bluuebunny/crossref_metadata_2025 huggingface dataset:
PslD Is a Secreted Protein Required for Biofilm Formation by
Pseudomonas aeruginosa
Authors: Campisano Andrea , Schroeder Christine , Schemionek Mirle , Overhage Joerg , Rehm Bernd H. A.
Published: April 2006

The function of
pslD
, which is part of the
psl
operon from
Pseudomonas aeruginosa
, was investigated in this study. The
psl
operon is involved in exopolysaccharide biosynthesis and biofilm formation. An isogenic marker-free
pslD
deletion mutant of
P. aeruginosa
PAO1 which was deficient in the formation of differentiated biofilms was generated. Expression of only the
pslD
gene coding region restored the wild-type phenotype. A C-terminal, hexahistidine tag fusion enabled the identification of PslD. LacZ and PhoA translational fusions with PslD indicated that PslD is a secreted protein required for biofilm formation, presumably via its role in exopolysaccharide export.
Arsenic Resistance in
Campylobacter
spp. Isolated from Retail Poultry Products
Authors: Sapkota Amy R. , Price Lance B. , Silbergeld Ellen K. , Schwab Kellogg J.
Published: April 2006

Organoarsenicals are commonly used for growth promotion in U.S. poultry production. Susceptibilities to arsenite, arsenate, and the organoarsenical roxarsone were measured in 251
Campylobacter
isolates from conventional and antimicrobial-free retail poultry products. Isolates from conventional poultry products had significantly higher roxarsone MICs (
z
= 8.22;
P
< 0.0001).
Absence of VanA- and VanB-Containing Enterococci in Poultry Raised on Nonintensive Production Farms in Brazil
Authors: Batista Xavier Diego , Moreno Bernal Francisco Ernesto , Titze-de-Almeida Ricardo
Published: April 2006

We examined cloacal samples from poultry raised on nonintensive production farms in Brazil for the presence of vancomycin-resistant enterococci. No VanA- or VanB-containing enterococci were identified in a total of 200 cloacal swabs. The most prevalent species were
Enterococcus gallinarum
(
vanC1
; 13.0%) and
E. casseliflavus
(
vanC2/3
; 5.5%).
Potential Impact of Conjugate Vaccine on the Incidence of Invasive Pneumococcal Disease among Children in Scotland
Authors: Clarke Stuart C. , Jefferies Johanna M. , Smith Andrew J. , McMenamin Jim , Mitchell Timothy J. , Edwards Giles F. S.
Published: April 2006

We sought to determine the potential impact of seven-valent pneumococcal conjugate vaccine on the incidence of invasive pneumococcal disease (IPD) among children in Scotland. Invasive pneumococci from blood and cerebrospinal fluid, isolated between 2000 and 2004 from all children aged less than 5 years in Scotland, were characterized by serotyping. Using reported efficacy data of the seven-valent pneumococcal conjugate vaccine (PCV7) along with likely coverage rates, we made an estimation of the potential impact on the incidence of IPD among children in Scotland. A total of 217 pneumococci were characterized into 22 different serogroups/types, the most common, in rank order, being 14, 19F, 6B, 18C, 23F, 9V, 4, 1, 19A, and 6A. Estimated serotype coverage for PCV7 was 76.5% in those aged less than 5 years of age but increased to 88.9% for those aged 1 year. By using serotype coverage and estimates of vaccine efficacy and uptake, the potential impact of the vaccine for those greater than 2 months of age, but less than 5 years, was estimated as 67.3%, leading to an average of 29 preventable cases per year. The introduction of PCV7 into the childhood immunization schedule would reduce the burden of pneumococcal disease in children, and the incidence would be particularly reduced in those children aged 1 year. Additional benefits may be gained in adults through herd protection. Continued surveillance of IPD is required before, during, and after the introduction of PCV7.
Description ofMycobacterium conceptionensesp. nov., aMycobacterium fortuitumGroup Organism Isolated from a Posttraumatic Osteitis Inflammation
Authors: Adékambi Toïdi , Stein Andréas , Carvajal Joseph , Raoult Didier , Drancourt Michel
Published: April 2006

A nonpigmented rapidly growing mycobacterium was isolated from wound liquid outflow, bone tissue biopsy, and excised skin tissue from a 31-year-old woman who suffered an accidental open right tibia fracture and prolonged stay in a river. The three isolates grew in 3 days at 24 to 37°C. 16S rRNA sequence analyses over 1,483 bp showed that they were identical and shared 99.7% (4-bp difference) sequence similarity with that ofMycobacterium porcinum, the most closely related species. PartialrpoB(723 bp) sequence analyses showed that the isolates shared 97.0% sequence similarity with that ofM. porcinum. Further polyphasic approaches, including biochemical tests, antimicrobial susceptibility analyses, andhsp65,sodA, andrecAgene sequence analysis, as well as % G+C determination and cell wall fatty acid composition analysis supported the evidence that these isolates were representative of a new species. Phylogenetic analyses showed the close relationship withM. porcinumin theMycobacterium fortuitumgroup. The isolates were susceptible to most antibiotics and exhibited evidence for penicillinase activity, in contrast toM. porcinum. We propose the nameMycobacterium conceptionensesp. nov. for this new species associated with posttraumatic osteitis. The type strain is D16T(equivalent to CIP 108544Tand CCUG 50187T).
Identification and Quantification of Archaea Involved in Primary Endodontic Infections
Authors: Vianna M. E. , Conrads G. , Gomes B. P. F. A. , Horz H. P.
Published: April 2006

Members of the domain
Archaea
, one of the three domains of life, are a highly diverse group of prokaryotes, distinct from bacteria and eukaryotes. Despite their abundance and ubiquity on earth, including their close association with humans, animals, and plants, so far no pathogenic archaea have been described. As some archaea live in close proximity to anaerobic bacteria, for instance, in the human gut system and in periodontal pockets, the aim of our study was to assess whether archaea might possibly be involved in human endodontic infections, which are commonly polymicrobial. We analyzed 20 necrotic uniradicular teeth with radiographic evidence of apical periodontitis and with no previous endodontic treatment. Using real-time quantitative PCR based on the functional gene
mcrA
(encoding the methyl coenzyme M reductase, specific to methanogenic archaea) and on archaeal 16S rRNA genes, we found five cases to be positive. Direct sequencing of PCR products from both genes showed that the archaeal community was dominated by a
Methanobrevibacter oralis
-like phylotype. The size of the archaeal population at the diseased sites ranged from 1.3 × 10
5
to 6.8 × 10
5
16S rRNA gene target molecule numbers and accounted for up to 2.5% of the total prokaryotic community (i.e., bacteria plus archaea). Our findings show that archaea can be intimately connected with infectious diseases and thus support the hypothesis that members of the domain
Archaea
may have a role as human pathogens.
Presence of E6 and E7 mRNA from Human Papillomavirus Types 16, 18, 31, 33, and 45 in the Majority of Cervical Carcinomas
Authors: Kraus Irene , Molden Tor , Holm Ruth , Lie A. Kathrine , Karlsen Frank , Kristensen Gunnar B. , Skomedal Hanne
Published: April 2006

The oncogenic potential of the human papillomavirus (HPV) early genes E6 and E7 is well established and a source of interest with regard to HPV testing for cervical carcinoma. Here we present a study performed with 204 histologically confirmed invasive cervical squamous cell carcinomas (SCCs) in which we evaluated the HPV E6 and E7 mRNA detection assay PreTect HPV-Proofer for detection of high-risk HPV types 16, 18, 31, 33, and 45. For further evaluation, detection of E6 and E7 mRNA from HPV types 35, 52, and 58 by real-time multiplex nucleic acid sequence-based amplification was also included. For comparison and to assess the overall prevalence of various HPV types, samples were also tested for HPV DNA by both consensus and type-specific PCR, reverse line blotting, sequencing, and in situ hybridization. The overall prevalence of HPV was 97%. HPV E6 and E7 transcripts were detected in 188 of 204 (92%) biopsy specimens, of which 181 contained one of the following HPV types: 16, 18, 31, 33, or 45. Consensus PCR and type-specific PCR detected HPV in 187 of 204 and 188 of 204 (92%) specimens, respectively. In conclusion, this study verifies the presence of HPV E6 and E7 mRNA in SCCs and demonstrates that HPV infections among Norwegian women with SCCs are limited mainly to the five high-risk types, 16, 18, 31, 33, and 45. This, together with the fact that PreTect HPV-Proofer detects the HPV oncogenic transcripts, suggests that the assay is a valuable approach in the field of HPV detection in cervical carcinoma.
Occurrence of Overlooked Zoonotic Tuberculosis: Detection of
Mycobacterium bovis
in Human Cerebrospinal Fluid
Authors: Shah N. P. , Singhal A. , Jain A. , Kumar P. , Uppal S. S. , Srivatsava M. V. P. , Prasad H. K.
Published: April 2006

The paucibacillary nature of the cerebrospinal fluid (CSF) has been a major obstacle in the diagnosis of human tuberculous meningitis (TBM). This study shows that with molecular techniques direct precise determination to the species level of mycobacterial pathogens can be made. The present report describes the utility of a nested PCR (N-PCR) assay (A. Mishra, A. Singhal, D. S. Chauhan, V. M. Katoch, K. Srivastava, S. S. Thakral, S. S. Bharadwaj, V. Sreenivas, and H. K. Prasad, J. Clin. Microbiol. 43:5670-5678, 2005) in detecting
M. tuberculosis
and
M. bovis
in human CSF. In 2.8% (6/212) of the samples,
M. tuberculosis
was detected, and in 17% (36/212),
M. bovis
was detected. Mixed infection was observed in 22 samples. Comparative analysis of clinical diagnosis, smear microscopy, and N-PCR in 69 patients (TBM, 25; non-TBM, 44) showed that the sensitivity of N-PCR (61.5%) was greater than that of smear microscopy (38.4%). Determination to the species level is important from the viewpoint of determining the prevalence of these mycobacteria in a community and would influence strategies currently adopted for the prevention of tuberculosis.
Rapid Detection of Norovirus from Fecal Specimens by Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay
Authors: Fukuda Shinji , Takao Shinichi , Kuwayama Masaru , Shimazu Yukie , Miyazaki Kazuo
Published: April 2006

In this study, we developed a one-step, single-tube genogroup-specific reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the detection of norovirus (NoV) genomes targeting from the C terminus of the RNA-dependent RNA polymerase gene to the capsid N-terminal/shell domain region. This is the first report on the development of an RT-LAMP assay for the detection of NoV genomes. Because of the diversity of NoV genotypes, we used 9 and 13 specially designed primers containing mixed bases for genogroup I (GI) and II (GII), respectively. The RT-LAMP assay had the advantages of rapidity, simplicity, specificity, and selectively and could obtain results within 90 min, generally even within 60 min, under isothermal conditions at 62°C. The detection limits for NoV genomes were between 10
2
and 10
3
copies/tube for GI and GII with differentiation by genotype, and no cross-reactions among NoV GI and GII and other gastroenteritis viruses, such as sapovirus, human astrovirus, adenovirus type 40 and 41, and group A and C rotavirus, were found. In the evaluation tests with fecal specimens obtained from gastroenteritis outbreaks, the sensitivity and specificity of the RT-LAMP assay with regard to RT-PCR were 100 and 94% for GI and 100 and 100% for GII, respectively. These findings establish that the RT-LAMP assay is potentially useful for the rapid detection of NoV genomes from fecal specimens in outbreaks of food-borne and person-to-person-transmitted gastroenteritis.
Use of PCR and Reverse Line Blot Hybridization Assay for Rapid Simultaneous Detection and Serovar Identification of
Chlamydia trachomatis
Authors: Xiong Likuan , Kong Fanrong , Zhou Hua , Gilbert Gwendolyn L.
Published: April 2006

The aim of this study was to develop and evaluate multiplex and nested PCR-reverse line blot (RLB) hybridization assays for detection and serovar identification of
Chlamydia trachomatis
. Two sets of primers targeting the VD2 region of the
omp1
gene and one set targeting the cryptic plasmid were designed for use in multiplex (both targets) and nested PCR (
omp1
only). For the RLB assay, labeled
omp1
amplicons were hybridized to a membrane containing probes specific for 15
C. trachomatis
serovars. The assays were used to test 429 clinical specimens, which had been previously tested for
C. trachomatis
using the COBAS AMPLICOR system. Specimens were tested without knowledge of the COBAS AMPLICOR result. Of 205 specimens that were positive by COBAS AMPLICOR, 201 (98%) were positive by multiplex PCR-RLB and 188 (92%) were also positive by
omp1
nested PCR-RLB. In addition, three of 224 COBAS AMPLICOR-negative specimens were positive by
omp1
nested PCR-RLB. One hundred sixty-six of 191 (87%) specimens in which
C. trachomatis
serovars were identified contained only one serovar and 25 (13%) contained two or three serovars. Serovars D, E, and F were found in 31 (16%), 83 (43%), and 51 (27%) specimens, respectively. Serovar E (41%) was the most commonly identified single serovar. Serovars J and K were found alone uncommonly (<2% each), but 18 of 25 (72%) specimens with multiple
C. trachomatis
serovars contained one or both (10 specimens) of these serovars. The nested (
ompI
) PCR-RLB is a specific and sensitive method for simultaneous detection and serovar identification of
C. trachomatis
, which can reliably identify mixed
C. trachomatis
serovars. It is suitable for use in epidemiological studies.
Use of a Serotype-Specific DNA Microarray for Identification of Group B
Streptococcus
(
Streptococcus agalactiae
)
Authors: Wen Linyan , Wang Quan , Li Yayue , Kong Fanrong , Gilbert Gwendolyn L. , Cao Boyang , Wang Lei , Feng Lu
Published: April 2006

Group B
Streptococcus
(GBS;
Streptococcus agalactiae
) is an important cause of sepsis and meningitis. Nine GBS serotypes, based on capsular polysaccharide (CPS) antigens, have been described. Their distribution varies worldwide and needs to be monitored to understand the epidemiology of GBS disease and inform the development of vaccines. In this study, we sequenced
cpsH
of GBS serotype II (
cpsHII
) and compared it with that of the other eight serotypes to identify serotype-specific regions. We then developed a DNA microarray based on the
cpsH
gene and used it to test 88 GBS isolates—9 serotype reference strains and 79 clinical isolates—and 7 other bacterial and fungal species which are commonly present in the vagina flora. The microarray was shown to be specific and reproducible. This is the first report of a microarray which can identify the nine GBS serotypes. The use of a microarray has advantages over traditional serotyping methods and will be of practical value in both reference and diagnostic laboratories.
Can the
fliC
PCR-Restriction Fragment Length Polymorphism Technique Replace Classic Serotyping Methods for Characterizing the H Antigen of Enterotoxigenic
Escherichia coli
Strains?
Authors: Ramos Moreno Ana Carolina , Cabilio Guth Beatriz Ernestina , Baquerizo Martinez Marina
Published: April 2006

In this study, we performed
fliC
PCR-restriction fragment length polymorphism (RFLP) to investigate whether this technique would be better than classic serotyping for the characterization of the H antigen in enterotoxigenic
Escherichia coli
(ETEC) strains. We showed that the
fliC
genes from ETEC strains can be characterized by restriction analysis of their polymorphism. Only one allele of the
fliC
gene from ETEC strains was found for each flagellar antigen, with the exception of H21. Nonmotile strains could also be characterized using this molecular technique. Moreover, determination of the somatic antigen was guided by the identification of the flagellar antigen from previously unknown serotypes of ETEC strains by PCR-RFLP, thus reducing the number of anti-antigen O sera used. The PCR-RFLP technique proved to be faster than classic serotyping for the characterization of the
E. coli
H antigen, taking 2 days to complete instead of the 7 or more days using classic serotyping. In conclusion, the H molecular typing for
Enterobacteriaceae
members may become an important epidemiological tool for the characterization of the H antigen of
E. coli
pathotypes. The PCR-RFLP technique is capable of guiding the determination of the H antigen and could partially replace seroagglutination. With the determination of the molecular profiles of alleles from strains obtained in epidemiological studies, new patterns will be described for ETEC strains or other
E. coli
pathotypes, thus permitting widespread use of this technique to characterize
fliC
genes and determine the H antigen of
E. coli
strains.
Characterization of Human Metapneumovirus Infections in Israel
Authors: Regev Liora , Hindiyeh Musa , Shulman Lester M. , Barak Asher , Levy Virginia , Azar Roberto , Shalev Yael , Grossman Zehava , Mendelson Ella
Published: April 2006

Respiratory tract infections are a leading cause of morbidity and mortality worldwide. Even with the advancement of diagnostic tools, the causative agent of 20 to 30% of upper respiratory tract infections go undiagnosed. Recently, a newly identified human respiratory virus, human metapneumovirus (hMPV), was discovered in young children in The Netherlands. To study the prevalence of hMPV infections in Israeli children, respiratory specimens from 388 hospitalized children less than 5 years of age were evaluated for the presence of hMPV RNA, which was present in 42 (10.8%) of these samples. All hMPV-positive samples were negative for respiratory syncytial virus (RSV), influenza viruses (Flu) A and B, adenovirus, and parainfluenza viruses 1, 2, and 3. Conversely, hMPV RNA was not detected in 76 RSV-positive and 38 Flu A- or B-positive samples. Most hMPV activity was between the months February and April. Sequence analysis of 20 positive samples revealed that both of the hMPV genotypes (groups 1 and 2) have circulated in central Israel during the study period. Moreover, three of the four known hMPV subgroups (1A, 1B, and 2B) were detected among the tested samples. Seroprevalence of hMPV in 204 patients from the central part of Israel revealed that 100% of the children are hMPV seropositive by the age of 5 years old. We conclude that hMPV is a common respiratory pathogen in Israel, while mixed infections of hMPV with RSV or Flu in hospitalized children are apparently rare.
Quantification of Hepatitis B Virus (HBV) DNA with a TaqMan HBV Analyte-Specific Reagent following Sample Processing with the MagNA Pure LC Instrument
Authors: Germer Jeffrey J. , Qutub Mohammed O. , Mandrekar Jayawant N. , Mitchell P. Shawn , Yao Joseph D. C.
Published: April 2006

TaqMan hepatitis B virus (HBV) analyte-specific reagent (ASR; Roche Molecular Systems, Inc., Branchburg, NJ) is designed for the quantification of HBV DNA in serum or plasma. The performance characteristics of TaqMan HBV ASR following automated sample processing with the MagNA Pure LC instrument (MP; Roche Applied Science, Indianapolis, IN) were evaluated in this study. Analytical sensitivity and precision were assessed with commercially available HBV standards, while clinical serum specimens from HBsAg-seropositive patients and healthy blood donors were used to determine clinical sensitivity, specificity, and correlation with other commercially available assays. Analytical studies yielded a limit of detection of 2.4 IU/ml, with good linearity and correlation (
R
2
= 0.9958) with expected HBV DNA titers over a wide range (6.0 × 10
0
to 2.1 × 10
8
IU/ml). Clinical sensitivity and specificity of the assay combined with automated sample processing were both 100%. Comparison of TaqMan HBV ASR and VERSANT HBV DNA 3.0 assay (bDNA; Bayer HealthCare LLC, Tarrytown, NY) results among clinical specimens yielded good correlation (
R
2
= 0.9237), with a mean difference in titer of −0.213 log
10
IU/ml (95% confidence interval, −0.678 to 1.10 log
10
IU/ml). The overall test failure rate was 2.0% among 204 clinical serum specimens tested. Total time required for MP sample processing and automated postelution handling of 24 samples was 224 min, with 57 min of actual hands-on time. MP is a reliable, labor-saving platform suitable for use with TaqMan HBV ASR, providing sensitive and accurate quantification of HBV DNA levels over a range of 8 log
10
IU/ml.
Differentiation of
Escherichia coli
Pathotypes by Oligonucleotide Spotted Array
Authors: Palaniappan Raghavan U. M. , Zhang Yu , Chiu David , Torres Alfonso , DebRoy Chobi , Whittam Thomas S. , Chang Yung-Fu
Published: April 2006

To accurately determine the pathotypes of
Escherichia coli
strains, a comprehensive assessment of each strain that targets multiple genes is required. A new approach to the identification and characterization of
E. coli
pathotypes was developed by constructing gene-specific probes (70-mers) for not only the virulence genes associated with each
E. coli
pathotype but also the O157-, CFT073-, and K-12-specific and common genes of each pathotype. Analysis of oligonucleotide probes with reference and clinical isolates of
E. coli
pathotypes indicated that the array could differentiate the pathotypes on the basis of their virulence and specific gene patterns. Probes targeting common genes of
E. coli
were present in all the reference and clinical strains.
Salmonella enterica
subsp.
enterica
-specific genes and
Salmonella
core genes were used as negative controls. The entire
E. coli
pathotype showed reactivity to only 4 of the 81
Salmonella
-specific gene probes. Characterization of the genetic and virulence profiles of a single strain by using probes for virulence factors and specific and common genes in the spotted array is an ideal diagnostic tool for determination of
E. coli
pathotypes and could also have a significant impact on the epidemiological analysis of
E. coli
infections.
A Sensor To Detect the Early Stages in the Development of CrystallineProteus mirabilisBiofilm on Indwelling Bladder Catheters
Authors: Stickler D. J. , Jones S. M. , Adusei G. O. , Waters M. G.
Published: April 2006

A simple sensor has been developed to detect the early stages of urinary catheter encrustation and avoid the clinical crises induced by catheter blockage. In laboratory models of colonization byProteus mirabilis, the sensor signaled encrustation at an average time of 43 h before catheters were blocked with crystalline biofilm.
Molecular Investigation of Human Immunodeficiency Virus Type 2 Subtype A Cases in South Korea
Authors: Nam Jeong-Gu , Kim Gab Jung , Baek Jin Young , Suh Soon Duck , Kee Mee Kyung , Lee Joo-Shil , Kim Sung Soon
Published: April 2006

We investigated the molecular characteristics of human immunodeficiency virus type 2 (HIV-2) subtype A isolates to clarify the transmission mode of HIV-2 within Korea. These findings indicated that the viruses from the six patients infected within Korea formed a distinct subcluster in the phylogenetic tree and might have been transmitted from one source.
Compliance of Clinical Microbiology Laboratories in the United States with Current Recommendations for Processing Respiratory Tract Specimens from Patients with Cystic Fibrosis
Authors: Zhou Juyan , Garber Elizabeth , Desai Manisha , Saiman Lisa
Published: April 2006

Respiratory tract specimens from patients with cystic fibrosis (CF) require unique processing by clinical microbiology laboratories to ensure detection of all potential pathogens. The present study sought to determine the compliance of microbiology laboratories in the United States with recently published recommendations for CF respiratory specimens. Microbiology laboratory protocols from 150 of 190 (79%) CF care sites were reviewed. Most described the use of selective media for
Burkholderia cepacia
complex (99%),
Staphylococcus aureus
(82%), and
Haemophilus influenzae
(89%) and identified the species of all gram-negative bacilli (87%). Only 52% delineated the use of agar diffusion assays for susceptibility testing of
Pseudomonas aeruginosa
. Standardizing laboratory practices will improve treatment, infection control, and our understanding of the changing epidemiology of CF microbiology.
Clonal Distribution and Differential Occurrence of the Enterotoxin Gene Cluster,
egc
, in Carriage- versus Bacteremia-Associated Isolates of
Staphylococcus aureus
Authors: van Belkum Alex , Melles Damian C. , Snijders Susan V. , van Leeuwen Willem B. , Wertheim Heiman F. L. , Nouwen Jan L. , Verbrugh Henri A. , Etienne Jerome
Published: April 2006

The
Staphylococcus aureus
enterotoxin gene cluster,
egc
, was detected in isolates from healthy individuals and in those from patients with bacteremia. The
egc
genes cooccur and are slightly enriched in strains from healthy carriers (present in 63.7% of carriage-associated isolates versus 52.9% of invasive isolates;
P
= 0.03). Multilocus sequence typing revealed that successful staphylococcal clones usually harbor the
egc
locus.
Early Detection of Hepatitis C Virus Infection by Use of a New Combined Antigen-Antibody Detection Assay: Potential Use for High-Risk Individuals
Authors: Schnuriger A. , Dominguez S. , Valantin M.-A. , Tubiana R. , Duvivier C. , Ghosn J. , Simon A. , Katlama C. , Thibault V.
Published: April 2006

The performance of a new combined antigen-antibody assay (Monolisa HCV Ag-Ab Ultra; Bio-Rad Laboratories) was evaluated in the context of acute hepatitis C in human immunodeficiency virus-infected patients. The combined assay became positive as early as the first PCR and earlier than a third-generation enzyme-linked immunosorbent assay in 65% of the cases. Reduction of the long period of HCV seronegativity should improve the diagnosis of hepatitis C infection, especially in high-risk populations.
Human Herpesvirus 6 Chromosomal Integration in Immunocompetent Patients Results in High Levels of Viral DNA in Blood, Sera, and Hair Follicles
Authors: Ward Katherine N. , Leong Hoe Nam , Nacheva Elisabeth P. , Howard Julie , Atkinson Claire E. , Davies Nicholas W. S. , Griffiths Paul D. , Clark Duncan A.
Published: April 2006

Six immunocompetent patients with human herpesvirus 6 (HHV-6) chromosomal integration had HHV-6 and β-globin DNA quantified in various samples by PCR. The mean HHV-6 DNA concentration (log
10
copies/milliliter) in blood was 7.0 (≥1 HHV-6 DNA copies/leukocyte), and in serum it was 5.3 (≥1 HHV-6 DNA copies/lysed cell). The mean HHV-6 DNA load (log
10
copies)/hair follicle was 4.2 (≥1 copies/hair follicle cell), demonstrating that viral integration is not confined to blood cells. The characteristically high HHV-6 DNA levels in chromosomal integration may confound laboratory diagnosis of HHV-6 infection and should be given due consideration.
Molecular Characterization and Drug Resistance Testing of
Mycobacterium tuberculosis
Isolates from Chad
Authors: Diguimbaye Colette , Hilty Markus , Ngandolo Richard , Mahamat Hassane H. , Pfyffer Gaby E. , Baggi Franca , Tanner Marcel , Schelling Esther , Zinsstag Jakob
Published: April 2006

The molecular characterizations of the first 40
Mycobacterium tuberculosis
isolates from Chad revealed a high proportion of isolates of the Cameroon family (33%), of which one isolate showed a monodrug resistance. In total, 9/33 (27%) isolates were resistant to isoniazid. The implications of these findings are discussed.
A 24-Hour Screening Protocol for Identification of Vancomycin-Resistant
Enterococcus faecium
Authors: Drews Steven J. , Johnson Grant , Gharabaghi Farhad , Roscoe Margaret , Matlow Anne , Tellier Raymond , Richardson Susan E.
Published: April 2006

We describe a 24-h protocol for the identification of patients who are positive for vancomycin-resistant
Enterococcus faecium
(VRE), using stool and rectal swab samples and VRE screening broth, automated DNA extraction, and real-time PCR for
vanA
and
vanB
genes. Compared to conventional screening methods, this protocol had a high sensitivity and specificity and a negative predictive value.
Fulminant, Acyclovir-Resistant, Herpes Simplex Virus Type 2 Hepatitis in an Immunocompetent Woman
Authors: Czartoski Todd , Liu Charlie , Koelle David M. , Schmechel Stephen , Kalus Andrea , Wald Anna
Published: April 2006

We report an immunocompetent woman with multisystem organ failure following herpes simplex virus type 2 (HSV-2) hepatitis. After she initially responded to intravenous acyclovir, she was switched to oral valacyclovir. She developed respiratory failure and opportunistic infections and died. Autopsy confirmed disseminated HSV infection, and lung tissue grew acyclovir-resistant HSV-2.
Fluoroquinolone Resistance in
Ureaplasma parvum
in the United States
Authors: Duffy Lynn , Glass John , Hall Geraldine , Avery Robin , Rackley Raymond , Peterson Scott , Waites Ken
Published: April 2006

We report the first case of naturally occurring fluoroquinolone resistance in
Ureaplasma
spp. from the United States. Resistance in this case probably developed as a result of mutations in the
gyrA
and
parC
genes of the DNA gyrase/topoisomerase IV complex that occurred in the presence of antimicrobial selective pressure.
The patient with supine hypertension and orthostatic hypotension: a clinical dilemma
Authors: Naschitz J E , Slobodin G , Elias N , Rosner I
Published: April 2006

Coexistent supine hypertension and orthostatic hypotension (SH-OH) pose a particular therapeutic dilemma, as treatment of one aspect of the condition may worsen the other. Studies of SH-OH are to be found by and large on patients with autonomic nervous disorders as well as patients with chronic arterial hypertension. In medical practice, however, the aetiologies and clinical presentation of the syndrome seem to be more varied. In the most typical cases the diagnosis is straightforward and the responsible mechanism evident. In those patients with mild or non-specific symptoms, the diagnosis is more demanding and the investigation may benefit from results of the tilt test, bedside autonomic tests as well as haemodynamic assessment. Discrete patterns of SH-OH may be recognisable. This review focuses on the management of the patient with coexistent SH-OH.
Recognition of acute illness in people with chronic neurological disability
Authors: Bakheit A M O
Published: April 2006

Coincidental acute illness is common in patients with chronic progressive or stable neurological disability. Although the presentation of acute illness in these patients is often similar to that in people without pre-existing disability, sometimes it may be atypical. In addition, an adequate medical history is often difficult to obtain in patients with advanced neurological disease, for example because of the presence of aphasia, severe dysarthria, or cognitive impairment. In these circumstances early and accurate diagnosis of acute medical and surgical emergencies requires both clinical acumen and a high index of suspicion. The aim of this article is to describe the main atypical manifestations of acute illness in patients with chronic disabling neurological disorders.
COX-2 inhibitors and the heart: are all coxibs the same?
Authors: Sooriakumaran P
Published: April 2006

The selective COX-2 inhibitors (coxibs) were originally developed to minimise the adverse effects of conventional non-steroidal anti-inflammatory drugs (NSAIDs) while maintaining the same analgesic and anti-inflammatory properties. Many large studies confirmed the improved gastric side effect profile of coxibs compared with non-selective NSAIDs; however, reports of increased cardiovascular morbidity and mortality followed, and the manufacturer Merck was forced to withdraw rofecoxib (Vioxx) from the market. Other coxibs have also either perished or had restrictions placed on their use. However, there seem to be significant differences between coxibs regarding their cardiovascular profiles, and the evidence for a class effect is dubious. In this paper, the current body of knowledge regarding the cardiovascular toxicities of coxibs is reviewed. The take home message for prescribing NSAIDs and those coxibs still on the market seems to be one of caution rather than contraindication, except in patients with significant cardiovascular risk factors.
Management of inflammatory bowel diseases in Eastern Europe
Authors: Lakatos L , Lakatos P L
Published: April 2006

Limited data are available on the management of inflammatory bowel diseases (IBD) in East European countries. The diagnostic tools and most treatment options are also available in Eastern Europe. The diagnostic procedures commonly used became more sophisticated in the past few years, with a greater use of computed tomography/magnetic resonance imaging and serology testing; however, double contrast barium enema, enteroclysis, and endoscopy remained standard. The medical therapy and surgical strategies are also somewhat different from those applied in Western countries. In ulcerative colitis, besides mesalazine, the use of sulphasalazine is still frequent, while azathioprine is only used in a minority of patients. The use of conventional corticosteroids is common and the rate of non-colorectal cancer associated colectomies is low. In contrast, 5-aminosalicylates are still used for maintenance in Crohn's disease and azathioprine is generally less frequently given compared with Western Europe. Biological agents have also become available about five years ago, yet their use is restricted mainly to specialised centres.
INVARIANCE OF WEYL ORDERING OF FERMI OPERATORS UNDER SIMILAR TRANSFORMATIONS
Authors: FAN HONG-YI
Published: April 2006

A Weyl–Wigner quantization scheme for Fermi system which is parallel to the bosonic case is established. We prove a new theorem: Weyl ordering of Fermi operators is invariant under fermionic similar transformations. Comparing with the preceding paper,1 we see the Bose–Fermi supersymmetry in this aspect. As an application of the theorem, we construct a generalized fermionic squeezed state.
Late Carboniferous plutonism within the pre-Alpine basement of the External Hellenides (Kithira, Greece): evidence from U–Pb zircon dating
Authors: Xypolias P. , Dörr W. , Zulauf G.
Published: April 2006

In memory of Theodor Doutsos, for his invaluable contribution to the understanding of geology of Greece. U–Pb zircon dating of three granitic orthogneisses, sampled from Kithira island, SW Greece, provide the first reliable evidence for Carboniferous plutonism within the pre-Alpine basement of the External Hellenides. Concordant zircons from two samples yielded ages at 324 ± 2 Ma and 323 ±3 Ma, whereas the third sample yielded a lower intercept age at 320 ± 1.2 Ma. The concordant ages are interpreted to date the emplacement of the igneous protolith. Ages of inherited zircons of two orthogneisses range from 2.5 to 2.4 Ga, indicating Late Archaean–Early Proterozoic crustal components. In combination with published ages for other Aegean metagranitoids, the new U–Pb ages provide additional evidence of a temporally restricted period of plutonism in the pre-Alpine Aegean region from 325 to 300 Ma. Comparing the post-Neoproterozoic evolution of the investigated basement with that of the Cycladic and Pelagonian basements and of the continental massifs of NE Greece and Turkey, we argue that all these crustal blocks were part of the Gondwana-derived Cimmerian terrane. Based on the spatial and temporal distribution of Late Carboniferous Aegean plutonism, we suggest that this period of magmatic events results from the southward subduction of Palaeotethys beneath the marginal fragments of northern Gondwana.
Subglacial imprints associated with the isolation and decay of an ice mass in the Lower Lough Erne basin, Co. Fermanagh, NW Ireland
Authors: Lafferty B. , Quinn R. , Breen C.
Published: April 2006

High-resolution Chirp sub-bottom profiler and side-scan sonar data record the final phases of ice margin activity in the Lower Lough Erne basin at the end of the last cycle of Quaternary glaciations in Ireland. Relative to the terrestrial glacial landforms the features in the Lough are smaller in scale and are considered to represent local ice mass dynamics that followed regional-scale events. Four phases are identified. (1) After the last ice sheet-wide readvance associated with the Killard Point Stadial (between 15.0 and 14.1
14
C ka bp), stagnation zone retreat resulted in isolation of a residual ice block in the Lower Lough Erne basin. (2) Proglacial waters developed coeval with retreat of the western margin of the ice block. Drawdown induced localized surging and the generation of push features and lineations. (3) Squeeze-up features, reflecting a heavily crevassed ice margin, mark the quiescent phase of the local surge cycle. (4) Iceberg grounding pits and keel marks record calving and rapid disintegration of the Lough Erne ice margin. The well-preserved glacigenic features observed in the lake basin suggest formation by a succession of mechanisms that were too short lived to obliterate the underlying evidence of ice margin dynamics.
Extensional fault control on the sedimentation patterns in a continental rift basin: El Castellar Formation, Galve sub-basin, Spain
Authors: Liesa Carlos L. , Soria Ana R. , Meléndez Nieves , Meléndez Alfonso
Published: April 2006

In the Galve sub-basin, the sedimentary record of the Upper Hauterivian–Lower Barremian El Castellar Formation is divided into two stages by a marlstone interval with gypsum. Stage 1 shows a great variety of subenvironments and facies (alluvial, palustrine and lacustrine) whereas in stage 2 an extensive, shallow carbonate lake developed. Sedimentation was controlled by a system of south-dipping, ENE–WSW listric normal faults, laterally bounded by NNW–SSE steeper transfer faults. Faults controlled sedimentation from a basin scale (basin margins and main characteristics and evolution of sediments) to a regional and a local scale (thickness and facies distribution of the synrift series as well as the location and evolution of lakes and minor alluvial fans). The changes between stages 1 and 2 are related to the passing from an independent movement of faults to the movement of all the extensional faults as a whole, at a sole detachment level. The interval with gypsum was caused by underground water flow changes associated with the interrelation and connection of the faults in the transition period. These changes have been correlated with the transition from the rift initial stage to the rift climax stage, which took place in the Hauterivian–Barremian transition.
Heterogeneous reworking of Grampian metamorphic complexes during Scandian thrusting in the Scottish Caledonides: insights from the structural setting and U–Pb geochronology of the Strath Halladale Granite
Authors: Kocks H. , Strachan R.A. , Evans J.A.
Published: April 2006

The Neoproterozoic Moine Supergroup in the Caledonides of northern Scotland is disposed in a series of thrust nappes. The Skinsdale Thrust in East Sutherland separates Moine rocks of the Loch Coire Migmatite Complex, which underwent anatexis during the Ordovician Grampian phase of the Caledonian orogeny, from the generally unmigmatized Scaraben succession. The Strath Halladale Granite occurs as an east-dipping sheeted complex that cuts discordantly at a low angle across the thrust. The pluton carries a magmatic foliation that was reworked by solid-state deformation at high to moderate temperatures; shear zones within the pluton display top-to-the-NW sense of shear similar to that deduced for the Skinsdale Thrust. Granite sheets are deformed by curvilinear folds that developed during ductile thrusting. Field and petrographic evidence is consistent with a thrust-related mode of emplacement. U–Pb monazite geochronology yields a crystallization age for the granite of 426 ± 2 Ma. This is interpreted to broadly date displacement along the Skinsdale Thrust, which therefore is of similar age to other NW-directed Silurian ductile thrusts within this sector of the Caledonides. These findings substantiate a model involving Ordovician (Grampian) deformation and metamorphism followed by Silurian (Scandian) thrust-related reworking and emplacement of nappes onto the Laurentian foreland.
The Pennsylvanian tropical biome reconstructed from the Joggins Formation of Nova Scotia, Canada
Authors: Falcon-Lang H.J. , Benton M.J. , Braddy S.J. , Davies S.J.
Published: April 2006

The Pennsylvanian (Langsettian) Joggins Formation contains a diverse fossil assemblage, first made famous by Lyell and Dawson in the mid-19th century. Collector curves based on
c
. 150 years of observation suggest that the Joggins fossil record is relatively complete. A key feature of the site is that fossils occur in (par)autochthonous assemblages within a narrow time interval (<1 Ma). Analysis of co-occurring taxa within a precise facies context permits ecosystem reconstruction, and three main communities are recognized in this study. Brackish seas, the distal extension of European marine bands, were populated by Foraminifera, molluscs, annelids, arthropods, fishes, and aquatic tetrapods. Poorly drained coastal plains were covered by rainforests of lycopsids, calamiteans, ferns, pteridosperms, and cordaitaleans, inhabited by a terrestrial fauna of molluscs, annelids, arthropods, and tetrapods, including the earliest known reptiles. Well-drained alluvial plains were covered by fire-prone cordaitalean scrub containing a low-diversity fauna of molluscs, arthropods, and tetrapods, locally preserved in waterholes. These three environments repeatedly interchanged with one another in response to base-level fluctuations forced by tectonism and glacioeustasy. Located further inland than other well-studied Pennsylvanian tropical sites, the Joggins Formation is significant because it contains a record of intra-continental terrestrial ecosystems.
Calcitonin gene-related peptide inhibits interleukin-1β-induced endogenous monocyte chemoattractant protein-1 secretion in type II alveolar epithelial cells
Authors: Li Wenjing , Wang Tengke , Ma Chenming , Xiong Tingting , Zhu Yi , Wang Xian
Published: April 2006

As important multifunctional cells in the lung, alveolar epithelial type II (AEII) cells secrete numerous chemokines on various stimuli. Our previous data showed that AEII cells also express the neuropeptide calcitonin gene-related peptide (CGRP) and the proinflammatory factor interleukin (IL)-1β induces CGRP secretion in the A549 human AEII cell line. In the present study, the CGRP-1 receptor antagonist human (h)CGRP8–37(0.1–1 nM) greatly amplified the production of IL-1β-induced monocyte chemoattractant protein (MCP)-1. The inhibition of CGRP expression by small interfering RNA significantly increased MCP-1 secretion on IL-1β stimulation. However, exogenous hCGRP (10–100 nM) suppressed IL-1β-evoked MCP-1 secretion in MCP-1 promoter activity, and CGRP gene stably transfected cell clones significantly inhibited both the mRNA and protein levels of MCP-1 induced by IL-1β. These data imply that AEII-derived CGRP suppressed IL-1β-induced MCP-1 secretion in an autocrine/paracrine mode. Subsequent investigation revealed that CGRP inhibited IL-1β-evoked NF-κB activity by suppressing IκBα phosphorylation and degradation. Moreover, CGRP attenuated IL-1β-induced reactive oxygen species (ROS) formation, the early event in proinflammatory factor signaling. We previously showed that the CGRP inhibitory effect was mediated by elevated intracellular cAMP and show here that analogs of cAMP, 8-bromoadenosine 3′,5′-cyclic monophosphothioate and the Sp isomer of adenosine 3′,5′-cyclic monophosphothioate, mimicked the CGRP suppressive effect on IL-1β-induced ROS formation, NF-κB activation, and MCP-1 secretion. Thus increased endogenous CGRP secretion in lung inflammatory disease might eliminate the excessive response by elevating the cAMP level through inhibiting the ROS-NF-κB-MCP-1 pathway.
Mechanism of induction of pancreatic acinar cell apoptosis by hydrogen sulfide
Authors: Cao Yang , Adhikari Sharmila , Ang Abel Damien , Moore Philip K. , Bhatia Madhav
Published: April 2006

The present study investigated the mechanism of mouse pancreatic acinar cell apoptosis induced by H2S in an in vitro system, using isolated pancreatic acini. Treatment of pancreatic acini with 10 μM NaHS (a donor of H2S) for 3 h caused phosphatidylserine externalization as shown by annexin V binding, an indicator of early stages of apoptosis. This treatment also resulted in the activation of the caspase cascade and major changes at the mitochondrial level. Caspase-3, -8, and -9 activities were stimulated by H2S treatment. Treatment with inhibitors of caspase-3, -8, and -9 significantly inhibited H2S-induced phosphatidylserine externalization as shown by reduced annexin V staining. The mitochondrial membrane potential was collapsed in H2S-treated acini as evidenced by fluorescence microscopy and quantitative analysis. Furthermore, the treatment of acini with H2S caused the release of cytochrome c by the mitochondria. To investigate the mechanism underlying pancreatic acinar cell apoptosis, we also characterized the protein expression of a range of molecules that are each known to influence the apoptotic pathway. Among proapoptotic proteins, Bax expression was activated in H2S-treated cells but not Bid, and the antiapoptotic proteins Bcl-XL and Bcl-2 did not show any activation in pancreatic acinar cell apoptosis. The death effector domain-containing protein Flip is downregulated in H2S-treated acini. These results demonstrate the induction of pancreatic acinar cell apoptosis in vitro by H2S and the involvement of both mitochondrial and death receptor pathways in the process of apoptosis.
Thermosensitive TRP ion channels mediate cytosolic calcium response in human synoviocytes
Authors: Kochukov Mikhail Y. , McNearney Terry A. , Fu Yibing , Westlund Karin N.
Published: April 2006

The transient receptor potential (TRP) channels are important membrane sensors, responding to thermal, chemical, osmotic, or mechanical stimuli by activation of calcium and sodium fluxes. In this study, three distinct TRP channels were detected and their role established in mediating cytosolic free calcium concentration ([Ca2+]cyt) response in tumor-derived SW982 synoviocytes and primary cultures of human synovial cells from patients with inflammatory arthropathies. As shown by fura-2 ratio measurements while cells were incubated in a temperature-regulated chamber, significant [Ca2+]cyt elevation was elicited by rapid changes in bath temperature, application of TRPV1 receptor agonists capsaicin and resiniferatoxin, or a cold receptor stimulator, icilin. Temperature thresholds for calcium response were determined to be 12 ± 1°C for cold and 28 ± 2°C for heat activation. Temperature increases or decreases beyond these thresholds resulted in a significant rise in the magnitude of [Ca2+]cyt spikes. Observed changes in [Ca2+]cyt were completely abolished in calcium-free medium and thus resulted from direct calcium entry through TRP channels rather then by activation of voltage-dependent calcium channels. Two heat sensitive channels, TRPV1 and TRPV4, and a cold-sensitive channel, TRPA1, were detected by RT-PCR. Minimal mRNA for TRPV3 or TRPM8 was amplified. The RT-PCR results support the data obtained with the [Ca2+]cyt measurements. We propose that the TRP channels are functionally expressed in human synoviocytes and may play a critical role in adaptive or pathological changes in articular surfaces during arthritic inflammation.