphoma or tuberculosis. We investigate the use of two B. henselae proteins – GroEL and 17-kDa – to develop a serological-based ELISA, showing promising results with the potential for further development as an effective tool for the differential diagnosing of B. henselae infection.
The chronic use of beta‐blockers and proton pump inhibitors may affect the rate of bacterial infections in cirrhosis
Authors: Merli Manuela , Lucidi Cristina , Di Gregorio Vincenza , Giannelli Valerio , Giusto Michela , Ceccarelli Giancarlo , Riggio Oliviero , Venditti Mario 
Published: May 2014

AbstractBackground & AimsBacterial infections are among the most common and life‐threatening complications in cirrhosis. Qualitative and quantitative modifications of the gut microbiota, dysfunction of the intestinal barrier and multiple immune defects are factors that contribute to a pathological ‘bacterial translocation’ (BT), leading to a higher susceptibility to infections in cirrhotic patients. Long‐term therapies, commonly adopted in cirrhotic patients, may influence BT and modify the risk of infection in these patients. To investigate the influence of chronic therapies on the prevalence and microbiological characteristics of infections in cirrhosis.MethodsConsecutive cirrhotic patients hospitalised from 2008 to 2013 were enrolled. All previous treatments were carefully recorded. Infections were actively sought out, patients were actively monitored for infection, and possible risk factors were evaluated.ResultsFour hundred cirrhotic patients were included. The most frequent therapies were proton pump inhibitors (PPIs) (67%), non‐absorbable‐disaccharides (44%), beta‐blockers (BBs) (39%) and non‐absorbable‐antibiotics (10%). Child‐Pugh C (P < 0.001; OR 5; 95%CI: 2.6–9.9) and PPI therapy (P = 0.008; OR 2; 95% CI: 1.2–3.2) were found to be independent predictors of infection, and the use of BBs was a protective factor (P = 0.001; OR 0.46; 95%CI: 0.3–0.7). Cirrhotic patients with bacterial infection showed lower morbidity and mortality when taking BBs.ConclusionsProton pump inhibitors increase the risk of infection in cirrhosis and should not be prescribed in these patients without specific indications. In contrast, the use of BBs is associated with a lower rate of infection and attenuates the consequences of infections in cirrhotic patients.
No Virological Evidence for an Influenza A ‐ like Virus in European Bats
Authors: Fereidouni S. , Kwasnitschka L. , Balkema Buschmann A. , Müller T. , Freuling C. , Schatz J. , Pikula J. , Bandouchova H. , Hoffmann R. , Ohlendorf B. , Kerth G. , Tong S. , Donis R. , Beer M. , Harder T. 
Published: May 2014

SummaryNew members of the influenza A virus genus have been detected recently in bats from South America. By molecular investigations, using a generic real‐time RT‐PCR (RT‐qPCR) that detects all previously known influenza A virus subtypes (H1‐H16) and a newly developed RT‐qPCR specific for the South American bat influenza‐like virus of subtype H17, a total of 1571 samples obtained from 1369 individual bats of 26 species from Central Europe were examined. No evidence for the occurrence of such influenza viruses was found. Further attempts towards a more comprehensive evaluation of the role of bats in the ecology and epidemiology of influenza viruses should be based on more intense monitoring efforts. However, given the protected status of bats, not only in Europe, such activities need to be embedded into existing pathogen‐monitoring programs.
Impact of endothelin blockade on acute exercise‐induced changes in blood flow and endothelial function in type 2 diabetes mellitus
Authors: Schreuder Tim H. A. , van Lotringen Jaap H. , Hopman Maria T. E. , Thijssen Dick H. J. 
Published: May 2014

New Findings
What is the central question of this study?
Doesendothelin receptor blockade increase exercise‐induced blood flow and post‐exercise endothelial function in men with type 2 diabetes?
What is the main finding and its importance?
Despite increasing exercise‐induced blood flow, endothelin receptor blockade did not impact post‐exercise endothelial function. The increase in blood flow may have beneficial effects in repeated exercise training.
Positive vascular effects of exercise training are mediated by acute increases in blood flow. Type 2 diabetes patients show attenuated exercise‐induced increases in blood flow, possibly mediated by the endothelin pathway, preventing an optimal stimulus for vascular adaptation. We examined the impact of endothelin receptor blockade (bosentan) on exercise‐induced blood flow in the brachial artery and on pre‐ and postexercise endothelial function in type 2 diabetes patients (n = 9, 60 ± 7 years old) and control subjects (n = 10, 60 ± 5 years old). Subjects reported twice to the laboratory to perform hand‐grip exercise in the presence of endothelin receptor blockade or placebo. We examined brachial artery endothelial function (via flow‐mediated dilatation) before and after exercise, as well as blood flow during exercise. Endothelin receptor blockade resulted in a larger increase in blood flow during exercise in type 2 diabetes patients (P = 0.046), but not in control subjects (P = 0.309). Exercise increased shear rate across the exercise protocol, unaffected by endothelin receptor blockade. Exercise did not alter brachial artery diameter in either group, but endothelin receptor blockade resulted in a larger brachial artery diameter in type 2 diabetes patients (P = 0.033). Exercise significantly increased brachial artery flow‐mediated dilatation in both groups, unaffected by endothelin receptor blockade. Endothelin receptor blockade increased exercise‐induced brachial artery blood flow in type 2 diabetes patients, but not in control subjects. Despite this effect of endothelin receptor blockade on blood flow, we found no impact on baseline or post‐exercise endothelial function in type 2 diabetes patients or control subjects, possibly related to normalization of the shear stimulus during exercise. The successful increase in blood flow during exercise in type 2 diabetes patients through endothelin receptor blockade may have beneficial effects in repeated exercise training.
Extracellular acidosis contracts coronary but neither renal nor mesenteric artery via modulation of H+,K+‐ATPase, voltage‐gated K+ channels and L‐type Ca2+ channels
Authors: Niu Longgang , Liu Yu , Hou Xiaomin , Cui Lijuan , Li Jiangtao , Zhang Xuanping , Zhang Mingsheng 
Published: May 2014

New Findings
What is the central question of this study?
We asked whether there are any differences in responses to extracellular acidosis among arteries of different origin and whether K+ channels, Ca2+ channels and H+,K+‐ATPase are involved in extracellular acidosis‐induced coronary myogenic responses.
What is the main finding and its importance?
This report is the first to demonstrate that extracellular acidosis alkalinizes the cytosol of arterial smooth muscle cells freshly isolated from rat coronary artery and contracts the artery exclusively and to suggest that modulation of voltage‐gated K+ channels, voltage‐gated Ca2+ channels and H+,K+‐ATPase are involved in the contraction.
Extracellular acidosis (EA) jeopardizes the heart, whereas mild extracellular alkalinization is cardioprotective, but it remains elusive how the coronary artery (CA) responses to EA. In the present study, EA was demonstrated to induce contraction in rat coronary artery (RCA) in a manner dependent on extracellular pH (pHo, 7.2–6.6), whereas it did not affect the resting tone of either rat renal interlobe artery (RIA) or mesenteric artery (MA). The amplitude of contraction provoked by pHo 6.8 was approximately equal to that induced by 60 mmol l−1 KCl at pHo 7.4. Blockade of L‐type voltage‐gated Ca2+ channels and inhibition of H+,K+‐ATPase attenuated the contraction, whereas inhibition of nitric oxide synthesis and endothelial denudation augmented it. A molecular probe study showed that EA acidified the cytosol of arterial smooth muscle cells (ASMCs) in RIA and MA, but alkalinized it in RCA. Extracellular acidosis elevated the intracellular Ca2+ concentration exclusively in RCA ASMCs. Patch‐clamp studies showed that EA enhanced L‐type voltage‐gated Ca2+ channel currents in RCA ASMCs, but depressed the currents in MA ASMCs and did not affect the currents in RIA ASMCs. Extracellular acidosis depressed voltage‐gated K+ channel (KV) currents only in RCA ASMCs. Lansoprazole blunted all these observed effects of EA on RCA. Taken together, the present results demonstrate that the responses of RCA to EA are different from those of RIA and MA and suggest that activation of L‐type voltage‐gated Ca2+ channels and H+,K+‐ATPase as well as depression of KV may, at least partly, underlie the EA‐induced contraction in RCA.
Use of Yeast Spores for Microencapsulation of Enzymes
Authors: Shi Libing , Li Zijie , Tachikawa Hiroyuki , Gao Xiao-Dong , Nakanishi Hideki 
Published: May 2014

ABSTRACT

Here, we report a novel method to produce microencapsulated enzymes using
Saccharomyces cerevisiae
spores. In sporulating cells, soluble secreted proteins are transported to the spore wall. Previous work has shown that the spore wall is capable of retaining soluble proteins because its outer layers work as a diffusion barrier. Accordingly, a red fluorescent protein (RFP) fusion of the α-galactosidase, Mel1, expressed in spores was observed in the spore wall even after spores were subjected to a high-salt wash in the presence of detergent. In vegetative cells, however, the cell wall cannot retain the RFP fusion. Although the spore wall prevents diffusion of proteins, it is likely that smaller molecules, such as sugars, pass through it. In fact, spores can contain much higher α-galactosidase activity to digest melibiose than vegetative cells. When present in the spore wall, the enzyme acquires resistance to environmental stresses including enzymatic digestion and high temperatures. The outer layers of the spore wall are required to retain enzymes but also decrease accessibility of the substrates. However, mutants with mild spore wall defects can retain and stabilize the enzyme while still permitting access to the substrate. In addition to Mel1, we also show that spores can retain the invertase. Interestingly the encapsulated invertase has significantly lower activity toward raffinose than toward sucrose. This suggests that substrate selectivity could be altered by the encapsulation.
Multilocus Sequence Typing Confirms Wild Birds as the Source of a Campylobacter Outbreak Associated with the Consumption of Raw Peas
Authors: Kwan Patrick S. L. , Xavier Catherine , Santovenia Monica , Pruckler Janet , Stroika Steven , Joyce Kevin , Gardner Tracie , Fields Patricia I. , McLaughlin Joe , Tauxe Robert V. , Fitzgerald Collette 
Published: May 2014

ABSTRACT

From August to September 2008, the Centers for Disease Control and Prevention (CDC) assisted the Alaska Division of Public Health with an outbreak investigation of campylobacteriosis occurring among the residents of Southcentral Alaska. During the investigation, pulsed-field gel electrophoresis (PFGE) of
Campylobacter jejuni
isolates from human, raw pea, and wild bird fecal samples confirmed the epidemiologic link between illness and the consumption of raw peas contaminated by sandhill cranes for 15 of 43 epidemiologically linked human isolates. However, an association between the remaining epidemiologically linked human infections and the pea and wild bird isolates was not established. To better understand the molecular epidemiology of the outbreak,
C. jejuni
isolates (
n
= 130; 59 from humans, 40 from peas, and 31 from wild birds) were further characterized by multilocus sequence typing (MLST). Here we present the molecular evidence to demonstrate the association of many more human
C. jejuni
infections associated with the outbreak with raw peas and wild bird feces. Among all sequence types (STs) identified, 26 of 39 (67%) were novel and exclusive to the outbreak. Five clusters of overlapping STs (
n
= 32 isolates; 17 from humans, 2 from peas, and 13 from wild birds) were identified. In particular, cluster E (
n
= 7 isolates; ST-5049) consisted of isolates from humans, peas, and wild birds. Novel STs clustered closely with isolates typically associated with wild birds and the environment but distinct from lineages commonly seen in human infections. Novel STs and alleles recovered from human outbreak isolates allowed additional infections caused by these rare genotypes to be attributed to the contaminated raw peas.
Gene Discovery for Enzymes Involved in Limonene Modification or Utilization by the Mountain Pine Beetle-Associated Pathogen Grosmannia clavigera
Authors: Wang Ye , Lim Lynette , Madilao Lina , Lah Ljerka , Bohlmann Joerg , Breuil Colette 
Published: May 2014

ABSTRACT

To successfully colonize and eventually kill pine trees,
Grosmannia clavigera
(Gs cryptic species), the main fungal pathogen associated with the mountain pine beetle (
Dendroctonus ponderosae
), has developed multiple mechanisms to overcome host tree chemical defenses, of which terpenoids are a major component. In addition to a monoterpene efflux system mediated by a recently discovered ABC transporter, Gs has genes that are highly induced by monoterpenes and that encode enzymes that modify or utilize monoterpenes [especially (+)-limonene]. We showed that pine-inhabiting Ophiostomale fungi are tolerant to monoterpenes, but only a few, including Gs, are known to utilize monoterpenes as a carbon source. Gas chromatography-mass spectrometry (GC-MS) revealed that Gs can modify (+)-limonene through various oxygenation pathways, producing carvone,
p
-mentha-2,8-dienol, perillyl alcohol, and isopiperitenol. It can also degrade (+)-limonene through the C-1-oxygenated pathway, producing limonene-1,2-diol as the most abundant intermediate. Transcriptome sequencing (RNA-seq) data indicated that Gs may utilize limonene 1,2-diol through beta-oxidation and then valine and tricarboxylic acid (TCA) metabolic pathways. The data also suggested that at least two gene clusters, located in genome contigs 108 and 161, were highly induced by monoterpenes and may be involved in monoterpene degradation processes. Further, gene knockouts indicated that limonene degradation required two distinct Baeyer-Villiger monooxygenases (BVMOs), an epoxide hydrolase and an enoyl coenzyme A (enoyl-CoA) hydratase. Our work provides information on enzyme-mediated limonene utilization or modification and a more comprehensive understanding of the interaction between an economically important fungal pathogen and its host's defense chemicals.
Novel Outer Membrane Protein Involved in Cellulose and Cellooligosaccharide Degradation by Cytophaga hutchinsonii
Authors: Ji Xiaofei , Wang Ying , Zhang Cong , Bai Xinfeng , Zhang Weican , Lu Xuemei 
Published: May 2014

ABSTRACT

Cytophaga hutchinsonii
is an aerobic cellulolytic soil bacterium which was reported to use a novel contact-dependent strategy to degrade cellulose. It was speculated that cellooligosaccharides were transported into the periplasm for further digestion. In this study, we reported that most of the endoglucanase and β-glucosidase activity was distributed on the cell surface of
C. hutchinsonii
. Cellobiose and part of the cellulose could be hydrolyzed to glucose on the cell surface. However, the cell surface cellulolytic enzymes were not sufficient for cellulose degradation by
C. hutchinsonii
. An outer membrane protein, CHU_1277, was disrupted by insertional mutation. Although the mutant maintained the same endoglucanase activity and most of the β-glucosidase activity, it failed to digest cellulose, and its cellooligosaccharide utilization ability was significantly reduced, suggesting that CHU_1277 was essential for cellulose degradation and played an important role in cellooligosaccharide utilization. Further study of cellobiose hydrolytic ability of the mutant on the enzymatic level showed that the β-glucosidase activity in the outer membrane of the mutant was not changed. It revealed that CHU_1277 played an important role in assisting cell surface β-glucosidase to exhibit its activity sufficiently. Studies on the outer membrane proteins involved in cellulose and cellooligosaccharide utilization could shed light on the mechanism of cellulose degradation by
C. hutchinsonii
.
Comparative Genomic and Morphological Analyses of Listeria Phages Isolated from Farm Environments
Authors: Denes Thomas , Vongkamjan Kitiya , Ackermann Hans-Wolfgang , Moreno Switt Andrea I. , Wiedmann Martin , den Bakker Henk C. 
Published: May 2014

ABSTRACT

The genus
Listeria
is ubiquitous in the environment and includes the globally important food-borne pathogen
Listeria monocytogenes
. While the genomic diversity of
Listeria
has been well studied, considerably less is known about the genomic and morphological diversity of
Listeria
bacteriophages. In this study, we sequenced and analyzed the genomes of 14
Listeria
phages isolated mostly from New York dairy farm environments as well as one related
Enterococcus faecalis
phage to obtain information on genome characteristics and diversity. We also examined 12 of the phages by electron microscopy to characterize their morphology. These
Listeria
phages, based on gene orthology and morphology, together with previously sequenced
Listeria
phages could be classified into five orthoclusters, including one novel orthocluster. One orthocluster (orthocluster I) consists of large-genome (∼135-kb) myoviruses belonging to the genus “Twort-like viruses,” three orthoclusters (orthoclusters II to IV) contain small-genome (36- to 43-kb) siphoviruses with icosahedral heads, and the novel orthocluster V contains medium-sized-genome (∼66-kb) siphoviruses with elongated heads. A novel orthocluster (orthocluster VI) of
E. faecalis
phages, with medium-sized genomes (∼56 kb), was identified, which grouped together and shares morphological features with the novel
Listeria
phage orthocluster V. This new group of phages (i.e., orthoclusters V and VI) is composed of putative lytic phages that may prove to be useful in phage-based applications for biocontrol, detection, and therapeutic purposes.
Environmental Surveillance of Human Enteroviruses in Shandong Province, China, 2008 to 2012: Serotypes, Temporal Fluctuation, and Molecular Epidemiology
Authors: Wang Haiyan , Tao Zexin , Li Yan , Lin Xiaojuan , Yoshida Hiromu , Song Lizhi , Zhang Yong , Wang Suting , Cui Ning , Xu Wenbo , Song Yanyan , Xu Aiqiang 
Published: May 2014

ABSTRACT
Environmental surveillance is an effective approach in investigating the circulation of polioviruses (PVs) and other human enteroviruses (EVs) in the population. The present report describes the results of environmental surveillance conducted in Shandong Province, China, from 2008 to 2012. A total of 129 sewage samples were collected, and 168 PVs and 1,007 nonpolio enteroviruses (NPEVs) were isolated. VP1 sequencing and typing were performed on all isolates. All PV strains were Sabin-like, with the numbers of VP1 substitutions ranging from 0 to 7. The NPEVs belonged to 19 serotypes, and echovirus 6 (E6), E11, coxsackievirus B3 (CVB3), E3, E12, and E7 were the six main serotypes, which accounted for 18.3%, 14.8%, 14.5%, 12.9%, 9.0%, and 5.7% of NPEVs isolated, respectively. Typical summer-fall peaks of NPEV were observed in the monthly distribution of isolation, and an epidemic pattern of annual circulation was revealed for the common serotypes. Phylogenetic analysis was performed on environmental CVB3 and E3 strains with global reference strains and local strains from aseptic meningitis patients. Shandong strains formed distinct clusters, and a close relationship was observed between local environmental and clinical strains. As an EV-specific case surveillance system is absent in China and many other countries, continuous environmental surveillance should be encouraged to investigate the temporal circulation and phylogeny of EVs in the population.