bluuebunny/crossref_metadata_2025_split huggingface dataset:
AN ANTIBODY MODIFIED AUTOMATED ENZYME‐LINKED IMMUNOSORBENT ASSAY‐BASED METHOD FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN*
Authors: BENNETT REGINALD W. 
Published: December 2008

ABSTRACT Studies were conducted with an automated enzyme‐linked immunosorbent assay (ELISA)‐based method (Vidas, Staph enterotoxin‐II [SET‐II]), exhibiting an antibody capture that had undergone modification by removal of the Fc fragment on the antibody. Raw liquid or shell eggs containing a nontoxin component with an attraction to the staphylococcal antienterotoxins were studied. Prior to ELISA testing, the eggs were homogenized and extracts collected by centrifugation. Studies showed that regardless of the ELISA‐based method used that utilized the unmodified antibodies with both the Fab1 + Fc fragments intact, positive (false positive) ELISA responses occurred with fertilized egg yolks and fertilized whole liquid or whole shell eggs. Conversely, when modified (Fab1) antibodies were used, the automated SET‐II enzyme‐linked fluorescent immunoassay was negative.PRACTICAL APPLICATIONSMost enzyme‐linked immunosorbent assays as well as other serological systems use the whole antibody that has not undergone modification for the detection of the staphylococcal enterotoxins. The use of whole antibody (Fab1 + Fc fragments) has on occasion produced false positive results. However, the antibody in which the Fc fragment has been removed leaving the Fab1 fragment provides a more specific antibody for the identification of this microbial toxin. The use of modified antibody (Fab1 fragment) represents significant improvement in antibody quality thus ensuring a greater degree of specificity without sacrificing the sensitivity of serological methods for the detection of staphylococcal enterotoxin in foods.
Health Care Reform: Lessons from the Past, Lessons for the Future
Authors: Wilensky Gail R. 
Published: December 2008

We are well into the political season that guarantees the election of a new president. Actually, this season, the election cycle began in November 2006, as soon as the off-year election ended. Not surprisingly, health care and reforming health care are major issues for the election — although somewhat less important than they were before late 2007.I use the phrase “not surprisingly” because there are easily understandable reasons why health care tends to be an election issue whenever we are electing a new president. One obvious reason is that, for several decades, we have been plagued by sizeable numbers of people without health insurance and even more importantly, by unsustainable increases in health care spending and substantial numbers of people experiencing patient safety and clinical appropriateness problems. I regard the unsustainable spending and safety and quality problems as especially important because they affect all of us — not just the 15% of the population that is without health insurance.
Influence of Water Temperature on Morphological Deformities in Cultured Larvae of Japanese Eel, Anguilla japonica, at Completion of Yolk Resorption
Authors: Kurokawa Tadahide , Okamoto Takuma , Gen Koichiro , Uji Susumu , Murashita Koji , Unuma Tatsuya , Nomura Kazuharu , Matsubara Hajime , Kim Shin‐Kwon , Ohta Hiromi , Tanaka Hideki 
Published: December 2008

Abstract The occurrence of morphological deformities under different rearing water temperatures (18, 20, 22, 24, 26, 28, and 30 C) was examined in Japanese eel larvae. The rates of hatching and survival until yolk resorption at 22–26 C were higher than those at other water temperatures. Fertilized eggs never hatched at 18 and 30 C. The rates of occurrence of abnormal larvae reared at the water temperatures 24–28 C were lower than those at 20 or 22 C. Pericardial edema and lower jaw deformities occurred most frequently at lower temperatures (20 and 22 C). In contrast, the incubation temperature did not significantly affect the relative frequency of some neurocranial deformities and of spinal curvature. These results imply that the optimal temperatures for rearing Japanese eel eggs and embryos are 24–26 C from the viewpoints of survival and deformity.
Effects of Fish Size and Water Temperature on the Acute Toxicity of Copper for Japanese Flounder, Paralichthys olivaceus, and Red Sea Bream, Pagrus major
Authors: Furuta Takeshi , Iwata Nakahiro , Kikuchi Kotaro 
Published: December 2008

Abstract The acute toxicities of copper were examined for Japanese flounder, Paralichthys olivaceus, and red sea bream, Pagrus major, in terms of fish size and water temperature. Artificial seawater of low pH of 5.4–6.7 was used as testing water to keep dissolved copper concentration at 0.04–41 mg Cu/L. Japanese flounder of 0.3–17 g and red sea bream of 0.5–13 g were exposed to different concentrations of copper for 96 h at 20 C under semistatic condition. Median‐lethal concentration for 96 h of Japanese flounder and red sea bream were 8.7–12.2 and 2.0–5.2 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and fish size for Japanese flounder, while the value decreased significantly with increasing fish size for red sea bream. Effect of water temperature on the acute toxicity was examined for Japanese flounder of 0.3 and 0.4 g at 10, 15, 20, and 25 C and red sea bream of 0.5 and 1.0 g at 12, 15, 20, and 25 C. Ninety‐six‐hour median‐lethal concentrations for Japanese flounder and red sea bream were 5.1–11.2 and 1.0–5.3 mg Cu/L, respectively. No significant relationships were observed between median‐lethal concentrations for 96 h and water temperature for both fish species.
Multinormal In Vitro Distribution Model Suitable for the Distribution ofPlasmodium falciparumChemosusceptibility to Doxycycline
Authors: Briolant Sébastien , Baragatti Meili , Parola Philippe , Simon Fabrice , Tall Adama , Sokhna Cheikh , Hovette Philippe , Mamfoumbi Modeste Mabika , Koeck Jean-Louis , Delmont Jean , Spiegel André , Castello Jacky , Gardair Jean Pierre , Trape Jean Francois , Kombila Maryvonne , Minodier Philippe , Fusai Thierry , Rogier Christophe , Pradines Bruno 
Published: December 2008

ABSTRACTThe distribution and range of 50% inhibitory concentrations (IC50s) of doxycycline were determined for 747 isolates obtained between 1997 and 2006 from patients living in Senegal, Republic of the Congo, and Gabon and patients hospitalized in France for imported malaria. The statistical analysis was designed to answer the specific question of whetherPlasmodium falciparumhas different phenotypes of susceptibility to doxycycline. A triple normal distribution was fitted to the data using a Bayesian mixture modeling approach. The IC50geometric mean ranged from 6.2 μM to 11.1 μM according to the geographical origin, with a mean of 9.3 μM for all 747 parasites. The values for all 747 isolates were classified into three components: component A, with an IC50mean of 4.9 μM (±2.1 μM [standard deviation]); component B, with an IC50mean of 7.7 μM (±1.2 μM); and component C, with an IC50mean of 17.9 μM (±1.4 μM). According to the origin of theP. falciparumisolates, the triple normal distribution was found in each subgroup. However, the proportion of isolates predicted to belong to component B was most important in isolates from Gabon and Congo and in isolates imported from Africa (from 46 to 56%). In Senegal, 55% of theP. falciparumisolates were predicted to be classified as component C. The cutoff of reduced susceptibility to doxycycline in vitro was estimated to be 35 μM.
Immunity to the Bacteriocin Sublancin 168 Is Determined by the SunI (YolF) Protein of
Bacillus subtilis
Authors: Dubois Jean-Yves F. , Kouwen Thijs R. H. M. , Schurich Anna K. C. , Reis Carlos R. , Ensing Hendrik T. , Trip Erik N. , Zweers Jessica C. , van Dijl Jan Maarten 
Published: December 2008

ABSTRACT

Bacillus subtilis
strain 168 produces the extremely stable lantibiotic sublancin 168, which has a broad spectrum of bactericidal activity. Both sublancin 168 production and producer immunity are determined by the SPβ prophage. While the
sunA
and
sunT
genes for sublancin 168 production have been known for several years, the genetic basis for sublancin 168 producer immunity has remained elusive. Therefore, the present studies were aimed at identifying an SPβ gene(s) for sublancin 168 immunity. By systematic deletion analysis, we were able to pinpoint one gene, named
yolF
, as the sublancin 168 producer immunity gene. Growth inhibition assays performed using plates and liquid cultures revealed that YolF is both required and sufficient for sublancin 168 immunity even when heterologously produced in the sublancin-sensitive bacterium
Staphylococcus aureus
. Accordingly, we propose to rename
yolF
to
sunI
(for
su
blanci
n i
mmunity). Subcellular localization studies indicate that the SunI protein is anchored to the membrane with a single N-terminal membrane-spanning domain that has an N
out
-C
in
topology. Thus, the bulk of the protein faces the cytoplasm of
B. subtilis
. This topology has not yet been reported for known bacteriocin producer immunity proteins, which implies that SunI belongs to a novel class of bacteriocin antagonists.
Candidate Polyanionic Microbicides Inhibit Human T-Cell Lymphotropic Virus Type 1 Receptor Interactions, Cell-Free Infection, and Cell-Cell Spread
Authors: Romer Daniela , Brighty David W. , Robson Cynthia L. , Sattentau Quentin J. 
Published: December 2008

ABSTRACTThe human T-cell lymphotropic virus type 1 (HTLV-1) is the cause of adult T-cell leukemia and inflammatory diseases including HTLV-1-associated myelopathy/tropical spastic paraparesis. HTLV-1 can be transmitted through sexual contact, mother-to-child transmission, and exposure to contaminated blood. Microbicides are agents that interfere with microbial infectivity at mucous membranes, and candidates are under development for use against sexually transmitted viruses such as human immunodeficiency virus type 1. We previously demonstrated that cell surface polyanionic heparan sulfate proteoglycans bind the HTLV-1 envelope glycoprotein surface subunit gp46, facilitating cell-cell and cell-free virus spread in vitro. We now show, using assays for Env-receptor binding inhibition, Env-induced cell-cell fusion, cell-cell virus spread, and pseudotype HTLV-1 infectivity, that the soluble polyanions PRO 2000 and dextran sulfate are potent inhibitors of HTLV-1 spread in vitro, with PRO 2000 being the more promising candidate. The results of these studies suggest that candidate topical microbicides may be of use in reducing HTLV-1 sexual transmission.
Optimized Method for Preparation of DNA from Pathogenic and Environmental Mycobacteria
Authors: Käser Michael , Ruf Marie-Thérèse , Hauser Julia , Marsollier Laurent , Pluschke Gerd 
Published: December 2008

ABSTRACT
Genomic studies on pathogenic and environmental mycobacteria are of growing interest for understanding of their evolution, distribution, adaptation, and host-pathogen interaction. Since most mycobacteria are slow growers, material from in vitro cultures is usually scarce. The robust mycobacterial cell wall hinders both experimental cell lysis and efficient DNA extraction. Here, we compare elements of several DNA preparation protocols and describe a method that is economical and practical and reliably yields large amounts—usually 10-fold increased compared to earlier protocols—of highly pure genomic DNA for sophisticated downstream applications. This method was optimized for cultures of a variety of pathogenic and environmental mycobacterial species and proven to be suitable for direct mycobacterial DNA extraction from infected insect specimens.
Analysis of Dibenzothiophene Desulfurization in a Recombinant
Pseudomonas putida
Strain
Authors: Calzada Javier , Zamarro María T. , Alcón Almudena , Santos Victoria E. , Díaz Eduardo , García José L. , Garcia-Ochoa Felix 
Published: December 2008

ABSTRACT

Biodesulfurization was monitored in a recombinant
Pseudomonas putida
CECT5279 strain. DszB desulfinase activity reached a sharp maximum at the early exponential phase, but it rapidly decreased at later growth phases. A model two-step resting-cell process combining sequentially
P. putida
cells from the late and early exponential growth phases was designed to significantly increase biodesulfurization.
Antimicrobial Resistance in
Escherichia coli
Isolates from Swine and Wild Small Mammals in the Proximity of Swine Farms and in Natural Environments in Ontario, Canada
Authors: Kozak Gosia K. , Boerlin Patrick , Janecko Nicol , Reid-Smith Richard J. , Jardine Claire 
Published: December 2008

ABSTRACT

Wild animals not normally exposed to antimicrobial agents can acquire antimicrobial agent-resistant bacteria through contact with humans and domestic animals and through the environment. In this study we assessed the frequency of antimicrobial resistance in generic
Escherichia coli
isolates from wild small mammals (mice, voles, and shrews) and the effect of their habitat (farm or natural area) on antimicrobial resistance. Additionally, we compared the types and frequency of antimicrobial resistance in
E. coli
isolates from swine on the same farms from which wild small mammals were collected. Animals residing in the vicinity of farms were five times more likely to carry
E. coli
isolates with tetracycline resistance determinants than animals living in natural areas; resistance to tetracycline was also the most frequently observed resistance in isolates recovered from swine (83%). Our results suggest that
E. coli
isolates from wild small mammals living on farms have higher rates of resistance and are more frequently multiresistant than
E. coli
isolates from environments, such as natural areas, that are less impacted by human and agricultural activities. No
Salmonella
isolates were recovered from any of the wild small mammal feces. This study suggests that close proximity to food animal agriculture increases the likelihood that
E. coli
isolates from wild animals are resistant to some antimicrobials, possibly due to exposure to resistant
E. coli
isolates from livestock, to the resistance genes of these isolates, or to antimicrobials through contact with animal feed.
Simultaneous Analysis of Bacterioferritin Gene Expression and Intracellular Iron Status in
Pseudomonas putida
KT2440 by Using a Rapid Dual Luciferase Reporter Assay
Authors: Chen Shicheng , Bleam William F. , Hickey William J. 
Published: December 2008

ABSTRACT

A dual luciferase reporter (DLR) system utilizing firefly and
Renilla
luciferases was developed and tested in a model rhizobacterium,
Pseudomonas putida
KT2440. The DLR was applied to simultaneously analyze expression of three putative bacterioferritin genes (
bfr
α,
bfr
β, and
bfr
) and assess the cellular iron status of strain KT2440 by monitoring expression of the Fur-regulated
fepA
-
fes
promoter. The DLR proved to be reproducible and sensitive. Expression of
bfr
α (PP0482) and
bfr
β (PP1082) was consistent with expectations for bacterioferritin and varied directly with the iron level. However, expression of
bfr
(PP4856) was inversely related to the iron concentration and it was thus more likely to encode a Dps-like protein rather than a bacterioferritin.
Phage Lysin LysK Can Be Truncated to Its CHAP Domain and Retain Lytic Activity against Live Antibiotic-Resistant Staphylococci
Authors: Horgan Marianne , O’Flynn Gary , Garry Jennifer , Cooney Jakki , Coffey Aidan , Fitzgerald Gerald F. , Ross R. Paul , McAuliffe Olivia 
Published: December 2008

ABSTRACT

A truncated derivative of the phage endolysin LysK containing only the CHAP (cysteine- and histidine-dependent amidohydrolase/peptidase) domain exhibited lytic activity against live clinical staphylococcal isolates, including methicillin-resistant
Staphylococcus aureus
. This is the first known report of a truncated phage lysin which retains high lytic activity against live staphylococcal cells.
Characterization of Serine Proteinase Expression in
Agaricus bisporus
and
Coprinopsis cinerea
by Using Green Fluorescent Protein and the
A. bisporus SPR1
Promoter
Authors: Heneghan Mary N. , Porta Claudine , Zhang Cunjin , Burton Kerry S. , Challen Michael P. , Bailey Andy M. , Foster Gary D. 
Published: December 2008

ABSTRACT

The
Agaricus bisporus
serine proteinase 1 (SPR1) appears to be significant in both mycelial nutrition and senescence of the fruiting body. We report on the construction of an SPR promoter::green fluorescent protein (GFP) fusion cassette, pGreen_hph1_SPR_GFP, for the investigation of temporal and developmental expression of SPR1 in homobasidiomycetes and to determine how expression is linked to physiological and environmental stimuli. Monitoring of
A. bisporus
pGreen_hph1_SPR_GFP transformants on media rich in ammonia or containing different nitrogen sources demonstrated that SPR1 is produced in response to available nitrogen. In
A. bisporus
fruiting bodies, GFP activity was localized to the stipe of postharvest senescing sporophores. pGreen_hph1_SPR_GFP was also transformed into the model basidiomycete
Coprinopsis cinerea
. Endogenous
C. cinerea
proteinase activity was profiled during liquid culture and fruiting body development. Maximum activity was observed in the mature cap, while activity dropped during autolysis. Analysis of the
C. cinerea
genome revealed seven genes showing significant homology to the
A. bisporus SPR1
and
SPR2
genes. These genes contain the aspartic acid, histidine, and serine residues common to serine proteinases. Analysis of the promoter regions revealed at least one CreA and several AreA regulatory motifs in all sequences. Fruiting was induced in
C. cinerea
dikaryons, and fluorescence was determined in different developmental stages. GFP expression was observed throughout the life cycle, demonstrating that serine proteinase can be active in all stages of
C. cinerea
fruiting body development. Serine proteinase expression (GFP fluorescence) was most concentrated during development of young tissue, which may be indicative of high protein turnover during cell differentiation.
Comparative Genomics Reveal Extensive Transposon-Mediated Genomic Plasticity and Diversity among Potential Effector Proteins within the GenusCoxiella
Authors: Beare Paul A. , Unsworth Nathan , Andoh Masako , Voth Daniel E. , Omsland Anders , Gilk Stacey D. , Williams Kelly P. , Sobral Bruno W. , Kupko John J. , Porcella Stephen F. , Samuel James E. , Heinzen Robert A. 
Published: December 2008

ABSTRACTGenetically distinct isolates ofCoxiella burnetii, the cause of human Q fever, display different phenotypes with respect to in vitro infectivity/cytopathology and pathogenicity for laboratory animals. Moreover, correlations betweenC. burnetiigenomic groups and human disease presentation (acute versus chronic) have been described, suggesting that isolates have distinct virulence characteristics. To provide a more-complete understanding ofC. burnetii's genetic diversity, evolution, and pathogenic potential, we deciphered the whole-genome sequences of the K (Q154) and G (Q212) human chronic endocarditis isolates and the naturally attenuated Dugway (5J108-111) rodent isolate. Cross-genome comparisons that included the previously sequenced Nine Mile (NM) reference isolate (RSA493) revealed both novel gene content and disparate collections of pseudogenes that may contribute to isolate virulence and other phenotypes. WhileC. burnetiigenomes are highly syntenous, recombination between abundant insertion sequence (IS) elements has resulted in genome plasticity manifested as chromosomal rearrangement of syntenic blocks and DNA insertions/deletions. The numerous IS elements, genomic rearrangements, and pseudogenes ofC. burnetiiisolates are consistent with genome structures of other bacterial pathogens that have recently emerged from nonpathogens with expanded niches. The observation that the attenuated Dugway isolate has the largest genome with the fewest pseudogenes and IS elements suggests that this isolate's lineage is at an earlier stage of pathoadaptation than the NM, K, and G lineages.
Genome-Wide Transposon Mutagenesis in Pathogenic
Leptospira
Species
Authors: Murray Gerald L. , Morel Viviane , Cerqueira Gustavo M. , Croda Julio , Srikram Amporn , Henry Rebekah , Ko Albert I. , Dellagostin Odir A. , Bulach Dieter M. , Sermswan Rasana W. , Adler Ben , Picardeau Mathieu 
Published: December 2008

ABSTRACT

Leptospira interrogans
is the most common cause of leptospirosis in humans and animals. Genetic analysis of
L. interrogans
has been severely hindered by a lack of tools for genetic manipulation. Recently we developed the
mariner
-based transposon
Himar1
to generate the first defined mutants in
L. interrogans
. In this study, a total of 929 independent transposon mutants were obtained and the location of insertion determined. Of these mutants, 721 were located in the protein coding regions of 551 different genes. While sequence analysis of transposon insertion sites indicated that transposition occurred in an essentially random fashion in the genome, 25 unique transposon mutants were found to exhibit insertions into genes encoding 16S or 23S rRNAs, suggesting these genes are insertional hot spots in the
L. interrogans
genome. In contrast, loci containing notionally essential genes involved in lipopolysaccharide and heme biosynthesis showed few transposon insertions. The effect of gene disruption on the virulence of a selected set of defined mutants was investigated using the hamster model of leptospirosis. Two attenuated mutants with disruptions in hypothetical genes were identified, thus validating the use of transposon mutagenesis for the identification of novel virulence factors in
L. interrogans
. This library provides a valuable resource for the study of gene function in
L. interrogans
. Combined with the genome sequences of
L. interrogans
, this provides an opportunity to investigate genes that contribute to pathogenesis and will provide a better understanding of the biology of
L. interrogans
.
Crystal Structure of ORF12 fromLactococcus lactisPhage p2 Identifies a Tape Measure Protein Chaperone
Authors: Siponen Marina , Sciara Giuliano , Villion Manuela , Spinelli Silvia , Lichière Julie , Cambillau Christian , Moineau Sylvain , Campanacci Valérie 
Published: December 2008

ABSTRACTWe report here the characterization of the nonstructural protein ORF12 of the virulent lactococcal phage p2, which belongs to theSiphoviridaefamily. ORF12 was produced as a soluble protein, which forms large oligomers (6- to 15-mers) in solution. Using anti-ORF12 antibodies, we have confirmed that ORF12 is not found in the virion structure but is detected in the second half of the lytic cycle, indicating that it is a late-expressed protein. The structure of ORF12, solved by single anomalous diffraction and refined at 2.9-Å resolution, revealed a previously unknown fold as well as the presence of a hydrophobic patch at its surface. Furthermore, crystal packing of ORF12 formed long spirals in which a hydrophobic, continuous crevice was identified. This crevice exhibited a repeated motif of aromatic residues, which coincided with the same repeated motif usually found in tape measure protein (TMP), predicted to form helices. A model of a complex between ORF12 and a repeated motif of the TMP of phage p2 (ORF14) was generated, in which the TMP helix fitted exquisitely in the crevice and the aromatic patches of ORF12. We suggest, therefore, that ORF12 might act as a chaperone for TMP hydrophobic repeats, maintaining TMP in solution during the tail assembly of the lactococcal siphophage p2.
Nanoarchaeal Origin of Histone H3?
Authors: Friedrich-Jahn Ulrike , Aigner Johanna , Längst Gernot , Reeve John N. , Huber Harald 
Published: December 2008

ABSTRACTNEQ288, one of two archaeal histones inNanoarchaeum equitans, has a unique four-residue insertion that closely resembles an insertion in the eukaryotic histone H3 lineage. NEQ288 bound DNA but did not compact DNA in vitro in the absence of NEQ348, the secondN. equitansarchaeal histone. The properties of NEQ288 suggest an intermediate between the archaeal and H3 histone lineages and an evolutionary step toward the now-mandatory assembly of eukaryotic histones into heterodimers.