bluuebunny/crossref_metadata_2025 huggingface dataset:
Land cover change in Europe between 1950 and 2000 determined employing aerial photography
Authors: Gerard F. , Petit S. , Smith G. , Thomson A. , Brown N. , Manchester S. , Wadsworth R. , Bugar G. , Halada L. , Bezák P. , Boltiziar M. , De badts E. , Halabuk A. , Mojses M. , Petrovic F. , Gregor M. , Hazeu G. , Mücher C.A. , Wachowicz M. , Huitu H. , Tuominen S. , Köhler R. , Olschofsky K. , Ziese H. , Kolar J. , Sustera J. , Luque S. , Pino J. , Pons X. , Roda F. , Roscher M. , Feranec J.
Published: March 2010

BIOPRESS (‘Linking Pan-European Land Cover Change to Pressures on Biodiversity’), a European Commission funded ‘Global Monitoring for Environment and Security’ project, produced land cover change information (1950—2000) for Europe from aerial photographs and tested the suitability of this for monitoring habitats and biodiversity. The methods and results related to the land cover change work are summarized. Changes in land cover were established through 73 window and 59 transect samples distributed across Europe. Although the sample size was too small and biased to fully represent the spatial variability observed in Europe, the work highlighted the importance of method consistency, the choice of nomenclature and spatial scale. The results suggest different processes are taking place in different parts of Europe: the Boreal and Alpine regions are dominated by forest management; abandonment and intensification are mainly encountered in the Mediterranean; urbanization and drainage are more characteristic of the Continental and Atlantic regions.
Clinical Follow-up of Professional Baseball Players Undergoing Ulnar Collateral Ligament Reconstruction Using the New Kerlan-Jobe Orthopaedic Clinic Overhead Athlete Shoulder and Elbow Score (KJOC Score)
Authors: Domb Benjamin G. , Davis J. T. , Alberta Frank G. , Mohr Karen J. , Brooks Adam G. , ElAttrache Neal S. , Yocum Lewis M. , Jobe Frank W.
Published: March 2010

Background: There are no validated outcome measures consistently used in the literature to report results of ulnar collateral ligament reconstruction in overhead athletes.

Hypothesis: The Kerlan-Jobe Orthopaedic Clinic Overhead Athlete Shoulder and Elbow score (KJOC score) will correlate with other validated scores for upper extremity assessment but will be more accurate in evaluating ulnar collateral ligament reconstruction outcomes in professional baseball players.

Study Design: Cohort study (diagnosis); Level of evidence, 2.

Methods: Fifty-five professional baseball players who underwent ulnar collateral ligament reconstruction were asked to complete the KJOC score, the Disabilities of the Arm, Shoulder and Hand (DASH) score, and the DASH sports module. Players were separated into 3 categories—(1) playing without pain, (2) playing with pain, and (3) not playing because of pain—and compared with 123 asymptomatic throwers. Pearson (parametric) and Spearman rank (nonparametric) correlations among the 3 systems were conducted to validate the KJOC score. Means across categories were compared using a Wilcoxon rank-sum test, and a threshold score separating categories 1 and 3 was determined using receiver operator characteristic discrimination analysis.

Results: Significant correlations were found between the KJOC score and the DASH (−.693, P < .0001), and the DASH sports module (−0.804, P < .0001). Only the KJOC score was able to discriminate between categories 2 and 3, as well as category 1 and the uninjured population. In addition, the KJOC score was the most sensitive and accurate method of discriminating category 1 from category 3, with a threshold score of 81.3.

Conclusion: The results of this study validate the use of the KJOC score for evaluation of overhead athletes undergoing ulnar collateral ligament reconstruction. The KJOC score is the most sensitive score for detecting subtle changes in performance in the throwing athlete.
Distal Semitendinosus Ruptures in Elite-Level Athletes
Authors: Cooper Daniel E. , Conway John E.
Published: March 2010

Background: No case series of isolated complete rupture of the distal semitendinosus tendon have been reported previously.

Purpose: This study was undertaken to increase awareness and report the authors’ treatment experience, particularly the less than favorable results of nonoperative initial treatment.

Study Design: Case series; Level of evidence, 4.

Methods: The authors identified 25 cases of distal semitendinosus tendon rupture over a 14-year period (1991-2005). All players were male professional (20), collegiate (4), or high-level amateur (1) athletes. Follow-up of 17 cases averaged 13 months (range, 4-55), and 8 patients were lost to follow-up. Eight Major League Baseball, 8 National Football League, and 1 National Hockey League athletes were included in this study. Early treatment experience always involved nonoperative treatment, including rest, modalities, and rehabilitation exercises, followed by functional progression. “Recovery” was defined by clinical criteria including clearance to return to play. Failure to improve with nonoperative treatment, and thus requiring surgical treatment, was deemed a failure of nonoperative treatment. There were 12 players who had initial nonoperative treatment. The authors had later experience with 5 players who had surgery early in the acute phase in hopes of speeding return to competition.

Results: In the nonoperative treatment group (12), 7 players recovered at an average of 10.4 weeks (range, 3-35). Five of these players (42%) failed initial nonoperative treatment (mean, 16.8 weeks) and subsequently had surgery to resect the torn tendon and surrounding scar tissue. These 5 players recovered at an average of 12.8 weeks postoperatively. In the acute surgery group, 5players had surgery to resect the torn tendon and scar tissue within 4 weeks of injury. The acute-phase group had an average recovery of 6.8 weeks after surgery.

Conclusion: Distal semitendinosus ruptures frequently (42%) do not recover after nonoperative treatment. Acute surgical resection of the completely ruptured semitendinosus tendon may speed recovery when the athlete has a tender mass and difficulty extending the knee fully in the stance phase of gate. Future investigation is warranted to compare the long-term outcome of nonoperative treatment with that after acute surgery.
Biomechanical Comparison of a Modified Weaver-Dunn and a Free-Tissue Graft Reconstruction of the Acromioclavicular Joint Complex
Authors: Michlitsch Michael G. , Adamson Gregory J. , Pink Marilyn , Estess Allyson , Shankwiler James A. , Lee Thay Q.
Published: March 2010

Background: Most surgical reconstructions of the separated acromioclavicular joint do not address the injured ligaments and capsule of the acromioclavicular joint.

Purpose: This study was undertaken to compare the biomechanical characteristics of a modified Weaver-Dunn reconstruction and an intramedullary acromioclavicular joint reconstruction that uses a free-tissue graft for reconstruction of both the coracoclavicular and acromioclavicular ligaments.

Study Design: Controlled laboratory study.

Methods: Each pair of 6 matched pairs of cadaveric shoulders was randomly selected for a modified Weaver-Dunn reconstruction on 1 side and the contralateral side was used for free-tissue graft reconstruction of the coracoclavicular and acromioclavicular ligamentous complexes. Anterior-posterior and superior-inferior acromioclavicular joint translation (in millimeters) was measured with acromioclavicular joint compressions of 10, 20, and 30 N, and with translational loads of 10 and 15 N both before and after acromioclavicular joint reconstruction. Load-to-failure testing was then performed for each construct. Repeated-measures analysis of variance (translational testing) and Wilcoxon signed rank test (load-to-failure testing), both with P = .05, were used for statistical analysis.

Results: Mean anterior-posterior and superior-inferior translation of the intramedullary acromioclavicular joint reconstruction was significantly less than that of the modified Weaver-Dunn under all loading conditions ( P < .001 and P = .001, respectively), but was not significantly different from that of the intact state ( P = .656 and P = .173, respectively). Although the mean ultimate and yield loads and linear stiffness for the intramedullary acromioclavicular reconstruction were greater than that of the modified Weaver-Dunn reconstruction, this did not reach statistical significance ( P = .625, P = .625, and P = .625, respectively).

Conclusion: Acromioclavicular joint reconstruction with free-tissue graft for both the coracoclavicular and acromioclavicular ligamentous complexes demonstrates initial stability significantly better than a modified Weaver-Dunn and similar to that of intact specimens.

Clinical Relevance: This acromioclavicular joint reconstruction provides the surgeon with a relatively nondestructive option.
Screening of Late-Onset Pompe Disease in a Sample of Mexican Patients With Myopathies of Unknown Etiology: Identification of a Novel Mutation in the Acid α-Glucosidase Gene
Authors: Alcántara-Ortigoza Miguel Angel , González-del Angel Ariadna , Barrientos-Ríos Rehotbevely , Cupples Courtney , Garrido-García Luis Martín , de León-Bojorge Beatríz , Alva-Chaire Adriana del Carmen
Published: March 2010

Pompe disease or glycogen-storage disease type 2 (GSD2, OMIM 232300) is an autosomal recessive disorder caused by mutations in the acid α-glucosidase gene. Late-onset GSD2 resembles some limb-girdle and Becker muscular dystrophies. The screening of GSD2 through the measurement of acid α-glucosidase activity in dried blood spots was applied to a selected sample of 5 Mexican patients with proximal myopathies of unknown etiology. Only 1 male patient showed a low level of acid α-glucosidase activity and a compound heterozygote genotype for the c.-32-13T>G splicing mutation present in most white late-onset Pompe disease cases and the novel mutation p.C558S. To our knowledge, this is the first report of a Mexican patient with late-onset GSD2. The identification of c.-32-13T>G in our patient could reflect the genetic contribution of European ancestry to the Mexican population. The enzymatic screening of GSD2 could be justified in patients with myopathies of unknown etiology.
Domain I of β2-glycoprotein I: its role as an epitope and the potential to be developed as a specific target for the treatment of the antiphospholipid syndrome
Authors: Ioannou Y. , Rahman A.
Published: March 2010

Antiphospholipid syndrome (APS) represents one of the most common acquired causes of thrombophilia and recurrent miscarriages. The only treatment of proven benefit is anticoagulation, often required at high intensity and life-long duration. This therapy can be associated with side effects such as bleeding and is not always effective. Hence, there remains a need for safer, targeted and ideally more effective therapies. Antiphospholipid antibodies (aPL) are pathogenic and promote thrombosis. Independent groups, including our own, have show that the major epitopes that pathogenic aPL targets lie within domain I of the protein β2-glycoprotein I (β2GPI). This review focuses on the evidence presented thus far which characterizes the immunodominant epitopes within domain I, demonstrating that the epitope is a conformational one centred around residues R39—G43 and also involving other residues within domain I, such as residues D8 and D9. The hypothesis is proposed that a recombinant domain I molecule, and a recombinant mutant domain I with enhanced aPL binding properties, may be used as an inhibitor of aPL binding and thus inhibit aPL-induced pathogenicity. In vivo proof-of-concept studies within the murine femoral vein injury model are presented supporting this hypothesis, and the rationale as well as potential benefits and problems of employing such an approach to treat APS are discussed.
Autoimmunity, infection and adjuvants
Authors: Rose NR
Published: March 2010

The effect of infection in initiating autoimmune disease has been debated for many years. There are, even now, few instances of a human autoimmune disease clearly caused by prior infection, probably due to the frequent separation in time and space from the clinical outcomes. As our understanding of the immunologic consequences of the infectious process has deepened, we can re-think some of the issues by focusing attention on the varied adjuvant effects of microbial products. We are now able to distinguish some of the critical steps in progression from virus infection to benign autoimmunity to autoimmune disease in an experimental model of myocarditis. Immune regulators, such as cytokines and costimulatory molecules, serve as signposts in the process. The lessons learned may be broadly applicable to autoimmune disorders.
Pathophysiology of β2-glycoprotein I in antiphospholipid syndrome
Authors: Matsuura E. , Shen L. , Matsunami Y. , Quan N. , Makarova M. , Geske FJ , Boisen M. , Yasuda S. , Kobayashi K. , Lopez LR
Published: March 2010

Since β2-glycoprotein I (β2GPI) was described as the major antigenic target for antiphospholipid antibodies, many studies have focused their attention to the physiological role of β2GPI and anti-β2GPI antibodies on autoimmune-mediated thrombosis. Studies reporting the physiological role of β2GPI have been numerous, but the exact mechanism of action(s) has yet to be completely determined. β2GPI’s epitopes for anti-β2GPI autoantibodies have been characterized, however, not all of the heterogeneous anti-β2GPI antibodies are pathogenic. The pathophysiologic role of β2GPI has been reported in the fields of coagulation, fibrinolysis, angiogenesis, and atherosclerosis. Our understanding of the impact of β2GPI, its metabolites and autoantibodies to β2GPI on these physiological functions may contribute to the development of better therapeutic strategies to treat and prevent autoimmune-mediated atherothrombotic vascular disease. Lupus (2010) 19, 379—384.
Antiprothrombin antibody: why do we need more assays?
Authors: Atsumi T. , Koike T.
Published: March 2010

Anticardiolipin antibodies (aCL), anti-β2-glycoprotein I (β2GPI) antibodies and lupus anticoagulant (LA) are the only laboratory tests considered within the revised criteria for the classification of the antiphospholipid syndrome (APS). Recently, the significance to assay the antibodies against phosphatidylserine—prothrombin complex (aPS/PT) has been discussed, and these antibodies, rather than antibodies against prothrombin alone, are closely associated with APS and LA. The sensitivity and specificity of aPS/PT for the diagnosis of APS were assessed in a population of patients with a variety of autoimmune disorders. The aCL and aPS/PT have similar diagnostic value for APS, and most of APS patients with aPS/PT had positive LA. Therefore, aPS/PT should be further explored, not only for research purposes, but also as a candidate for one of the enzyme-linked immunosorbent assay (ELISA)-based confirmatory test for APS associated LA.
Strains of Epstein-Barr virus infecting multiple sclerosis patients
Authors: Brennan RM , Burrows JM , Bell MJ , Bromham L. , Csurhes PA , Lenarczyk A. , Sverndal J. , Klintenstedt J. , Pender MP , Burrows SR
Published: March 2010

Both epidemiological and experimental studies have indicated that the ubiquitous herpesvirus Epstein—Barr virus (EBV) plays a role in the pathogenesis of multiple sclerosis (MS). Some features of MS epidemiology, such as the decline in risk among migrants from high to low MS prevalence areas, suggest the presence of variant EBV strains that increase MS risk. The objective of this study was to investigate whether genetic variability in EBV is associated with MS. Genes encoding for two EBV antigens (EBNA1 and BRRF2) were sequenced in EBV isolates from 40 MS patients and a similar number of control subjects. These viral antigens were chosen for analysis because they are known to stimulate atypical immune responses in MS. Extensive sequence polymorphism was observed within the EBNA1 and BRRF2 genes in isolates from both MS patients and controls. Interestingly, several single nucleotide polymorphisms within the EBNA1 gene, and one within the BRRF2 gene, were found to occur at marginally different frequencies in EBV strains infecting MS patients versus controls. Although this study does not find a simple causal relationship between EBV strains and the occurrence of MS, the existence of haplotypes that occur at different frequencies in MS patients versus controls may provide an area for future study of the role of EBV strain variation in multiple sclerosis.
Time domain and spectral domain optical coherence tomography in multiple sclerosis: a comparative cross-sectional study
Authors: Bock Markus , Brandt Alexander Ulrich , Dörr Jan , Pfueller Caspar F , Ohlraun Stephanie , Zipp Frauke , Paul Friedemann
Published: March 2010

Conventional time domain optical coherence tomography has been established for the in vivo assessment of retinal axonal loss in multiple sclerosis. The innovative spectral domain imaging is superior to the conventional technique with respect to data acquisition speed, resolution and reproducibility. However, until now comparability of the two techniques has not been investigated in multiple sclerosis. In this study involving 55 multiple sclerosis patients, data obtained using both techniques (Stratus time domain optical coherence tomography and Cirrus spectral domain optical coherence tomography, Carl Zeiss Meditec) showed an excellent correlation (Pearson’s r = 0.926, p < 0.001). However, owing to considerable differences in absolute retinal nerve fibre layer measurements (mean ± standard deviation 8.1 µm ± 6.2, range -12 to 23 µm), results from the two devices are not interchangeable.
Moral Psychology Must Not Be Based on Faith and Hope: Commentary on Narvaez (2010)
Authors: Haidt Jonathan
Published: March 2010

Narvaez (2010, this issue) calls for a moral psychology in which reasoning and intuitions are equal partners. But empirical research on the power of implicit processes and on the weakness of everyday reasoning indicates that the partnership is far from equal. The ancient rationalist faith that good reasoning can be taught and that it will lead to improved behavior is no longer justified. The social intuitionist model (Haidt, 2001) is a more realistic portrayal of the ways that moral intuition and reasoning work together.
A comparative study of the modified Sauvé–Kapandji procedure for rheumatoid wrist with and without stabilization of the proximal ulnar stump
Authors: Kawabata A. , Egi T. , Hashimoto H. , Masada K. , Saito S.
Published: March 2010

We compared the clinical and radiological results of the modified Sauvé–Kapandji procedure for 41 of 86 operated rheumatoid wrists with ( n = 22) and without ( n = 19) stabilization of the proximal ulnar stump with a slip of half the extensor carpi ulnaris tendon. Gender, age, and follow-up period were similar in the two groups. We found no difference clinically or on radiographs between the two groups other than better early postoperative pain relief in those stabilized. Stabilization of the proximal ulnar stump may not be necessary in the modified Sauvé–Kapandji procedure for rheumatoid wrists.
Review: Functional dyspepsia
Authors: Brun Rita , Kuo Braden
Published: March 2010

Dyspepsia is a common term used for a heterogeneous group of abdominal symptoms. Functional dyspepsia (FD) is the focus of this review. The 2006 Rome III criteria defined FD and its subgroups, postprandial distress syndrome (PDS) and epigastric pain syndrome (EPS). FD is a very common condition with a high prevalence throughout the world, adversely affecting the quality of life of patients. The pathophysiology of FD has been under investigation during the past two decades. Multiple mechanisms such as abnormal gastric emptying, visceral hypersensitivity, impaired gastric accommodation, and central nervous system factors are likely involved. Several tests are available for the assessment of various physiologic functions possibly involved in the pathogenesis of FD, and some of these could be used in clinical practice, helping to understand the abnormalities underlining patients’ complaints. Currently, the possibilities of pharmacological therapy for FD are still limited, however, experience of using prokinetics, tricyclic antidepressants, selective serotonin-reuptake inhibitors (SSRIs), proton-pump inhibitors (PPIs), and several alternative techniques has been accumulated. The different combinations of alterations in physiologic gastrointestinal and central nervous system functions result in the very heterogeneous nature of FD so combined approaches to these patients could be beneficial in challenging cases.
Efficacy of Osseous Abnormalities Correction with Arthroscopic Surgery in Femoroacetabular Impingement
Authors: Camacho Alvarez Daniel R. , Mardones Rodrigo M.
Published: March 2010

Femoroacetabular impingement (FAI) is a clinical syndrome characterized by subtle abnormal morphology of the proximal femur and/or the acetabulum that leads to abnormal contact between the femoral neck and the acetabular rim during the hip range of motion. Traditionally, FAI has been managed safely and effectively with surgical hip dislocation; less invasive arthroscopic techniques are now being used to an increasing extent, trying to emulate the results of the open technique. The purpose of this study was to evaluate the radiographic results of arthroscopic acetabular rim trimming and femoral osteochondroplasty in FAI. This was a retrospective analysis of preoperative and postoperative plain radiographs of 80 patients treated for FAI with arthroscopic surgery between April 2007 and December 2008. We evaluated 2 parameters: the Wiberg angle (center-edge angle) (normal, 25°-35°), and the anterior/posterior relation of femoral head-neck offset (normal, 0.8-1). Of 80 hips, 10 (12.5%) were pincer-type impingement, 17 (21.25%) were cam type, and 53 (66.25%) were mixed type. The preoperative Wiberg average was 39° (range, 25°-51°), and the postoperative Wiberg average was 32° (range, 25°-42°). The preoperative anterior/posterior femoral offset relation average was 0.42 (range, -0.38 to 1), and the postoperative anterior/posterior femoral offset relation average was 0.94 (range, 0.61-1.2). Our results show that it is possible to obtain an anatomical correction of the osseous abnormalities with arthroscopic surgery in FAI. Level of evidence: level III.
TRAF6 inhibits Th17 differentiation and TGF-β–mediated suppression of IL-2
Authors: Cejas Pedro J. , Walsh Matthew C. , Pearce Erika L. , Han Daehee , Harms Gretchen M. , Artis David , Turka Laurence A. , Choi Yongwon
Published: March 2010

Transforming growth factor-β (TGF-β) has an essential role in the generation of inducible regulatory T (iTreg) and T helper 17 (Th17) cells. However, little is known about the TGF-β–triggered pathways that drive the early differentiation of these cell populations. Here, we report that CD4+ T cells lacking the molecular adaptor tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) exhibit a specific increase in Th17 differentiation in vivo and in vitro. We show that TRAF6 deficiency renders T cells more sensitive to TGF-β–induced Smad2/3 activation and proliferation arrest. Consistent with this, in TRAF6-deficient T cells, TGF-β more effectively down-regulates interleukin-2 (IL-2), a known inhibitor of Th17 differentiation. Remarkably, TRAF6-deficient cells generate normal numbers of Foxp3-expressing cells in iTreg differentiation conditions where exogenous IL-2 is supplied. These findings show an unexpected role for the adaptor molecule TRAF6 in Smad-mediated TGF-β signaling and Th17 differentiation. Importantly, the data also suggest that a main function of TGF-β in early Th17 differentiation may be the inhibition of autocrine and paracrine IL-2–mediated suppression of Th17 cell generation.
Chronic coumarin treatment is associated with increased extracoronary arterial calcification in humans
Authors: Rennenberg Roger J. M. W. , van Varik Bernard J. , Schurgers Leon J. , Hamulyak Karly , ten Cate Hugo , Leiner Tim , Vermeer Cees , de Leeuw Peter W. , Kroon Abraham A.
Published: March 2010

Vascular calcification is a marker of increased cardiovascular risk. Vitamin K–dependent matrix Gla protein (MGP) is important in inhibiting calcification. Because MGP activation is vitamin K dependent, we performed a cross-sectional study investigating the relationship between the use of vitamin K antagonists and extracoronary vascular calcification. From the Dutch thrombosis services we selected 19 patients younger than 55 years who had no other cardiovascular risk factors and who had used coumarins for more than 10 years, and compared these to 18 matched healthy controls. MGP was measured, and a plain x-ray of the thighs was taken to assess femoral arterial calcifications. The odds ratio for calcification in patients versus controls was 8.5 (95% confidence interval [CI] 2.01-35.95). Coumarin use and MGP were associated with calcification, even after adjusting for other risk factors. We conclude that long-term use of coumarins is associated with enhanced extracoronary vascular calcification, possibly through the inhibition of MGP carboxylation.
Mechanisms of bradykinin-induced contraction in human fetal lung fibroblasts
Authors: Petecchia L. , Sabatini F. , Usai C. , Carnevali S. , Ognibene M. , Vanni C. , Eva A. , Fabbri L.M. , Rossi G.A. , Ricciardolo F.L.M.
Published: March 2010

Bradykinin (BK) induces fibroblast contraction but the structural changes and intracellular mechanisms involved have not been completely explored.

We stimulated HFL-1 fibroblasts with BK to assess: 1) fibroblast contractility; 2) the role of α-smooth muscle actin (SMA) in contraction by small interfering RNA (siRNA); 3) α-SMA protein expression; 4) α-SMA and F-actin structure; 5) intracellular calcium concentration ([Ca2+]i); and 6) phosphorylated myosin light-chain (pMLC) and MLC kinase (MLCK) expression.

BK triggered concentration- and time-dependent fibroblast gel contraction in conjunction with α-SMA over expression, but not in α-SMA-siRNA-treated cells. BK also increased α-SMA+and F-actin+cell number and stress fibre polymerisation (detectable at 5–60 min). These BK-induced changes were associated with an increase in [Ca2+]i, which peaked within 15 s, and activation of pMLC, which was detectable at 5–60 min. No MLCK content modification was observed. The different manifestations of the BK-induced fibroblast activation were downregulated at different levels (25–100%) by HOE140, a specific BK B2 receptor (B2R) antagonist and by the Ca2+chelator, EGTA.

Thus, BK-induced fibroblast contraction, associated with differentiation into α-SMA+myofibroblasts, is mediated through the activation of the B2R and involves the Ca2+/calmodulin pMLC-dependent pathway.
Effects of varying Notch1 signal strength on embryogenesis and vasculogenesis in compound mutant heterozygotes
Authors: Ge Changhui , Stanley Pamela
Published: March 2010

Background

Identifying developmental processes regulated by Notch1 can be addressed in part by characterizing mice with graded levels of Notch1 signaling strength. Here we examine development in embryos expressing various combinations of Notch1 mutant alleles. Mice homozygous for the hypomorphic Notch112fallele, which removes the single O-fucose glycan in epidermal growth factor-like repeat 12 (EGF12) of the Notch1 ligand binding domain (lbd), exhibit reduced growth after weaning and defective T cell development. Mice homozygous for the inactive Notch1
lbd
allele express Notch1 missing an ~20 kDa internal segment including the canonical Notch1 ligand binding domain, and die at embryonic day ~E9.5. The embryonic and vascular phenotypes of compound heterozygous Notch112f/lbdembryos were compared with Notch1+/12f, Notch112f/12f, and Notch1lbd/lbdembryos. Embryonic stem (ES) cells derived from these embryos were also examined in Notch signaling assays. While Notch1 signaling was stronger in Notch112f/lbdcompound heterozygotes compared to Notch1lbd/lbdembryos and ES cells, Notch1 signaling was even stronger in embryos carrying Notch112fand a null Notch1 allele.

Results

Mouse embryos expressing the hypomorphic Notch112fallele, in combination with the inactive Notch1
lbd
allele which lacks the Notch1 ligand binding domain, died at ~E11.5-12.5. Notch112f/lbdES cells signaled less well than Notch112f/12fES cells but more strongly than Notch1lbd/lbdES cells. However, vascular defects in Notch112f/lbdyolk sac were severe and similar to Notch1lbd/lbdyolk sac. By contrast, vascular disorganization was milder in Notch112f/lbdcompared to Notch1lbd/lbdembryos. The expression of Notch1 target genes was low in Notch112f/lbdyolk sac and embryo head, whereas Vegf and Vegfr2 transcripts were increased. The severity of the compound heterozygous Notch112f/lbdyolk sac phenotype suggested that the allelic products may functionally interact. By contrast, compound heterozygotes with Notch112fin combination with a Notch1 null allele (Notch1tm1Con) were capable of surviving to birth.

Conclusions

Notch1 signaling in Notch112f/lbdcompound heterozygous embryos is more defective than in compound heterozygotes expressing a hypomorphic Notch112fallele and a Notch1 null allele. The data suggest that the gene products Notch1lbd and Notch112f interact to reduce the activity of Notch112f.
A 2cM genome-wide scan of European Holstein cattle affected by classical BSE
Authors: Murdoch Brenda M , Clawson Michael L , Laegreid William W , Stothard Paul , Settles Matthew , McKay Stephanie , Prasad Aparna , Wang Zhiquan , Moore Stephen S , Williams John L
Published: March 2010

Background

Classical bovine spongiform encephalopathy (BSE) is an acquired prion disease that is invariably fatal in cattle and has been implicated as a significant human health risk. Polymorphisms that alter the prion protein of sheep or humans have been associated with variations in transmissible spongiform encephalopathy susceptibility or resistance. In contrast, there is no strong evidence that non-synonymous mutations in the bovine prion gene (PRNP) are associated with classical BSE disease susceptibility. However, two bovine PRNP insertion/deletion polymorphisms, one within the promoter region and the other in intron 1, have been associated with susceptibility to classical BSE. These associations do not explain the full extent of BSE susceptibility, and loci outside of PRNP appear to be associated with disease incidence in some cattle populations. To test for associations with BSE susceptibility, we conducted a genome wide scan using a panel of 3,072 single nucleotide polymorphism (SNP) markers on 814 animals representing cases and control Holstein cattle from the United Kingdom BSE epidemic.

Results

Two sets of BSE affected Holstein cattle were analyzed in this study, one set with known family relationships and the second set of paired cases with controls. The family set comprises half-sibling progeny from six sires. The progeny from four of these sires had previously been scanned with microsatellite markers. The results obtained from the current analysis of the family set yielded both some supporting and new results compared with those obtained in the earlier study. The results revealed 27 SNPs representing 18 chromosomes associated with incidence of BSE disease. These results confirm a region previously reported on chromosome 20, and identify additional regions on chromosomes 2, 14, 16, 21 and 28. This study did not identify a significant association near the PRNP in the family sample set. The only association found in the PRNP region was in the case-control sample set and this was not significant after multiple test correction. The genome scan of the case-control animals did not identify any associations that passed a stringent genome-wide significance threshold.

Conclusions

Several regions of the genome are statistically associated with the incidence of classical BSE in European Holstein cattle. Further investigation of loci on chromosomes 2, 14, 16, 20, 21 and 28 will be required to uncover any biological significance underlying these marker associations.
Testes and brain gene expression in precocious male and adult maturing Atlantic salmon (Salmo salar)
Authors: Guiry Aoife , Flynn Denis , Hubert Sophie , O'Keeffe Allan M , LeProvost Olivier , White Samantha L , Forde Patrick F , Davoren Pamela , Houeix Benoit , Smith Terry J , Cotter Deirdre , Wilkins Noel P , Cairns Michael T
Published: March 2010

Background

The male Atlantic salmon generally matures in fresh water upon returning after one or several years at sea. Some fast-growing male parr develop an alternative life strategy where they sexually mature before migrating to the oceans. These so called 'precocious' parr or 'sneakers' can successfully fertilise adult female eggs and so perpetuate their line. We have used a custom-built cDNA microarray to investigate gene expression changes occurring in the salmon gonad and brain associated with precocious maturation. The microarray has been populated with genes selected specifically for involvement in sexual maturation (precocious and adult) and in the parr-smolt transformation.

Results

Immature and mature parr collected from a hatchery-reared stock in January were significantly different in weight, length and condition factor. Changes in brain expression were small - never more than 2-fold on the microarray, and down-regulation of genes was much more pronounced than up-regulation. Significantly changing genes included isotocin, vasotocin, cathepsin D, anamorsin and apolipoprotein E. Much greater changes in expression were seen in the testes. Among those genes in the testis with the most significant changes in expression were anti-Mullerian hormone, collagen 1A, and zinc finger protein (Zic1), which were down-regulated in precocity and apolipoproteins E and C-1, lipoprotein lipase and anti-leukoproteinase precursor which were up-regulated in precocity. Expression changes of several genes were confirmed in individual fish by quantitative PCR and several genes (anti-Mullerian hormone, collagen 1A, beta-globin and guanine nucleotide binding protein (G protein) beta polypeptide 2-like 1 (GNB2L1) were also examined in adult maturing testes. Down-regulation of anti-Mullerian hormone was judged to be greater than 160-fold for precocious males and greater than 230-fold for November adult testes in comparison to July testes by this method. For anti-Mullerian hormone and guanine nucleotide binding protein beta polypeptide 2-like 1 expression changes in precocious males mirrored mature adults (November) but for collagen 1A and beta-globin the pattern was more complex.

Conclusions

Expression changes in the fish brain during the process of precocious sexual maturation were small compared to those in the testes. Microarray analysis suggested down-regulation of housekeeping functions and up-regulation of a small number of specific processes. Transcriptional changes in the testes were much more pronounced with anti-Mullerian hormone playing a major role. Expression profiles for mature parr and maturing adult testes indicate subtle differences in gene expression between these two related groups.
Structural and functional studies of STAT1 from Atlantic salmon (Salmo salar)
Authors: Skjesol Astrid , Hansen Tom , Shi Cheng-Yin , Thim Hanna L , Jørgensen Jorunn B
Published: March 2010

Background

Type I and type II interferons (IFNs) exert their effects mainly through the JAK/STAT pathway, which is presently best described in mammals. STAT1 is involved in signaling pathways induced by both types of IFNs. It has a domain-like structure including an amino-terminus that stabilizes interaction between STAT dimers in a promoter-binding situation, a coiled coil domain facilitating interactions to other proteins, a central DNA-binding domain, a SH2 domain responsible for dimerization of phosphorylated STATs and conserved phosphorylation sites within the carboxy terminus. The latter is also the transcriptional activation domain.

Results

A salmon (Salmo salar) STAT1 homologue, named ssSTAT1a, has been identified and was shown to be ubiquitously expressed in various cells and tissues. The ssSTAT1a had a domain-like structure with functional motifs that are similar to higher vertebrates. Endogenous STAT1 was shown to be phosphorylated at tyrosine residues both in salmon leukocytes and in TO cells treated with recombinant type I and type II IFNs. Also ectopically expressed ssSTAT1 was phosphorylated in salmon cells upon in vitro stimulation by the IFNs, confirming that the cloned gene was recognized by upstream tyrosine kinases. Treatment with IFNs led to nuclear translocation of STAT1 within one hour. The ability of salmon STAT1 to dimerize was also shown.

Conclusions

The structural and functional properties of salmon STAT1 resemble the properties of mammalian STAT1.
Adjuvant glycerol is not beneficial in experimental pneumococcal meningitis
Authors: Blaser Cornelia , Klein Matthias , Grandgirard Denis , Wittwer Matthias , Peltola Heikki , Weigand Michael , Koedel Uwe , Leib Stephen L
Published: March 2010

Background

Bacterial meningitis in children causes high rates of mortality and morbidity. In a recent clinical trial, oral glycerol significantly reduced severe neurological sequelae in paediatric meningitis caused by Haemophilus influenzae type b, and a tendency towards a benefit of adjunctive glycerol was seen in pneumococcal meningitis.

Methods

Here we examined the effects of glycerol in pneumococcal meningitis of infant rats and adult mice. All animals received ceftriaxone, and glycerol or placebo. Brain damage, hearing loss, and inflammatory parameters were assessed.

Results

Clinically and by histopathology, animals treated with glycerol or placebo did not differ. While both groups showed equally high levels of matrix metalloproteinase-9 at 24 h after infection, a significant difference in favour of glycerol was observed at 40 h after infection. However, this difference in matrix metalloproteinase-9 in late disease did not result in an improvement of histopathologic parameters.

Conclusion

No benefit of adjunctive glycerol was found in these models of pneumococcal meningitis.
Low incidence of limb-girdle muscular dystrophy type 2C revealed by a mutation study in Japanese patients clinically diagnosed with DMD
Authors: Okizuka Yo , Takeshima Yasuhiro , Itoh Kyoko , Zhang Zhujun , Awano Hiroyuki , Maruyama Koichi , Kumagai Toshiyuki , Yagi Mariko , Matsuo Masafumi
Published: March 2010

Background

Limb-girdle muscular dystrophy type 2C (LGMD2C) is an autosomal recessive muscle dystrophy that resembles Duchenne muscular dystrophy (DMD). Although DMD is known to affect one in every 3500 males regardless of race, a widespread founder mutation causing LGMD2C has been described in North Africa. However, the incidence of LGMD2C in Japanese has been unknown because the genetic background remains uncharacterized in many patients clinically diagnosed with DMD.

Methods

We enrolled 324 patients referred to the Kobe University Hospital with suspected DMD. Mutations in the dystrophin or the SGCG genes were analyzed using not only genomic DNA but also cDNA.

Results

In 322 of the 324 patients, responsible mutations in the dystrophin were successfully revealed, confirming DMD diagnosis. The remaining two patients had normal dystrophin expression but absence of γ-sarcoglycan in skeletal muscle. Mutation analysis of the SGCG gene revealed homozygous deletion of exon 6 in one patient, while the other had a novel single nucleotide insertion in exon 7 in one allele and deletion of exon 6 in the other allele. These mutations created a stop codon that led to a γ-sarcoglycan deficiency, and we therefore diagnosed these two patients as having LGMD2C. Thus, the relative incidence of LGMD2C among Japanese DMD-like patients can be calculated as 1 in 161 patients suspected to have DMD (2 of 324 patients = 0.6%). Taking into consideration the DMD incidence for the overall population (1/3,500 males), the incidence of LGMD2C can be estimated as 1 per 560,000 or 1.8 per million.

Conclusions

To the best of our knowledge, this is the first study to demonstrate a low incidence of LGMD2C in the Japanese population.
Allelic variants of IL1R1gene associate with severe hand osteoarthritis
Authors: Näkki Annu , Kouhia Sanna T , Saarela Janna , Harilainen Arsi , Tallroth Kaj , Videman Tapio , Battié Michele C , Kaprio Jaakko , Peltonen Leena , Kujala Urho M
Published: March 2010

Background

In search for genes predisposing to osteoarthritis (OA), several genome wide scans have provided evidence for linkage on 2q. In this study we targeted a 470 kb region on 2q11.2 presenting the locus with most evidence for linkage to severe OA of distal interphalangeal joints (DIP) in our genome wide scan families.

Methods

We genotyped 32 single nucleotide polymorphisms (SNPs) in this 470 kb region comprising six genes belonging to the interleukin 1 superfamily and monitored for association with individual SNPs and SNP haplotypes among severe familial hand OA cases (material extended from our previous linkage study; n = 134), unrelated end-stage bilateral primary knee OA cases (n = 113), and population based controls (n = 436).

Results

Four SNPs in the IL1R1 gene, mapping to a 125 kb LD block, provided evidence for association with hand OA in family-based and case-control analysis, the strongest association being with SNP rs2287047 (p-value = 0.0009).

Conclusions

This study demonstrates an association between severe hand OA and IL1R1 gene. This gene represents a highly relevant biological candidate since it encodes protein that is a known modulator of inflammatory processes associated with joint destruction and resides within a locus providing consistent evidence for linkage to hand OA. As the observed association did not fully explain the linkage obtained in the previous study, it is plausible that also other variants in this genome region predispose to hand OA.
The percentage of CD133+ cells in human colorectal cancer cell lines is influenced by Mycoplasma hyorhinis infection
Authors: Mariotti Elisabetta , Gemei Marica , Mirabelli Peppino , D'Alessio Francesca , Di Noto Rosa , Fortunato Giuliana , Del Vecchio Luigi
Published: March 2010

Background

Mollicutes contamination is recognized to be a critical issue for the cultivation of continuous cell lines. In this work we characterized the effect of Mycoplasma hyorhinis contamination on CD133 expression in human colon cancer cell lines.

Methods

MycoAlert® and mycoplasma agar culture were used to detect mycoplasma contamination on GEO, SW480 and HT-29 cell lines. Restriction fragment length polymorphism assay was used to determine mycoplasma species. All cellular models were decontaminated by the use of a specific antibiotic panel (Enrofloxacin, Ciprofloxacin, BM Cyclin 1 and 2, Mycoplasma Removal Agent and MycoZap®). The percentage of CD133 positive cells was analyzed by flow cytometry on GEO, SW480 and HT-29 cell lines, before and after Mycoplasma hyorhinis eradication.

Results

Mycoplasma hyorhinis infected colon cancer cell lines showed an increased percentage of CD133+ cells as compared to the same cell lines rendered mycoplasma-free by effective exposure to antibiotic treatment. The percentage of CD133 positive cells increased again when mycoplasma negative cells were re-infected by Mycoplasma hyorhinis.

Conclusions

Mycoplasma hyorhinis infection has an important role on the quality of cultured human colon cancer cell lines giving a false positive increase of cancer stem cells fraction characterized by CD133 expression. Possible explanations are (i) the direct involvement of Mycoplasma on CD133 expression or (ii) the selective pressure on a subpopulation of cells characterized by constitutive CD133 expression.

In keeping with United Kingdom Coordinating Committee on Cancer Research (UKCCCR) guidelines, the present data indicate the mandatory prerequisite, for investigators involved in human colon cancer research area, of employing mycoplasma-free cell lines in order to avoid the production of non-reproducible or even false data.
Analysis of the effectiveness of interventions used during the 2009 A/H1N1 influenza pandemic
Authors: Halder Nilimesh , Kelso Joel K , Milne George J
Published: March 2010

Background

Following the emergence of the A/H1N1 2009 influenza pandemic, public health interventions were activated to lessen its potential impact. Computer modelling and simulation can be used to determine the potential effectiveness of the social distancing and antiviral drug therapy interventions that were used at the early stages of the pandemic, providing guidance to public health policy makers as to intervention strategies in future pandemics involving a highly pathogenic influenza strain.

Methods

An individual-based model of a real community with a population of approximately 30,000 was used to determine the impact of alternative interventions strategies, including those used in the initial stages of the 2009 pandemic. Different interventions, namely school closure and antiviral strategies, were simulated in isolation and in combination to form different plausible scenarios. We simulated epidemics with reproduction numbers R0of 1.5, which aligns with estimates in the range 1.4-1.6 determined from the initial outbreak in Mexico.

Results

School closure of 1 week was determined to have minimal effect on reducing overall illness attack rate. Antiviral drug treatment of 50% of symptomatic cases reduced the attack rate by 6.5%, from an unmitigated rate of 32.5% to 26%. Treatment of diagnosed individuals combined with additional household prophylaxis reduced the final attack rate to 19%. Further extension of prophylaxis to close contacts (in schools and workplaces) further reduced the overall attack rate to 13% and reduced the peak daily illness rate from 120 to 22 per 10,000 individuals. We determined the size of antiviral stockpile required; the ratio of the required number of antiviral courses to population was 13% for the treatment-only strategy, 25% for treatment and household prophylaxis and 40% for treatment, household and extended prophylaxis. Additional simulations suggest that coupling school closure with the antiviral strategies further reduces epidemic impact.

Conclusions

These results suggest that the aggressive use of antiviral drugs together with extended school closure may substantially slow the rate of influenza epidemic development. These strategies are more rigorous than those actually used during the early stages of the relatively mild 2009 pandemic, and are appropriate for future pandemics that have high morbidity and mortality rates.
Peer substance use overestimation among French university students: a cross-sectional survey
Authors: Riou Franca Lionel , Dautzenberg Bertrand , Falissard Bruno , Reynaud Michel
Published: March 2010

Background

Normative misperceptions have been widely documented for alcohol use among U.S. college students. There is less research on other substances or European cultural contexts. This study explores which factors are associated with alcohol, tobacco and cannabis use misperceptions among French college students, focusing on substance use.

Methods

12 classes of second-year college students (n = 731) in sociology, medicine, nursing or foreign language estimated the proportion of tobacco, cannabis, alcohol use and heavy episodic drinking among their peers and reported their own use.

Results

Peer substance use overestimation frequency was 84% for tobacco, 55% for cannabis, 37% for alcohol and 56% for heavy episodic drinking. Cannabis users (p = 0.006), alcohol (p = 0.003) and heavy episodic drinkers (p = 0.002), are more likely to overestimate the prevalence of use of these consumptions. Tobacco users are less likely to overestimate peer prevalence of smoking (p = 0.044). Women are more likely to overestimate tobacco (p < 0.001) and heavy episodic drinking (p = 0.007) prevalence. Students having already completed another substance use questionnaire were more likely to overestimate alcohol use prevalence (p = 0.012). Students exposed to cannabis prevention campaigns were more likely to overestimate cannabis (p = 0.018) and tobacco use (p = 0.022) prevalence. Other identified factors are class-level use prevalences and academic discipline.

Conclusions

Local interventions that focus on creating realistic perceptions of substance use prevalence could be considered for cannabis and alcohol prevention in French campuses.