bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cell Adsorption and Selective Desorption for Separation of Microbial Cells by Using Chitosan-Immobilized Silica
Authors: Kubota Munehiro , Matsui Masayoshi , Chiku Hiroyuki , Kasashima Nobuyuki , Shimojoh Manabu , Sakaguchi Kengo 
Published: December 2005

ABSTRACT

Cell adsorption and selective desorption for separation of microbial cells were conducted by using chitosan-immobilized silica (CIS). When chitosan was immobilized onto silica surfaces with glutaraldehyde, bacterial cells adsorbed well and retained viability. Testing of the adsorption and desorption ability of CIS using various microbes such as
Escherichia coli
,
Aeromonas hydrophila
,
Pseudomonas aeruginosa
,
Bacillus subtilis
,
Micrococcus luteus
,
Staphylococcus aureus
,
Staphylococcus epidermidis
,
Lactobacillus casei
,
Streptococcus mutans
,
Streptococcus sobrinus
,
Streptococcus salivarius
,
Saccharomyces cerevisiae
,
Saccharomyces ludwigii
, and
Schizosaccharomyces pombe
revealed that most microbes could be adsorbed and selectively desorbed under different conditions. In particular, recovery was improved when
l
-cysteine was added. A mixture of two bacterial strains adsorbed onto CIS could also be successfully separated by use of specific solutions for each strain. Most of the desorbed cells were alive. Thus, quantitative and selective fractionation of cells is readily achievable by employing chitosan, a known antibacterial material.
Plasmids for Nicotine-Dependent and -Independent Gene Expression in
Arthrobacter nicotinovorans
and Other
Arthrobacter
Species
Authors: Sandu Cristinel , Chiribau Calin-Bogdan , Sachelaru Paula , Brandsch Roderich 
Published: December 2005

ABSTRACT

The first inducible
Arthrobacter
overexpression system, based on the promoter/operator and the repressor of the 6-
d
-hydroxynicotine oxidase gene of
Arthrobacter nicotinovorans
, is described here. Nicotine-dependent overproduction and affinity purification of recombinant proteins are presented. The system will allow the production of complex enzymes and genetic complementation studies in
Arthrobacter
species.
Heterologous Production of Functional Forms of
Rhizopus oryzae
Lipase in
Escherichia coli
Authors: Di Lorenzo Mirella , Hidalgo Aurelio , Haas Michael , Bornscheuer Uwe T. 
Published: December 2005

ABSTRACT

To date, expression of the lipase from
Rhizopus oryzae
(ROL) in
Escherichia coli
always led to the formation of inclusion bodies and inactive protein. However, the production of active ROL and its precursor ProROL in soluble form was achieved when
E. coli
Origami(DE3) and pET-11d were used as expression systems.
Growth and Multiplexed Analysis of Microorganisms on a Subdivided, Highly Porous, Inorganic Chip Manufactured from Anopore
Authors: Ingham Colin J. , van den Ende Maaike , Pijnenburg Dirk , Wever Peter C. , Schneeberger Peter M. 
Published: December 2005

ABSTRACT

A highly porous inorganic material (Anopore) was shown to be an effective support for culturing and imaging a wide range of microorganisms. An inert barrier grid was printed on the rigid surface of Anopore to create a “living chip” of 336 miniaturized compartments (200/cm
2
) with broad applications in microbial culture.
Rapid Detection of Subtype-Selective Nuclear Hormone Receptor Binding with Bacterial Genetic Selection
Authors: Skretas Georgios , Wood David W. 
Published: December 2005

ABSTRACT

Subtype-selective nuclear hormone receptor modulators could potentially allow the development of valuable tissue-specific therapeutics. A simple biosensor that allows subtype-specific nuclear hormone receptor binding to be reflected by the growth phenotype of
Escherichia coli
cells has been constructed. This system will potentially enable the facile detection or evolution of subtype-selective hormone analogues.
Results from the ARTEMIS DISK Global AntifungalSurveillance Study: a 6.5-Year Analysis of Susceptibilitiesof
Candida
and Other Yeast Species to Fluconazole andVoriconazole by Standardized Disk Diffusion Testing
Authors: Pfaller M. A. , Diekema D. J. , Rinaldi M. G. , Barnes R. , Hu B. , Veselov A. V. , Tiraboschi N. , Nagy E. , Gibbs D. L. 
Published: December 2005

ABSTRACT

Fluconazole
in vitro susceptibility test results for 140,767 yeasts were collected
from 127 participating investigators in 39 countries from June 1997
through December 2003. Data were collected on 79,343 yeast isolates
tested with voriconazole from 2001 through 2003. All investigators
tested clinical yeast isolates by the CLSI (formerly NCCLS) M44-A disk
diffusion method. Test plates were automatically read and results were
recorded with the BIOMIC Vision Image Analysis System. Species, drug,
zone diameter, susceptibility category, and quality control results
were collected quarterly via e-mail for analysis. Duplicate (the same
patient, same species, and same susceptible-resistant biotype profile
during any 7-day period) and uncontrolled test results were not
analyzed. The 10 most common species of yeasts all showed less
resistance to voriconazole than to fluconazole.
Candida krusei
showed the largest difference, with over 70% resistance to fluconazole
and less than 8% to voriconazole. All species of yeasts tested were
more susceptible to voriconazole than to fluconazole, assuming proposed
interpretive breakpoints of ≥17 mm (susceptible) and≤
13 mm (resistant) for voriconazole. MICs reported in this
study were determined from the zone diameter in millimeters from the
continuous agar gradient around each disk, which was calibrated with
MICs determined from the standard CLSI M27-A2 broth dilution method by
balanced-weight regression analysis. The results from this
investigation demonstrate the broad spectrum of the azoles for most of
the opportunistic yeast pathogens but also highlight several areas
where resistance may be progressing and/or where previously rare
species may be“
emerging.”
Use of Immunoglobulin G Avidity Assays for Differentiation of Primary from Previous Infections with West Nile Virus
Authors: Levett P. N. , Sonnenberg K. , Sidaway F. , Shead S. , Niedrig M. , Steinhagen K. , Horsman G. B. , Drebot M. A. 
Published: December 2005

ABSTRACTSince its introduction in 1999, West Nile virus (WNV) infections have spread rapidly across the North American continent. Diagnosis of acute WNV infection by detection of WNV-specific immunoglobulin M (IgM) is complicated by the persistence of detectable IgM for more than 1 year in some patients. IgG antibody avidity testing was assessed as a supplemental assay in the diagnosis of current infections. Three groups of serum samples were assayed in parallel by two different IgG avidity test systems (indirect immunofluorescence test [IIFT] and prototype enzyme-linked immunosorbent assay [ELISA]; EUROIMMUN, Luebeck, Germany). Group I (40 sera taken between 2 and 9 days after the onset of influenza-like symptoms) and group II (40 sera taken between 10 and 43 days after onset) were acute and convalescent specimens from patients with a positive anti-WNV IgM test (ELISA; Focus Diagnostics, Cypress, CA). Group III consisted of 43 patient sera collected between 6 and 12 months after infection. IgG antibodies specific for WNV were detected in 38% (ELISA) and 50% (IIFT) of group I sera, in 90% (ELISA and IIFT) of group II sera, and in 100% (ELISA and IIFT) of group III sera. Low-avidity IgG antibodies were demonstrated in 86% (ELISA) and 95% (IIFT) of IgG-positive patient samples taken between 2 and 43 days after the onset of symptoms (groups I and II). High-avidity IgG antibodies were detected in 100% of group III sera obtained 6 months or more after the onset of symptoms (ELISA and IIFT). IgG avidity tests for WNV infections are rapid and simple to perform. The determination of IgG avidity provides additional diagnostic certainty in differentiating between recently acquired and previous infections with WNV.
Genotyping of
Toxoplasma gondii
Strains from Immunocompromised Patients Reveals High Prevalence of Type I Strains
Authors: Khan A. , Su C. , German M. , Storch G. A. , Clifford D. B. , Sibley L. David 
Published: December 2005

ABSTRACT

Toxoplasma gondii
is an important food- and waterborne opportunistic pathogen that causes severe disease in immunocompromised patients.
T. gondii
has an unusual clonal population structure consisting of three widespread lineages known as I, II, and III. To establish the genotypes of strains of
T. gondii
associated with human toxoplasmosis, we have developed a set of four highly sensitive and polymorphic nested PCR markers. Multiplex nested PCR analysis was used to genotype parasites in cerebral spinal fluid samples from 8 of 10 human immunodeficiency virus-positive patients. Remarkably, a majority of these patients had infections with type I strains or strains containing type I alleles, despite the fact that this lineage is normally uncommon in humans and animals. Multiplex analysis of these four unlinked makers was able to distinguish all three common genotypes and also detected two strains with mixed genotypes. Further analysis based on sequencing of a polymorphic intron revealed that one of these recombinant strains was an exotic lineage distinct from the archetypal clonal lineages. The multiplex nested PCR analysis described here will be useful for analyzing the contribution of parasite genotype to toxoplasmosis.
Use of Several Inducer and Substrate Antibiotic Combinations in a Disk Approximation Assay Format To Screen for AmpC Induction in Patient Isolates of
Pseudomonas aeruginosa
,
Enterobacter
spp.,
Citrobacter
spp., and
Serratia
spp
Authors: Dunne W. Michael , Hardin Daniel J. 
Published: December 2005

ABSTRACT

Two-hundred consecutive, single patient isolates of
Enterobacter
spp.,
Serratia
spp.,
Citrobacter
spp., and
Pseudomonas aeruginosa
were evaluated for AmpC production using a variety of inducer-substrate antibiotic combinations in a disk approximation format. The combinations examined included cefoxitin-piperacillin, imipenem-cefotaxime, imipenem-ceftazidime, imipenem-piperacillin-tazobactam, and imipenem-cefoxitin. All isolates were also screened for the presence of extended-spectrum β-lactamase (ESBL) activity. In total, 85.5% of isolates were shown to be inducible for the production of AmpC by one or more inducer/substrate combinations and 11% of all isolates were stably derepressed for the expression of AmpC. Of all of the combinations, imipenem/piperacillin-tazobactam provided the greatest sensitivity (97.1%). All combinations were 100% specific when a positive test was observed. Given this background among these organisms in our institution, it is reasonable to develop an antibiotic reporting strategy that favors the selection of agents for therapy of these organisms that do not serve as labile substrates of AmpC.
Comparison of Two Human Immunodeficiency Virus (HIV) RNA Surrogate Assays to the Standard HIV RNA Assay
Authors: Jennings Cheryl , Fiscus Susan A. , Crowe Suzanne M. , Danilovic Aleksandra D. , Morack Ralph J. , Scianna Salvatore , Cachafeiro Ada , Brambilla Donald J. , Schupbach Jorg , Stevens Wendy , Respess Richard , Varnier Oliviero E. , Corrigan Gary E. , Gronowitz J. Simon , Ussery Michael A. , Bremer James W. 
Published: December 2005

ABSTRACT

Human immunodeficiency virus (HIV) RNA testing is the gold standard for monitoring antiretroviral therapy in HIV-infected patients. However, equipment and reagent costs preclude widespread use of the assay in resource-limited settings. The Perkin-Elmer Ultrasensitive p24 assay and the Cavidi Exavir Load assay both offer potentially simpler, less costly technologies for monitoring viral load. These assays were compared to the Roche Amplicor HIV-1 Monitor Test, v1.5, using panels of clinical samples (subtype B) from HIV-positive subjects and HIV-spiked samples (subtypes A, C, D, CRF_01AE, CRF_02AG, and F). The Ultrasensitive p24 assay detected 100% of the spiked samples with virus loads of >250,000copies/ml and 61% of the clinical samples with virus loads of 219 to 288,850 copies/ml. Detection rates were improved substantially if an external lysis buffer was added to the procedure. The Cavidi assay detected 54 to 100% of spiked samples with virus loads >10,000 copies/ml and 68% of the clinical samples. These detection rates were also greatly improved with a newly implemented version of this kit. Coefficients of variation demonstrate good reproducibility for each of these kits. The results from the Cavidi v1.0, Cavidi v2.0, and Perkin-Elmer, and the Perkin-Elmer Plus external buffers all correlated well with the results from the Roche Monitor Test (
r
= 0.83 to 0.96,
r
= 0.84 to 0.99,
r
= 0.58 to 0.67, and
r
= 0.59 to 0.95, respectively). Thus, the use of these two assays for monitoring patients, together with less-frequent confirmation testing, offers a feasible alternative to frequent HIV RNA testing in resource-limited settings.
Analysis of Genomic Downsizing on the Basis of Region-of-Difference Polymorphism Profiling of
Mycobacterium tuberculosis
Patient Isolates Reveals Geographic Partitioning
Authors: Rao K. Rajender , Kauser Farhana , Srinivas Sriramula , Zanetti Stefania , Sechi Leonardo A. , Ahmed Niyaz , Hasnain Seyed E. 
Published: December 2005

ABSTRACT

Mycobacterium tuberculosis
, the etiological agent of tuberculosis, has lost many coding and noncoding regions in its genome during the course of evolution. We performed region-of-difference (RD) analysis using PCR-based genotyping of 131
M. tuberculosis
clinical isolates obtained from four different countries, namely, India, Peru, Libya, and Angola. Our studies revealed that RD patterns are often distinct for strains circulating in specific geographical regions and can be used to trace the descent and spread of an isolate from its original reservoir. We describe our findings, which show that no single isolate from the four countries (
n
= 131) had all the 15 RDs either deleted or retained. Tuberculosis-specific deletion 1 (TbD1) was found to be conserved in 23% of the Indian isolates, indicating their possible ancient origin. RD9 was the most conserved region, RD11 was predominantly deleted, and RD6 was the most variable among the isolates in our collection irrespective of their geographic region. In contrast to earlier reports, our results demonstrate that the deletion of RD1 does not correlate with a decrease in the virulence potential of
M. tuberculosis
, as Indian isolates (
n
= 30) examined by us were from diseased individuals and yet had lost the RD1 region. Our results further illustrated that the intactness of the RD5 region may be associated with increased virulence of the organism. This study highlights that the RDs in
M. tuberculosis
genomes are geographically distributed and specific and may possibly be associated with virulence spectrum.
Multilocus Sequence Typing of
Neisseria meningitidis
Directly from Clinical Samples and Application of the Method to the Investigation of Meningococcal Disease Case Clusters
Authors: Birtles Andrew , Hardy Katie , Gray Stephen J. , Handford Suzanne , Kaczmarski Edward B. , Edwards-Jones Valerie , Fox Andrew J. 
Published: December 2005

ABSTRACT

Infections associated with
Neisseria meningitidis
are a major public health problem in England, Wales, and Northern Ireland. Currently, over 40% of cases are confirmed directly from clinical specimens using PCR-based methodologies without an organism being isolated. A nested/seminested multilocus sequence typing (MLST) system was developed at the Health Protection Agency Meningococcal Reference Unit to allow strain characterization beyond the serogroup for cases confirmed by PCR only. This system was evaluated on a panel of 20 meningococcus-positive clinical specimens (3 cerebrospinal fluid and 17 blood samples) from different patients containing various concentrations of meningococcal DNA that had corresponding
N. meningitidis
isolates. In each case, the sequence type generated from the clinical specimens matched that produced from the corresponding
N. meningitidis
isolate; the sensitivity of the MLST system was determined to be less than 12 genome copies per PCR. The MLST system was then applied to 15 PCR meningococcus-positive specimens (2 cerebrospinal fluid and 13 blood samples), each from a different patient, involved in three case clusters (two serogroup B and one serogroup W135) for which no corresponding
N. meningitidis
organisms had been isolated. In each case, an MLST sequence type was generated, allowing the accurate assignment of individual cases within each of the case clusters. In summary, the adaptation of the
N. meningitidis
MLST to a sensitive nested/seminested format for strain characterization directly from clinical specimens provides an important tool for surveillance and management of meningococcal infection.
Molecular Fingerprinting of
Leishmania infantum
Strains following an Outbreak of Visceral Leishmaniasis in Central Israel
Authors: Nasereddin Abedelmajeed , Baneth Gad , Schönian Gabriele , Kanaan Moein , Jaffe Charles L. 
Published: December 2005

ABSTRACT

Human and canine visceral leishmaniasis caused by
Leishmania infantum
emerged in central Israel after an absence of over 30 years. The origin of this outbreak was investigated by examining genetic polymorphisms in 37 strains isolated from dogs and patients with visceral leishmaniasis in the continuously active northern Israeli and West Bank foci and in a new Israeli focus using DNA fingerprinting with the human multilocus minisatellite probe 33.15. Analysis of the patterns obtained by DNA fingerprinting separated the strains geographically into northern (clade B) and central (clades A and C) genotypic groups. These results suggest that the emergence of visceral leishmaniasis in central Israel is due not to parasite spread from northern Israel to the new focus but rather to increased parasite transmission in central Israel and the West Bank coupled with changes in the ecoepidemiology of this region.
Bacterial Characteristics in Relation to Clinical Source of
Escherichia coli
Isolates from Women with Acute Cystitis or Pyelonephritis and Uninfected Women
Authors: Johnson James R. , Owens Krista , Gajewski Abby , Kuskowski Michael A. 
Published: December 2005

ABSTRACT

Characteristics differentiating
Escherichia coli
strains that cause cystitis or pyelonephritis from fecal
E. coli
remain incompletely defined, particularly among adult women in the United States. Accordingly, phylogenetic group, O antigens, and virulence factors (VFs) were analyzed among 329
E. coli
isolates from the mid-to-late 1990s from women in the United States with acute pyelonephritis (
n
= 170), cystitis (
n
= 83), or no infection (fecal;
n
= 76). Compared with fecal and cystitis isolates, pyelonephritis isolates exhibited a greater prevalence of phylogenetic group B2, most virulence-associated O antigens, and most VFs and had higher VF scores. In contrast, cystitis and fecal isolates differed minimally. By stepwise multivariable logistic regression, significant (
P
≤ 0.015) predictors of cystitis and/or pyelonephritis (versus fecal) included
afa/dra
(Dr-binding adhesins),
ibeA
(invasion of brain endothelium),
iha
(putative adhesin-siderophore),
malX
(pathogenicity island marker), the O75 antigen,
papEF
(P fimbriae),
papG
allele II (P adhesin variant), group B2, and
sfa
/
foc
(S and F1C fimbriae). However, virulence profiles overlapped considerably among source groups and varied greatly within each group.
E. coli
“clonal group A” (CGA) and the O2:K5/K7:H1 and O75:K+ clonal groups were significantly associated with cystitis and/or pyelonephritis. These findings identify potential vaccine targets, suggest that urovirulence is multiply determined, and confirm the urovirulence of specific
E. coli
clonal groups, including recently recognized CGA.
Molecular Evolution of Human Influenza A/H3N2 Virus in Asia and Europe from 2001 to 2003
Authors: Chi X. Sherry , Bolar Trentice V. , Zhao Ping , Tam John S. , Rappaport Ruth , Cheng Sheau-Mei 
Published: December 2005

ABSTRACT
Hemagglutinin sequences of 146 human influenza A/H3N2 strains identified in respiratory specimens from Asia and Europe during the 2001-2003 influenza seasons were analyzed by DNA sequencing. Our results suggest that four amino acid substitutions, L25I, H75Q, H155T, and Q156H, led to the antigenic conversion of the previously predominant A/Panama/2007/99-like strains to the more recent A/Fujian/411/2002-like strains.
Comparison of the Binax NOW Flu A Enzyme Immunochromatographic Assay and R-Mix Shell Vial Culture for the 2003-2004 Influenza Season
Authors: Fader Robert C. 
Published: December 2005

ABSTRACT
The Binax NOW Flu A enzyme immunochromatographic assay was compared to viral culture with R-Mix shell vials for 455 nasal-wash or nasal-aspirate specimens. The overall sensitivity, specificity, positive predictive value, and negative predictive value of the assay were 64.9%, 98.4%, 89.3%, and 93.2%, respectively. However, the assay sensitivity decreased significantly with increasing patient age.
Development of a Triplex Real-Time PCR Assay for Detection of Panton-Valentine Leukocidin Toxin Genes in Clinical Isolates of Methicillin-Resistant
Staphylococcus aureus
Authors: McDonald Ryan R. , Antonishyn Nick A. , Hansen Toni , Snook Laelie A. , Nagle Evelyn , Mulvey Michael R. , Levett Paul N. , Horsman Greg B. 
Published: December 2005

ABSTRACT

Community-associated methicillin-resistant
Staphylococcus aureus
harboring Panton-Valentine leukocidin (PVL) genes is an emerging pathogen. A novel real-time PCR assay for identification of MRSA isolates containing PVL was developed. The PVL assay was used in a triplex format allowing simultaneous amplification of
mecA
,
nuc
, and PVL genes in 614 clinical isolates. This assay facilitates the rapid identification of PVL-positive isolates of MRSA.
Detection and Serotyping of
Streptococcus pneumoniae
from Nasopharyngeal Samples by PCR-Based Multiplex Assay
Authors: Moreno Jaime , Hernández Elkin , Sanabria Olga , Castañeda Elizabeth 
Published: December 2005

ABSTRACT

We developed a multiplex PCR-based methodology for nasopharyngeal samples maintained in egg thioglycolate antibiotic and skim milk-tryptone-glucose-glycerol media to identify and serotype the most important serotypes of
Streptococcus pneumoniae
that cause invasive disease in children. This technique can be used to study the epidemiology of pneumococcal colonization and the effect of conjugate vaccines.
Molecular Epidemiology of an Outbreak Caused by Methicillin-Resistant
Staphylococcus aureus
in a Health Care Ward and Associated Nursing Home
Authors: Kerttula Anne-Marie , Lyytikäinen Outi , Vuopio-Varkila Jaana , Ibrahem Salha , Agthe Niina , Broas Markku , Jägerroos Henrik , Virolainen Anni 
Published: December 2005

ABSTRACT

Our point-prevalence survey followed an outbreak of methicillin-resistant
Staphylococcus aureus
(MRSA) in a long-term care facility and identified five MRSA strains, of which two possessed an outbreak genotype not encountered previously and three had another profile. All of them possessed SCC
mec
type V. Six methicillin-sensitive
S. aureus
strains were genotypically related to the epidemic strains.
Frequent Importation of Enterovirus 71 from Surrounding Countries into the Local Community of Yamagata, Japan, between 1998 and 2003
Authors: Mizuta K. , Abiko C. , Murata T. , Matsuzaki Y. , Itagaki T. , Sanjoh K. , Sakamoto M. , Hongo S. , Murayama S. , Hayasaka K. 
Published: December 2005

ABSTRACT
Phylogenetic analysis of 45 enterovirus 71 (EV71) isolates for 6 years in Yamagata, Japan, clarified that the annual outbreak of hand-foot-and-mouth disease was due to four genetically distinct subgenogroups, including a novel “B5.” Our results suggest that the importation of EV71 from surrounding countries has had a major epidemiological impact on the local community used in our study.