bluuebunny/crossref_metadata_2025 huggingface dataset:
Toxin-Producing Ability among
Bacillus
spp. Outside the
Bacillus cereus
Group
Authors: From Cecilie , Pukall Rudiger , Schumann Peter , Hormazábal Víctor , Granum Per Einar
Published: March 2005

A total of 333
Bacillus
spp. isolated from foods, water, and food plants were examined for the production of possible enterotoxins and emetic toxins using a cytotoxicity assay on Vero cells, the boar spermatozoa motility assay, and a liquid chromatography-mass spectrometry method. Eight strains produced detectable toxins; six strains were cytotoxic, three strains produced putative emetic toxins (different in size from cereulide), and one strain produced both cytotoxin(s) and putative emetic toxin(s). The toxin-producing strains could be assigned to four different species,
B. subtilis
,
B. mojavensis
,
B. pumilus
, or
B. fusiformis
, by using a polyphasic approach including biochemical, chemotaxonomic, and DNA-based analyses. Four of the strains produced cytotoxins that were concentrated by ammonium sulfate followed by dialysis, and two strains produced cytotoxins that were not concentrated by such a treatment. Two cultures maintained full cytotoxic activity, two cultures reduced their activity, and two cultures lost their activity after boiling. The two most cytotoxic strains (both
B. mojavensis
) were tested for toxin production at different temperatures. One of these strains produced cytotoxin at growth temperatures ranging from 25 to 42°C, and no reduction in activity was observed even after 24 h of growth at 42°C. The strains that produced putative emetic toxins were tested for the influence of time and temperature on the toxin production. It was shown that they produced putative emetic toxin faster or just as fast at 30 as at 22°C. None of the cytotoxic strains produced
B. cereus
-like enterotoxins as tested by PCR or by immunological methods.
Nodulation ofLupinus albusby Strains ofOchrobactrum lupinisp. nov
Authors: Trujillo Martha E. , Willems Anne , Abril Adriana , Planchuelo Ana-María , Rivas Raúl , Ludeña Dolores , Mateos Pedro F. , Martínez-Molina Eustoquio , Velázquez Encarna
Published: March 2005

The nodulation of legumes has for more than a century been considered an exclusive capacity of a group of microorganisms commonly known as rhizobia and belonging to the α-Proteobacteria. However, in the last 3 years four nonrhizobial species, belonging to α and β subclasses of theProteobacteria, have been described as legume-nodulating bacteria. In the present study, two fast-growing strains, LUP21 and LUP23, were isolated from nodules ofLupinus honoratus.The phylogenetic analysis based on the 16S and 23S rRNA gene sequences showed that the isolates belong to the genusOchrobactrum. The strains were able to reinfectLupinusplants. A plasmid profile analysis showed the presence of three plasmids. ThenodDandnifHgenes were located on these plasmids, and their sequences were obtained. These sequences showed a close resemblance to thenodDandnifHgenes of rhizobial species, suggesting that thenodDandnifHgenes carried by strain LUP21Twere acquired by horizontal gene transfer. A polyphasic study including phenotypic, chemotaxonomic, and molecular features of the strains isolated in this study showed that they belong to a new species of the genusOchrobactrumfor which we propose the nameOchrobactrum lupinisp. nov. Strain LUP21T(LMG 20667T) is the type strain.
In Vitro Reconstitution of an NADPH-Dependent Superoxide Reduction Pathway from
Pyrococcus furiosus
Authors: Grunden Amy M. , Jenney Francis E. , Ma Kesen , Ji Mikyoung , Weinberg Michael V. , Adams Michael W. W.
Published: March 2005

A scheme for the detoxification of superoxide in
Pyrococcus furiosus
has been previously proposed in which superoxide reductase (SOR) reduces (rather than dismutates) superoxide to hydrogen peroxide by using electrons from reduced rubredoxin (Rd). Rd is reduced with electrons from NAD(P)H by the enzyme NAD(P)H:rubredoxin oxidoreductase (NROR). The goal of the present work was to reconstitute this pathway in vitro using recombinant enzymes. While recombinant forms of SOR and Rd are available, the gene encoding
P. furiosus
NROR (PF1197) was found to be exceedingly toxic to
Escherichia coli
, and an active recombinant form (rNROR) was obtained via a fusion protein expression system, which produced an inactive form of NROR until cleavage. This allowed the complete pathway from NAD(P)H to the reduction of SOR via NROR and Rd to be reconstituted in vitro using recombinant proteins. rNROR is a 39.9-kDa protein whose sequence contains both flavin adenine dinucleotide (FAD)- and NAD(P)H-binding motifs, and it shares significant similarity with known and putative Rd-dependent oxidoreductases from several anaerobic bacteria, both mesophilic and hyperthermophilic. FAD was shown to be essential for activity in reconstitution assays and could not be replaced by flavin mononucleotide (FMN). The bound FAD has a midpoint potential of −173 mV at 23°C (−193 mV at 80°C). Like native NROR, the recombinant enzyme catalyzed the NADPH-dependent reduction of rubredoxin both at high (80°C) and low (23°C) temperatures, consistent with its proposed role in the superoxide reduction pathway. This is the first demonstration of in vitro superoxide reduction to hydrogen peroxide using NAD(P)H as the electron donor in an SOR-mediated pathway.
Characterization of Hydrogenase and Reductive Dehalogenase Activities of
Dehalococcoides ethenogenes
Strain 195
Authors: Nijenhuis Ivonne , Zinder Stephen H.
Published: March 2005

Dehalococcoides ethenogenes
strain 195 reductively dechlorinates tetrachloroethene (PCE) and trichloroethene (TCE) to vinyl chloride and ethene using H
2
as an electron donor. PCE- and TCE-reductive dehalogenase (RD) activities were mainly membrane associated, whereas only about 20% of the hydrogenase activity was membrane associated. Experiments with methyl viologen (MV) were consistent with a periplasmic location for the RDs or a component feeding electrons to them. The protonophore uncoupler tetrachlorosalicylanilide did not inhibit reductive dechlorination in cells incubated with H
2
and PCE and partially restored activity in cells incubated with the ATPase inhibitor
N
,
N
′-dicyclohexylcarbodiimide. Benzyl viologen or diquat (
E
o
′ ≈ −360 mV) supported reductive dechlorination of PCE or TCE at rates comparable to MV (−450 mV) in cell extracts.
Comparison of Specimen Processing and Nucleic Acid Extraction by the Swab Extraction Tube System versus the MagNA Pure LC System for Laboratory Diagnosis of Herpes Simplex Virus Infections by LightCycler PCR
Authors: Issa N. C. , Espy M. J. , Uhl J. R. , Harmsen W. S. , Mandrekar J. N. , Gullerud R. E. , Davis M. D. , Smith T. F.
Published: March 2005

A total of 563 specimens (234 dermal and 329 genital swabs) from patients suspected of having herpes simplex virus (HSV) infections were processed using two different extraction methods (the MagNA Pure LC system and the swab extraction tube system [SETS]); HSV DNA was amplified by LightCycler PCR. HSV DNA was detected in 157 of 563 specimens (27.9%) processed by the MagNA Pure LC system and in 179 of 563 specimens (31.8%) processed by SETS (
P
< 0.0001). There was no specimen processed by the MagNA Pure LC extraction method that was positive only for HSV DNA. Of 157 specimens positive by both methods, HSV DNA copy levels were higher (using cycle crossover points [cycle threshold {

C
T

}]) with SETS (mean

C
T

, 25.9 cycles) than with the MagNA Pure LC system (mean

C
T

, 32.0 cycles) (
P
< 0.0001). The time to process 32 samples was longer with the MagNA Pure LC extraction system (90 min) than with SETS (35 min). HSV DNA extraction using SETS is faster, less expensive, and more sensitive than the MagNA Pure LC system and could replace the latter for the laboratory diagnosis of HSV infections using LightCycler PCR.
Rapid Identification of the Coxsackievirus A24 Variant by Molecular Serotyping in an Outbreak of Acute Hemorrhagic Conjunctivitis
Authors: Park Sang-Won , Lee Chang-Seop , Jang Hee-Chang , Kim Eui-Chong , Oh Myoung-don , Choe Kang-Won
Published: March 2005

We evaluated the clinical applicability of a molecular serotyping method for determination of the cause of epidemic acute hemorrhagic conjunctivitis. Seventy conjunctival swab specimens from individuals involved in a nationwide acute hemorrhagic conjunctivitis outbreak were tested. Viral culture and a molecular biology-based assay were compared by directly using clinical specimens. On the one hand, virus culture was done to isolate the enteroviruses, and serotyping was done by a coxsackievirus A24 variant-specific PCR. On the other hand, the original clinical specimens were directly screened for enterovirus by reverse transcription (RT)-PCR with panenterovirus-specific primers. Enterovirus screening-positive specimens were subjected to RT-PCR for detection of the VP1 region of enterovirus, and the amplicons were sequenced. Molecular serotyping was done by calculating the pairwise identity scores for the sequences with the maximum identities to the sequences of known prototype enteroviruses. Thirty-two specimens (45.7%) were culture positive, whereas 37 specimens (52.8%) were screening PCR positive (
P
< 0.001). The VP1 regions were amplified from 21 of the 37 specimens (56.8%), and the products amplified from 9 specimens were appropriately sequenced. These nine sequences were homologous with the sequence of the coxsackievirus A24 variant. Molecular serotyping by direct use of clinical specimens without cell culture could be applied for the rapid identification of the causative agent of epidemic acute hemorrhagic conjunctivitis.
Multiprefectural Spread of Gastroenteritis Outbreaks Attributable to a Single Genogroup II Norovirus Strain from a Tourist Restaurant in Nagasaki, Japan
Authors: Hirakata Yoichi , Arisawa Kokichi , Nishio Osamu , Nakagomi Osamu
Published: March 2005

A series of gastroenteritis outbreaks caused by noroviruses (NVs) among tourist groups from several prefectures was associated with eating a lunch prepared by a restaurant in Nagasaki City, Japan, on 18 and 19 November 2003. A retrospective cohort study was performed to estimate the magnitude of the outbreak and identify the source of infection. Epidemiological information was obtained through the local public health centers in the areas where the illness occurred. Stool and vomit specimens and food and environmental samples were analyzed by reverse transcription-PCR with genogroup-specific primers. Positive samples were sequenced and analyzed phylogenetically. Of 1,492 tourists who ate a lunch prepared by the restaurant during the 2-day period, 660 (44.2%) developed illness, with an average incubation time of 31.2 h. Whereas NVs were not detected in any food samples, identical sequences most closely related to the Mexico genotype of genogroup II NV were found in specimens from case patients, restaurant staff, and the kitchen table. Food handlers were concluded to be the source of the outbreak as a result of the contamination of several meals. The series of outbreaks described here exemplifies the role of tourism as a contemporary way to distribute a single infectious agent to multiple and geographically remote areas.
Novel Antibody-Lectin Enzyme-Linked Immunosorbent Assay That Distinguishes Prion Proteins in Sporadic and Variant Cases of Creutzfeldt-Jakob Disease
Authors: Pan Tao , Li Ruliang , Wong Boon-Seng , Kang Shin-Chung , Ironside James , Sy Man-Sun
Published: March 2005

We used different anti-prion protein (anti-PrP) monoclonal antibodies to capture either full-length or truncated PrP species and then used biotinylated lectin to compare the nature of the glycans on bound PrP species present in control, sporadic Creutzfeldt-Jakob disease (sCJD), or variant CJD (vCJD) brains. When full-length PrP species in these three groups were compared, no significant difference in the binding of concanavalin A orAleuria aurantialectin was detected. However, the binding ofRicinus communisagglutinin I (RCA) to sCJD and vCJD samples was significantly increased. In contrast, when only truncated PrP species were compared, only vCJD samples had more RCA binding activity. Therefore, while most of the RCA binding activity in sCJD is restricted to the full-length PrP species, the RCA binding activity in vCJD is associated with truncated and full-length PrP species. Furthermore, the RCA binding activity in sCJD and vCJD samples is mostly associated with proteinase K-resistant PrP species, a known signature of infectious prion. Therefore, PrP species in sCJD and vCJD have dissimilar lectin immunoreactivity, which reflects differences in their N-linked glycans. These differences may account for the distinct phenotypes of sCJD and vCJD.
Ascariasis Is a Zoonosis in Denmark
Authors: Nejsum Peter , Parker E. Davis , Frydenberg Jane , Roepstorff Allan , Boes Jaap , Haque Rashidul , Astrup Ingrid , Prag Jørgen , Skov Sørensen Uffe B.
Published: March 2005

A preliminary epidemiological survey indicated an association between
Ascaris
infections in Danish patients and contact with pigs or pig manure. In the present study, we compared
Ascaris
worms collected from humans and
Ascaris
worms collected from pigs by amplified fragment length polymorphism (AFLP) analysis, a technique for whole-genome fingerprinting, and by PCR-linked restricted fragment length polymorphism (PCR-RFLP) analysis of the internal transcribed spacer region of nuclear rDNA. The AFLP data were analyzed by distance- and model-based clustering methods. These results assigned
Ascaris
worms from Danish patients to a cluster different from that for worms from humans in other geographic areas. In contrast, worms from humans and pigs in Denmark were assigned to the same cluster. These results were supported by the PCR-RFLP results. Thus, all of the examined Danish patients had acquired
Ascaris
infections from domestic pigs; ascariasis may therefore be considered a zoonotic disease in Denmark.
Rapid Identification of Staphylococcal Strains from Positive-Testing Blood Culture Bottles by Internal Transcribed Spacer PCR Followed by Microchip Gel Electrophoresis
Authors: Fujita Shin-Ichi , Senda Yasuko , Iwagami Thikako , Hashimoto Takuma
Published: March 2005

PCR analysis of the 16S-23S rRNA gene internal transcribed spacer (ITS) followed by microchip gel electrophoresis (MGE) was evaluated for its usefulness in identification of staphylococci. Forty ITS PCR patterns were demonstrated among 228 isolated colonies of
Staphylococcus aureus
: 26 patterns for methicillin-susceptible
S. aureus
(MSSA; 91 strains), 11 patterns for methicillin-resistant
S. aureus
(MRSA; 99 strains), and 3 patterns for both MSSA and MRSA (38 strains). Thirty-seven control strains of coagulase-negative staphylococci (CNS) representing 16 species showed unique ITS PCR patterns (24 patterns) at the species and subspecies levels: two patterns for
S. caprae
,
S. cohnii
,
S. haemolyticus
, and
S. saprophyticus
; three patterns for
S. lugdunensis
; four patterns for
S. capitis
; and one pattern for each of the other CNS species. The combined PCR-MGE method was prospectively adapted to the positive blood culture bottles, and this method correctly identified MSSA and MRSA in 102 (89%) of 114 blood cultures positive for
S. aureus
on the basis of the ITS PCR patterns. Eight ITS PCR patterns were demonstrated from 166 blood culture bottles positive for CNS. The most frequent CNS species isolated from blood cultures were
S. epidermidis
(76%),
S. capitis
(11%), and
S. hominis
(8%). Overall, all 280 blood culture bottles shown to contain a single
Staphylococcus
species by routine phenotypic methods were correctly identified by the PCR-MGE method at the species level, whereas the organism failed to be identified in 8 culture bottles (3%) with mixed flora. The PCR-MGE method is useful not only for rapid identification (∼1.5 h) of staphylococci in positive blood culture bottles, but also for strain delineation of
S. aureus
.
Potential Limitations of the 16S-23S rRNA Intergenic Region for Molecular Detection of
Bartonella
Species
Authors: Maggi Ricardo G. , Breitschwerdt Edward B.
Published: March 2005

PCR targeting the 16S-23S rRNA gene intergenic transcribed spacer (ITS) region has been proposed as a rapid and reliable method for the detection of
Bartonella
species DNA in clinical samples. Because of variation in ITS sequences among
Bartonella
species, a single PCR amplification can be used to detect different species within this genus. Therefore, by targeting the ITS region, multiple PCRs or additional sample-processing steps beyond the primary amplification can be avoided when attempting to achieve molecular diagnostic detection of
Bartonella
species. Although PCR amplification targeting this region is considered highly sensitive, amplification specificity obviously depends on primer design. We report evidence of nonspecific PCR amplification of
Mesorhizobium
species with previously published primers that were designed to amplify the
Bartonella
consensus ITS region. Use of these or other, less species-specific, primers could lead to a false-positive diagnostic test result when evaluating clinical samples. We also report the presence of
Mesorhizobium
species DNA as a contaminant in molecular-grade water, a series of homologous sequences in the ITS region that are common to
Bartonella
and
Mesorhizobium
species, the amplification of
Mesorhizobium
DNA with unpublished primers designed in our laboratory targeting the ITS region, and the subsequent design of unambiguous ITS primers that avoid nonspecific amplification of
Mesorhizobium
species. Our results define some potential limitations associated with the molecular detection of
Bartonella
species in patient samples and indicate that primer specificity is of critical importance if the ITS region is used as a diagnostic target for detection of
Bartonella
species.
Clinical Evaluation of the DiversiLab Microbial Typing System Using Repetitive-Sequence-Based PCR for Characterization of
Staphylococcus aureus
Strains
Authors: Shutt Cheryl K. , Pounder June I. , Page Sam R. , Schaecher Barbara J. , Woods Gail L.
Published: March 2005

The DiversiLab System, which includes microfluidics-based detection, reagent kits, and software for data processing and analysis, is an automated method using repetitive sequence-based PCR (rep-PCR) for microbial strain typing. To assess the reliability of the DiversiLab System for strain characterization of
Staphylococcus aureus
, we tested clinical isolates sent to ARUP Laboratories for typing and compared results to those of pulsed field electrophoresis (PFGE) aided by the cluster analysis provided by BioNumerics software.
spa
typing was performed when the results of these two methods for an outbreak were not concordant. The study included 89
S. aureus
isolates (65
mecA
positive, 24
mecA
negative) from 19 outbreaks (2 to 11 isolates/outbreak). The DiversiLab and PFGE-BioNumerics results were concordant for 15 of the 19 outbreaks. For the remaining four outbreaks, there was partial concordance between the two methods.
spa
typing results were the same as or more similar to rep-PCR results for three of those outbreaks and were more similar to PFGE results for one. With regard to performance, the DiversiLab system was considerably less labor intensive than PFGE and provided results in less than 24 h, compared with 2 to 3 days for PFGE. Additionally, the Web-based DiversiLab software provides standardized comparisons among isolates almost instantaneously and generates user-friendly, customized reports.
Seroepidemiology of Human Metapneumovirus (hMPV) on the Basis of a Novel Enzyme-Linked Immunosorbent Assay Utilizing hMPV Fusion Protein Expressed in Recombinant Vesicular Stomatitis Virus
Authors: Leung Jessica , Esper Frank , Weibel Carla , Kahn Jeffrey S.
Published: March 2005

Human metapneumovirus (hMPV) is a newly identified human respiratory virus now recognized as a major respiratory pathogen of infants and children. To define the seroepidemiology of hMPV, we developed a novel enzyme-linked immunosorbent assay (ELISA) based on expression of the fusion protein of hMPV (hMPV F) in recombinant vesicular stomatitis virus (VSV). Western blot analysis using an hMPV F-specific antiserum confirmed the expression of hMPV in recombinant VSV. The ELISA is specific for hMPV F; antibody specific for the most closely related human paramyxovirus, respiratory syncytial virus, does not bind to hMPV F. Overall, 216 serum specimens were tested. The percentages of seropositive individuals were 89.1% in children ≤5 months old, 55.0% in children 6 to 11 months old, 36.0% in children 12 to 23 months old, 45.0% in children 24 to 47 months old, 77.3% in children 48 to 59 months old, 91.3% in children 5 to 10 years old, and 95.5% for individuals 11 to 20 years old. This is the first seroepidemiological survey of hMPV in the United States and the first analysis to determine the prevalence of antibody to a specific hMPV protein. The data suggest that exposure to hMPV is common in childhood and that hMPV F is an antigenic determinant of hMPV.
Systematic Molecular Characterization of Multidrug-Resistant
Mycobacterium tuberculosis
Complex Isolates from Spain
Authors: Samper S. , Iglesias M. J. , Rabanaque M. J. , Gómez L. I. , Lafoz M. C. , Jiménez M. S. , Ortega A. , Lezcano M. A , Van Soolingen D. , Martín C.
Published: March 2005

We used spoligotyping and restriction fragment length polymorphism (RFLP) of the IS
6110
-insertion sequence to study the molecular epidemiology of multidrug-resistant (MDR) tuberculosis in Spain. We analyzed 180
Mycobacterium tuberculosis
complex isolates collected between January 1998 and December 2000. Consecutive isolates from the same patients (
n
= 23) always had identical genotypes, meaning that no cases of reinfection occurred. A total of 105 isolates (58.3%) had unique RFLP patterns, whereas 75 isolates (41.7%) were in 20 different RFLP clusters. Characterization of the
katG
and
rpoB
genes showed that 14 strains included in the RFLP clusters did not actually cluster. Only 33.8% of the strains isolated were suggestive of MDR transmission, a frequency lower than that for susceptible strains in Spain (46.6%). We found that the Beijing/W genotype, which is prevalent worldwide, was significantly associated with immigrants. The 22 isolates in the largest cluster corresponded to the
Mycobacterium bovis
strain responsible for two nosocomial MDR outbreaks in Spain.
Analysis of the Influence of Tween Concentration, Inoculum Size, Assay Medium, and Reading Time on Susceptibility Testing of
Aspergillus
spp
Authors: Gomez-Lopez Alicia , Aberkane Amel , Petrikkou Eva , Mellado Emilia , Rodriguez-Tudela Juan Luis , Cuenca-Estrella Manuel
Published: March 2005

The influence of several test variables on susceptibility testing of
Aspergillus
spp. was assessed. A collection of 28 clinical isolates was tested against amphotericin B, itraconazole, voriconazole, and terbinafine. Inoculum size (10
4
CFU/ml versus 10
5
CFU/ml) and glucose supplementation (0.2% versus 2%) did not have significant effects on antifungal susceptibility testing results and higher inoculum size and glucose concentration did not falsely elevate MICs. In addition, antifungal susceptibility testing procedure with an inoculum size of 10
5
CFU/ml distinctly differentiated amphotericin B or itraconazole-resistant
Aspergillus
strains in vivo from the susceptible ones. Time of incubation significantly affected the final values of MICs, showing major increases (two to six twofold dilutions,
P
< 0.01 by analysis of variance) between MIC readings at 24 and 48 h, but no differences were observed between antifungal susceptibility testing results obtained at 48 h and at 72 h. Significantly higher MICs were uniformly associated with higher concentrations of Tween (
P
< 0.01), used as a dispersing agent in the preparation of inoculum suspensions. The geometric mean MICs showed increases of between 1.5- and 10-fold when the Tween concentration varied from 0.1% (the geometric means for amphotericin B, itraconazole, voriconazole, and terbinafine were 1.29, 0.69, 1.06, and 0.64 μg/ml, respectively) to 5% (the geometric means for amphotericin B, itraconazole, voriconazole, and terbinafine were 1.97, 5.79, 1.60, and 4.66 μg/ml, respectively). The inhibitory effect of Tween was clearly increased with inoculum sizes of 10
5
CFU/ml and was particularly dramatic for itraconazole, terbinafine, and
Aspergillus terreus
. The inoculum effect was not observed when the Tween concentration was below 0.5% (
P
> 0.01).
Distribution ofMycobacterium aviumComplex Isolates in Tissue Samples of Pigs Fed Peat Naturally Contaminated with Mycobacteria as a Supplement
Authors: Matlova Ludmila , Dvorska Lenka , Ayele Wuhib Yayo , Bartos Milan , Amemori Takashi , Pavlik Ivo
Published: March 2005

In early 1999, there was an increased incidence of tuberculous lesions in the lymph nodes of slaughtered pigs in the Czech Republic. In part 1 of this study, tuberculous lesions were detected in 140 (62%) tissue samples collected from pigs coming from 15 farms in 15 districts at routine veterinary meat inspections in abattoirs. Mycobacteria were isolated from 37 (16%) tissue samples: 34Mycobacterium aviumsubsp.hominissuisisolates and three environmentally derived mycobacteria. In search of infection sources,M. aviumsubsp.hominissuiswas isolated from 38 (79%) samples of peat used as a feed supplement. In part 2 of our study, the head, mesenteric, and inguinal lymph nodes of 117 randomly selected slaughtered pigs from one farm with young piglets fed peat as a supplement were investigated for mycobacterial infection. From 65 (56%) pigs, a total of 76 mycobacterial isolates were identified (56M. aviumsubsp.hominissuisisolates, 5M. aviumsubsp.aviumisolates, 3M. intracellulareisolates, and 12 environmentally derived mycobacterial isolates). IS1245restriction fragment length polymorphism (RFLP) types with >20 bands of 45 distinct RFLP types were found in 49M. aviumsubsp.hominissuisisolates from pigs (n= 31) and peat (n= 18). Identical RFLP types were found in only four pig isolates. Five randomly selected isolates from pigs and peat were subcultured to six independent clones or colonies. Among the IS1245RFLP types of 30 clones, identical RFLP types obtained from pigs and peat were identified, which confirmed the hypothesis that peat contaminated with mycobacteria represents a significant source of mycobacterial infection for pigs.
Molecular Epidemiology of Macrolide-Resistant
Streptococcus pneumoniae
Isolates in Europe
Authors: Reinert Ralf René , Ringelstein Adrian , van der Linden Mark , Cil Murat Yücel , Al-Lahham Adnan , Schmitz Franz-Josef
Published: March 2005

In many European countries, the level of pneumococcal resistance to macrolides has now passed the level of resistance to penicillin G. A total of 82 erythromycin A-resistant isolates of
Streptococcus pneumoniae
were collected by 11 laboratories in seven European countries. All of the isolates were tested for antimicrobial susceptibility, analyzed for clonal relatedness by multilocus sequence typing, and characterized for macrolide resistance genotypes. The prevalence of the macrolide resistance genotypes varied substantially between countries. In France (87.5% of all strains), Spain (77.3%), Switzerland (80%), and Poland (100%), strains were predominantly
erm
(B) positive, whereas higher levels of
mef
(A)-positive strains were reported from Greece (100%) and Germany (33.3%). Macrolide resistance was caused by the oligoclonal spread of some multilocus sequence types, but significant differences in clonal distribution were noted between France and Spain, countries from which high levels of macrolide resistance have been reported. Overall, sequence type 81 (Spain23F-1 clone) was by far the most widespread. The mainly
erm
(B)-positive serotype 14 clone (sequence type 143), first reported in Poland in the mid-1990s, is now widespread in France.
Evaluation of a Point-of-Care Test, BVBlue, and Clinical and Laboratory Criteria for Diagnosis of Bacterial Vaginosis
Authors: Bradshaw C. S. , Morton A. N. , Garland S. M. , Horvath L. B. , Kuzevska I. , Fairley C. K.
Published: March 2005

Bacterial vaginosis (BV) remains the most common cause of abnormal vaginal discharge in women of reproductive age and is associated with increased susceptibility to human immunodeficiency virus and sexually transmitted infections and preterm delivery. Present diagnostic methods require access to microscopy and laboratory expertise; however, the majority of women, particularly those in populations with a high prevalence of BV, do not have access to clinical services with on-site microscopy capabilities. We evaluated a point-of-care test for the diagnosis of BV, the BVBlue test, with 288 women attending a sexual health service with symptoms of abnormal vaginal discharge and/or odor. The BVBlue test performed well compared with conventional diagnostic methods for the assessment of women with symptoms suggestive of BV at the bedside and significantly better than other simple tests, such as vaginal pH determination and the amine test, that do not require microscopy. The BVBlue test was sensitive (88%; 95% confidence interval [CI], 81 to 93%) and specific (95%; 95% CI, 91 to 98%) compared to the method of Nugent et al. (R. P. Nugent, M. A. Krohn, and S. L. Hillier, J. Clin. Microbiol. 29:297-301, 1991) and performed well compared with the method of Amsel et al. (R. Amsel, P. A. Totten, C. A. Spiegel, K. C. Chen, D. Eschenbach, and K. K. Holmes, Am. J. Med. 74:14-22, 1983), with a sensitivity of 88% (95% CI, 81 to 93%) and a specificity of 91% (95% CI, 85 to 94%). The BVBlue test is a simple, rapid, and objective test for the diagnosis of BV and has the potential to facilitate prompt diagnosis and appropriate treatment of BV in the absence of microscopy. The majority of women at the greatest risk for the sequelae of BV are not in settings where the conventional diagnostic methods are either practical or possible, and they would greatly benefit from access to rapid and reliable point-of-care tests to improve the diagnosis and management of BV.
Systemic Disease in Vaal Rhebok (Pelea capreolus) Caused by Mycoplasmas in the Mycoides Cluster
Authors: Nicolas Melissa M. , Stalis Ilse H. , Clippinger Tracy L. , Busch Martin , Nordhausen Robert , Maalouf Gabriel , Schrenzel Mark D.
Published: March 2005

In the winter of 2002, an outbreak of mycoplasma infection in Vaal rhebok (Pelea capreolus) originating from South Africa occurred 15 weeks after their arrival in San Diego, Calif. Three rhebok developed inappetence, weight loss, lethargy, signs related to pulmonary or arthral dysfunction, and sepsis. All three rhebok died or were euthanized. Primary postmortem findings were erosive tracheitis, pleuropneumonia, regional cellulitis, and necrotizing lymphadenitis. Mycoplasmas were detected in numerous tissues by electron microscopy, immunohistochemistry, and PCR. The three deceased rhebok were coinfected with ovine herpesvirus-2, and two animals additionally had a novel gammaherpesvirus. However, no lesions indicative of herpesvirus were seen microscopically in any animal. The rheboks' mycoplasmas were characterized at the level of the 16S rRNA gene, the 16S-23S intergenic spacer region, and the fructose biphosphate aldolase gene. Denaturing gradient gel electrophoresis was carried out to address the possibility of infection with multiple strains. Two of the deceased rhebok were infected with a single strain ofMycoplasma capricolumsubsp.capricolum, and the third animal had a single, unique strain most closely related toMycoplasma mycoidessubsp.mycoides large-colony. A PCR survey of DNA samples from 46 other ruminant species demonstrated the presence of several species of mycoplasmas in the mycoides cluster, including a strain ofM. capricolumsubsp.capricolumidentical to that found in two of the rhebok. These findings demonstrate the pervasiveness of mycoplasmas in the mycoides cluster in small ruminants and the potential for interspecies transmission and disease when different animal taxa come in contact.
Molecular Epidemiology of Meningococci Isolated in Niger in 2003 Shows Serogroup A Sequence Type (ST)-7 and Serogroup W135 ST-11 or ST-2881 Strains
Authors: Nicolas P. , Djibo S. , Moussa A. , Tenebray B. , Boisier P. , Chanteau S.
Published: March 2005

In 2003, in the Zinder and Maradi regions (Niger), epidemics were due to serogroup A:4:P1.9 meningococci belonging to sequence type 7 (ST-7). In Niamey, only sporadic cases were reported: 55% of the meningococcus strains were in serogroup A, and 38% were in serogroup W135 and could be placed in ST-11, identical to the 2002 Burkina Faso epidemic clone, and in ST-2881, a new ST.
Rate and Incidence Estimates of Recent Human Immunodeficiency Virus Type 1 Infections among Pregnant Women in São Paulo, Brazil, from 1991 to 2002
Authors: de Freitas Oliveira Carmem A. , Ueda Mirthes , Yamashiro Rosemeire , Rodrigues Rosângela , Sheppard Haynes W. , de Macedo Brígido Luís Fernando
Published: March 2005

The serological testing algorithm for recent human immunodeficiency virus (HIV) seroconversion (STARHS) was employed to estimate HIV incidence among pregnant women from São Paulo, Brazil. A cross-sectional study (1999 to 2002) showed an incidence of infection of 0.2 per 100 pregnant women per year (95% confidence interval, 0.041 to 0.608). Western blot profiles suggested an association between results of the STARHS analysis and gp41/gp31 bands.
Adenovirus Type F Subtype 41 Causing Disseminated Disease following Bone Marrow Transplantation for Immunodeficiency
Authors: Slatter Mary A. , Read Steven , Taylor Clive E. , Crooks Bruce N. A. , Abinun Mario , Flood Terence J. , Cant Andrew J. , Wright Christopher , Gennery Andrew R.
Published: March 2005

Adenovirus causes disseminated disease following bone marrow transplantation (BMT). We report a child who underwent T-cell-depleted BMT. Adenovirus subgenus F serotype 41 was detected antemortem by PCR in cerebrospinal fluid and postmortem in other tissues. Serotypes 40 and 41, associated with gastrointestinal disease, have not previously been implicated in disseminated disease.
Endophthalmitis Caused by
Enterococcus mundtii
Authors: Higashide Tomomi , Takahashi Mami , Kobayashi Akira , Ohkubo Shinji , Sakurai Mayumi , Shirao Yutaka , Tamura Toshihiro , Sugiyama Kazuhisa
Published: March 2005

Enterococcus mundtii
has rarely been isolated from environmental or human sources. We report the identification of
E. mundtii
as a pathogen of human infectious disease by DNA sequencing of 16S rRNA and
sodA
genes in a case of endophthalmitis developed in a 66-year-old immunocompetent gardener.
Risk Factors Associated with Colonization by Pneumococci with Reduced Susceptibility to Fluoroquinolones in Adult Outpatients
Authors: Jiménez M. Raquel Marín , Bellido Juan Luis Muñoz , García Rodríguez José Ángel
Published: March 2005

We developed a case-control study in order to identify risk factors associated with pharyngeal colonization by
Streptococcus pneumoniae
with reduced susceptibility to fluoroquinolones (ciprofloxacin MIC, ≥4 μg/ml). A total of 400 patients were studied for colonization by quinolone-nonsusceptible
S. pneumoniae
(QNSP) isolates and risk factors for this colonization. Isolate susceptibility was determined by the agar dilution method. Forty patients were colonized by QNSP (case patients), and 360 patients were not colonized by QNSP (control patients). The MIC range of ciprofloxacin for QNSP isolates was 4 to 8 μg/ml. No isolates were resistant to levofloxacin and moxifloxacin. Risk factors significantly associated with QNSP colonization, according to univariate analysis, were recent hospitalizations (odds ratio [OR], 3.43; 95% confidence interval [CI], 1.6 to 7.2;
P
< 0.01) and prior exposure to fluoroquinolones (OR, 6.04; 95% CI, 3.0 to 12.0;
P
< 0.01). Other factors such as chronic obstructive pulmonary disease (OR, 1.94; 95% CI; 0.7 to 5.0), prior exposure to penicillins (OR, 1,68; 95% CI, 0.8 to 3.3) and prior exposure to macrolides (OR 2; 95% CI, 0.6 to 6.2) were more frequent among patients colonized with QNSP, but there was no statistical significance. Multivariate analysis showed that exposure to fluoroquinolones was the only independent factor associated with colonization by QNSP (OR, 4.2; 95% CI, 1.8 to 9.4;
P
< 0.01). Throat colonization by QNSP is becoming frequent, though most of these isolates (all the isolates in this case) remain susceptible to newer fluoroquinolones. Previous treatment with fluoroquinolones seems to be the main risk factor associated with colonization by QNSP.
Medical intelligence in Sweden. Vitamin B12: oral compared with parenteral?
Authors: Nilsson M , Norberg B , Hultdin J , Sandström H , Westman G , Lökk J
Published: March 2005

Background

Sweden is the only country in which oral high dose vitamin B12 has gained widespread use in the treatment of deficiency states.

Objective

The aim of the study was to describe prescribing patterns and sales statistics of vitamin B12 tablets and injections in Sweden 1990–2000.

Design, setting, and sources

Official statistics of cobalamin prescriptions and sales were used.

Results

The use of vitamin B12 increased in Sweden 1990–2000, mainly because of an increase in the use of oral high dose vitamin B12 therapy. The experience, in statistical terms a “total investigation”, comprised 1 000 000 patient years for tablets and 750 000 patient years for injections. During 2000, 13% of residents aged 70 and over were treated with vitamin B12, two of three with the tablet preparation. Most patients in Sweden requiring vitamin B12 therapy have transferred from parenteral to oral high dose vitamin B12 since 1964, when the oral preparation was introduced.

Conclusion

The findings suggest that many patients in other post-industrial societies may also be suitable for oral vitamin B12 treatment.
Colonoscopy in the very old: why bother?
Authors: Yoong K K Y , Heymann T
Published: March 2005

Objectives

To evaluate the use of colonoscopy in patients aged at least 85 years. Does the ideal of an ageism free service apply?

Design

A retrospective audit.

Setting

Department of gastroenterology that carries out about 1000 colonoscopies annually in a district general hospital serving a population of about 320 000.

Subjects

All patients aged at least 85 years who underwent colonoscopy over five years to 2003.

Main outcome measures

The indications for colonoscopy and its findings. The outcome of patients found to have colonic cancers.

Results

Colonoscopy was completed in 219 cases (69%). The main reasons for failure were poor bowel preparation and severe diverticular disease. Normal findings occurred in 65 (30%) of the 219 cases that had had a complete examination. Colonoscopy identified a problem that explained the patient’s symptoms in 116 (37%) cases. Polyps were found in 45 (14.2%) cases and malignancy in 28 (8.8%).

Conclusions

The absence of significant complications and comparatively high yield of colonic malignancies and polyps reinforces the value of colonoscopy as a diagnostic tool even after 85 years of age and despite the technical challenges of the procedure in this age group that limited completion. Increasing age alone should not preclude a patient from colonoscopy.
Intestinal adaptation after massive intestinal resection
Authors: Weale A R , Edwards A G , Bailey M , Lear P A
Published: March 2005

Patients with short bowel syndrome require long term parenteral nutrition support. However, after massive intestinal resection the intestine undergoes adaptation and nutritional autonomy may be obtained. Given that the complications of parenteral nutrition may be life threatening or result in treatment failure and the need for intestinal transplantation, a more attractive option is to wean patients off nutrition support by optimising the adaptive process. The article examines the evidence that after extensive small bowel resection adaptation occurs in humans and focuses on the factors that influence adaptation and the strategies that have been used to optimise this process. The review is based on an English language Medline search with secondary references obtained from key articles. There is evidence that adaptation occurs in humans. Adaptation is a complex process that results in response to nutrient and non-nutrient stimuli. Successful and reproducible strategies to improve adaptation remain elusive despite an abundance of experimental data. Nevertheless given the low patient survival and quality of life associated with other treatments for irreversible intestinal failure it is imperative that clinical research continues into the optimisation of the adaptation.
Placebo
Authors: McQuay H J , Moore R A
Published: March 2005

Placebo, used here to mean an inert treatment given as if it was a real treatment, means lots of different things to different people. The structure of the article is that it begins by talking about the technical use of placebos in clinical trials, and the extent of the placebo response, then about the mechanism—“How does the placebo work?”—and last about the ethics of placebo in the contexts of research and in everyday practice.
Acute liver failure
Authors: O’Grady J G
Published: March 2005

Acute liver failure is a complex multisystemic illness that evolves quickly after a catastrophic insult to the liver leading to the development of encephalopathy. The underlying aetiology and the pace of progression strongly influence the clinical course. The commonest causes are paracetamol, idiosyncratic drug reactions, hepatitis B, and seronegative hepatitis. The optimal care is multidisciplinary and up to half of the cases receive liver transplants, with survival rates around 75%–90%. Artificial liver support devices remain unproven in efficacy in acute liver failure.
Supernumerary phantom limb after stroke
Authors: Bakheit A M O , Roundhill S
Published: March 2005

The perception of a phantom limb is commonly reported after amputations. However, only a few cases have been described after a stroke. This article presents a patient who reported a supernumerary phantom limb (pseudopolymelia) after spontaneous intracerebral haemorrhage and discusses the possible underlying mechanisms for this rare phenomenon.
Chromatin supraorganization, DNA fragmentation, and cell death in snake erythrocytes
Authors: Miyamoto Maristela , Vidal Benedicto C , Mello Maria Luiza S
Published: March 2005

In nucleate erythrocytes of several vertebrate groups, the frequency and intensity of DNA fragmentation associated with programed cell death vary considerably. Although hemoglobin efficiency may be related to erythrocyte life span, and hemoglobin types and erythrocyte life spans are assumed to vary in reptiles, no data on DNA fragmentation and chromatin organization as related to cell death exist for snakes. In the present study, chromatin supraorganization, DNA fragmentation, and cell death were investigated in four snake species (Crotalus durissus terrificus, Bothrops jararaca, Bothrops alternatus, and Bothrops neuwiedii), which differ in their geographical distribution and habitats, by using image analysis of Feulgen hydrolysis kinetics, the TUNEL assay, single-cell gel electrophoresis, and transmission electron microscopy. Relatively few circulating erythrocytes were found to be simultaneously committed to cell death, although there was some variation among the snake species. Conspicuous nuclear and cytoplasmic organelles suggestive of metabolic activity were seen ultrastructurally in most snake erythrocytes. The DNA of the snake erythrocyte chromatin was much more resistant to Feulgen acid hydrolysis (DNA depurination and breakdown) than that of young adult bullfrog erythrocytes, which had a high frequency and intensity of DNA fragmentation. Of the species studied, B. neuwiedii and C. d. terrificus showed the greatest resistance to Feulgen acid hydrolysis and to the DNA fragmentation, revealed by the TUNEL assay. Although B. neuwiedii also showed the lowest frequency of cells with more damaged DNA in the single-cell gel electrophoresis assay, C. d. terrificus had the highest frequency of damaged cells, possibly because of the abundance of alkaline-sensitive DNA sites. The results for DNA fragmentation and cell death in erythrocytes of B. jararaca and B. alternatus generally differed from those for C. d. terrificus and B. neuwiedii and may reflect differences in the biology of these species selected under different geographical habitats. The differences in erythrocyte cell biology reported here may be related to hemoglobin variants selected in the mentioned snake species and that would lead the cells to different resistances to unfavorable environmental conditions.Key words: erythrocytes, snakes, DNA fragmentation, chromatin supraorganization, cell death.
Tocotrienol-induced cytotoxicity is unrelated to mitochondrial stress apoptotic signaling in neoplastic mammary epithelial cells
Authors: Shah Sumit J , Sylvester Paul W
Published: March 2005

Tocotrienols and tocopherols represent the 2 subgroups within the vitamin E family of compounds, but tocotrienols display significantly greater apoptotic activity against a variety of cancer cell types. However, the exact mechanism mediating tocotrienol-induced apoptosis is not understood. Studies were conducted to determine the effects of tocotrienols on mitochondrial-stress-mediated apoptotic signaling in neoplastic +SA mammary epithelial cells grown in vitro. Exposure for 24 h to 0–20 µmol/L γ-tocotrienol resulted in a dose–responsive increase in +SA cells undergoing apoptosis, as determined by flow cytometric analysis of Annexin V staining. However, tocotrienol-induced apoptosis was not associated with a disruption or loss of mitochondrial membrane potential, or the release of mitochondrial cytochrome c into the cytoplasm, as determined by JC-1 flow cytometric staining and ELISA assay, respectively. Interestingly, apoptotic +SA cells showed a paradoxical decrease in mitochondrial levels of pro-apoptotic proteins Bid, Bax, and Bad, and a corresponding increase in mitochondrial levels of anti-apoptotic proteins, Bcl-2 and Bcl-xL, suggesting that mitochondrial membrane stability and integrity might actually be enhanced for a limited period of time following acute tocotrienol exposure. In summary, these findings clearly demonstrate that tocotrienol-induced apoptosis occurs independently of mitochondrial stress apoptotic signaling in neoplastic +SA mammary epithelial cells.Key words: breast cancer, tocotrienols, apoptosis, mitochondria, Bcl-2.
NONDIFFERENTIABLE SECOND-ORDER SYMMETRIC DUALITY
Authors: AHMAD I. , HUSAIN Z.
Published: March 2005

A pair of Mond–Weir type nondifferentiable second-order symmetric primal and dual problems in mathematical programming is formulated. Weak duality, strong duality, and converse duality theorems are established under η-pseudobonvexity assumptions. Symmetric minimax mixed integer primal and dual problems are also investigated. Moreover, the self duality theorem is also discussed.
THE KNOT GROUP AND THE FUNDAMENTAL GROUP OF THE EMBEDDING 3-MANIFOLD
Authors: LOPES PEDRO , MORALES C. A.
Published: March 2005

We propose a conjecture concerning 3-manifolds (which implies the ℚ conjecture of Myers') and construct a countable sequence of examples which supports it.
AFFINE LATTICE MODELS
Authors: ALBANESE CLAUDIO , KUZNETSOV ALEXEY
Published: March 2005

We introduce a new class of lattice models based on a continuous time Markov chain approximation scheme for affine processes, whereby the approximating process itself is affine. A key property of this class of lattice models is that the location of the time nodes can be chosen in a payoff dependent way and one has the flexibility of setting them only at the relevant dates. Time stepping invariance relies on the ability of computing node-to-node discounted transition probabilities in analytically closed form. The method is quite general and far reaching and it is introduced in this article in the framework of the broadly used single-factor, affine short rate models such as the Vasiček and CIR models. To illustrate the use of affine lattice models in these cases, we analyze in detail the example of Bermuda swaptions.