bluuebunny/crossref_metadata_2025 huggingface dataset:
Serodiagnosis Using Recombinant Nipah Virus Nucleocapsid Protein Expressed in
Escherichia coli
Authors: Yu Fuxun , Khairullah Nor Shahidah , Inoue Shingo , Balasubramaniam Vijayamalar , Berendam Stella Joan , Teh Leok Kin , Ibrahim Nik Shamsiah Wan , Abdul Rahman Sohayati , Hassan Sharifah Syed , Hasebe Futoshi , Sinniah Mangalam , Morita Kouichi
Published: September 2006

Nipah virus nucleocapsid (NiV-N) protein was expressed in
Escherichia coli
and purified by histidine tag-based affinity chromatography. An indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) for human and swine sera and an IgM capture ELISA for human sera were established using the recombinant NiV-N protein as an antigen. One hundred thirty-three suspected patient sera and 16 swine sera were used to evaluate the newly established ELISA systems in comparison with the CDC inactivated-virus-based ELISA systems. For the human sera, the NiV-N protein-based indirect IgG ELISA had a sensitivity of 98.6% and a specificity of 98.4%, and the NiV-N protein-based IgM capture ELISA had a sensitivity of 91.7% and a specificity of 91.8%, with reference to the CDC ELISA systems. The NiV-N-based IgM ELISA was found to be more sensitive than the inactivated-virus-based ELISA in that it captured eight additional cases. For the swine sera, the two test systems were in 100% concordance. Our data indicate that the Nipah virus nucleocapsid protein is a highly immunogenic protein in human and swine infections and a good target for serodiagnosis. Our NiV-N protein-based ELISA systems are useful, safe, and affordable tools for diagnosis of Nipah virus infection and are especially fit to be used in large-scale epidemiological investigations and to be applied in developing countries.
Serological Detection of Varicella-Zoster Virus-Specific Immunoglobulin G by an Enzyme-Linked Immunosorbent Assay Using Glycoprotein Antigen
Authors: Sauerbrei A. , Wutzler P.
Published: September 2006

Since the introduction of varicella vaccination in several countries, there has been an urgent need for commercially available test procedures that allow highly sensitive and specific quantitative determination of the varicella-zoster virus (VZV)-specific immune status, including immunity postimmunization. This study compared the performance of two enzyme-linked immunosorbent assays (ELISAs) for the sensitive and specific determination of VZV-specific immunoglobulin G (IgG) in seronegative and latently infected persons, as well as in vaccinees. One ELISA is based on the detection of antibody to VZV-specific envelope glycoproteins (gp), and the other comprises the whole antigen extract prepared from VZV-infected cells. A modified standard fluorescent-antibody-to-membrane-antigen (FAMA) assay was used as a reference. An excellent sensitivity (100%) in relation to FAMA was demonstrated for the gpELISA (Virion\Serion), while the non-gpELISA (Dade Behring) had a lower sensitivity (83%) when sera from latently infected persons were tested. After postvaccinal immunity was measured, a sensitivity of 87% was achieved with gpELISA, whereas the ELISA incorporating antigen extract of VZV-infected cells had a sensitivity of 78%. Excellent specificity (100%) was calculated for both the gpELISA and the non-gpELISA. In conclusion, SERION ELISA
classic
VZV IgG is useful for the sensitive and specific quantitative determination of VZV immune status after natural infection. The test can also be recommended for measuring antibody response after varicella vaccination, particularly after the cutoff value was optimized.
Molecular Epidemiology of Multidrug-ResistantMycobacterium bovisIsolates with the Same Spoligotyping Profile as Isolates from Animals
Authors: Romero Beatriz , Aranaz Alicia , Juan Lucía de , Álvarez Julio , Bezos Javier , Mateos Ana , Gómez-Mampaso Enrique , Domínguez Lucas
Published: September 2006

PCR-based characterization techniques have been adopted in most laboratories forMycobacterium bovistyping. We report a molecular characterization of human multidrug-resistantM. bovisisolates and three bovine isolates that share the spoligotyping profile. The analysis of the direct repeat region showed that both groups differed in the presence of spacers not included in the current membrane. They were also distinguished by two out of the nine mycobacterial interspersed repetitive unit variable-number tandem repeat loci tested, indicating that the human infection was not acquired from the cattle from which isolates were obtained. These results highlight that a combination of techniques is required for appropriate discrimination, even for those spoligotypes that have a low frequency.
Peptide Nucleic Acid Fluorescence In Situ Hybridization-Based Identification of
Candida albicans
and Its Impact on Mortality and Antifungal Therapy Costs
Authors: Forrest G. N. , Mankes K. , Jabra-Rizk M. A. , Weekes E. , Johnson J. K. , Lincalis D. P. , Venezia R. A.
Published: September 2006

The impact of rapid identification of
Candida albicans
blood isolates by peptide nucleic acid fluorescence in situ hybridization (PNA FISH) on the selection and expenditure of antifungal therapy was evaluated. PNA FISH was 100% sensitive and specific in the rapid identification of 31 out of 72 candidemias as
C. albicans
and resulted in a significant reduction of caspofungin usage, with an overall cost savings of $1,729 per patient.
Prevalence of Newer β-Lactamases in Gram-Negative Clinical Isolates Collected in the United States from 2001 to 2002
Authors: Moland Ellen S. , Hanson Nancy D. , Black Jennifer A. , Hossain Ashfaque , Song Wonkeun , Thomson Kenneth S.
Published: September 2006

Newer β-lactamases such as extended-spectrum β-lactamases (ESBLs), transferable AmpC β-lactamases, and carbapenemases are associated with laboratory testing problems of false susceptibility that can lead to inappropriate therapy for infected patients. Because there appears to be a lack of awareness of these enzymes, a study was conducted during 2001 to 2002 in which 6,421 consecutive, nonduplicate clinical isolates of aerobically growing gram-negative bacilli from patients at 42 intensive care unit (ICU) and 21 non-ICU sites across the United States were tested on-site for antibiotic susceptibility. From these isolates, 746 screen-positive isolates (11.6%) were referred to a research facility and investigated to determine the prevalence of ESBLs in all gram-negative isolates, transferable AmpC β-lactamases in
Klebsiella pneumoniae
, and carbapenemases in
Enterobacteriaceae
. The investigations involved phenotypic tests, isoelectric focusing, β-lactamase inhibitor studies, spectrophotometric assays, induction assays, and molecular analyses. ESBLs were detected only in
Enterobacteriaceae
(4.9% of all
Enterobacteriaceae
) and were found in species other than those currently recommended for ESBL testing by the CLSI (formerly NCCLS). These isolates occurred at 74% of the ICU sites and 43% of the non-ICU sites. Transferable AmpC β-lactamases were detected in 3.3% of
K. pneumoniae
isolates and at 16 of the 63 sites (25%) with no difference between ICU and non-ICU sites. Three sites submitted isolates that produced class A carbapenemases. No class B or D carbapenemases were detected. In conclusion, organisms producing ESBLs and transferable AmpC β-lactamases were widespread. Clinical laboratories must be able to detect important β-lactamases to ensure optimal patient care and infection control.
Molecular Differentiation ofTreponema pallidumSubspecies
Authors: Centurion-Lara Arturo , Molini Barbara J. , Godornes Charmie , Sun Eileen , Hevner Karin , Van Voorhis Wesley C. , Lukehart Sheila A.
Published: September 2006

Treponema pallidumincludes three subspecies of antigenically highly related treponemes. These organisms cause clinically distinct diseases and cannot be distinguished by any existing test. In this report, genetic signatures are identified in twotprgenes which, in combination with the previously published signature in the 5′ flanking region of the tpp15 gene, can differentiate theT. pallidumsubspecies, as well as a simian treponeme.
External Quality Assessment of Molecular Typing of
Staphylococcus aureus
Isolates by a Network of Laboratories
Authors: Deplano A. , De Mendonça R. , De Ryck R. , Struelens M. J.
Published: September 2006

A network of laboratories designated Centres for Molecular Diagnosis was funded in 2000 by Belgian National Health Insurance to provide clinically relevant molecular diagnostic tests. These included typing of nosocomial pathogens as a service to local hospital infection control programs. Two external quality assessment (EQA) surveys were performed in 2001 and 2003 to evaluate the proficiencies of the laboratories at
Staphylococcus aureus
typing. EQA panels included
S. aureus
isolates with either indistinguishable, clonally related, or unrelated pulsed-field gel electrophoresis (PFGE) patterns. A hypothetical hospital outbreak problem was also submitted for analysis. Typeability, reproducibility, discrimination (
D
) index, and epidemiological concordance were evaluated. Ten centers participated in each survey. Seven centers performed PFGE analysis, while others used repetitive-element or randomly amplified polymorphic DNA PCR, amplified fragment length polymorphism, or
spa
typing. Full typeability (100%) was achieved by all centers, and all but one showed 100% reproducibility. Discrimination was appropriate (
D
index, ≥96%) for centers performing PFGE analysis but not for all those using other methods (
D
index range, 72% to 97%). Correct answers to the epidemiological questions were provided by 7/10 and 10/10 centers in 2001 and 2003, respectively. Individual feedback of results was provided to each center together with specific technical recommendations for improving performance. Our findings indicate that surveys of lab proficiency are useful for validation and optimization of molecular typing services to local hospital infection control programs.
Molecular Epidemiology of Rabies Virus Isolates in India
Authors: Nagarajan T. , Mohanasubramanian B. , Seshagiri E. V. , Nagendrakumar S. B. , Saseendranath M. R. , Satyanarayana M. L. , Thiagarajan D. , Rangarajan P. N. , Srinivasan V. A.
Published: September 2006

In India, rabies is enzootic and is a serious public health and economic problem. India has a large population of stray dogs which, together with a lack of effective control strategies, might have led to the persistence of rabies virus (RV) in the canine population. Our objective was to study the molecular epidemiology of RV isolates in India based on nucleotide sequence analysis of 29 RV isolates originating from different species of animals in four states. Here we have analyzed two sets of sequence data based upon a 132-nucleotide region of the cytoplasmic domain (CD) of the G gene (G-CD) and a 549-nucleotide region (Psi-L) that combines the noncoding G-L intergenic region (Psi) and a fragment of the polymerase gene (L). Phylogenetic analysis revealed that the RV isolates belong to genotype 1 and that they were related geographically but were not related according to host species. Five different genetic clusters distributed among three geographical regions were identified. Comparison of the deduced amino acid sequences of G-CD between RV isolates revealed three amino acid changes (amino acid 462G [aa462G], aa465H, and aa468K) that distinguished the Indian RVs from RV isolates in other parts of the world. Analysis of the data indicated that the dog rabies virus variants are the major circulating viruses in India that transmit the disease to other domestic animals and humans as well.
Development of a New Method for Diagnosis of Rubella Virus Infection by Reverse Transcription-Loop-Mediated Isothermal Amplification
Authors: Mori Nobuo , Motegi Yoshie , Shimamura Yasushi , Ezaki Takashi , Natsumeda Tomo , Yonekawa Toshihiro , Ota Yoshinori , Notomi Tsugunori , Nakayama Tetsuo
Published: September 2006

We developed a useful method for the detection of rubella virus genome RNA by reverse transcription loop-mediated isothermal amplification (RT-LAMP) and compared the sensitivity of RT-LAMP with that of other virological tests: reverse transcription-PCR (RT-PCR) and virus isolation. The rubella virus genome was amplified by RT-LAMP from clinical isolates obtained between 1987 and 2004 with similar sensitivities to the Takahashi vaccine strain. The detection limit of RT-LAMP was compared with that of RT-PCR using the Takahashi vaccine strain. We detected rubella virus genome material corresponding to 30 PFU/ml in a culture fluid sample by RT-LAMP within 60 min after the extraction of RNA with equal sensitivity to RT-nested PCR. The positive result rates of RT-LAMP, RT-PCR, and virus isolation were also compared using throat swabs obtained from patients who were clinically diagnosed with acute rubella virus infection in 2004 in Tochigi, Japan. Among nine patients with clinical rubella, the positive result rates were three/nine (33.3%) for virus isolation, six/nine (66.7%) for RT-PCR, and seven/nine (77.8%) for RT-LAMP. Consequently, RT-LAMP for rubella virus would be expected to be a reliable rapid diagnostic tool in the clinical setting.
Nosocomial Spread of
Enterococcus faecium
Resistant to Vancomycin and Linezolid in a Tertiary Care Medical Center
Authors: Dobbs Thomas E. , Patel Mukesh , Waites Ken B. , Moser Stephen A. , Stamm Alan M. , Hoesley Craig J.
Published: September 2006

In May 2004 our institution encountered its first clinical isolate of linezolid-resistant, vancomycin-resistant
Enterococcus faecium
(LRVRE). Between October 2004 and July 2005, 40 patients from whom LRVRE organisms were recovered in clinical specimens were characterized. Epidemiologic investigation and pulsed-field gel electrophoresis patterns indicated a clonal outbreak related to nosocomial spread.
Aspergillus
Galactomannan Enzyme-Linked Immunosorbent Assay Cross-Reactivity Caused by Invasive
Geotrichum capitatum
Authors: Giacchino Mareva , Chiapello Nadia , Bezzio Stefania , Fagioli Franca , Saracco Paola , Alfarano Alda , Martini Vincenza , Cimino Giuseppe , Martino Pietro , Girmenia Corrado
Published: September 2006

We report three cases of invasive
Geotrichum capitatum
infection in patients with acute leukemia for which an enzyme-linked immunosorbent assay (ELISA) for
Aspergillus
galactomannan was positive, with no evidence of aspergillosis. Supernatants obtained from suspensions of 17
G. capitatum
strains gave positive reactions with the
Aspergillus
galactomannan ELISA. These clinical and laboratory data seem to suggest that
G. capitatum
produces a soluble antigen that is cross-reactive with
Aspergillus
galactomannan.
Capsule Production in
Bacillus cereus
Strains Associated with Severe Pneumonia
Authors: Sue David , Hoffmaster Alex R. , Popovic Tanja , Wilkins Patricia P.
Published: September 2006

We identified three encapsulated
Bacillus cereus
strains, isolated from patients with severe pneumonia, in a collection of
B. cereus
isolates associated with human illness. We found that the extent of capsule expression was influenced by culturing conditions. Our findings highlight consequent clinical and laboratory diagnostic challenges posed by such isolates.
Staphylococcus aureus
Throat Colonization Is More Frequent than Colonization in the Anterior Nares
Authors: Nilsson Peter , Ripa Torvald
Published: September 2006

The aim of this study was to determine the frequency and persistence of
Staphylococcus aureus
carriage in the throat in relation to anterior naris carriage. By use of a sensitive enrichment broth,
S. aureus
was cultured from the two sites from 259 patients upon admission to an orthopedic ward and from 87 staff members of the same ward. The throat was the most common carriage site in both groups. Forty percent of the patients and 54% of the staff were positive for
S. aureus
in the throat, compared to 31% and 36%, respectively, in the anterior nares. To determine the persistence of carriage, 67 individuals were repeatedly sampled from the anterior nares and the throat over 2 years (5 to 10 sampling occasions; mean, 7.8). The majority, 58% (39/67), were defined as persistent carriers of
S. aureus
, considering culture results from both sites. Of the 39 persistent carriers, 15 individuals were culture positive from only the throat on more than half of the sampling occasions (these are called preferential throat carriers) while only 5% (two individuals) were preferential anterior naris carriers by use of the same definition. Typing of the collected
S. aureus
isolates by pulsed-field gel electrophoresis revealed that the same strain of
S. aureus
was present, over time, in the throat of an individual at least to the same extent as in the anterior nares. Throat carriage was at least as persistent as carriage in the anterior nares.
Effectiveness of Cephalosporins in the Sputum of Patients with Nosocomial Bronchopneumonia
Authors: Klekner Almos , Bagyi Kinga , Bognar Laszlo , Gaspar Attila , Andrasi Melinda , Szabo Judit
Published: September 2006

Nosocomial bronchopneumonia is a frequent complication in patients with chronic intratracheal intubation. Despite targeted antibiotic treatment, production of abundant bronchial secretion containing pathogen bacteria often tends to be chronic, and so mortality drastically increases. This problem led to an investigation of the penetration of five cephalosporin antibiotics into the sputum. Serum and sputum were collected from 24 chronically intubated patients having purulent nosocomial bronchopneumonia treated in an intensive care unit (ICU). Patients received the following doses intravenously every 24 h: five received 70 mg/kg of body weight cefuroxime, four received 110 mg/kg cefamandole, six received 80 mg/kg ceftriaxone, four received 80 mg/kg ceftazidime, and five received 80 mg/kg cefepime. Antibiotic concentrations in the serum and sputum were evaluated by capillary electrophoresis. MICs were determined for bacteria isolated from the purulent bronchial secretions. The mean levels of the cephalosporins in the sputum did not reach the MICs for the bacteria isolated from the same samples. Ceftriaxone was the only one of the investigated five cephalosporins that had a measurable concentration in the sputum (1.4 ± 1.2 mg/liter). The low concentration of antibiotics in the purulent tracheobronchial secretion can be one of the many reasons for ineffective therapy of nosocomial bronchopneumonia in intubated patients in the ICUs. In the case of intubated or mechanically ventilated patients having chronic bronchopneumonia, determination of drug concentration in the bronchial secretion might be considered when selecting an antibiotic for treatment.
Real-Time Reverse Transcription-PCR for Detection of Rotavirus and Adenovirus as Causative Agents of Acute Viral Gastroenteritis in Children
Authors: Logan Catriona , O'Leary John J. , O'Sullivan Niamh
Published: September 2006

Viral pathogens are the most common cause of gastroenteritis in developed countries. Human rotavirus and adenovirus infections are major causes of acute outbreaks and sporadic cases of gastroenteritis, occurring primarily among children less than 2 years of age. Patient hospitalization is often required, with enormous infection control implications. This work describes the development of real-time PCR assays for the detection of group F adenovirus, rotavirus A, and rotavirus C from stool specimens. Two hundred twenty stool samples from pediatric patients exhibiting symptoms of diarrhea and/or vomiting were examined. PCR results were compared with those of virus detection by electron microscopy and latex agglutination antigen detection. The incorporation of an internal-control RNA that was spiked into individual stool extracts functioned as an internal validation for the reporting of PCR-negative results. Rotavirus C was not detected by real-time PCR in the patient stool samples examined. Real-time reverse transcription-PCR resulted in 175% and 111% increases in the rates of detection of adenovirus F and rotavirus A, respectively, compared with latex agglutination testing. Molecular detection increased the number of stool specimens in which causative agents of gastroenteritis were identified by 155% compared to electron microscopy. Genotyping of a proportion of the rotavirus and adenovirus strains identified only genotype G1 rotavirus and both adenovirus genotypes 40 and 41 in circulation within the patient cohort examined. The results highlight the significance of rapid molecular methods for the routine screening of stool samples in hospital laboratories to provide rapid definitive diagnoses.
Identification of Genotype 1 Hepatitis E Virus in Samples from Swine in Cambodia
Authors: Caron M. , Enouf V. , Than S. C. , Dellamonica L. , Buisson Y. , Nicand E.
Published: September 2006

Hepatitis E virus (HEV) is a major enterically transmitted pathogen in many developing countries, where it causes outbreaks and sporadic cases of acute hepatitis. A study conducted with pigs from several livestock farms in Cambodia identified one swine genotype 1 HEV isolate as being associated with prevalent swine genotype 3 HEV.
A Modified Christensen's Urea and CLSI Broth Microdilution Method for Testing Susceptibilities of SixMalasseziaSpecies to Voriconazole, Itraconazole, and Ketoconazole
Authors: Rincón S. , Cepero de García M. C. , Espinel-Ingroff A.
Published: September 2006

Two supplemented broths (Christensen's urea with 0.1% Tween 80 and 0.5% Tween 40 and RPMI 1640 with 1% glycerol, 1% peptone, 1.8% glucose, and 0.05% Tween 80) were evaluated to determine voriconazole, itraconazole, and ketoconazole MICs for 200Malasseziasp. isolates.Malassezia globosaandM. restrictawere the least susceptible species (MICs at which 90% of the isolates tested were inhibited, 1 to ≥8 μg/ml versus 0.25 to 1 μg/ml).
Invader Plus Method Detects Herpes Simplex Virus in Cerebrospinal Fluid and Simultaneously Differentiates Types 1 and 2
Authors: Allawi Hatim T. , Li Haijing , Sander Tamara , Aslanukov Azamat , Lyamichev Victor I. , Blackman Amondrea , Elagin Slava , Tang Yi-Wei
Published: September 2006

We report here on the development and validation of a prototype Invader Plus method for the qualitative detection of herpes simplex virus types 1 and 2 in cerebrospinal fluid (CSF). The method combines PCR and Invader techniques in a single, closed-tube, continuous-reaction format that gives an analytical sensitivity of approximately 10 copies per reaction. The clinical sensitivity and specificity were 100.0% and 98.6%, respectively, when the results of the method were validated against the results obtained with a PCR colorimetric microtiter plate system by use of clinical CSF specimens.
Antibiotic Susceptibility and Molecular Diversity of
Bacillus anthracis
Strains in Chad: Detection of a New Phylogenetic Subgroup
Authors: Maho Angaya , Rossano Alexandra , Hächler Herbert , Holzer Anita , Schelling Esther , Zinsstag Jakob , Hassane Mahamat H. , Toguebaye Bhen S. , Akakpo Ayayi J. , Van Ert Matthew , Keim Paul , Kenefic Leo , Frey Joachim , Perreten Vincent
Published: September 2006

We genotyped 15
Bacillus anthracis
isolates from Chad, Africa, using multiple-locus variable-number tandem repeat analysis and three additional direct-repeat markers. We identified two unique genotypes that represent a novel genetic lineage in the A cluster. Chadian isolates were susceptible to 11 antibiotics and free of 94 antibiotic resistance genes.
Molecular Epidemiology of Adenovirus Isolates from Patients Diagnosed with Intussusception in Melbourne, Australia
Authors: Selvaraj Gowri , Kirkwood Carl , Bines Julie , Buttery Jim
Published: September 2006

Twenty-one intussusception (IS)-associated and 59 temporally linked adenoviral isolates from clinical infections were compared. Species C (15/21 IS- and 32/59 non-IS-associated isolates) dominated. Of these, serotype 2 (AdV-2) (7/15 IS-associated isolates) and serotype 1 (AdV-1) (16/32 non-IS-associated isolates) were the most commonly identified serotypes. DNA restriction analysis of AdV-2 isolates identified six genomic types; of these, type D2 (3/7 IS- and 8/11 non-IS-associated isolates) was the dominant type after BamHI and SmaI digestion. IS-associated isolates are similar to circulating non-IS-associated strains.
Evaluation of Real-Time and Conventional PCR Targeting Complex 85 Genes for Detection of
Mycobacterium leprae
DNA in Skin Biopsy Samples from Patients Diagnosed with Leprosy
Authors: Martinez Alejandra N. , Britto Constança F. P. C. , Nery José A. C. , Sampaio Elizabeth P. , Jardim Márcia R. , Sarno Euzenir N. , Moraes Milton O.
Published: September 2006

In spite of the decrease in the number of registered leprosy patients, the number of new cases diagnosed each year (400,000) has remained essentially unchanged. Leprosy diagnosis is difficult due to the low sensitivity of current methodologies to identify new cases. In this study, conventional and TaqMan real-time PCR assays for detection of
Mycobacterium leprae
DNA were compared to current classification based on clinical, bacteriological, and histological evaluation.
M. leprae
DNA was extracted from frozen skin biopsy specimens from 69 leprosy patients enrolled in the study and was amplified using specific primers for either the antigen 85B-coding gene or the 85A-C intergenic region by using conventional and real-time PCR. The detection rate was 100% among multibacillary (MB) patients and ranged from 62.5% to 79.2% among paucibacillary (PB) patients according to the assay used. The TaqMan system for 85B gene amplification showed the highest sensitivity, although conventional PCR using the 85A-C gene as a target was also efficient. The cycle threshold (

C
T

) values obtained using the TaqMan system were able to statistically (
P
< 0.0001) differentiate MB (mean

C
T

, 28.06; standard deviation [SD], 4.51) from PB (mean

C
T

, 33.06; SD, 2.24) patients. Also, there was a correlation between

C
T

values and the bacteriological index for MB patients (Pearson's
r
, −0.444;
P
= 0.008). Within the PB patients' group, we tested normal skin from six patients exhibiting the pure neuritic form of leprosy (PNL). Five out of six PNL patients were positive for the presence of
M. leprae
DNA, even in the absence of skin lesions. In conclusion, the TaqMan real-time PCR developed here seems to be a useful tool for rapidly detecting and quantifying
M. leprae
DNA in clinical specimens in which bacilli were undetectable by conventional histological staining.
Pulmonary Cholera Due to Infection with a Non-O1
Vibrio cholerae
Strain
Authors: Shannon Jack D. , Kimbrough Robert C.
Published: September 2006

We present 2 cases of primary pulmonary non-O1
Vibrio cholerae
infection. We believe that these are the first documented cases of primary pulmonary infection due to this organism from a freshwater source.
Loss of Catabolic Function in
Streptococcus agalactiae
Strains and Its Association with Neonatal Meningitis
Authors: Domelier Anne-Sophie , van der Mee-Marquet Nathalie , Grandet Adeline , Mereghetti Laurent , Rosenau Agnès , Quentin Roland
Published: September 2006

The abilities of 151
Streptococcus agalactiae
strains to oxidize 95 carbon sources were studied using the Biolog system. Two populations were constituted: one with a high risk of causing meningitis (HR group; 63 strains), and the other with a lower risk of causing meningitis (LR group; 46 strains). Strains belonging to the HR group were significantly less able to use four carbon sources, i.e., α-
d
-glucose-1-phosphate,
d
-ribose, β-methyl-
d
-glucoside, and
d
,
l
-α-glycerol phosphate, than strains from the LR group (
P
≤ 0.004). Moreover, strains in the HR group significantly more frequently possessed one of several mobile genetic elements or genome deletions previously shown to be associated with strains responsible for neonatal meningitis than strains in the LR group (
P
< 0.001). These findings suggest that genetic disruption might have occurred in virulent clones of
S. agalactiae
. Fifteen biotypes (B1 to B15) were identified from the results of oxidation of the four carbon sources, of which six (B1 to B6) included 92% of the isolates belonging to the HR group. Strains of biotypes B1 to B6 are thus 13 times more likely to be able to invade the central nervous system of neonates than strains of biotypes B7 to B15. In addition, 86% of strains recently associated with neonatal meningitis (42 strains studied) were identified as being of biotypes B1 to B6. Identification of particular
S. agalactiae
biotypes may therefore be one of the criteria to assist clinicians in assessing the level of risk of neonatal meningitis when a mother and/or her neonate is colonized with
S. agalactiae
.
As a Bacterial Culture Medium, Citrated Sheep Blood Agar Is a Practical Alternative to Citrated Human Blood Agar in Laboratories of Developing Countries
Authors: Russell F. M. , Biribo S. S. N. , Selvaraj G. , Oppedisano F. , Warren S. , Seduadua A. , Mulholland E. K. , Carapetis J. R.
Published: September 2006

Human blood agar (HuBA) is widely used in developing countries for the isolation of bacteria from clinical specimens. This study compared citrated sheep blood agar (CSBA) and HuBA with defibrinated horse blood agar and defibrinated sheep blood agar (DSBA) for the isolation and antibiotic susceptibility testing of reference and clinical strains of
Streptococcus pneumoniae
,
Streptococcus pyogenes
, and
Staphylococcus aureus
. Reference and clinical strains of all organisms were diluted in brain heart infusion and a clinical specimen of cerebrospinal fluid and cultured on all agars. Viable counts, colony morphology, and colony size were recorded. Susceptibility testing for
S. pneumoniae
and
S. pyogenes
was performed on defibrinated sheep blood Mueller-Hinton agar, citrated sheep blood Mueller-Hinton agar (CSB MHA), and human blood Mueller-Hinton agar plates. For all organisms, the colony numbers were similar on all agars. Substantially smaller colony sizes and absent or minimal hemolysis were noted on HuBA for all organisms. Antibiotic susceptibility results for
S. pneumoniae
were similar for the two sheep blood agars; however, larger zone sizes were displayed on HuBA, and quality control for the reference strain failed on HuBA. For
S. pyogenes
, larger zone sizes were demonstrated on HuBA and CSBA than on DSBA. Poor hemolysis made interpretation of the zone sizes difficult on HuBA. CSBA is an acceptable alternative for the isolation of these organisms. The characteristic morphology is not evident, and hemolysis is poor on HuBA; and so HuBA is not recommended for use for the isolation or the susceptibility testing of any of these organisms. CSB MHA may be suitable for use for the susceptibility testing of
S. pneumoniae
.
Effect of Primer Selection on Estimates of GB Virus C (GBV-C) Prevalence and Response to Antiretroviral Therapy for Optimal Testing for GBV-C Viremia
Authors: Souza I. E. , Allen J. B. , Xiang J. , Klinzman D. , Diaz R. , Zhang S. , Chaloner K. , Zdunek D. , Hess G. , Williams C. F. , Benning L. , Stapleton J. T.
Published: September 2006

GB virus C (GBV-C; also called hepatitis G virus) is a common cause of infection associated with prolonged survival among HIV-infected individuals. The prevalences of GBV-C viremia vary widely in different studies, and there has been poor agreement among different laboratories performing GBV-C RNA detection in quality control studies. To determine the optimal method of measuring GBV-C RNA in clinical samples, samples obtained from 939 HIV-infected subjects were studied using reverse transcription (RT)-PCR methods amplifying four separate regions of the GBV-C genome. Primers amplifying the E2 coding region were 100% specific; however, their sensitivity was only 76.6%. In contrast, primers amplifying three additional conserved regions of the GBV-C genome (the 5′ nontranslated region and the nonstructural protein-coding regions 3 and 5A) were more sensitive but produced higher rates of false-positive results. Using low-specificity primer sets influenced the significance of association between GBV-C viremia and response to antiretroviral therapy. Using a quantitative GBV-C RNA method, the GBV-C RNA concentration did not correlate with baseline or set point HIV RNA levels; however, a correlation between negative, low, and high GBV-C RNA levels and increasing reduction in HIV RNA following antiretroviral therapy was observed. Subjects with both GBV-C E2 antibody and viremia had significantly lower GBV-C RNA levels than did viremic subjects without E2 antibody. These studies demonstrate that accurate detection of GBV-C RNA by nested RT-PCR requires the use of primers representing multiple genome regions. Analyses based on testing with single primers do not lead to reliable conclusions about the association between GBV-C infection and clinical outcomes.
Detection and Characterization of Bovine Coronaviruses in Fecal Specimens of Adult Cattle with Diarrhea during the Warmer Seasons
Authors: Park Su-Jin , Jeong Cheol , Yoon Soon-Seek , Choy Hyoun E. , Saif Linda J. , Park Sung-Hee , Kim You-Jung , Jeong Jae-Ho , Park Sang-Ik , Kim Ha-Hyun , Lee Bong-Joo , Cho Ho-Seong , Kim Sang-Ki , Kang Mun-Il , Cho Kyoung-Oh
Published: September 2006

Bovine coronavirus (BCoV) is an etiological agent associated with winter dysentery (WD), prevalent in adult cattle during the winter. Although we previously detected, isolated, and characterized BCoV strains from adult cattle with WD (WD-BCoV strains) during the winter in South Korea, the precise epidemiology, as well as the causative agent of diarrhea in adult cattle in the warmer seasons, has not been examined. We examined 184 diarrheic fecal specimens collected from 75 herds of adult cattle from seven provinces during the spring (warm), autumn (warm), and summer (hot) seasons. Bovine coronavirus-positive reactions were detected for 107 (58.2%) diarrheic fecal samples (in 47/75 herds). Of these 107 positive samples, 90 fecal samples from 33 herds tested positive for BCoV alone and 17 fecal samples from 14 herds also tested positive for other pathogens. Biological comparisons between the 9 BCoV strains isolated in this study and the 10 previously isolated WD-BCoV strains revealed that there was no receptor-destroying enzyme (RDE) activity against mouse erythrocytes in the 9 BCoV strains but the 10 WD-BCoV strains had high RDE activity. Phylogenetic analysis of the spike (S) and hemagglutinin/esterase (HE) proteins revealed that all the Korean BCoVs clustered together regardless of season and were distinct from the other known BCoVs, suggesting a distinct evolutionary pathway for the Korean BCoVs. These and previous results revealed a high prevalence and widespread geographical distribution of BCoV, suggesting that this virus is endemic in adult cattle with diarrhea in all seasons in South Korea.
Potential role of dendritic cells for progression of atherosclerotic lesions
Authors: Ranjit S , Dazhu L
Published: September 2006

Atherosclerosis is a lipid related chronic inflammatory disease in which immune mechanisms play a pivotal part. Its lesion is filled with large numbers of immune cells. In 1995 dendritic cells (DCs) were identified in atherosclerotic plaques and thought to play an important part in atherogenesis. DCs express MHCI and ll, HLA-DR, CD1a, ICAM-1 and VCAM1 on their surfaces, and this explains their unique ability to activate naive T cells. The risk factors for atherosclerosis are the factors for DCs’ activation and migration. Mature DCs are capable of presenting antigen to T cells, which play an important part in progression of disease. Statin and diltiazem have been shown to protect endothelial function by suppressing the function of DCs and play an important part in preventing atherosclerosis.
Ageing changes in the eye
Authors: Salvi S M , Akhtar S , Currie Z
Published: September 2006

Ageing changes occur in all the structures of the eye causing varied effects. This article attempts to review the parameters of what is considered within the “normal limits” of ageing so as to be able to distinguish those conditions from true disease processes. Improving understanding of the ageing changes will help understand some of the problems that the ageing population faces.
The Foundation Programme assessment tools: An opportunity to enhance feedback to trainees?
Authors: Carr S
Published: September 2006

The recent change in working patterns of doctors in training has meant that the traditional systems of education are under increasing pressure and that there is the need to maximise new opportunities for learning. One new opportunity may arise after the introduction of the mandatory assessment systems (Mini-CEX, DOPPS, Multi-source feedback, and Case based discussion) in the Foundation Programmes. In this review the new assessment procedures for the Foundation Programmes are outlined and the potential of these assessments (using Mini-CEX as main example) as an opportunity to give feedback to trainees discussed. The importance of feedback in professional development and some of the techniques available for giving feedback are described. The Foundation Programme assessments will occupy a significant amount of trainees’ and trainers’ time and it is important that opportunity for feedback and learning is maximised.
Extrinsic allergic alveolitis: comparative study of the bronchoalveolar lavage profiles and radiological presentation
Authors: Sterclova M , Vasakova M , Dutka J , Kalanin J
Published: September 2006

Introduction

Extrinsic allergic alveolitis (EAA) is an immunologically mediated interstitial lung disease. The abnormalities in the bronchoalveolar lavage (BAL) fluid cell counts are almost always seen in patients with EAA according to the stage of the disease. The aim of this retrospective study was to find out how the BAL lymphocyte count, percentage of lymphocytes expressing HLA-DR, CD4/CD8 T cell ratio in BAL fluid, and the concentration of immunoglobulin G in serum correspond to the inflammatory activity of the disease.

Methods

The study included 14 patients with EAA. BAL fluid samples were obtained and processed for cytological and cytometric analysis. Immunoglobulin G serum concentrations were measured. High resolution computed tomography (HRCT) scoring system modified by Gay was used for establishing the alveolar and interstitial score in each patient.

Conclusions

It was found that subjects with normal value of CD4/CD8 ratio in BAL fluid had higher interstitial HRCT score. Clinical presentation, continuous exposure to the causative antigens, and BAL lymphocyte count positively correlated with the alveolar HRCT score. It is proposed that the increased BAL lymphocyte count could be the predictor of the inflammatory activity of the disease, especially in people with lasting exposure to the offending antigen.
Secular trends in the medical discharge summary in an acute medical hospital
Authors: Chow K M , Szeto C C
Published: September 2006

Background

Although recent research has reported an evolution in the level of medical care, little is known about secular trends in the medical discharge summary.

Methods

The audit evaluated the evolution of discharge summary abstract over a decade in an acute medicine hospital among three successive periods of 1994, 2000, and 2005.

Results

Of the 140 random samples of discharge summaries, significant between-cohort difference existed in the logarithm transformed length of discharge summary; the median length of discharge summary increased from 11 lines in 1994 to 26 lines in 2005 (p<0.001 corrected for multiple comparison). The closest univariate associations of discharge summary length were positively with the year of hospitalisation, length of stay, length of wordings about discharge plan and duplication of previous discharge summary, and negatively with clinical or planned admission (all p<0.01). In a multiple linear regression analysis, the year of patient admission correlated significantly with the length of discharge summary (p<0.001).

Conclusions

Over the 10 year study period, significant secular trends were seen in the discharge summary length and discharge plan documentation. The causes and implications for these trends deserve further investigation.
Management of thyroid disorders
Authors: Premawardhana L D K E , Lazarus J H
Published: September 2006

Autoimmune thyroid disease is the predominant form of thyroid dysfunction in the developed world. Although its precise cause is currently unclear, principles of management have been established. There is a vigorous debate about the management of the increasingly commonly recognised subclinical forms of thyroid dysfunction despite recent recommendations. Nodular thyroid disease and thyroid carcinoma have received wide attention. The effects of drugs and pregnancy on thyroid function have also been investigated widely. This short review attempts to give an overview and clarify the current management of common thyroid disorders.
Micronutrients in health and disease
Authors: Shenkin A
Published: September 2006

Micronutrients play a central part in metabolism and in the maintenance of tissue function. An adequate intake therefore is necessary, but provision of excess supplements to people who do not need them may be harmful. Single micronutrient deficiency states are comparatively easily recognised and treated. Subclinical deficiency, often of multiple micronutrients, is more difficult to recognise, and laboratory assessment is often complicated by the acute phase response. Clinical benefit is most likely in those people who are severely depleted and at risk of complications, and is unlikely if this is not the case. There is little evidence for supplements leading to a reduction in the incidence of infections in the elderly population, in coronary artery disease, or in malignant disease. The best evidence for benefit is in critical illness, and in children in developing countries consuming a deficient diet. More clinical trials are required with good clinical outcomes to optimise intake in prevention and treatment of disease.
Identification of potential organisms of relevance to Canadian boreal forest and northern lands for testing of contaminated soils
Authors: Römbke J , Jänsch S , Scroggins R
Published: September 2006

A literature review has been performed with the aim of identifying potential soil invertebrate species, culturing techniques, and testing methodologies of relevance to Canadian boreal forest, taiga, and tundra ecozones. This was achieved through a review of European and Canadian ecological and ecotoxicological literature. The criteria for candidate species selection included habitat, frequency and abundance, origin, taxonomy, practicability, and stress tolerance. Nineteen species or genera common to both Northern Europe and Canada were identified as proposed test organisms in ecotoxicological test methodologies. Some of these species are currently in culture at various European or Canadian laboratories. Three earthworm and one collembolan species were recommended for further method development, while two more collembolan species as well as a yet to be identified Canadian species of four genera of enchytraeids, nematods, predatory mites, and carabid beetles were recommended but need further research. A number of issues with the recommended species were identified that would need resolution through focused research. Also, additional potential test species were identified based on their frequency of occurrence in boreal forests and northern lands for possible further method development consideration. Available information and current research activities on Canadian soil invertebrates relevant to the boreal forest, taiga, and tundra ecozones should be compiled using a similar classification and short-list selection process as used for this contribution to help identify additional candidate test species. The recommended species could be used as part of a test battery covering litter layer and mineral soils of Canadian boreal forests and northern lands.Key words: Carabidae, Collembola, Enchytraeidae, Gamasida, Lumbricidae, Nematoda.
Metal contamination of natural surface soils from long-range atmospheric transport: Existing and missing knowledge
Authors: Steinnes Eiliv , Friedland Andrew J
Published: September 2006

This review focuses on the long-range atmospheric transport of metals to organic-rich surface soils (mostly 50–90% organic matter) in the temperate, coniferous, and boreal zones of North America and Europe. From various air-pollution related measurements (air, precipitation, moss, peat cores) Pb and Zn are known to be transported long distances in the air in large amounts. Arsenic, Cd, Hg, Sb, and Se are also typical representatives of long-range transported air masses, and there is evidence that Ag, Bi, In, Mo, Tl, and W belong to this group of elements. Through the use of “environmental archives” such as ice and peat cores it has become evident that long-range transport of pollutants and associated contamination of natural surfaces is not just a recent phenomenon. There is compelling evidence for widespread enrichment of surface soil horizons in Pb from long-range transport, and many studies support enrichment of Zn. Mercury is also generally elevated by anthropogenic emissions over natural levels in organic-rich surface soils, whereas results for Cd are less conclusive. There is evidence that As, Se, Ag, Mo, In, Sb, W, Tl, and Bi all are subject to some enrichment in organic-rich surface soils from long-range atmospheric transport, but studies are still few for most of these elements. With the exception of Pb, little is known about residence times of the elements in the organic-rich surface horizon, and more research is needed on this topic. Further studies are desirable on the temporal and spatial trends in supply of the above elements, which are poorly known in large parts of the northern temperate zone.Key words: natural soils, metals, long-range atmospheric transport, organic matter, lead, zinc, cadmium, mercury.
Red light inhibition of spore germination in Ophioglossum crotalophoroides
Authors: Whittier Dean P.
Published: September 2006

Spores of Ophioglossum crotalophoroides Walt., which give rise to subterranean, nonphotosynthetic, mycorrhizal gametophytes, germinate in the dark and not in the light. Red light, like white light, prevents the germination of these spores. Germination occurs after exposure to far-red. The effects of far-red light can be reversed by red light and those of red light can be partly reversed by far-red light, confirming the involvement of phytochrome. With the spores of O. crotalophoroides, the active form of phytochrome, Pfr, prohibits germination. The photoinhibition of germination by white or red light insures that these spores germinate underground in nature. Hypogean germination improves the chances for adequate soil moisture and for the young gametophytes to be colonized by mycorrhizal fungi.
Genetic diversity of bluebunch wheatgrass (Pseudoroegneria spicata) in the Thompson River valley of British Columbia
Authors: Fu Yong-Bi , Thompson Don
Published: September 2006

Bluebunch wheatgrass ( Pseudoroegneria spicata (Pursh) A. Löve) is a cool-season perennial grass native to semi-arid regions of western North America and has been used for habitat restoration. However, the genetic diversity of this species is poorly understood. A total of 172 expressed sequence tag-derived simple sequence repeat (eSSR) primer pairs that had been developed for wheat were characterized for genetic diversity studies of bluebunch wheatgrass. Of these, 12 eSSR primer pairs were found to be informative and were applied to screen 216 plants collected from six locations with two different elevations in the Thompson River valley of British Columbia. These analyses revealed a total of 106 eSSR polymorphic alleles (or bands) scorable for each sample. The number of polymorphic bands per primer pair ranged from 2 to 17 with a mean of 8.8. The frequencies of these bands ranged from 0.005 to 0.995 and averaged 0.146. Most (92.6%) of the eSSR variation detected was present within the 12 populations assessed. The between-population eSSR variability was significantly associated with their geographic distances, but not with their elevations. These findings are useful for genetic diversity and genetic mapping studies of this grass species and should facilitate the sampling and development of bluebunch wheatgrass germplasm for germplasm conservation and habitat restoration.