bluuebunny/crossref_metadata_2025_split huggingface dataset:
Microsatellite loci isolation in the Canarian common chaffinch (Fringilla coelebs) and their utility in other Canarian finches
Authors: SUÁREZ N. M. , BETANCOR E. , PESTANO J. J. 
Published: February 2009

AbstractThe taxonomic classification of the common chaffinch (Fringilla coelebs) that inhabits the Canary Islands has been under debate for decades, mainly due to the absence of nuclear DNA analyses. In this study we describe the isolation and characterization of ten microsatellite loci (AAAG, AAAT and GT) from a La Palma specimen using an enrichment protocol. Two loci were monomorphic in the populations analysed (La Gomera and La Palma), but the remaining ones presented 2 or more alleles, with an average of 11.63 alleles per locus and an average observed heterozygosity of 0.735 (n = 44). All loci were tested for their utility in other Canarian populations and other finch species.
Treatment of high‐energy tibial shaft fractures with internal fixation and early prophylactic NovaBone grafting
Authors: Sun Jun‐ying , Hao Si‐chun , Sun Rong‐bin , Yang Yu‐sheng 
Published: February 2009

Objective:  To evaluate the safety and efficacy of early NovaBone grafting in accelerating healing of high‐energy tibial shaft fractures.Methods:  In a prospective, randomized, controlled study 78 patients with open or closed tibial shaft fractures were treated with reamed interlocking nails, combined with NovaBone grafting. Forty patients were randomly assigned to the experimental group (with NovaBone grafting at fracture site), and 38 to the control group (without NovaBone grafting).Results:  All patients were followed up for an average of 8 months (range 6–18 months). The average fracture healing time was 12 weeks (range 8–16 weeks) in the experimental group, and 19 weeks (range 12–36 weeks) in the control group (P < 0.01). No delayed union or nonunion was found in the experimental group, but four cases (10.5%) in the control group were found to heal spontaneously between 7 and 8 months. When the data were analyzed at week 12 and week 26 according to the criteria of Johner and Wruhs, 88% and 100% of the cases in the experimental group were rated as excellent and good respectively, but in the control group, 50% and 89% were rated as excellent and good respectively, these being significant differences (P < 0.05).Conclusion:  Internal fixation supplemented with early prophylactic NovaBone grafting of high‐energy tibial shaft fractures is a safe and effective treatment which accelerates fracture healing.
Analysis of the relationship between morphology of intervertebral disc and some correlated factors following discography in patients with chronic low back pain
Authors: Zhang Ji‐dong , Xia Qun , Hu Yong‐cheng , Miao Jun , Feng Qi‐jin 
Published: February 2009

Objective:  To evaluate the diagnostic effectiveness of discography in discogenic low back pain (LBP).Methods:  Ninety‐six cases of chronic LBP with or without referred thigh pain were enrolled in this study. All these cases received CT scan following discography once conservative treatment for at least 6 months had failed. There were 42 men and 54 women, aged from 24 to 67 years (average 46.4). Discography was performed on 218 discs. The positive discs were classified morphologically according to the Dallas Discogram Description (DDD).Results:  (i) The 56 cases (58.3%) which were positive on discography were divided into two subgroups of age less or more than 50 years. Positive rates for the two subgroups were 33.3% and 66.7%, respectively; (ii) one hundred and twenty‐two discs, of which 62 (50.8%) were positive on discography, showed morphologic abnormality, whereas all the discography positive discs showed morphologic abnormality. No complication related to discography was found in any case.Conclusion:  (i) Compared with the younger patients, older LBP patients have a lower positive rate of discography despite the presence of more serious degenerative disc changes; (ii) outer layer disruption of the annulus fibrous correlates with positive discography; (iii) MRI intensity changes are not specific in diagnosing discogenic pain. Additional discography is needed to identify the painful disc; and (iv) the contrast volume injected into discs can be affected by a variety of factors which restrict its diagnostic value.
Artificial nucleus replacement: surgical and clinical experience
Authors: Zhang Zhong‐min , Zhao Liang , Qu Dong‐bin , Jin Da‐di 
Published: February 2009

Objective:  To assess long‐term clinical results of prosthetic disc nucleus (PDN) replacement for the treatment of lumbar disc herniation.Methods:  Seventy‐two patients with lumbar disc herniation were implanted with a single PDN device from March 2002 to December 2003. Fifty‐eight (80.6%) patients attended clinical, functional, and radiographic follow‐up examinations for more than 48 months. Independent analysis was performed by careful review of the interviews, operative reports, preoperative and postoperative radiographs, and computed tomography scans or magnetic resonance imaging (MRI).Results:  After implantation, a significant proportion of patients experienced pain relief. Improvements were noted in pain intensity, walking distance, neurological weakness, Oswestry and Prolo scores, intervertebral disc height and lumbar mobility. Intervertebral disc height was not well maintained, compared with the preoperative height it decreased 18% (P < 0.001). Several complications were associated with the implantation of PDN, including transient low‐back pain, implant dislocation, malposition of the implant, damage to the end plates and subsidence of implant.Conclusion:  The clinical data show that PDN is preferred and can effectively increase the range of lumbar motion in patients with lumbar disc herniation. However, this study only represents cases in our centre.
Trabecular bone micro‐architecture and bone mineral density in adolescent idiopathic and congenital scoliosis
Authors: Zhu Feng , Qiu Yong , Yeung Hiu Yan , Lee Kwong Man , Cheng Chun‐yiu Jack 
Published: February 2009

Objective:  To investigate the microstructure of trabecular bone in adolescent idiopathic scoliosis (AIS) and age‐matched congenital scoliosis (CS), and to evaluate the bone mineral status of CS patients compared with normal controls and AIS patients.Methods:  This study included 15 AIS and 16 CS female patients and 35 healthy female adolescents. Corrective surgery was indicated for the AIS and CS patients, from whom iliac crest biopsies were collected during autograft harvesting, and scanned by micro‐computer tomography. Bone mineral status was assessed at the lumbar and hip areas in every patient by dual energy X‐ray absorptiometry (DEXA).Results:  Significantly lower lumbar and femoral neck bone mineral density (BMD) was found in AIS patient compared with normal controls. All BMD and bone mineral content (BMC) parameters were significantly lower in CS patients compared with age‐matched normal controls. Under DEXA assessment significant associations between bone volume/tissue volume (BV/TV) and BMD values were observed. In the 3D model, BV/TV was significantly higher in AIS (19.9% ± 3.4%) than in CS (13.3% ± 3.0%, P < 0.05). Significant differences between AIS and CS were also found in trabecular thickness (Tb.Th) and bone surface/bone volume (BS/BV) (155.5 ± 54.9 µm vs. 108.1 ± 17.4 µm and 16.4% ± 3.3% vs. 22.0% ± 3.4% respectively, P < 0.05 in both).Conclusion:  Lower bone mineral status and weak trabecular bone structure observed in AIS and CS justify further investigation of the bone mineral status in scoliosis of various etiologies.
Low dose erythropoietin‐beta improves anemia and maintains ribavirin dose in chronic hepatitis C patients receiving combination therapy with ribavirin plus pegylated interferon Alfa‐2b
Authors: Tseng Kuo‐Chih , Chen Li‐Hsuan , Chen Chi‐Yi , Chang Ting‐Tsung , Chou An‐Liang , Wu I‐Chin , Cheng Pin‐Nan 
Published: February 2009

Aim: Anemia during combination therapy with pegylated interferon alfa‐2b plus ribavirin (RBV) for chronic hepatitis C virus (HCV) patients usually leads to RBV dose reduction or discontinuation. This study evaluated the effect of erythropoietin‐beta (EPO‐β) to maintain RBV dose and hemoglobin (Hb) level in chronic HCV patients treated with antiviral combination therapy.Methods: Eighty‐eight chronic HCV patients who developed anemia during therapy were enrolled into this retrospective study: 55 in the EPO‐β group and 33 in the untreated group. The study endpoints were to assess the RBV maintenance and the changes in Hb.Results: A higher percentage of patients with RBV maintenance was observed in the EPO‐β group compared with the untreated group (nadir Hb level <10.5 g/dL; 70% vs. 38%,P = 0.020; nadir Hb < 10 g/dL; 62% vs. 27%,P = 0.046). The mean Hb change from week 12 to week 20 was higher in the EPO‐β group when compared with the untreated group, especially for patients receiving a total EPO‐β dose of more than 16 000 U (+0.70 g/dL vs. −0.32 g/dL,P = 0.023) and of 10 000 U‐14 000 U (+0.60 g/dL vs. −0.32 g/dL,P = 0.023).Conclusions: Low‐dose EPO‐β can maintain RBV dose and increase Hb levels in anemic chronic HCV patients receiving combination therapy.
Special stain and X‐ray probe microanalysis of livers with Wilson disease
Authors: Hayashi Hisao , Fujita Yoshikazu , Tatsumi Yasuaki , Hattori Ai , Hayashi Kazuhiko , Katano Yoshiaki , Wakusawa Shinya , Yano Motoyoshi , Itoh Masafumi , Mizutani Naoki , Goto Hidemi 
Published: February 2009

Aim:  Primary copper toxicosis due to Wilson disease is clinically complex, often leading to delayed diagnosis. Because the metabolic disorder is frequently complicated by iron overload due to hypoceruloplasminemia, either a special stain or microanalysis has been recommended for liver biopsy specimens.Methods:  Liver biopsy was performed in three patients in whom Wilson disease was highly suspected. Light microscopic study included rubeanic acid stain for copper and Berlin blue stain for iron. To improve the resolution of ultra‐structures and preservation of toxic metals, short‐term fixation with a 0.1% osmic acid solution was applied for X‐ray probe microanalysis. Their diagnosis was confirmed by genetic study and copper chelation therapy.Results:  Two patients at the age of 17 and 23 years, respectively, demonstrated cirrhotic livers surrounded by fibrous septa, while a 7‐year‐old patient demonstrated fatty liver with mildly expanded portal tracts. Both copper grains stained with rubeanic acid and cuprothionein by microanalysis were found in the cirrhotic livers of aged patients. However, either morphological method failed to detect copper deposition in fatty liver tissues from the young patient. Iron deposits were also found in the cirrhotic livers of aged patients. The molecular basis of Wilson disease was confirmed by gene analysis. All patients responded to copper chelation therapy.Conclusion:  A morphological method of special staining or microanalysis improved with a new fixative may be unreliable for detecting diffusely distributed copper in the early stage of Wilson liver disease.
Comparison of quantitative immunoturbidimetric and semiquantitative latex‐agglutination assays for D‐dimer measurement in canine plasma
Authors: Boutet Philippe , Heath Fred , Archer Joy , Villiers Elizabeth 
Published: February 2009

Background: D‐dimer measurement in dogs is considered the most reliable test for detecting disseminated intravascular coagulation or thromboembolism.Objectives: The purposes of this study were to compare 2 D‐dimer assays, a quantitative immunoturbidimetric and a semiquantitative latex agglutination assay, and to assess the effect of hemolysis and storage conditions on D‐dimer concentration using the quantitative assay.Methods: The immunoturbidimetric assay was validated using canine citrated plasma samples containing different concentrations of D‐dimer. The effect of storage at various temperatures and times was assessed. Hemolysis was produced by adding lysed RBCs to the samples for a final hemoglobin concentration of 0.35 g/dL.Results: For clinically relevant values (>250 μg/L), intra‐assay and interassay coefficients of variation were 6.8% and 7.2%. The assay was linear (r2=1.00), and the tests had good agreement (κ=0.685, P<.001). Storage at 4 °C and −20 °C and hemolysis had no significant effect on D‐dimer concentrations. In hemolyzed samples stored at room temperature for ≥48 hours, fine clots were noted and often resulted in falsely increased D‐dimer concentrations.Conclusions: Our findings suggest that the immunoturbidimetric assay validated in this study is reliable and accurate for the measurement of D‐dimer in canine plasma. Samples can be stored for up to 1 month at −20 °C and moderate hemolysis does not significantly affect the D‐dimer concentration in frozen or refrigerated samples.
Comparison of flow cytometry with the Sysmex XT2000iV automated analyzer for the detection of reticulated platelets in dogs
Authors: Pankraz Alexander , Bauer Natali , Moritz Andreas 
Published: February 2009

Background: Immature (reticulated) platelets (r‐PLT) are not routinely assessed by hematology analyzers, but may be useful in the evaluation of the bone marrow response to thrombocytopenia.Objective: The aim of this study was to compare the Sysmex XT2000iV hematology analyzer with standard flow cytometry for the determination of r‐PLT percentage in dogs.Methods: Blood samples were obtained from 40 healthy dogs, 12 thrombocytopenic dogs, and 6 dogs with normal platelet counts but with disorders associated with increased thrombopoiesis. The percentage of r‐PLT was determined with a FACscan flow cytometer (r‐PLT[F]) using CD61‐phycoerythrin antibody and thiazole orange, and with the PLT‐O channel of the Sysmex analyzer (r‐PLT[S]). Mean platelet volume, platelet distribution width, and platelet large cell ratio were also determined on the Sysmex. Repeatability (intra‐assay precision) and effect of storage were tested for the automated analyzer.Results: The reference interval (mean±1.96 X SD) for r‐PLT(F) was 1.91±1.29% (range 0.78–3.68%) and for r‐PLT(S) was 0.56±0.82% (range 0.11–2.16%). For both flow cytometry and the Sysmex, the patient group had a significantly higher mean percentage of r‐PLT compared with the control group (P<.0001, unpaired Student's t‐tests). Fair correlation (r=0.71; Spearman's regression analysis) was found for r‐PLT results between the 2 methods, and a negative proportional systematic bias of −6.26 was found for the Sysmex (Bland–Altman analysis). Based on receiver operating characteristic curves and a cut‐off of ≥0.975%, a sensitivity of 94.7% and a specificity of 85.7% were obtained for detecting r‐PLT on the Sysmex, using flow cytometry as the reference method. Blood samples stored at 4 °C and 25 °C had a significant increase in the percentage of r‐PLT after 24 and 48 hours, respectively.Conclusions: The PLT‐O channel of the Sysmex XT2000iV is capable of detecting immature platelets in healthy, thrombocytopenic, and nonthrombocytopenic ill dogs.
Recombinant Vesicular Stomatitis Virus Expressing Influenza Nucleoprotein Induces CD8 T-Cell Responses That Enhance Antibody-Mediated Protection after Lethal Challenge with Influenza Virus
Authors: Barefoot Brice E. , Sample Christopher J. , Ramsburg Elizabeth A. 
Published: February 2009

ABSTRACTLive attenuated vaccine vectors based on recombinant vesicular stomatitis viruses (rVSVs) expressing foreign antigens are highly effective vaccines in animal models. In this study, we report that an rVSV expressing influenza nucleoprotein (VSV NP) from the first position of the VSV genome induces robust anti-NP CD8 T cells in immunized mice. These CD8 T cells are phenotypically similar to those induced by natural influenza infection and are cytotoxic in vivo. Animals immunized with an rVSV expressing the influenza hemagglutinin (rVSV HA) were protected but still exhibited considerable morbidity after challenge. Animals receiving a cocktail vaccine of rVSV NP and rVSV HA had reduced pulmonary viral loads, less weight loss, and reduced clinical signs of illness after influenza virus challenge, relative to those vaccinated with rVSV HA alone. Influenza NP is a highly conserved antigen, and induction of protective anti-NP responses may be a productive strategy for generating heterologous protection against divergent influenza strains.
Species-Specific Identification of a Wide Range of Clinically Relevant Fungal Pathogens by Use of Luminex xMAP Technology
Authors: Landlinger C. , Preuner S. , Willinger B. , Haberpursch B. , Racil Z. , Mayer J. , Lion T. 
Published: February 2009

ABSTRACT

In immunocompromised patients suffering from invasive fungal infection, rapid identification of the fungal species is a prerequisite for selection of the most appropriate antifungal treatment. We present an assay permitting reliable identification of a wide range of clinically relevant fungal pathogens based on the high-throughput Luminex microbead hybridization technology. The internal transcribed spacer (ITS2) region, which is highly variable among genomes of individual fungal species, was used to generate oligonucleotide hybridization probes for specific identification. The spectrum of pathogenic fungi covered by the assay includes the most commonly occurring species of the genera
Aspergillus
and
Candida
and a number of important emerging fungi, such as
Cryptococcus, Fusarium, Trichosporon, Mucor, Rhizopus, Penicillium, Absidia, and Acremonium
. Up to three different probes are employed for the detection of each fungal species. The redundancy in the design of the assay should ensure unambiguous fungus identification even in the presence of mutations in individual target regions. The current set of hybridization oligonucleotides includes 75 species- and genus-specific probes which had been carefully tested for specificity by repeated analysis of multiple reference strains. To provide adequate sensitivity for clinical application, the assay includes amplification of the ITS2 region by a seminested PCR approach prior to hybridization of the amplicons to the probe panel using the Luminex technology. A variety of fungal pathogens were successfully identified in clinical specimens that included peripheral blood, samples from biopsies of pulmonary infiltrations, and bronchotracheal secretions derived from patients with documented invasive fungal infections. Our observations demonstrate that the Luminex-based technology presented permits rapid and reliable identification of fungal species and may therefore be instrumental in routine clinical diagnostics.
Mitochondrion-Enriched Anionic Phospholipids Facilitate Flock House Virus RNA Polymerase Membrane Association
Authors: Stapleford Kenneth A. , Rapaport Doron , Miller David J. 
Published: February 2009

ABSTRACT

One characteristic of all positive-strand RNA viruses is the necessity to assemble viral RNA replication complexes on host intracellular membranes, a process whose molecular details are poorly understood. To study viral replication complex assembly we use the established model system of Flock House virus (FHV), which assembles its replication complexes on the mitochondrial outer membrane. The FHV RNA-dependent RNA polymerase, protein A, is the only viral protein necessary for genome replication in the budding yeast
Saccharomyces cerevisiae
. To examine the host components involved in protein A-membrane interactions, an initial step of FHV RNA replication complex assembly, we established an in vitro protein A membrane association assay. Protein A translated in vitro rapidly and specifically associated with mitochondria isolated from yeast, insect, and mammalian cells. This process was temperature dependent but independent of protease-sensitive mitochondrial outer membrane components or the host mitochondrial import machinery. Furthermore, lipid-binding studies revealed that protein A preferentially bound to specific anionic phospholipids, in particular the mitochondrion-specific phospholipid cardiolipin. These studies implicate membrane phospholipids as important host determinants for FHV RNA polymerase membrane association and provide evidence for the involvement of host phospholipids in positive-strand RNA virus membrane-specific targeting.
Temporal Regulation of the Mre11-Rad50-Nbs1 Complex during Adenovirus Infection
Authors: Karen Kasey A. , Hoey Peter J. , Young C. S. H. , Hearing Patrick 
Published: February 2009

ABSTRACT
Adenovirus infection induces a cellular DNA damage response that can inhibit viral DNA replication and ligate viral genomes into concatemers. It is not clear if the input virus is sufficient to trigger this response or if viral DNA replication is required. Adenovirus has evolved two mechanisms that target the Mre11-Rad50-Nbs1 (MRN) complex to inhibit the DNA damage response. These include E4-ORF3-dependent relocalization of MRN proteins and E4-ORF6/E1B-55K-dependent degradation of MRN components. The literature suggests that degradation of the MRN complex due to E4-ORF6/E1B-55K does not occur until after viral DNA replication has begun. We show that, by the time viral DNA accumulates, the MRN complex is inactivated by either of the E4-induced mechanisms and that, with E4-ORF6/E1B-55K, this inactivation is due to MRN degradation. Our data are consistent with the conclusion that input viral DNA is sufficient to induce the DNA damage response. Further, we demonstrate that when the DNA damage response is active in E4 mutant virus infections, the covalently attached terminal protein is not cleaved from viral DNAs, and the viral origins of replication are not detectably degraded at a time corresponding to the onset of viral replication. The sequences of concatemeric junctions of viral DNAs were determined, which supports the conclusion that nonhomologous end joining mediates viral DNA ligation. Large deletions were found at these junctions, demonstrating nucleolytic procession of the viral DNA; however, the lack of terminal protein cleavage and terminus degradation at earlier times shows that viral genome deletion and concatenation are late effects.
Primary B-Cell Infection with a ΔBALF4 Epstein-Barr Virus Comes to a Halt in the Endosomal Compartment yet Still Elicits a Potent CD4-Positive Cytotoxic T-Cell Response
Authors: Neuhierl Bernhard , Feederle Regina , Adhikary Dinesh , Hub Birgit , Geletneky Karsten , Mautner Josef , Delecluse Henri-Jacques 
Published: February 2009

ABSTRACT

Epstein-Barr virus (EBV) infection is mediated by several viral envelope glycoproteins. We have assessed gp110's functions during the virus life cycle using a mutant that lacks BALF4 (ΔBALF4). Exposure of various cell lines and primary cell samples of epithelial or lymphoid lineages to the ΔBALF4 mutant failed to establish stable infections. The ΔBALF4 virus, however, did not differ from wild-type EBV in its ability to bind and become internalized into primary B cells, in which it elicited a potent T-cell-specific immune reaction against virion constituents. These findings show that ΔBALF4 viruses can reach the endosome-lysosome compartment and dovetail nicely with the previously identified contribution of gp110 to virus-cell fusion. Other essential steps of the virus life cycle were unaffected in the viral mutant; DNA lytic replication and viral titers were not altered in the absence of gp110, and ΔBALF4 viruses complemented in
trans
transformed infected B cells with an efficiency indistinguishable from that observed with wild-type viruses. All of the steps of virus maturation could be observed in lytically induced 293/ΔBALF4 cells. Induction of lymphoblastoid cells generated with transiently complemented ΔBALF4 virus led to the production of rare mature virions. We therefore infer that gp110 is not required for virus maturation and egress in 293 cells or in B cells. The ΔBALF4 virus's phenotypic traits, an inability to infect human cells coupled with potent antigenicity, potentially qualify this mutant as a live vaccine. It will provide a useful tool for the detailed study of EBV-cell interactions in a physiological context.