bluuebunny/crossref_metadata_2025 huggingface dataset:
Ursodeoxycholic acid reduces CpG‐induced IgM production in patients with primary biliary cirrhosis
Authors: Kikuchi Kentaro , Hsu Willy , Hosoya Naomi , Moritoki Yuki , Kajiyama Yusuke , Kawai Toshihiro , Takai Atsuko , Hayami Eriko , Selmi Carlo , Gershwin M. Eric , Miyakawa Hiroshi
Published: December 2008

Aim:  Ursodeoxycholic acid (UDCA) treatment reduces IgM serum levels in patients with primary biliary cirrhosis (PBC) without affecting serum antimitochondrial antibody (AMA) titers. We previously reported that PBC‐associated hyper‐IgM is secondary to a disease‐specific hyperproduction following bacterial stimulation by B cells.

Methods:  We isolated peripheral blood mononuclear cells (PBMC) from patients with PBC and controls and evaluated whether bacterial CpG challenge in the presence of UDCA at concentrations consistent with those achieved in treated patients led to changes in total IgM, IgG‐AMA, and IgM‐AMA production. Further, p65 phosphorylation and CD38 cell expression were analyzed as measures of activation of the NF‐kB signaling pathway and B cell subsets, respectively.

Results:  UDCA significantly reduced CpG‐induced total IgM and IgM‐AMA production, but had no impact on IgG‐AMA production. UDCA also significantly reduced the activation ofnaïve and IgM memory, but not IgG memory, B cells, as represented by CD38 expression levels. Further, p65 phosphorylation was significantly reduced in the presence of UDCA.

Conclusion:  UDCA reduces total and IgM‐AMA production in PBMC from patients with PBC by downregulating B cell activation and NF‐kB signaling. These data ultimately suggest novel mechanisms of action for UDCA in chronic autoimmune cholestasis.
Immunosuppressive effect of IDO on T cells in patients with chronic hepatitis B*
Authors: Chen Yong‐bing , Li Si‐dan , He Yi‐ping , Shi Xian‐jie , Chen Yong , Gong Jian‐ping
Published: December 2008

Aim:  Recently indoleamine 2,3‐dioxygenase (IDO) has drawn considerable attention as a mechanism of immune regulation. Our study was to observe the role of IDO in immune tolerance of chronic hepatitis B (CHB), so as to provide a novel approach for reestablishment of active immunity.

Methods:  Peripheral venous blood samples were taken from 50 CHB patients and HBV viral load, T lymphocyte subsets as well as the mRNA, protein and activity of IDO were detected. The correlations between HBV viral load, T lymphocyte subsets and IDO were statistically analyzed. Blood samples from 50 healthy people were tested as a control group.

Results:  In CHB patients, the mRNA, protein and activity of IDO were all significantly more than those in control group (mRNA:[2.11 ± 0.615] × 103 vs. [0.143 ± 0.026] × 103; protein: 0.22 ± 0.06 vs. 0.02 ± 0.0017; activity: 26.07 ± 8.12 vs. 4.98 ± 1.65; P < 0.05) and IDO mRNA was positively correlated with HBV viral load (r = 0.502, P < 0.001) and alanine aminotransferase (ALT) (r = 0.65, P < 0.01). Furthermore, IDO mRNA, protein and activity were negatively correlated with CD4(+) T cells (r = −0.622, −0.682, −0.549 respectively, P < 0.05), CD8 (+) T cells (r = −0.487, −367, −294 respectively, P < 0.05) and the ratio of CD4/CD8 (r = −0.426, −0.533, −0.397 respectively, P < 0.05).

Conclusion:  IDO closely correlates with HBV viral load and is responsible for immunotolerance against HBV. Suppression of IDO could be a novel approach to break tolerance in CHB.
Hematologic abnormalities and flow cytometric immunophenotyping results in dogs with hematopoietic neoplasia: 210 cases (2002–2006)
Authors: Tasca Silvia , Carli Erika , Caldin Marco , Menegazzo Lucia , Furlanello Tommaso , Gallego Laia Solano
Published: December 2008

Background:Growing interest in veterinary oncohematology has facilitated the recent development and advancement of new techniques, such as flow cytometry, for immunophenotyping hematopoietic neoplasia in animals.

Objective:The aim of this retrospective study was to characterize hematologic abnormalities and flow cytometric immunophenotyping (FCI) results in cases of hematopoietic neoplasia in dogs.

Methods:Signalment, CBC data, and FCI results were obtained for 210 dogs with blood samples submitted to our laboratory. Immunophenotyping was carried out using an Epics XL‐MCL flow cytometer and a panel of 10 antibodies (CD45, CD3, CD4, CD8, CD79, CD21, CD14, CD34, CD41/61, CD61). The prevalence and severity of hematologic abnormalities was determined for the different types of hematopoietic neoplasms.

Results:Based on cell morphology and phenotype, cases were classified as: acute lymphoblastic leukemia (ALL,n=51), acute myeloid leukemia (AML,n=33), chronic lymphocytic leukemia (CLL,n=61), and leukemic high‐grade lymphoma (L‐HGL,n=65). Most cases of ALL (47/51) and L‐HGL (41/65) had a B‐cell phenotype, while most cases of CLL (54/61) had a T‐cell phenotype, with a high prevalence of the large granular lymphocyte subtype (49/61). Anemia was found in 85% of all cases and was significantly more severe in ALL and AML compared with CLL and L‐HGL. Neutropenia was seen in 64–78% of acute leukemias (AML and ALL) in contrast to no cases of CLL and 11% of L‐HGL. Thrombocytopenia was seen in 88–90% of acute leukemias in contrast to 15% of CLL and 40% of L‐HGL. Thrombocytopenia was more prevalent (71% vs 22%) and significantly more severe in T‐cell vs B‐cell L‐HGL.

Conclusion:A standard CBC is useful in suggesting the type of hemoproliferative disorder and may also help to predict the phenotype, especially in cases of L‐HGL.
Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis
Authors: Riond Barbara , Wenger‐Riggenbach Bettina , Hofmann‐Lehmann Regina , Lutz Hans
Published: December 2008

Background:Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and α‐, β‐, and γ‐globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported.

Objectives:The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm‐blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined.

Methods:Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi‐automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within‐run and within‐assay precision. Data from warm‐blooded and draught horses were compared using the Mann–WhitneyUtest.

Results:Within‐run and within‐assay CVs were <5% for all protein fractions. No significant difference was found between warm‐blooded and heavy draught horses and so combined reference intervals (2.5–97.5%) were calculated for total protein (51.0–72.0 g/L), albumin (29.6–38.5 g/L), α1‐globulin (1.9–3.1 g/L), α2‐globulin (5.3–8.7 g/L), β1‐globulin (2.8–7.3g/L), β2‐globulin (2.2–6.0 g/L), and γ‐globulin (5.8–12.7 g/L) concentrations, and albumin/globulin ratio (0.93–1.65).

Conclusion:Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.
Presumptive increase in protein‐bound serum calcium in a dog with multiple myeloma
Authors: Tripp Chelsea D. , Bryan Jeffrey N. , Wills Tamara B.
Published: December 2008

Abstract: An 11‐year‐old male castrated Australian Shepherd was presented with a history of lethargy, panting, and weight loss for 1 month. Physical examination revealed a moderately enlarged spleen. Laboratory abnormalities included thrombocytopenia and marked hypercalcemia, with hyperglobulinemia, hypoalbuminemia, and a monoclonal spike in the β‐globulin region on serum protein electrophoresis. Serum total calcium concentration was markedly increased (16.5 mg/dL, reference interval 8.9–11.4 mg/dL) but ionized calcium concentration (1.39 mmol/L) was within the reference interval (1.25–1.45 mmol/L). Isosthenuria was noted, but the dog was not polyuric or polydipsic. Serum parathyroid hormone concentration was within reference limits and parathyroid hormone‐related peptide concentration was 0 pmol/L. Radiographic findings were largely unremarkable. Results of cytologic evaluation of a fine‐needle aspirate specimen from the spleen indicated plasma cell neoplasia. Based on the results of the electrophoresis, splenic aspirates, radiographs, and hypercalcemia, a diagnosis of splenic multiple myeloma was made. The marked hypercalcemia, normal ionized calcium and parathyroid hormone concentrations, and lack of osteolytic lesions indicated a presumptive increase in protein‐bound serum calcium, likely due to binding to molecules of the paraprotein (M protein). Protein binding of calcium in dogs with multiple myeloma should be considered as a potential mechanism of elevated total serum calcium concentration.
Clinical Evaluation of a Novel Liquid Formulation of L‐Thyroxine for Once Daily Treatment of Dogs with Hypothyroidism
Authors: Le Traon G. , Brennan S.F. , Burgaud S. , Daminet S. , Gommeren K. , Horspool L.J.I. , Rosenberg D. , Mooney C.T.
Published: December 2008

Background:A liquid solution of levothyroxine (L‐T4) is available for treatment of canine hypothyroidism.

Hypothesis:Once daily oral administration of a liquid L‐T4 solution is effective and safe for controlling hypothyroidism in dogs.

Animals:Thirty‐five dogs with naturally occurring hypothyroidism.

Methods:Dogs received L‐T4 solution PO once daily at a starting dosage of 20 μg/kg body weight (BW). The dose was adjusted every 4 weeks, based on clinical signs and peak serum total T4 (tT4) concentrations. Target peak serum tT4 and thyroid stimulating hormone (TSH) concentrations, 4–6 hours posttreatment, were 35–95 nmol/L and < 0.68 ng/mL, respectively. Dogs were followed for up to 22 weeks after establishment of the maintenance dose.

Results:Clinical signs of hypothyroidism improved or resolved in 91% of dogs after 4 weeks of L‐T4 treatment at 20 μg/kg once daily. The maintenance dose was established in 76, 94, and 100% of dogs after 4, 8, and 12 weeks of treatment, respectively. This was 20 μg L‐T4/kg BW for 79% of the dogs, 30 μg/kg BW for 15%, and 10–15 μg/kg BW in the remaining 6%, once daily. Thereafter, median peak tT4 and TSH concentrations were 51 nmol/L and 0.18 ng/mL, respectively, and remained stable during the 22‐week follow‐up; clinical signs did not recur.

Conclusions and Clinical Importance:All of the hypothyroid dogs had rapid clinical and hormonal responses to supplementation with the PO‐administered L‐T4 solution. The starting dosage of 20 μg L‐T4/kg BW once daily was suitable for 79% of dogs.
Serum Cardiac Troponin I Concentration in Retired Racing Greyhounds
Authors: LaVecchio D. , Marin L.M. , Baumwart R. , Iazbik M.C. , Westendorf N. , Couto C.G.
Published: December 2008

Background:Cardiac troponin I (cTnI) is a polypeptide found specifically in cardiac muscle tissue that has been used as a diagnostic and prognostic indicator of cardiomyopathy. Increases in cTnI are associated with myocardial pathologic processes. However, high serum cTnI concentrations have been observed in normal Greyhounds.

Hypothesis:We hypothesized that Greyhounds have cTnI concentrations higher than non‐Greyhound dogs, and that a separate reference range should be established for Greyhounds.

Animals:Blood samples were collected from the jugular vein from a group of 20 healthy Greyhound blood donors.

Methods:Analysis of serum cTnI was performed with an immunoassay system with a detection level of 0.01 ng/mL, as described previously. The Greyhound values were compared with 2 groups of Boxers with and without arrhythmogenic right ventricular cardiomyopathy (ARVC), and to a group of non‐Boxer control dogs from a previous study.

Results:The mean cTnI concentration in Greyhounds was significantly higher (P< .0001) than that in non‐Greyhound control dogs, although not significantly different from normal Boxers (P= .50), or Boxers with ARVC (P= .58). Greyhound serum cTnI concentrations were in the range found in Boxers with ARVC. The proposed reference range for cTnI in Greyhounds is 0.05–0.16 ng/mL.

Conclusions and Clinical Importance:Greyhounds have a reference range for serum cTnI concentrations that differs from that of other previously published reference ranges for dogs of other breeds. Until a broader database and more precise reference range can be established, caution should be exercised in interpreting serum cTnI concentrations in Greyhounds with suspected cardiac disease.
Adenohypophyseal Function in Dogs with Primary Hypothyroidism and Nonthyroidal Illness
Authors: Diaz‐Espiñeira M.M. , Mol J.A. , Rijnberk A. , Kooistra H.S.
Published: December 2008

Background:A recent study of dogs with induced primary hypothyroidism (PH) demonstrated that thyroid hormone deficiency leads to loss of thyrotropin (TSH) hypersecretion, hypersomatotropism, hypoprolactinemia, and pituitary enlargement with large vacuolated “thyroid deficiency” cells that double‐stained for growth hormone (GH) and TSH, indicative of transdifferentiation of somatotropes to thyrosomatropes.

Hypothesis:Similar functional changes in adenohypophyseal function occur in dogs with spontaneous PH as do in dogs with induced PH, but not in dogs with nonthyroidal illness (NTI).

Animals:Fourteen dogs with spontaneous PH and 13 dogs with NTI.

Methods:Adenohypophyseal function was investigated by combined intravenous administration of 4 hypophysiotropic releasing hormones (4RH test), followed by measurement of plasma concentrations of ACTH, GH, luteinizing hormone (LH), prolactin (PRL), and TSH. In the PH dogs this test was repeated after 4 and 12 weeks of thyroxine treatment.

Results:In 6 PH dogs, the basal TSH concentration was within the reference range. In the PH dogs, the TSH concentrations did not increase with the 4RH test. However, TSH concentrations increased significantly in the NTI dogs. Basal and stimulated GH and PRL concentrations indicated reversible hypersomatotropism and hyperprolactinemia in the PH dogs, but not in the NTI dogs. Basal and stimulated LH and ACTH concentrations did not differ between groups.

Conclusions and Clinical Importance:Dogs with spontaneous PH hypersecrete GH but have little or no TSH hypersecretion. Development of hyperprolactinemia (and possible galactorrhea) in dogs with PH seems to occur only in sexually intact bitches. In this group of dogs with NTI, basal and stimulated plasma adenohypophyseal hormone concentrations were not altered.
Calprotectin in Myeloid and Epithelial Cells of Laminae from Horses with Black Walnut Extract‐Induced Laminitis
Authors: Faleiros R.R. , Nuovo G.J. , Belknap J.K.
Published: December 2008

Background:Laminar inflammation is one of the earliest events in equine laminitis. Calprotectin (CP), a Damage‐Associated Molecular Pattern protein, is overexpressed in inflammatory conditions of human skin.

Hypothesis:CP is overexpressed in the laminar epidermis of horses with black walnut extract (BWE)‐induced laminitis.

Animals:Twenty adult horses.

Methods:Experimental study. Horses were allocated to one of 4 groups. BWE was administered to horses in 3 groups, which were sampled 1.5, 3, and 12 hours (LAM) later. CP was visualized by immunohistochemistry. Laminar leukocyte counts and intensity of laminar epithelial staining were scored for all animals and statistically analyzed.

Results:Laminar epidermal CP signal was significantly increased (P= .02) at the LAM time point, compared with other groups. Rare leukocytes were detected in laminae with CP staining in CON group, but there were marked increases in number of leukocytes in BWE‐treated groups (P= .003). Sequential hematoxylin and eosin staining demonstrated that the majority of CP‐positive leukocytes were perivascular polymorphonuclear neutrophils (PMN) at each of the developmental time points. CP‐positive PMN and mononuclear cells were detected in perivascular locations and close to the epidermal basement membrane in the LAM group.

Conclusions and Clinical Importance:CP expression in the laminar epidermis occurs after extravasation of leukocytes, indicating that leukocyte emigration might be an initiating factor in laminar epithelial stress and inflammation in BWE‐induced laminitis. These results indicate a possible role of CP in laminitis pathophysiology and laminar failure.
Characterization of Reference Materials for Human Antiserum to Pertussis Antigens by an International Collaborative Study
Authors: Xing Dorothy , Wirsing von König Carl Heinz , Newland Penny , Riffelmann Marion , Meade Bruce D. , Corbel Michael , Gaines-Das Rose
Published: December 2008

Enzyme-linked immunosorbent assay (ELISA) has been widely used to evaluate antibody responses to pertussis vaccination and infection. A common reference serum is essential for the standardization of these assays. However, no internationally recognized reference serum is available. At the request of the Expert Committee on Biological Standardization (ECBS) of the World Health Organization (WHO), a set of four candidate international standards has been prepared. These candidate materials have been assessed for suitability and compared to the widely used U.S. reference pertussis antiserum (human) lot 3, lot 4, and lot 5 by 22 laboratories from 15 countries in an international collaborative study. Laboratories measured immunoglobulin G (IgG) and IgA antibodies to pertussis toxin (PT), filamentous hemagglutinin (FHA), pertactin (PRN), and fimbriae (Fim2&3) using their established immunoassays. The results of this study showed each of the four candidates to be suitable as an international standard. With the agreement of the participants, a recommendation has been made to the ECBS that the candidate material coded 06/140 be established as the First International Standard for pertussis antiserum (human), with the following assigned international units (IU): IgG anti-PT, 335 IU/ampoule; IgA anti-PT, 65 IU/ampoule; IgG anti-FHA, 130 IU/ampoule; IgA anti-FHA, 65 IU/ampoule; IgG anti-PRN, 65 IU/ampoule; and IgA anti-PRN, 42 IU/ampoule. No formal units have been proposed for anti-Fim2&3 because most assays used a mixture of fimbrial antigens. In addition, the candidate material coded 06/142 has been proposed as a WHO working preparation for characterization of assay systems.
Meningococcal Outer Membrane Vesicle Vaccines Derived from Mutant Strains Engineered To Express Factor H Binding Proteins from Antigenic Variant Groups 1 and 2
Authors: Koeberling Oliver , Giuntini Serena , Seubert Anja , Granoff Dan M.
Published: December 2008

Meningococcal outer membrane vesicle (OMV) vaccines, which are treated with detergents to decrease endotoxin activity, are safe and effective in humans. However, the vaccines elicit serum bactericidal antibody responses largely directed against PorA, which is antigenically variable. We previously prepared a native (non-detergent-treated) OMV vaccine from a mutant of group B strain H44/76 in which the
lpxL1
gene was inactivated, which resulted in penta-acylated lipid A with attenuated endotoxin activity. To enhance protection, we overexpressed factor H binding protein (fHbp) from the antigenic variant 1 group. The vaccine elicited broad serum bactericidal antibody responses in mice against strains with fHbp variant 1 (∼70% of group B isolates) but not against strains with variant 2 or 3. In the present study, we constructed a mutant of group B strain NZ98/254 with attenuated endotoxin that expressed both endogenous variant 1 and heterologous fHbp variant 2. A mixture of the two native OMV vaccines from the H44/76 and NZ98/254 mutants stimulated proinflammatory cytokine responses by human peripheral blood mononuclear cells similar to those stimulated by control, detergent-treated OMV vaccines from the wild-type strains. In mice, the mixture of the two native OMV vaccines elicited broad serum bactericidal antibody responses against strains with heterologous PorA and fHbp in the variant 1, 2, or 3 group. By adsorption studies, the principal bactericidal antibody target was determined to be fHbp. Thus, native OMV vaccines from mutants expressing fHbp variants have the potential to be safe for humans and to confer broad protection against meningococcal disease from strains expressing fHbp from each of the antigenic variant groups.
Clostridium glycolicum
Isolated from a Patient with Otogenic Brain Abscesses
Authors: Van Leer C. , Wensing A. M. J. , van Leeuwen J. P. , Zandbergen E. G. J. , Swanink C. M. A.
Published: December 2008

We describe a case of brain abscesses with gas formation following otitis media, for which the patient treated himself by placing clay in his ear. Several microorganisms, including
Clostridium glycolicum
, were cultured from material obtained from the patient. This is the first report of an infection in an immunocompetent patient associated with this microorganism.
Simple, Sensitive, and Swift Sequencing of Complete H5N1 Avian Influenza Virus Genomes
Authors: Höper Dirk , Hoffmann Bernd , Beer Martin
Published: December 2008

The spread of highly pathogenic avian influenza A virus (HPAIV) of subtype H5N1 demands fast and reliable methods for in-depth, full-length sequence analysis. For this purpose, we designed a simple and sensitive method for the preparation of sequencing libraries from H5N1 HPAIV diagnostic RNA samples for sequencing with the Genome Sequencer FLX instrument. The method presented seamlessly integrates high-throughput pyrosequencing with the Roche/454 instrument into diagnostics without the need for additional equipment or molecular biological techniques besides standard PCR and the Genome Sequencer FLX sample preparation and sequencing pipeline.
Population Structure of Human Isolates of
Streptococcus agalactiae
from Dakar and Bangui
Authors: Brochet Mathieu , Couvé Elisabeth , Bercion Raymond , Sire Jean-Marie , Glaser Philippe
Published: December 2008

Multilocus sequence types of 163 human
Streptococcus agalactiae
strains isolated in Bangui and Dakar were analyzed. We identified local specificities in the distribution of sequence types and capsular serotypes. However, the overall population structure is similar to that in the United States and Europe, suggesting that few specific clones colonize humans.
Detection and Toxin Typing ofClostridium perfringensin Formalin-Fixed, Paraffin-Embedded Tissue Samples by PCR
Authors: Wu Josephine , Zhang Wandi , Xie Boxun , Wu Maoxin , Tong Xiaodi , Kalpoe Jayant , Zhang David
Published: December 2008

Since current microbiology methods are not suitable to detectClostridium perfringensin formalin-fixed, paraffin-embedded tissue samples, we developed a PCR assay to detect toxin-encoding genes and the 16S rRNA gene ofC. perfringens. We successfully detected and genotypedC. perfringensin tissue sections from two autopsy cases.
Infective Endocarditis in a Dog and the Phylogenetic Relationship of the Associated “
Bartonella rochalimae
” Strain with Isolates from Dogs, Gray Foxes, and a Human
Authors: Henn Jennifer B. , Gabriel Mourad W. , Kasten Rickie W. , Brown Richard N. , Koehler Jane E. , MacDonald Kristin A. , Kittleson Mark D. , Thomas William P. , Chomel Bruno B.
Published: December 2008

The first case of canine endocarditis caused by “
Bartonella rochalimae
” is reported. By PCR-restriction fragment length polymorphism, sequence, and phylogenetic analyses,
Bartonella
isolates from a dog with endocarditis, 22 gray foxes, and three dogs, described as
B. clarridgeiae
like, were confirmed to belong to the new species “
B. rochalimae
,” suggesting canids as the natural reservoir.
Evolution of Central Nervous System Multidrug-Resistant
Mycobacterium tuberculosis
and Late Relapse of Cryptic Prosthetic Hip Joint Tuberculosis: Complications during Treatment of Disseminated Isoniazid-Resistant Tuberculosis in an Immunocompromised Host
Authors: Upton Arlo , Woodhouse Andrew , Vaughan Ross , Newton Sandie , Ellis-Pegler Rod
Published: December 2008

We report a case of disseminated isoniazid-resistant tuberculosis in an immunocompromised patient with evolution of rifampin (rifampicin) resistance in the central nervous system. This was cured with intraventricular and oral treatment but was followed by a late relapse of the original infection in a prosthetic hip joint. We provide drug levels in cerebrospinal fluid and serum.
Gordonia araii
Infection Associated with an Orthopedic Device and Review of the Literature on Medical Device-Associated
Gordonia
Infections
Authors: Jannat-Khah Deanna P. , Halsey Eric S. , Lasker Brent A. , Steigerwalt Arnold G. , Hinrikson Hans P. , Brown June M.
Published: December 2008

Gordonia
infections in humans are rare and usually affect immunocompromised patients. We present the first case of
Gordonia araii
infection associated with a medical device in an immunocompetent patient. Sequencing was required for conclusive identification. We compared our case to the 16
Gordonia
species-associated medical device infections reported to date.
Population Snapshot of
Streptococcus pneumoniae
Serotype 19A Isolates before and after Introduction of Seven-Valent Pneumococcal Vaccination for French Children
Authors: Mahjoub-Messai Farah , Doit Catherine , Koeck Jean-Louis , Billard Typhaine , Evrard Bénédicte , Bidet Philippe , Hubans Christine , Raymond Josette , Levy Corinne , Cohen Robert , Bingen Edouard
Published: December 2008

Serotype 19A
Streptococcus pneumoniae
strains are now more frequent in French children than before the introduction of a seven-valent conjugate vaccine (PCV7). By applying multilocus sequence typing to 144 serotype 19A isolates collected before and after beginning PCV7 vaccination, we detected clonal expansion of the preexisting penicillin-intermediate sequence type 276.
Clinical Predictive Value of Real-Time PCR Quantification of Human Cytomegalovirus DNA in Amniotic Fluid Samples
Authors: Goegebuer T. , Van Meensel B. , Beuselinck K. , Cossey V. , Van Ranst M. , Hanssens M. , Lagrou K.
Published: December 2008

The aim of this study was to evaluate the diagnostic reliability and prognostic significance of the quantification of cytomegalovirus (CMV) DNA in amniotic fluid (AF). We retrospectively reviewed the results for 282 amniotic fluid samples that had been tested for CMV by a quantitative real-time PCR. We observed three cases in which no CMV genomes were detected in the AF but in which the children were nevertheless congenitally infected. Hence, we conclude that a negative result by PCR for CMV in AF cannot rule out the possibility of congenital infection. No false-positive PCR results were observed. A correlation between the CMV viral load in AF and the fetal and neonatal outcomes could not be demonstrated in our study. Instead, a correlation was found between the CMV viral load and the gestational age at the time of amniocentesis.
Oral Abscess Caused by
Campylobacter rectus
: Case Report and Literature Review
Authors: Mahlen Steven D. , Clarridge Jill E.
Published: December 2008

Campylobacter rectus
was isolated under routine anaerobic conditions (no additional hydrogen gas in the atmosphere) from an oral, nonperiodontal abscess from a patient with gastroesophageal adenocarcinoma. We report the first case of a palate abscess caused by
C. rectus
and review the literature and atmospheric requirements of this organism.
Reversion of Influenza A (H3N2) Virus from Amantadine Resistant to Amantadine Sensitive by Further Reassortment in Japan during the 2006-to-2007 Influenza Season
Authors: Furuse Yuki , Suzuki Akira , Kamigaki Taro , Shimizu Midori , Fuji Naoko , Oshitani Hitoshi
Published: December 2008

In the 2006-to-2007 influenza season, amantadine-sensitive strains were found among the N-lineage influenza A (H3N2) viruses, which were previously believed to be associated with amantadine resistance. Whole-genome sequencing results indicated that this was due to a further reassortment event.
Practical Considerations for High-Throughput Influenza A Virus Surveillance Studies of Wild Birds by Use of Molecular Diagnostic Tests
Authors: Munster Vincent J. , Baas Chantal , Lexmond Pascal , Bestebroer Theo M. , Guldemeester Judith , Beyer Walter E. P. , de Wit Emmie , Schutten Martin , Rimmelzwaan Guus F. , Osterhaus Albert D. M. E. , Fouchier Ron A. M.
Published: December 2008

Influenza A virus surveillance studies of wild bird populations are essential to improving our understanding of the role of wild birds in the ecology of low-pathogenic avian influenza viruses and their potential contribution to the spread of H5N1 highly pathogenic avian influenza viruses. Whereas the primary results of such surveillance programs have been communicated extensively, practical considerations and technical implementation options generally receive little attention. In the present study, the data obtained from 39,490 samples were used to compare the impacts of variables such as the sampling procedure, storage and transport conditions, and the choice of molecular and classical diagnostic tests on the outcome of the results. Molecular diagnostic tests allowed estimation of the virus load in samples, which has implications for the ability to isolate virus. Virus isolation in embryonated eggs was more sensitive than virus isolation in cell cultures. Storage and transport conditions had less of an impact on diagnostics by the use of molecular tests than by the use of classical approaches. These findings indicate that molecular diagnostic tests are more sensitive and more reliable than classical tests. In addition, molecular diagnostic tests facilitated analyses in real time and allowed the discrimination of H5 influenza viruses with low and high pathogenicities without the need for virus isolation. Critical assessment of the methods used in large surveillance studies like this will facilitate comparison of the results between studies. Moreover, the lessons learned from current large-scale influenza A virus surveillance activities could be valuable for other pathogen surveillance programs in the future.
Comparative Molecular Analysis Substantiates Zoonotic Potential of Equine Methicillin-ResistantStaphylococcus aureus
Authors: Walther Birgit , Monecke Stefan , Ruscher Claudia , Friedrich Alexander W. , Ehricht Ralf , Slickers Peter , Soba Alexandra , Wleklinski Claus-G. , Wieler Lothar H. , Lübke-Becker Antina
Published: December 2008

Despite the increasing importance of methicillin-resistantStaphylococcus aureus(MRSA) in veterinary medicine, knowledge about the epidemiology of the pathogen in horses is still poor. The phylogenetic relationship of strains of human and equine origins has been addressed before, usually by analyzing results of common standard classification methods for MRSA. This work intends to go beyond the baseline of typing procedures in order to comparatively characterize equine and human MRSA strains with similar phylogenetic backgrounds. In addition to multilocus sequence typing, pulsed-field gel electrophoresis,spatyping, staphylococcal cassette chromosomemectyping, and a PCR for Panton-Valentine leukocidin gene detection, a microarray analysis of a total of 185 structural, virulence-associated, and resistance loci was applied. The results showed that clonal complex 8 (CC8) was absolutely predominant (16 strains) in 19 investigated equine MSRA strains. Of the CC8 strains, 13 belonged to sequence type 254 (ST254) and the other 3 to ST8. This genotype has been isolated from different equine patients in various regions over several years, substantiating the apparent predominance of CC8 STs in MRSA strains of horses worldwide. Furthermore, comparatively investigated human strains of ST254 displayed molecular-typing results indistinguishable from those for strains of equine origin. Two further equine strains (ST22 and ST1117) showed similarity to ST22 human strains (CC22). One equine strain belonged to ST398, a genotype recently described as being frequently isolated from specimens from pigs and pig farmers. These data provide evidence for the adaptation of certain MRSA genotypes to more than one mammalian species, reflecting their extended host spectra.
Unreliable Extended-Spectrum β-Lactamase Detection in the Presence of Plasmid-Mediated AmpC in
Escherichia coli
Clinical Isolates
Authors: Robberts F. J. L. , Kohner P. C. , Patel R.
Published: December 2008

The emergence of extended-spectrum β-lactamase (ESBL) and plasmid-mediated AmpC (pAmpC) enzymes in
Escherichia coli
raises concern regarding accurate laboratory detection and interpretation of susceptibility testing results. Twenty-six cefpodoxime ESBL screen-positive, cefoxitin-resistant
E. coli
clinical isolates were subjected to clavulanate ESBL confirmatory testing employing disk augmentation, Etest, and the BD Phoenix NMC/ID-132 panel. Phenotypic pAmpC production was assessed by boronic acid disk augmentation. ESBL and pAmpC genes were detected by gene amplification and sequencing. ESBL genes (SHV and/or CTX-M-type genes) were detected in only 7/26 ESBL screen-positive isolates. Of 23 aminophenylboronic acid screen-positive isolates, pAmpC genes were detected in 20 (CMY-2 or FOX-5 genes). High incidences of false-positive ESBL confirmatory results were observed for both clavulanate disk augmentation (9/19) and BD Phoenix (5/19). All were associated with the presence of pAmpC genes with or without TEM-1. Etest performed poorly, as the majority of interpretations were nondeterminable. In addition, false-negative ESBL confirmatory results were observed in isolates possessing concomitant ESBL and pAmpC genes for Etest (four of five), BD Phoenix (three of five), and disk augmentation (one of five). The results indicate poor performance of currently employed ESBL confirmatory methods in the setting of concomitant pAmpC. Some isolates with pAmpC and ESBL genes fell within the susceptible category to extended-spectrum cephalosporins, raising concern over currently employed breakpoints.
Neisseria gonorrhoeae
Endocarditis Confirmed by Nucleic Acid Amplification Assays Performed on Aortic Valve Tissue
Authors: Nielsen Ulla S. , Knudsen Jane B. , Pedersen Lisbeth N. , Møller Jens K.
Published: December 2008

The Aptima Combo 2 assay is proposed as a rapid method of diagnosing
Neisseria gonorrhoeae
endocarditis or other suspected disseminated gonococcal disease.
Use of the BacT/Alert MB Mycobacterial Blood Culture System for Detection of Mycobacteria in Sterile Body Fluids Other than Blood
Authors: Mattei Romano , Savarino Arnaldo , Fabbri Marco , Moneta Sara , Tortoli Enrico
Published: December 2008

The definitive diagnosis of extrapulmonary tuberculosis is made by a positive body fluid culture result. Conventional culture methods require centrifugation or filtration of body fluid (peritoneal, pleural, synovial, or pericardial fluid) in order to improve the sensitivity. The aim of the present study was to evaluate the feasibility of the direct inoculation, at the patient's bedside, of up to 5 ml of uncentrifuged fluid onto BacT/Alert MB culture bottles (bioMérieux, Durham, NC).
Value of Serial Quantification of Fungal DNA by a Real-Time PCR-Based Technique for Early Diagnosis of Invasive Aspergillosis in Patients with Febrile Neutropenia
Authors: Cuenca-Estrella Manuel , Meije Yolanda , Diaz-Pedroche Carmen , Gomez-Lopez Alicia , Buitrago Maria J. , Bernal-Martinez Leticia , Grande Carlos , Juan Rafael San , Lizasoain Manuel , Rodriguez-Tudela Juan L. , Aguado Jose M.
Published: December 2008

A study was designed to assess the reliability of the serial detection ofAspergillussp. DNA to diagnose invasive aspergillosis (IA) in patients with febrile neutropenia. Two blood and two serum samples were taken weekly from 83 patients. A total of 2,244 samples were analyzed by real-time quantitative PCR. Twelve (14.4%) patients were diagnosed with IA. Taking two consecutive positive results as the diagnostic criterion, PCR detected 11 cases, with 4 false positives, giving sensitivity, specificity, positive, and negative predictive values of 91.6%, 94.4%, 73.3%, and 98.5%, respectively. On analyzing in conjunction with high-resolution chest tomography (HRCT) and galactomannan (GM) testing, the combination of serial PCR and GM detected 100% of aspergillosis cases, with a positive predictive value of 75.1%. This diagnostic strategy presented, according to CART analysis, a receiver-operator curve with an area under the curve of 0.97 (95% confidence interval, 0.895 to 1.032;P< 0.01), with a relative risk of IA 6.92 times higher than the control population and with predictive success of 95.2%. As regards early diagnosis, the serial detection ofAspergillusDNA took on average 21 days less than HRCT and 68 days less than GM. The serial detection ofAspergillusDNA using real-time quantitative PCR has great diagnostic applicability, which increases when combined with GM quantification.
Outbreak of Acute Respiratory Disease in China Caused by B2 Species of Adenovirus Type 11
Authors: Zhu Zhen , Zhang Yong , Xu Songtao , Yu Pengbo , Tian Xiaoping , Wang Li , Liu Zhonghua , Tang Liuying , Mao Naiying , Ji Yixin , Li Chongshan , Yang Zhaohui , Wang Shiwen , Wang Jingjun , Li Dexin , Xu Wenbo
Published: December 2008

An outbreak of acute respiratory tract infection occurred in Shanxi Province, China, from March to April 2006. Of the 254 patients affected by this outbreak, 247 patients were students of a senior high school; 1 of these patients died during the outbreak. Serological tests and blood culture revealed no evidence of bacterial infection. The results of direct reverse transcription-PCR or PCR performed with clinical specimens collected from the patients, including the sole patient who died, were positive for human adenoviruses (HAdVs) but negative for influenza virus, measles virus, rubella virus, mumps virus, parainfluenza virus, respiratory syncytial virus, and human enteroviruses. These findings were confirmed by enzyme-linked immunosorbent assay for HAdV immunoglobulin A, the conventional neutralization test, and viral isolation and identification. Sequencing of the entire hexon gene revealed that HdAV type 11a (HAdV-11a) belonging to the B2 species of HAdV was the etiological agent responsible for the outbreak. However, both the analysis of the phylogenetic relationship and the similarity plot indicated that the sequence of the 3′ end of the hexon gene outside the hypervariable regions the HAdV-11a strain isolated in this outbreak may be a recombinant with the sequence of the HAdV-14 strain of species B2. Although isolates of HAdV species B2 seldom cause respiratory infections, they may pose a new global challenge with regard to acute respiratory diseases; this possibility cannot be overlooked and should be carefully considered. Hence, the need to establish and improve both epidemiological and virological surveillance of HAdV infections in China should be emphasized.
Molecular Epidemiology of Norovirus Gastroenteritis Investigated Using Samples Collected from Children in Tunisia during a Four-Year Period: Detection of the Norovirus Variant GGII.4 Hunter as Early as January 2003
Authors: Sdiri-Loulizi Khira , Ambert-Balay Katia , Gharbi-Khelifi Hakima , Sakly Nabil , Hassine Mouna , Chouchane Slaheddine , Guediche Mohamed Neji , Pothier Pierre , Aouni Mahjoub
Published: December 2008

Human noroviruses (NoVs) cause epidemic and endemic acute gastroenteritis in children and adults. To study the prevalence and genetic diversity of NoV in children in Tunisia, a total of 788 fecal samples were collected during a 4-year period in the region of Monastir, from children 12 years of age or younger, hospitalized or presenting in dispensaries with symptoms of acute gastroenteritis. NoV was detected by reverse transcription-PCR and confirmed by sequence analysis. This is the first report that describes the molecular epidemiology of NoV in Tunisian children: NoVs were characterized as the causative agent in 128 (16.2%) of the samples. Fourteen samples contained a mixture of two NoVs, and 33 samples were coinfected with additional enteric viruses. Eight distinct NoV genotypes were detected (GGI.2, GGI.4, GGII.1, GGII.4, GGII.8, GGII.14, GGIIb/GGII.2, and GGIIb/GGII.3). GGII.4 was the most prevalent genotype, accounting for 83 (64.8%) cases. Interestingly the GGII.4 variant Hunter, described as spreading all over the world in 2004, was found in Tunisia as early as January 2003. The delay of 1 year between the isolation in Tunisia and the worldwide emergence is somewhat surprising, considering the importance of the contacts between North Africa and Europe particularly. Nevertheless, this illustrates the idea that sporadic gastroenteritis cases may be a reservoir for emerging epidemic NoV strains.
Optical Fingerprinting in Bacterial Epidemiology: Raman Spectroscopy as a Real-Time Typing Method
Authors: Willemse-Erix Diana F. M. , Scholtes-Timmerman Maarten J. , Jachtenberg Jan-Willem , van Leeuwen Willem B. , Horst-Kreft Deborah , Bakker Schut Tom C. , Deurenberg Ruud H. , Puppels Gerwin J. , van Belkum Alex , Vos Margreet C. , Maquelin Kees
Published: December 2008

Hospital-acquired infections (HAI) increase morbidity and mortality and constitute a high financial burden on health care systems. An effective weapon against HAI is early detection of potential outbreaks and sources of contamination. Such monitoring requires microbial typing with sufficient reproducibility and discriminatory power. Here, a microbial-typing method is presented, based on Raman spectroscopy. This technique provides strain-specific optical fingerprints in a few minutes instead of several hours to days, as is the case with genotyping methods. Although the method is generally applicable, we used 118
Staphylococcus aureus
isolates to illustrate that the discriminatory power matches that of established genotyping techniques (numerical index of diversity [
D
] = 0.989) and that concordance with the gold standard (pulsed-field gel electrophoresis) is high (95%). The Raman clustering of isolates was reproducible to the strain level for five independent cultures, despite the various culture times from 18 h to 24 h. Furthermore, this technique was able to classify stored (−80°C) and recent isolates of a methicillin-resistant
Staphylococcus aureus
-colonized individual during surveillance studies and did so days earlier than established genotyping techniques did. Its high throughput and ease of use make it suitable for use in routine diagnostic laboratory settings. This will set the stage for continuous, automated, real-time epidemiological monitoring of bacterial infections in a hospital, which can then be followed by timely corrective action by infection prevention teams.
Inhibition of a Large Double-Stranded DNA Virus by MxA Protein
Authors: Netherton Christopher L. , Simpson Jennifer , Haller Otto , Wileman Thomas E. , Takamatsu Haru-Hisa , Monaghan Paul , Taylor Geraldine
Published: December 2008

Increasing evidence points to the importance of the interferon (IFN) response in determining the host range and virulence of African swine fever virus (ASFV). Infection with attenuated strains of ASFV leads to the upregulation of genes controlled by IFN pathways, including myxovirus resistance (Mx) genes that are potent effectors of the antiviral state. Mx gene products are known to inhibit the replication of many negative-sense single-stranded RNA viruses, as well as double-stranded RNA viruses, positive-sense single-stranded RNA viruses, and the reverse-transcribing DNA virus hepatitis B virus. Here, we provide data that extend the known range of viruses inhibited by Mx to include the large double-stranded DNA viruses. Stably transfected Vero cells expressing human MxA protein did not support ASFV plaque formation, and virus replication in these cells was reduced 100-fold compared with that in control cells. In contrast, ASFV replication in cells expressing MxB protein or a mutant MxA protein was similar to that in control Vero cells. There was a drastic reduction in ASFV late protein synthesis in MxA-expressing cells, correlating with the results of previous work on the effect of IFN on viral replication. Strikingly, the inhibition of ASFV replication was linked to the recruitment of MxA protein to perinuclear viral assembly sites, where the protein surrounded the virus factories. Interactions between ASFV and MxA were similar to those seen between MxA and different RNA viruses, suggesting a common inhibitory mechanism.
Sharing of Endogenous Lentiviral Gene Fragments among Leporid Lineages Separated for More than 12 Million Years
Authors: van der Loo W. , Abrantes J. , Esteves P. J.
Published: December 2008

Lentiviruses are causal agents of severe pathologies of a variety of mammals, including cattle and humans (e.g., AIDS and different types of lymphoma). While endogenous forms of lentivirus do not occur in these species, A. Katzourakis and coworkers (A. Katzourakis, M. Tristem, O. G. Pybus, and R. J. Gifford, Proc. Natl. Acad. Sci. USA 104:6261-6265, 2007) recently reported the presence in the genome of the European rabbit (
Oryctolagus cuniculus
) of multiple sequences defining a lentiviral subgroup elegantly referred to as RELIK (
r
abbit
e
ndogenous
l
ent
i
virus type
K
). Sequence comparisons indicated that the RELIK ancestor may have integrated into the rabbit lineage more than 7 million years ago. We have substantiated this by producing sequence data certifying the sharing of RELIK sequences among leporid lineages that diverged some 12 million years ago.
Severe Acute Respiratory Syndrome Coronavirus Triggers Apoptosis via Protein Kinase R but Is Resistant to Its Antiviral Activity
Authors: Krähling Verena , Stein David A. , Spiegel Martin , Weber Friedemann , Mühlberger Elke
Published: December 2008

In this study, infection of 293/ACE2 cells with severe acute respiratory syndrome coronavirus (SARS-CoV) activated several apoptosis-associated events, namely, cleavage of caspase-3, caspase-8, and poly(ADP-ribose) polymerase 1 (PARP), and chromatin condensation and the phosphorylation and hence inactivation of the eukaryotic translation initiation factor 2α (eIF2α). In addition, two of the three cellular eIF2α kinases known to be virus induced, protein kinase R (PKR) and PKR-like endoplasmic reticulum kinase (PERK), were activated by SARS-CoV. The third kinase, general control nonderepressible-2 kinase (GCN2), was not activated, but late in infection the level of GCN2 protein was significantly reduced. Reverse transcription-PCR analyses revealed that the reduction of GCN2 protein was not due to decreased transcription or stability of GCN2 mRNA. The specific reduction of PKR protein expression by antisense peptide-conjugated phosphorodiamidate morpholino oligomers strongly reduced cleavage of PARP in infected cells. Surprisingly, the knockdown of PKR neither enhanced SARS-CoV replication nor abrogated SARS-CoV-induced eIF2α phosphorylation. Pretreatment of cells with beta interferon prior to SARS-CoV infection led to a significant decrease in PERK activation, eIF2α phosphorylation, and SARS-CoV replication. The various effects of beta interferon treatment were found to function independently on the expression of PKR. Our results show that SARS-CoV infection activates PKR and PERK, leading to sustained eIF2α phosphorylation. However, virus replication was not impaired by these events, suggesting that SARS-CoV possesses a mechanism to overcome the inhibitory effects of phosphorylated eIF2α on viral mRNA translation. Furthermore, our data suggest that viral activation of PKR can lead to apoptosis via a pathway that is independent of eIF2α phosphorylation.