bluuebunny/crossref_metadata_2025_split huggingface dataset:
In Vitro Activity of CEM-101 against
Streptococcus pneumoniae
and
Streptococcus pyogenes
with Defined Macrolide Resistance Mechanisms
Authors: McGhee Pamela , Clark Catherine , Kosowska-Shick Klaudia M. , Nagai Kensuke , Dewasse Bonifacio , Beachel Linda , Appelbaum Peter C. 
Published: November 2009

ABSTRACT

CEM-101 had MIC ranges of 0.002 to 0.016 μg/ml against macrolide-susceptible pneumococci and 0.004 to 1 μg/ml against macrolide-resistant phenotypes. Only 3 strains with
erm
(B), with or without
mef
(A), had CEM-101 MICs of 1 μg/ml, and 218/221 strains had CEM-101 MICs of ≤0.5 μg/ml. CEM-101 MICs were as much as 4-fold lower than telithromycin MICs against all strains. For
Streptococcus pyogenes
, CEM-101 MICs ranged from 0.008 to 0.03 μg/ml against macrolide-susceptible strains and from 0.015 to 1 μg/ml against macrolide-resistant strains. Against
erm
(B) strains, erythromycin, azithromycin, and clarithromycin MICs were 32 to >64 μg/ml, while 17/19 strains had telithromycin MICs of 4 to 16 μg/ml; CEM-101 MICs were 0.015 to 1 μg/ml. By comparison,
erm
(A) and
mef
(A) strains had CEM-101 MICs of 0.015 to 0.5 μg/ml, clindamycin and telithromycin MICs of ≤1 μg/ml, and erythromycin, azithromycin, and clarithromycin MICs of 0.5 to >64 μg/ml. Pneumococcal multistep resistance studies showed that although CEM-101 yielded clones with higher MICs for all eight strains tested, seven of eight strains had clones with CEM-101 MICs that rose from 0.004 to 0.03 μg/ml (parental strains) to 0.06 to 0.5 μg/ml (resistant clones); for only one
erm
(B)
mef
(A) strain with a parental MIC of 1 μg/ml was there a resistant clone with a MIC of 32 μg/ml, with no detectable mutations in the L4, L22, or 23S rRNA sequence. Among two of five
S. pyogenes
strains tested, CEM-101 MICs rose from 0.03 to 0.25 μg/ml, and only for the one strain with
erm
(B) did CEM-101 MICs rise from 1 to 8 μg/ml, with no changes occurring in any macrolide resistance determinant. CEM-101 had low MICs as well as low potential for the selection of resistant mutants, independent of bacterial species or resistance phenotypes in pneumococci and
S. pyogenes
.
Echinocandin Susceptibility Testing ofCandidaSpecies: Comparison of EUCAST EDef 7.1, CLSI M27-A3, Etest, Disk Diffusion, and Agar Dilution Methods with RPMI and IsoSensitest Media
Authors: Arendrup Maiken Cavling , Garcia-Effron Guillermo , Lass-Flörl Cornelia , Lopez Alicia Gomez , Rodriguez-Tudela Juan-Luis , Cuenca-Estrella Manuel , Perlin David S. 
Published: November 2009

ABSTRACTThis study compared nine susceptibility testing methods and 12 endpoints for anidulafungin, caspofungin, and micafungin with the same collection of blindedFKShot spot mutant (n= 29) and wild-type isolates (n= 94). The susceptibility tests included EUCAST Edef 7.1, agar dilution, Etest, and disk diffusion with RPMI-1640 plus 2% glucose (2G) and IsoSensitest-2G media and CLSI M27A-3. Microdilution plates were read after 24 and 48 h. The following test parameters were evaluated:fkshot spot mutants overlapping the wild-type distribution, distance between the two populations, number of very major errors (VMEs;fksmutants misclassified as susceptible), and major errors (MEs; wild-type isolates classified as resistant) using a wild-type-upper-limit value (WT-UL) (two twofold-dilutions higher than the MIC50) as the susceptibility breakpoint. The methods with the lowest number of errors (given as VMEs/MEs) across the three echinocandins were CLSI (12%/1%), agar dilution with RPMI-2G medium (14%/0%), and Etest with RPMI-2G medium (8%/3%). The fewest errors overall were observed for anidulafungin (4%/1% for EUCAST, 4%/3% for CLSI, and 3%/9% for Etest with RPMI-2G). For micafungin, VME rates of 10 to 71% were observed. For caspofungin, agar dilution with either medium was superior (VMEs/MEs of 0%/1%), while CLSI, EUCAST with IsoSensitest-2G medium, and Etest were less optimal (VMEs of 7%, 10%, and 10%, respectively). Applying the CLSI breakpoint (S ≤ 2 μg/ml) for CLSI results, 89.2%fkshot spot mutants were classified as anidulafungin susceptible, 60.7% as caspofungin susceptible, and 92.9% as micafungin susceptible. In conclusion, no test was perfect, but anidulafungin susceptibility testing using the WT-UL to define susceptibility reliably identifiedfkshot spot mutants.
Identification of a Claudin-4 Residue Important for Mediating the Host Cell Binding and Action of
Clostridium perfringens
Enterotoxin
Authors: Robertson Susan L. , Smedley James G. , McClane Bruce A. 
Published: November 2009

ABSTRACT

The 24-member claudin protein family plays a key role in maintaining the normal structure and function of epithelial tight junctions. Previous studies with fibroblast transfectants and naturally sensitive Caco-2 cells have also implicated certain claudins (e.g., Claudin-4) as receptors for
Clostridium perfringens
enterotoxin (CPE). The present study first provided evidence that the second extracellular loop (ECL-2) of claudins is specifically important for mediating the host cell binding and cytotoxicity of native CPE. Rat fibroblast transfectants expressing a Claudin-4 chimera, where the natural ECL-2 was replaced by ECL-2 from Claudin-2, exhibited no CPE-induced cytotoxicity. Conversely, CPE bound to, and killed, CPE-treated transfectants expressing a Claudin-2 chimera with a substituted ECL-2 from Claudin-4. Site-directed mutagenesis was then used to alter an ECL-2 residue that invariably aligns as N in claudins known to bind native CPE but as D or S in claudins that cannot bind CPE. Transfectants expressing a Claudin-4
N149D
mutant lost the ability to bind or respond to CPE, while transfectants expressing a Claudin-1 mutant with the corresponding ECL-2 residue changed from D to N acquired CPE binding and sensitivity. Identifying carriage of this N residue in ECL-2 as being important for native CPE binding helps to explain why only certain claudins can serve as CPE receptors. Finally, preincubating CPE with soluble recombinant Claudin-4, or Claudin-4 fragments containing ECL-2 specifically blocked the cytotoxicity on Caco-2 cells. This result opens the possibility of using receptor claudins as therapeutic decoys to ameliorate CPE-mediated intestinal disease.
Norepinephrine Augments
Salmonella enterica
-Induced Enteritis in a Manner Associated with Increased Net Replication but Independent of the Putative Adrenergic Sensor Kinases QseC and QseE
Authors: Pullinger Gillian D. , Carnell Sonya C. , Sharaff Fathima F. , van Diemen Pauline M. , Dziva Francis , Morgan Eirwen , Lyte Mark , Freestone Primrose P. E. , Stevens Mark P. 
Published: November 2009

ABSTRACT

Stress has long been correlated with susceptibility to microbial infection. One explanation for this phenomenon is the ability of pathogens to sense and respond to host stress-related catecholamines, such as norepinephrine (NE). In Gram-negative enteric pathogens, it has been proposed that NE may facilitate growth by mediating iron supply, or it may alter gene expression by activating adrenergic sensor kinases. The aim of this work was to investigate the relative importance of these processes in a model in which NE alters the outcome of
Salmonella enterica
serovar Typhimurium infection. A bovine ligated ileal loop model was used to study the effect of NE on enteritis induced by
S
. Typhimurium and on the bacterial in vivo replication rate. Mutants lacking putative adrenergic receptor genes were assessed in the loop model, in a calf intestinal colonization model, and in vitro.
S
. Typhimurium-induced enteritis was significantly enhanced by addition of 5 mM NE. This effect was associated with increased net bacterial replication in the same model. Exogenous ferric iron also stimulated bacterial replication in the medium used but not transcription of enteritis-associated loci. The putative adrenergic sensors QseC and QseE were not required for NE-enhanced enteritis, intestinal colonization of calves, or NE-dependent growth in iron-restricted medium and did not influence expression or secretion of enteritis-associated virulence factors. Our findings support a role for stress-related catecholamines in modulating the virulence of enteric bacterial pathogens in vivo but suggest that bacterial adrenergic sensors may not be the vital link in such interkingdom signaling in
Salmonella
.
Characterisation of the liver progenitor cell niche in liver diseases: potential involvement of Wnt and Notch signalling
Authors: Spee Bart , Carpino Guido , Schotanus Baukje A , Katoonizadeh Azeam , Borght Sara Vander , Gaudio Eugenio , Roskams Tania 
Published: November 2009

BackgroundHepatic progenitor cells (HPCs) hold a great potential for therapeutic intervention for currently untreatable liver diseases. However, in human diseases molecular mechanisms involved in proliferation and differentiation of HPCs are poorly understood.Methods and resultsIn the present study activated HPCs and their microenvironment (niche) were investigated in acute and chronic human liver disease by gene-expression analysis and immunohistochemistry/immunofluorescence. Cryopreserved liver tissues were used from patients with parenchymal versus biliary diseases: acute necrotising hepatitis (AH), cirrhosis after hepatitis C infection, and primary biliary cirrhosis in order to study differentiation of HPCs towards hepatocytic versus biliary lineage. Keratin 7 positive HPCs/reactive ductules were captured by means of laser capture microdissection and gene-expression profiles were obtained by using a customised PCR array. Gene expression results were confirmed by immunohistochemistry and immunofluorescence double staining. In all disease groups, microdissected HPCs expressed progenitor cell markers such as KRT7, KRT19, NCAM, ABCG2, LIF, KIT, OCT4, CD44 and TERT. In AH, HPCs were most activated and showed a high expression of prominin-1 (CD133) and α-fetoprotein, and a strong activation of the Wnt pathway. In contrast to parenchymal diseases, HPCs in primary biliary cirrhosis (biliary differentiation) showed a high activation of Notch signalling.ConclusionA distinct pattern of HPC surface markers was found between acute and chronic liver diseases. Similar to what is known from animal experiments, strong evidence has been found signifying the role of Wnt signalling in proliferation of human HPCs whereas Notch signalling is involved in biliary differentiation. These pathways can be targeted in future therapies.
Acid and weakly acidic solutions impair mucosal integrity of distal exposed and proximal non-exposed human oesophagus
Authors: Farré Ricard , Fornari Fernando , Blondeau Kathleen , Vieth Michael , De Vos Rita , Bisschops Raf , Mertens Veerle , Pauwels Ans , Tack Jan , Sifrim Daniel 
Published: November 2009

BackgroundOesophageal mucosa dilated intercellular spaces (DIS) may be important for symptom perception in non-erosive reflux disease (NERD). Patients with NERD might have DIS even in the proximal oesophagus. We aimed to assess the effect of oesophageal perfusions with acid and weakly acidic solutions on ‘exposed’ and ‘non-exposed’ oesophageal mucosa and its relationship to symptoms in healthy subjects.Methods14 healthy volunteers underwent upper gastrointestinal endoscopy with biopsies at 3 and 13 cm proximal to the oesophagogastric junction (OGJ). In following sessions, subjects received 30 min perfusions with neutral, weakly acidic, acidic and acidic-bile acid solutions at 5 cm above the EGJ (separated 4 weeks). Biopsies were taken 20 min after perfusions. Electron microscopy was used to measure DIS. Subjects scored heartburn during perfusions using a visual analogue scale.Results(1) Oesophageal perfusion with acid solutions, with or without bile acids, provoked DIS in the ‘exposed’ oesophageal mucosa; (2) oesophageal perfusion with weakly acidic solutions provoked identical changes to those observed after perfusion with acid solutions; (3) distal oesophageal perfusions not only provoked changes in the ‘exposed’ but also in the more proximal ‘non-exposed’ mucosa; and (4) in spite of the presence of perfusion-induced DIS, most healthy subjects did not perceive heartburn during the experiments.ConclusionsThe human oesophageal mucosa is very sensitive to continuous exposure with acidic and weakly acidic solutions. In spite of the presence of intraluminal acid and DIS, healthy subjects did not experience heartburn, suggesting that NERD patients should have other critical factors underlying their symptoms.
Increased bladder cancer risk among workers exposed too-toluidine and aniline: a reanalysis
Authors: Carreón Tania , Hein Misty J , Viet Susan M , Hanley Kevin W , Ruder Avima M , Ward Elizabeth M 
Published: November 2009

IntroductionIn 1991, the US National Institute for Occupational Safety and Health (NIOSH) reported an increased bladder cancer risk in a cohort of 1749 workers potentially exposed too-toluidine and aniline at a chemical manufacturing plant. As additional information showed that workers in certain departments had been misclassified regardingo-toluidine exposure, we therefore conducted a reanalysis of the data using updated exposure categories.MethodsWe updated exposure categories based on information ascertained during a plant walkthrough, documents on file at the plant, interviews with current and former employees, and answers provided by company and union officials to specific questions. Bladder cancer incidence was determined through 31 December 1988 and mortality through 31 December 1994.ResultsThirteen cases of bladder cancer were observed versus 3.57 expected (New York State rates excluding New York City) (standardised incidence ratio (SIR) 3.64, 95% CI 1.94 to 6.23). Among workers classified as definitely exposed, increasing risks were observed for longer duration of employment (for ≥10 years, standardised rate ratio (SRR) 6.07, 95% CI 0.77 to 48.17) and time since first employment in the exposed departments (for ≥20 years, SRR 3.39, 95% CI 0.40 to 29.03). One bladder cancer death was observed among those definitely exposed.ConclusionsThese findings are comparable to the results reported earlier by NIOSH, and confirm that workers in this plant have an increased risk of bladder cancer.
Development and characterization of genic SSR markers inMedicago truncatulaand their transferability in leguminous and non-leguminous species
Authors: Gupta Sarika , Prasad Manoj 
Published: November 2009

Expressed sequence tag (EST)–derived simple sequence repeat (eSSR) markers are important resources for gene discovery and comparative mapping aimed at crop improvement. In this study, we developed eSSR markers for Medicago truncatula and assessed their cross-species transferability. We detected 36 847 non-redundant sequences (“unigenes”) from 198 642 M. truncatula EST sequences. Mining of microsatellites from the 36 847 unigene sequences (representing ∼25.8 Mb) revealed 14 637 eSSRs in 11 750 SSR-containing ESTs, and primer pairs were successfully designed for 4 636 (39.5%). Of the 14 637 eSSRs, 82.6% were mononucleotide repeats and the rest (in descending order of abundance) were tri-, di-, penta-, and tetranucleotide repeats. When less stringent SSR detection criteria were used, the frequency of dinucleotide repeat motifs increased more than twofold, and the frequencies of di- (11%) and trinucleotide motifs (10.6%) were almost equal. This demonstrates that the eSSR frequency and distribution were related to the choice of search criteria. Forty-one randomly selected primer pairs were validated, and their transferability in three leguminous and three non-leguminous species was assessed. The markers showed a high level of transferability in the leguminous (53%–71%) and non-leguminous (33%–44%) species. The validation studies thus demonstrate the utility of the Medicago eSSRs in assessing genomic relationships in both leguminous and non-leguminous species.
Genome size variation inBegonia
Authors: Dewitte Angelo , Leus Leen , Eeckhaut Tom , Vanstechelman Ives , Van Huylenbroeck Johan , Van Bockstaele Erik 
Published: November 2009

The genome sizes of a Begonia collection comprising 37 species and 23 hybrids of African, Asiatic, Middle American, and South American origin were screened using flow cytometry. Within the collection, 1C values varied between 0.23 and 1.46 pg DNA. Genome sizes were, in most cases, not positively correlated with chromosome number, but with pollen size. A 12-fold difference in mean chromosome size was found between the genotypes with the largest and smallest chromosomes. In general, chromosomes from South American genotypes were smaller than chromosomes of African, Asian, or Middle American genotypes, except for B. boliviensis and B. pearcei. Cytological chromosome studies in different genotypes showed variable chromosome numbers, length, width, and total chromosome volume, which confirmed the diversity in genome size. Large secondary constrictions were present in several investigated genotypes. These data show that chromosome number and structure exhibit a great deal of variation within the genus Begonia, and likely help to explain the large number of taxa found within the genus.
Evaluation of degenerating germ cells in normal juvenile mice
Authors: Trifonova Anastassia , Moens Peter B. 
Published: November 2009

Absence of spermiogenesis in mice with meiotic defects complicates the staging of meiotic arrest using light microscopy. Consequently, new methodologies are required to establish accurate relationships among germ cells. In this study, we utilized a novel approach to analyze germ cell degeneration in juvenile mice. We used terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) in combination with meiosis-specific antibodies. Germ cell degeneration is a normal component of early spermatogenesis in juvenile mice. The incidence of germ cell death was monitored at various postnatal ages of mice using the TUNEL assay to quantify the incidence of apoptosis. Cell death occurred predominantly at 15.5 days after birth. It was found that groups of apoptotic cells were apparent in tubules containing two generations of spermatocytes that form in two progressive cohorts. Electron microscopic observations further illustrated that the majority of cells in the first cohort are in late pachytene, while groups of cells in the second cohort can degenerate in early pachytene. The methodology utilized in this study is significant because it allows one to accurately determine the point at which germ cells arrest. Consequently, we believe that these methods can be applied to study animals with meiotic defects that prevent spermiogenesis.
Presence of triploid cytotypes in the common fig (Ficus carica L.)
Authors: Falistocco E. 
Published: November 2009

Ficus carica (2n = 26) is one of the oldest fruit trees of the Mediterranean basin. Recently there has been increasing interest in this species, in particular for questions related to germplasm such as genetic diversity and cultivar identification. This study was undertaken to gain more knowledge of F. carica cytogenetics and provide data useful for the characterization of its germplasm. Karyomorphological analysis and physical mapping of 18S–25S and 5S rRNA genes by the FISH technique contributed to defining the basic traits of the chromosome complement of F. carica. However, the most interesting result was the discovery of triploid (2n = 39) cytotypes of the cultivated common fig. This result demonstrates the importance of cytogenetic investigations in studies of fig germplasm and emphasizes the role of cross-fertilization as a source of variability not only in wild populations but also in cultivated forms. The results of pollen analysis suggest spontaneous sexual polyploidization as a possible origin of triploid cytotypes. Further studies are necessary to clarify the origin and effective spreading of polyploidy, the presence of other ploidy levels, and their distribution in wild and cultivated forms.
The physiological effects of pre-event and midevent cooling during intermittent running in the heat in elite female soccer players
Authors: Price Michael J. , Boyd Craig , Goosey-Tolfrey Victoria L. 
Published: November 2009

The purpose of this study was to examine the effects of both pre-exercise and combined pre-exercise and midexercise cooling strategies during simulated match play in elite female soccer players in the heat. Eight elite female soccer players performed two 45 min periods of intermittent running separated by 15 min seated rest on 3 separate occasions (30.6 ± 0.2 °C, 63.4 ± 2.5% relative humidity). Participants undertook a no-cooling (CON) or ice-vest cooling for 20 min pre-exercise (PRE) or both pre-exercise and during the 15 min rest period (PRE+MID). Rectal temperature (Tre), skin temperatures, and heart rate were monitored continuously. Mean skin temperature (TMS) and heat storage were calculated. Significant interactions (trial × time) were observed for the change in Tre from rest, TMS, and heat storage (p < 0.05). The change in Tre from rest was greater during CON when compared with PRE and PRE+MID from 35 min until the end of exercise (p < 0.05). When compared with CON (p < 0.05), TMS was lower after precooling (PRE and PRE+MID) and during the 15 min rest period and the first 5 min of the second exercise bout for PRE+MID. Heat storage was also lower after precooling (PRE and PRE+MID) (p < 0.05) and from 60 min until the end of exercise for PRE+MID (p < 0.05) and until 85 min and again at 95 min during PRE (p < 0.05). The results of this study suggest that both cooling strategies were effective in reducing thermal strain during intermittent exercise in the heat. However, PRE+MID cooling was more effective than PRE cooling in offsetting heat storage.
The interleukin-6, serotonin transporter, and monoamine oxidase A genes and endurance performance during the South African Ironman Triathlon
Authors: de Milander Liesl , Stein Dan J. , Collins Malcolm 
Published: November 2009

Previous studies have identified an association of genetic variants believed to alter physiological and biochemical processes locally within the skeletal muscle and therefore performance in the Ironman triathlon. There is growing evidence that the serotonergic system and circulating interleukin (IL)-6 levels are also involved in mediating endurance capacity. Investigators have demonstrated that recombinant human IL-6 administration and serotonergic neurotransmission manipulation, with 5-hydroxytryptamine transporter (5-HTT) and monoamine oxidase A (MAO-A) inhibitors, prior to exercise, can alter running performance, consistent with a central governor hypothesis. The aim of this study was to investigate possible associations of functional polymorphisms within the IL-6 , 5-HTT , and MAO-A genes with endurance performance of Ironman triathletes. Four hundred sixty-eight male Caucasian triathletes who completed the 2000 and (or) 2001 South African Ironman Triathlon and 200 healthy Caucasian male controls were genotyped for the –174 IL-6 G/C, 5-HTT 40 base pair (bp) insertion–deletion and 30 bp variable number of tandem repeats (VNTR) MAO-A gene polymorphisms. There were no significant differences in the relative genotype distributions within the IL-6 (p = 0.636), 5-HTT (p = 0.659), and MOA-A (p = 0.227) polymorphisms when the fastest-fnishing, middle-finishing, and slowest-finishing triathletes, as well as the control groups, were compared. There were no direct associations between the IL-6 –174 G/C, 5-HTT 44 bp insertion–deletion, and MAO-A 30 bp VNTR gene polymorphisms and endurance performance in the 2000 and (or) 2001 South African Ironman Triathlons. The neurogenetic basis of the central governor requires further investigation.