bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of Incubation Temperature on the Detection of Thermophilic Campylobacter Species from Freshwater Beaches, Nearby Wastewater Effluents, and Bird Fecal Droppings
Authors: Khan Izhar U. H. , Hill Stephen , Nowak Eva , Edge Thomas A. 
Published: September 2013

ABSTRACT

This large-scale study compared incubation temperatures (37°C versus 42°C) to study the detection of thermophilic
Campylobacter
species, including
Campylobacter jejuni
,
C. coli
, and
C. lari
, in various surface water samples and bird fecal droppings around Hamilton Harbor, Lake Ontario. The putative culture isolates obtained from incubation temperatures of 37 and 42°C were confirmed by
Campylobacter
genus- and species-specific triplex PCR assays targeting the 16S rRNA gene and the 16S-23S rRNA gene internal transcribed spacer (ITS) region. A total of 759 water, wastewater, and bird fecal dropping samples were tested. Positive amplification reactions for the genus
Campylobacter
were found for 454 (60%) samples incubated at 37°C, compared to 258 (34%) samples incubated at 42°C.
C. jejuni
(16%) and
C. lari
(12%) were detected significantly more frequently at the 42°C incubation temperature than at 37°C (8% and 5%, respectively). In contrast, significantly higher rates of
C. coli
(14%) and other
Campylobacter
spp. (36%) were detected at the 37°C incubation temperature than at 42°C (8% and 7%, respectively). These results were consistent across surface water, wastewater, and bird fecal dropping samples. At times,
Campylobacter
spp. were recovered and detected at 37°C (3% for
C. jejuni
, 10% for
C. coli
, and 3% for
C. lari
) when the same samples incubated at 42°C were negative. A significantly higher rate of other
Campylobacter
spp. was detected only at 37°C (32%) than only at 42°C (3%). These results indicate that incubation temperature can significantly influence the culturability and detection of thermophilic and other fastidious
Campylobacter
spp. and that a comprehensive characterization of the
Campylobacter
spp. in surface water, wastewaters, or bird fecal droppings will require incubation at both 37 and 42°C.
Depth-Related Changes in Community Structure of Culturable Mineral Weathering Bacteria and in Weathering Patterns Caused by Them along Two Contrasting Soil Profiles
Authors: Huang Jing , Sheng Xia-Fang , Xi Jun , He Lin-Yan , Huang Zhi , Wang Qi , Zhang Zhen-Dong 
Published: September 2013

ABSTRACT
Bacteria play important roles in mineral weathering and soil formation. However, few reports of mineral weathering bacteria inhabiting subsurfaces of soil profiles have been published, raising the question of whether the subsurface weathering bacteria are fundamentally distinct from those in surface communities. To address this question, we isolated and characterized mineral weathering bacteria from two contrasting soil profiles with respect to their role in the weathering pattern evolution, their place in the community structure, and their depth-related changes in these two soil profiles. The effectiveness and pattern of bacterial mineral weathering were different in the two profiles and among the horizons within the respective profiles. The abundance of highly effective mineral weathering bacteria in the Changshu profile was significantly greater in the deepest horizon than in the upper horizons, whereas in the Yanting profile it was significantly greater in the upper horizons than in the deeper horizons. Most of the mineral weathering bacteria from the upper horizons of the Changshu profile and from the deeper horizons of the Yanting profile significantly acidified the culture media in the mineral weathering process. The proportion of siderophore-producing bacteria in the Changshu profile was similar in all horizons except in the Bg2 horizon, whereas the proportion of siderophore-producing bacteria in the Yanting profile was higher in the upper horizons than in the deeper horizons. Both profiles existed in different highly depth-specific culturable mineral weathering community structures. The depth-related changes in culturable weathering communities were primarily attributable to minor bacterial groups rather than to a change in the major population structure.
Cooperative Degradation of Chitin by Extracellular and Cell Surface-Expressed Chitinases from Paenibacillus sp. Strain FPU-7
Authors: Itoh Takafumi , Hibi Takao , Fujii Yutaka , Sugimoto Ikumi , Fujiwara Akihiro , Suzuki Fumiko , Iwasaki Yukimoto , Kim Jin-Kyung , Taketo Akira , Kimoto Hisashi 
Published: September 2013

ABSTRACT

Chitin, a major component of fungal cell walls and invertebrate cuticles, is an exceedingly abundant polysaccharide, ranking next to cellulose. Industrial demand for chitin and its degradation products as raw materials for fine chemical products is increasing. A bacterium with high chitin-decomposing activity,
Paenibacillus
sp. strain FPU-7, was isolated from soil by using a screening medium containing α-chitin powder. Although FPU-7 secreted several extracellular chitinases and thoroughly digested the powder, the extracellular fluid alone broke them down incompletely. Based on expression cloning and phylogenetic analysis, at least seven family 18 chitinase genes were found in the FPU-7 genome. Interestingly, the product of only one gene (
chiW
) was identified as possessing three S-layer homology (SLH) domains and two glycosyl hydrolase family 18 catalytic domains. Since SLH domains are known to function as anchors to the Gram-positive bacterial cell surface, ChiW was suggested to be a novel multimodular surface-expressed enzyme and to play an important role in the complete degradation of chitin. Indeed, the ChiW protein was localized on the cell surface. Each of the seven chitinase genes (
chiA
to
chiF
and
chiW
) was cloned and expressed in
Escherichia coli
cells for biochemical characterization of their products. In particular, ChiE and ChiW showed high activity for insoluble chitin. The high chitinolytic activity of strain FPU-7 and the chitinases may be useful for environmentally friendly processing of chitin in the manufacture of food and/or medicine.
Effect of Bacteriophage Application on Campylobacter jejuni Loads in Commercial Broiler Flocks
Authors: Kittler Sophie , Fischer Samuel , Abdulmawjood Amir , Glünder Gerhard , Klein Günter 
Published: September 2013

ABSTRACT

Campylobacteriosis is the most frequent food-borne human enteritis. The major source for infection with
Campylobacter
spp. is broiler meat. Risk assessments consider the reduction of
Campylobacter
in primary production to be most beneficial for human health. The aim of this study was to test the efficacy of a bacteriophage application under commercial conditions which had proved to be effective in previous noncommercial studies under controlled experimental conditions. A phage cocktail for
Campylobacter
reduction was tested on three commercial broiler farms each with a control and an experimental group. Colonization of
Campylobacter
was confirmed prior to phage application in fecal samples. Subsequently, a phage cocktail was applied via drinking water in the experimental group (log
10
5.8 to 7.5 PFU/bird). One day after phage application,
Campylobacter
counts of one experimental group were reduced under the detection limit (<50 CFU/g,
P
= 0.0140) in fecal samples. At slaughter, a significant reduction of >log
10
3.2 CFU/g cecal content compared to the control was still detected (
P
= 0.0011). No significant reduction was observed in the experimental groups of the other trials. However, a significant drop in cecal
Campylobacter
counts occurred in a phage-contaminated control. These results suggest that maximum reduction of
Campylobacter
at the slaughterhouse might be achieved by phage application 1 to 4 days prior to slaughter.
The Taz1p Transacylase Is Imported and Sorted into the Outer Mitochondrial Membrane via a Membrane Anchor Domain
Authors: Herndon Jenny D. , Claypool Steven M. , Koehler Carla M. 
Published: September 2013

ABSTRACT

Mutations in the mitochondrial transacylase tafazzin, Taz1p, in
Saccharomyces cerevisiae
cause Barth syndrome, a disease of defective cardiolipin remodeling. Taz1p is an interfacial membrane protein that localizes to both the outer and inner membranes, lining the intermembrane space. Pathogenic point mutations in Taz1p that alter import and membrane insertion result in accumulation of monolysocardiolipin. In this study, we used yeast as a model to investigate the biogenesis of Taz1p. We show that to achieve this unique topology in mitochondria, Taz1p follows a novel import pathway in which it crosses the outer membrane via the translocase of the outer membrane and then uses the Tim9p-Tim10p complex of the intermembrane space to insert into the mitochondrial outer membrane. Taz1p is then transported to membranes of an intermediate density to reach a location in the inner membrane. Moreover, a pathogenic mutation within the membrane anchor (V224R) alters Taz1p import so that it bypasses the Tim9p-Tim10p complex and interacts with the translocase of the inner membrane, TIM23, to reach the matrix. Critical targeting information for Taz1p resides in the membrane anchor and flanking sequences, which are often mutated in Barth syndrome patients. These studies suggest that altering the mitochondrial import pathway of Taz1p may be important in understanding the molecular basis of Barth syndrome.
Tetracycline-Related Transcriptional Regulation of the CTnDOT Mobilization Region
Authors: Waters Jillian L. , Wang Gui-Rong , Salyers Abigail A. 
Published: September 2013

ABSTRACT

CTnDOT is a 65-kb conjugative transposon (CTn) in
Bacteroides
spp. that confers resistance to the antibiotics erythromycin and tetracycline (Tc). Conjugative transfer of CTnDOT is regulated upon exposure of cells to Tc. In the absence of Tc, no transfer is detectable; however, a cascade of regulatory events results in the conjugative transfer of CTnDOT upon Tc induction. Previous studies addressing regulation of CTnDOT conjugative transfer focused primarily on the 13-kb transfer (
tra
) operon, which encodes the proteins required for assembly of the mating apparatus. We report here that the
mob
operon that encodes the relaxase and coupling proteins required for mobilization of CTnDOT are regulated at the transcriptional level upon Tc induction. The Xis2d and Exc excision proteins are required for the upregulation of
mob
transcription upon Tc induction, and yet a deletion of
xis2c
has no effect. We also show preliminary evidence suggesting that the integrase, IntDOT, may play a regulatory role, as pLYL72 transfer is not detectable when
intDOT
is provided in
trans
.
Phenotypic Profiling of Mycobacterium tuberculosis EspA Point Mutants Reveals that Blockage of ESAT-6 and CFP-10 Secretion
In Vitro
Does Not Always Correlate with Attenuation of Virulence
Authors: Chen Jeffrey M. , Zhang Ming , Rybniker Jan , Basterra Laetitia , Dhar Neeraj , Tischler Anna D. , Pojer Florence , Cole Stewart T. 
Published: September 2013

ABSTRACT

The EspA protein of
Mycobacterium tuberculosis
is essential for the type VII ESX-1 protein secretion apparatus, which delivers the principal virulence factors ESAT-6 and CFP-10. In this study, site-directed mutagenesis of EspA was performed to elucidate its influence on the ESX-1 system. Replacing Trp
55
(W55) or Gly
57
(G57) residues in the putative W-X-G motif of EspA with arginines impaired ESAT-6 and CFP-10 secretion
in vitro
and attenuated
M. tuberculosis
. Replacing the Phe
50
(F50) and Lys
62
(K62) residues, which flank the W-X-G motif, with arginine and alanine, respectively, destabilized EspA, abolished ESAT-6 and CFP-10 secretion
in vitro
, and attenuated
M. tuberculosis
. Likewise, replacing the Phe
5
(F5) and Lys
41
(K41) residues with arginine and alanine, respectively, also destabilized EspA and blocked ESAT-6 and CFP-10 secretion
in vitro
. However, these two particular mutations did not attenuate
M. tuberculosis
in cellular models of infection or during acute infection in mice. We have thus identified amino acid residues in EspA that are important for facilitating ESAT-6 and CFP-10 secretion and virulence. However, our data also indicate for the first time that blockage of
M. tuberculosis
ESAT-6 and CFP-10 secretion
in vitro
and attenuation are mutually exclusive.
Paramedic Initiated Lisinopril For Acute Stroke Treatment (PIL-FAST): results from the pilot randomised controlled trial
Authors: Shaw Lisa , Price Christopher , McLure Sally , Howel Denise , McColl Elaine , Younger Paul , Ford Gary A 
Published: September 2013

BackgroundHigh blood pressure (BP) during acute stroke is associated with poorer stroke outcome. Trials of treatments to lower BP have not resulted in improved outcome, but this may be because treatment commenced too late. Emergency medical service staff (paramedics) are uniquely placed to administer early treatment; however, experience of prehospital randomised controlled trials (RCTs) is very limited.MethodsWe conducted a pilot RCT to determine the feasibility of a definitive prehospital BP-lowering RCT in acute stroke. Paramedics were trained to identify, consent and deliver a first dose of lisinopril or placebo to adults with suspected stroke and hypertension while responding to the emergency call. Further treatment continued in hospital. Study eligibility, recruitment rate, completeness of receipt of study medication and clinical data (eg, BP) were collected to inform the design of a definitive RCT.ResultsIn 14 months, 14 participants (median age=73 years, median National Institute of Health Stroke Scale=4) were recruited and received the prehospital dose of medication. Median time from stroke onset (as assessed by paramedic) to treatment was 70 min. Four participants completed 7 days of study treatment. Of ambulance transported suspected stroke patients, 1% were both study eligible and attended by a PIL-FAST paramedic.ConclusionsIt is possible to conduct a paramedic initiated double-blind RCT of a treatment for acute stroke. However, to perform a definitive RCT in a reasonable timescale, a large number of trained paramedics across several ambulance services would be needed to recruit the number of patients likely to be required.Clinical trial registrationhttp://www.clinicaltrials.gov. Unique identifier:NCT01066572.
Assessment of non-clinical attributes in paramedicine using multiple mini-interviews
Authors: Tavares Walter , Mausz Justin 
Published: September 2013

BackgroundNon-clinical attributes are increasingly emphasised as an important factor in paramedic practice. However, the assessment of these attributes often lacks the evidence base to support it. Exploring the relationship between non-clinical attributes and clinical skills is also of theoretical and practical importance.ObjectiveTo first seek evidence of reliability and validity for the assessment of non-clinical attributes using the multiple mini-interview (MMI) in paramedic contexts and second, to explore the association between non-clinical attributes and clinical skills in paramedicine.MethodsEntry to practice level paramedic candidates completed a 10-station MMI to assess non-clinical attributes on day 1 and a 10-station simulation-based assessment (SBA) of clinical skills on day 2. Both were assessed using different global rating scales. Our primary outcomes included MMI inter-station reliability (calculated using generalisability theory) and Pearson's correlation between non-clinical attributes and clinically focused skills.Results30 trainees completed the MMI and 26 of the 30 completed the SBA. Inter-station reliability for the MMI reached 0.77. Pearson's correlations (disattenuated correlations in parentheses) between the overall MMI score and mean SBA global rating scores reached r=0.31 (r=0.48) and ranged by dimension from r=−0.11 (−0.17) (procedural skills) to r=0.54(r=0.83) (communication).ConclusionsThe MMI demonstrated evidence of reliability and validity for the assessment of non-clinical attributes in paramedic contexts. Non-clinical attributes and paramedic clinical skills are likely distinct but related constructs, each contributing to the construct of paramedic practice. Programmes of assessment should include both to ensure the construct of paramedic practice is adequately represented.
JMJD2B Promotes Epithelial–Mesenchymal Transition by Cooperating with β-Catenin and Enhances Gastric Cancer Metastasis
Authors: Zhao Li , Li Wenjuan , Zang Wen , Liu Zhifang , Xu Xia , Yu Han , Yang Qing , Jia Jihui 
Published: September 2013

Abstract
Purpose: This study investigated the role of histone demethylase Jumonji domain–containing protein 2B (JMJD2B) in promoting epithelial–mesenchymal transition (EMT) and underlying molecular mechanisms in the progression of gastric cancer.
Experimental Design: The induction of EMT by JMJD2B in gastric cancer cells and its underlying mechanisms were examined by a series of assays. In vivo and in vitro assays were performed to clarify invasive potential of JMJD2B in gastric cancer cells. The expression dynamics of JMJD2B were detected using immunohistochemistry in 101 cases of primary gastric cancer tissues.
Results: Inhibition of JMJD2B by specific siRNA suppresses EMT of gastric cancer cells, whereas ectopic expression of JMJD2B induces EMT. Importantly, JMJD2B is physically associated with β-catenin and enhances its nuclear localization and transcriptional activity. JMJD2B, together with β-catenin, binds to the promoter of the β-catenin target gene vimentin to increase its transcription by inducing H3K9 demethylation locally. JMJD2B inhibition attenuates migration and invasion of gastric cancer cells in vitro and metastasis in vivo. The expression of JMJD2B was positively correlated with tumor size (P = 0.017), differentiation status (P = 0.002), tumor invasion (P = 0.045), lymph node metastasis (P = 0.000), distant metastasis (P = 0.024), and tumor–node–metastasis (TNM) stage (P = 0.002) in patients with gastric cancer.
Conclusions: The data reveal a novel function of JMJD2B in promoting EMT and gastric cancer invasion and metastasis, implicating JMJD2B as a potential target for reversing EMT and intervention of the progression of gastric cancer. Clin Cancer Res; 19(23); 6419–29. ©2013 AACR.
Phase I Study of Vismodegib in Children with Recurrent or Refractory Medulloblastoma: A Pediatric Brain Tumor Consortium Study
Authors: Gajjar Amar , Stewart Clinton F. , Ellison David W. , Kaste Sue , Kun Larry E. , Packer Roger J. , Goldman Stewart , Chintagumpala Murali , Wallace Dana , Takebe Naoko , Boyett James M. , Gilbertson Richard J. , Curran Tom 
Published: September 2013

Abstract
Purpose: To investigate the safety, dose-limiting toxicities, and pharmacokinetics of the smoothened inhibitor vismodegib in children with refractory or relapsed medulloblastoma.
Experimental design: Initially, vismodegib was administered daily at 85 mg/m2 and escalated to 170 mg/m2. The study was then revised to investigate a flat-dosing schedule of 150 mg for patients with small body surface area (BSA, 0.67–1.32 m2) or 300 mg for those who were larger (BSA, 1.33–2.20 m2). Pharmacokinetics were performed during the first course of therapy, and the right knees of all patients were imaged to monitor bone toxicity. Immunohistochemical analysis was done to identify patients with Sonic Hedgehog (SHH)-subtype medulloblastoma.
Results: Thirteen eligible patients were enrolled in the initial study: 6 received 85 mg/m2 vismodegib, and 7 received 170 mg/m2. Twenty eligible patients were enrolled in the flat-dosing part of the study: 10 at each dosage level. Three dose-limiting toxicities were observed, but no drug-related bone toxicity was documented. The median (range) vismodegib penetration in the cerebrospinal fluid (CSF) was 0.53 (0.26–0.78), when expressed as a ratio of the concentration of vismodegib in the CSF to that of the unbound drug in plasma. Antitumor activity was seen in 1 of 3 patients with SHH-subtype disease whose tumors were evaluable, and in none of the patients in the other subgroups.
Conclusions: Vismodegib was well tolerated in children with recurrent or refractory medulloblastoma; only two dose-limiting toxicities were observed with flat dosing. The recommended phase II study dose is 150 or 300 mg, depending on the patient's BSA. Clin Cancer Res; 19(22); 6305–12. ©2013 AACR.
Proteomic Markers of DNA Repair and PI3K Pathway Activation Predict Response to the PARP Inhibitor BMN 673 in Small Cell Lung Cancer
Authors: Cardnell Robert J. , Feng Ying , Diao Lixia , Fan You-Hong , Masrorpour Fatemah , Wang Jing , Shen Yuqiao , Mills Gordon B. , Minna John D. , Heymach John V. , Byers Lauren A. 
Published: September 2013

Abstract
Purpose: Small cell lung carcinoma (SCLC) is an aggressive malignancy affecting nearly 30,000 people annually in the United States. We have previously identified elevated PARP1 levels in SCLC and demonstrated in vitro sensitivity to the PARP inhibitors AZD 2281 and AG014699. Here, we evaluate activity of a novel, potent PARP inhibitor, BMN 673, and identify markers of response as a basis for developing predictive markers for clinical application.
Experimental Design: Inhibition of SCLC proliferation by BMN 673 was assayed in vitro and effects on tumor growth were measured in SCLC xenograft models. Protein expression and pathway activation was assessed by reverse phase protein array and western blot analysis. PARP inhibition was confirmed using a PAR ELISA.
Results: We demonstrate striking, single agent activity of BMN 673 in SCLC cell lines and xenografts, with single agent BMN 673 exhibiting in vivo activity similar to cisplatin. Sensitivity to BMN 673 was associated with elevated baseline expression levels of several DNA repair proteins, whereas greater drug resistance was observed in SCLC models with baseline activation of the PI3K/mTOR pathway. Furthermore, we developed and confirmed these data with a novel “DNA repair score” consisting of a group of 17 DNA repair proteins.
Conclusions: Elevated expression of multiple DNA repair proteins, as well as a corresponding “DNA repair protein score,” predict response to BMN 673 in in vitro SCLC models. These observations complement recent work in which PI3K inhibition sensitizes breast cancer models to PARP inhibition, suggesting cooperation between DNA repair and PI3K pathways. Clin Cancer Res; 19(22); 6322–8. ©2013 AACR.