bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intron 4 polymorphism of the endothelial nitric oxide synthase eNOS gene and early microangiopathy in type 1 diabetes
Authors: Mamoulakis D. , Bitsori M. , Galanakis E. , Vazgiourakis V. , Panierakis C. , Goulielmos G. N. 
Published: May 2009

SummaryNitric oxide (NO) is an endogenous vasodilator involved in inflammatory and autoimmune response, and in the pathophysiology of diabetic vascular disease. Endothelium‐derived NO is formed from l‐arginine by endothelial NO synthase (eNOS), and earlier studies have provided evidence for altered NO metabolism and impaired endothelial function in diabetes, probably due to polymorphisms in eNOS gene. In the present study we investigated the association of the eNOS gene intron 4 a/b VNTR polymorphism with diabetic microangiopathy in 61 young individuals with type 1 diabetes (T1D), 35 male and 26 female, aged 5.0–29.1 (mean 15.6) years, and followed up for 3.24–11.4 (mean 7.44) years. Ten patients (16.4%) had developed microalbuminuria, three hypertension and two retinopathy. Wild‐type b/b homozygosity for eNOS gene intron 4 VNTR was found in 37 (60.7%) and a/b polymorphism in 24 (39.3%). No significant relationship was demonstrated between eNOS gene intron 4 polymorphisms and microalbuminuria, hypertension or retinopathy in these young individuals. Our findings suggest that a/b polymorphism of the intron 4 eNOS gene is not associated with early onset diabetic microangiopathy.
Comparative analysis of the impact of a free cysteine in tapasin on the maturation and surface expression of murine MHC class I allotypes
Authors: Wang X. , Simone L. C. , Tuli A. , Solheim J. C. 
Published: May 2009

SummaryTapasin is a key molecule in the major histocompatibility complex (MHC) class I peptide‐loading complex, interacting with several other proteins in the complex. An amino acid substitution at a free cysteine position in tapasin has been shown to disrupt the covalent association of tapasin with ERp57. In this study, we mutated the free cysteine in mouse tapasin, and analysed the effects on the cell surface expression of the mouse MHC class I molecules Kd and Kb. The C95S substitution in mouse tapasin increased the proportion of open forms relative to folded forms for both types of MHC class I molecules at the cell surface. Furthermore, the C95S substitution resulted in increased association of tapasin with folded Kd. Overall, our studies with these mouse MHC class I allotypes have revealed that the free cysteine 95 in mouse tapasin influences stable expression at the plasma membrane for both MHC class I allotypes, and have shown that tapasin's interaction with folded Kd is elevated by the C95S substitution in tapasin.
HLA class I and class II polymorphisms in Saudi patients with myasthenia gravis
Authors: Hajeer A. H. , Sawidan F. Al , Bohlega S. , Saleh S. , Sutton P. , Shubaili A. , Tahan A. Al , Al Jumah M. 
Published: May 2009

SummaryMyasthenia gravis (MG) is a rare autoimmune disease of the neuromuscular junction. MG has been shown to be associated with many HLA antigens in different populations. Here we have analysed the frequency of HLA‐A, B, DR and DQ in a group of Saudi MG patients and compared their results to a group of healthy controls. MG in Saudi patients is found to be associated with HLA‐A*23, B*08, B*18, DRB1*16 and DRB1*13. The strongest association was with HLA‐B*08, which was associated with young age at onset and female gender. Our results are in line with other published results from around the world and warrant fine mapping of the area using microsatellite to map the disease gene.
The CTLA4 +49 A/G polymorphism is not associated with susceptibility to type 1 diabetes mellitus in the Portuguese population
Authors: Lemos M. C. , Coutinho E. , Gomes L. , Bastos M. , Fagulha A. , Barros L. , Carrilho F. , Geraldes E. , Regateiro F. J. , Carvalheiro M. 
Published: May 2009

Summary CTLA4 genetic polymorphisms have been associated with type 1 diabetes. We genotyped 207 patients and 249 controls for the most frequently investigated polymorphism of the CTLA4 gene (+49A/G (rs231775)). No significant differences were observed, suggesting that this polymorphism is not strongly associated with type 1 diabetes in the Portuguese population.
Full‐length sequence and expression analysis of a myeloid differentiation factor 88 (MyD88) in half‐smooth tongue sole Cynoglossus semilaevis
Authors: Yu Y. , Zhong Q. W. , Zhang Q. Q. , Wang Z. G. , Li C. M. , Yan F. S. , Jiang L. M. 
Published: May 2009

SummaryMyeloid differentiation factor 88 (MyD88) is a universal and crucial adaptor protein, which plays an essential role in the intracellular signalling elicited by IL‐1R/TLR superfamily. In the present study, we report the full‐length sequence of MyD88 gene in half‐smooth tongue sole (Cynoglossus semilaevis). In the 2855 bp genomic sequence, five exons and four introns were identified. The cloned cDNA exhibited 110 bp of 5′ UTR, 576 bp of 3′ UTR and 858 bp of the entire open‐reading frame encoding a polypeptide of 285 amino acids. The protein sequence included a typical conserved cytosolic Toll/interleukin‐1 receptor (TIR) domain, an intermediate domain (ID) and a death domain (DD), and shared greater than 70% identity with Japanese flounder Paralichthys olivaceu ortholog. Real‐time polymerase chain reaction (RT‐PCR) analysis indicated a broad expression of csMyD88, especially in ovary and spleen. Quantitative RT‐PCR analysis indicated that the csMyD88 mRNA levels were significantly increased in the spleen and head kidney after inactive Vibrio anguillarum challenge and the expression of csMyD88 appeared to be developmentally regulated during C. semilaevis ontogeny. Although, species‐specific differences were present, the similarity between mammalian and piscine MyD88s suggested that the main function of MyD88 might be conserved across vertebrates.
Effects of disturbance by Siberian marmots (Marmota sibirica) on spatial heterogeneity of vegetation at multiple spatial scales
Authors: Yoshihara Yu , Ohkuro Toshiya , Bayarbaatar Buuveibaatar , Takeuchi Kazuhiko 
Published: May 2009

AbstractAn understanding of the relationship between vegetation spatial heterogeneity and disturbance and its application to the management are important for maintaining biodiversity and functions of ecosystems. We examined the effects of disturbance by Siberian marmots on the spatial heterogeneity of vegetation at three spatial scales (fine, intermediate and coarse) in a Mongolian grassland. We established a 50 m × 50 m plot around five marmot mounds and another plot in an area with no mounds. Each plot was subdivided into 625 adjacent 4‐m2 square quadrats and the plants in them were surveyed. Spatial heterogeneity was calculated as the mean dissimilarity in species composition among sample quadrats in each plot. The off‐colony plant community was dominated by graminoids, but a variety of plants, including graminoids, forbs and shrubs, were present in the on‐colony plot. The slightly greater fine‐scale heterogeneity in the on‐colony plot compared to the off‐colony plot reflected the presence of either disturbed or undisturbed patch within a single measurement unit, whereas the lower coarse‐scale heterogeneity in the on‐colony plot reflected the presence of both disturbed and undisturbed patches within a single measurement unit. Our results demonstrate the possibility of using the marmot as a conservation tool in our study site as far as a plant biodiversity is targeted.
Effects of land‐use change on chemical composition of soil organic matter in tropical lowland Bolivia
Authors: Abe Susumu S. , Mueller Carsten W. , Steffens Markus , Koelbl Angelika , Knicker Heike , Koegel‐Knabner Ingrid 
Published: May 2009

AbstractLand‐use change affects not only the amount of soil organic matter (SOM) but also its composition. We performed cross‐polarization magic angle spinning (CPMAS)13C and15N nuclear magnetic resonance (NMR) spectroscopy to investigate the chemical composition of bulk SOM in topsoils (0–15 cm) under different land use, namely native forest (NF), 27‐year cropland with wheat/soybean rotation (CL) and 27‐year rangelands with guineagrass (RG) and with bahiagrass (RB), in south‐east Bolivia. The findings of this study showed only a subtle alteration of composition of bulk SOM despite the large changes in carbon (C) content. Nevertheless, NF and RB showed a slightly lower abundance of aromatic C but a higher proportion of alkyl C compared to CL and RG where the loss of organic matter was substantial. This suggests that relatively stable components dominated by aromatic structures had relatively enriched during SOM decomposition under agricultural practices. A slight disparity of SOM composition observed between RG and RB (less O‐alkyl C but more aromatic C in RG than RB) suggests that grass species influenced SOM quality even under the same land use, namely the rangeland. On the other hand, organic N composition was less affected by land use or management practice than C forms.
Drugs Used in the Treatment of Rheumatoid Arthritis: Relationship between Current Use and Cardiovascular Risk Factors
Authors: Rho Young Hee , Oeser Annette , Chung Cecilia P. , Milne Ginger L. , Stein C. Michael 
Published: May 2009

ABSTRACTObjectives.  Drugs used for the treatment of rheumatoid arthritis (RA) have the potential to affect cardiovascular risk factors. There is concern that corticosteroids, non‐steroidal anti‐inflammatory drugs (NSAIDs) and COX‐2 inhibitors could affect cardiovascular risk adversely, while drugs such as the antimalarial, hydroxychloroquine, may have beneficial effects. However, there is limited information about cardiovascular risk factors in patients with RA receiving different drugs.Methods.  We measured cardiovascular risk factors including systolic and diastolic blood pressure, serum HDL and LDL cholesterol, glucose and homocysteine concentrations and urinary F2‐isoprostane excretion in 169 patients with RA. Risk factors were compared according to current use of corticosteroids, methotrexate, antimalarials, NSAIDs, COX‐2 inhibitors, leflunomide and TNF‐α blockers. Comparisons were adjusted for age, sex, race, disease activity (DAS28 score), current hypertension, diabetes, smoking status and statin use.Results.  No cardiovascular risk factor differed significantly among current users and non‐users of NSAIDs, COX‐2 inhibitors, methotrexate and TNF‐α blockers. Serum HDL cholesterol concentrations were significantly higher in patients currently receiving corticosteroids (42.2 ± 10.5 vs. 50.2 ± 15.3 mg/dL, adjusted P < 0.001). Diastolic blood pressure (75.9 ± 11.2 vs. 72.0 ± 9.1 mm Hg, adjusted P = 0.02), serum LDL cholesterol (115.6 ± 34.7 vs. 103.7 ± 27.8 mg/dL, adjusted P = 0.03) and triglyceride concentrations (157.7 ± 202.6 vs. 105.5 ± 50.5 mg/dL, adjusted P = 0.03) were significantly lower in patients taking antimalarial drugs. Plasma glucose was significantly lower in current lefunomide users (93.0 ± 19.2 vs. 83.6 ± 13.4 mg/dL, adjusted P = 0.006).Conclusions.  In a cross‐sectional setting drugs used to treat RA did not have major adverse effects on cardiovascular risk factors and use of antimalarials was associated with beneficial lipid profiles.
Cytometric Approach for Detection ofEncephalitozoon intestinalis, an Emergent Agent
Authors: Barbosa Joana , Rodrigues Acácio Gonçalves , Pina-Vaz Cidália 
Published: May 2009

ABSTRACTEncephalitozoon intestinalisis responsible for intestinal disease in patients with AIDS and immunocompetent patients. The infectious form is a small spore that is resistant to water treatment procedures. Its detection is very important, but detection is very cumbersome and time-consuming. Our main objective was to develop and optimize a specific flow cytometric (FC) protocol for the detection ofE. intestinalisin hospital tap water and human feces. To determine the optimal specific antibody (Microspor-FA) concentration, a known concentration ofE. intestinalisspores (Waterborne, Inc.) was suspended in hospital tap water and stool specimens with different concentrations of Microspor-FA, and the tap water and stool specimens were incubated under different conditions. The sensitivity limit and specificity were also evaluated. To study spore infectivity, double staining with propidium iodide (PI) and Microspor-FA was undertaken. Distinct approaches for filtration and centrifugation of the stool specimens were used.E. intestinalisspores stained with 10 μg/ml of Microspor-FA at 25°C overnight provided the best results. The detection limit was 5 × 104spores/ml, and good specificity was demonstrated. Simultaneous staining with Microspor-FA and PI ensured that theE. intestinalisspores were dead and therefore noninfectious. With the stool specimens, better spore recovery was observed with a saturated solution of NaCl and centrifugation at 1,500 ×gfor 15 min. A new approach for the detection ofE. intestinalisfrom tap water or human feces that ensures that the spores are not viable is now available and represents an important step for the prevention of this threat to public health.
Contributions of Immunoblotting, Real-Time PCR, and the Goldmann-Witmer Coefficient to Diagnosis of Atypical Toxoplasmic Retinochoroiditis
Authors: Talabani H. , Asseraf M. , Yera H. , Delair E. , Ancelle T. , Thulliez P. , Brézin A. P. , Dupouy-Camet J. 
Published: May 2009

ABSTRACT

Ocular toxoplasmosis is a major cause of posterior uveitis worldwide. The diagnosis is based mainly on ophthalmological examination. Biological diagnosis is necessary in atypical cases, and this requires aqueous humor sampling by anterior chamber paracentesis. We evaluated real-time PCR targeting the
Toxoplasma gondii
529-bp repeat element, the Goldmann-Witmer coefficient (GWC), and immunoblotting for the diagnosis of toxoplasmic retinochoroiditis in 54 patients with atypical uveitis. The results of these biological tests, applied to paired aqueous humor-serum samples, were compared to the clinical findings. Combining either PCR or the GWC with immunoblotting increased the sensitivity to 73% or 70%, respectively. Together, PCR and the GWC had 80% sensitivity. If feasible, sensitivity can be increased by combining the three methods (85% sensitivity). The interval between symptom onset and anterior chamber paracentesis strongly influenced the detection of specific intraocular antibody synthesis. The sensitivity of the GWC increased from 45% to 56% when sampling was performed 10 days after symptom onset, and that of immunoblotting increased from 53% to 72% when puncture was performed 30 days after symptom onset. PCR analysis of aqueous humor samples detected toxoplasmic DNA in 55% of patients. In contrast to the results of immunoblotting and the GWC, the results of PCR were not influenced by the interval between symptom onset and paracentesis. PCR was more informative than the GWC and immunoblotting for immunocompromised patients. Acute necrotizing retinal lesions were significantly larger in PCR-positive patients, with a mean of 3.5 optic disc diameters, than in PCR-negative patients, with a mean of 1.5 optic disc diameters.
Diversity of Group B Streptococcus Serotypes Causing Urinary Tract Infection in Adults
Authors: Ulett Kimberly B. , Benjamin William H. , Zhuo Fenglin , Xiao Meng , Kong Fanrong , Gilbert Gwendolyn L. , Schembri Mark A. , Ulett Glen C. 
Published: May 2009

ABSTRACT

Serotypes of group B streptococcus (GBS) that cause urinary tract infection (UTI) are poorly characterized. We conducted a prospective study of GBS UTI in adults to define the clinical and microbiological characteristics of these infections, including which serotypes cause disease. Patients who had GBS cultured from urine over a 1-year period were grouped according to symptoms, bacteriuria, and urinalysis. Demographic data were obtained by reviewing medical records. Isolates were serotyped by latex agglutination and multiplex PCR-reverse line blotting (mPCR/RLB). Antibiotic susceptibilities were determined by disc diffusion. GBS was cultured from 387/34,367 consecutive urine samples (1.1%): 62 patients had bacteriuria of >10
7
CFU/liter and at least one UTI symptom; of these patients, 31 had urinary leukocyte esterase and pyuria (others not tested), 50 (81%) had symptoms consistent with cystitis, and 12 (19%) had symptoms of pyelonephritis. Compared with controls (who had GBS isolated without symptoms), a prior history of UTI was an independent risk factor for disease. Increased age was also significantly associated with acute infection. Serotyping results were consistent between latex agglutination and mPCR/RLB for 331/387 (85.5%) isolates; 22 (5.7%) and 7 (1.8%) isolates were nontypeable with antisera and by mPCR/RLB, respectively; and 45/56 (80.4%) isolates with discrepant results were typed by mPCR/RLB as belonging to serotype V. Serotypes V, Ia, and III caused the most UTIs; serotypes II, Ib, and IV were less common. Nontypeable GBS was not associated with UTI. Erythromycin (39.5%) and clindamycin (26.4%) resistance was common. We conclude that a more diverse spectrum of GBS serotypes causes UTI than previously recognized, with the exception of nontypeable GBS.
Meningitis Due to a “
Bartonella washoensis
”-Like Human Pathogen
Authors: Probert Will , Louie Janice K. , Tucker James R. , Longoria Rose , Hogue Robin , Moler Silvia , Graves Margot , Palmer Heather J. , Cassady Joseph , Fritz Curtis L. 
Published: May 2009

ABSTRACT

We report the second human case of infection caused by an organism identified as the proposed
Bartonella
species, “
B
.
washoensis
.” The organism was isolated from a blood sample from a patient presenting with meningitis and early sepsis.
Oropsylla montana
fleas were implicated as the vector for disease transmission in this case.
Molecular Determination of
Mycobacterium leprae
Viability by Use of Real-Time PCR
Authors: Martinez Alejandra N. , Lahiri Ramanuj , Pittman Tana L. , Scollard David , Truman Richard , Moraes Milton O. , Williams Diana L. 
Published: May 2009

ABSTRACT

Mycobacterium leprae
, the etiological agent of leprosy, is noncultivable on axenic media. Therefore, the viability of
M. leprae
for clinical or experimental applications is often unknown. To provide new tools for
M. leprae
viability determination, two quantitative reverse transcriptase PCR (RT-PCR) assays were developed and characterized.
M. leprae sodA
mRNA and 16S rRNA were used as RNA targets, and
M. leprae
repetitive element (RLEP) DNA was used to determine relative bacterial numbers in the same purified bacterial preparations or from crude biological specimens. Results demonstrated that both assays were good predictors of
M. leprae
viability during short-term experiments (48 h) involving rifampin (rifampicin) treatment in axenic medium, within rifampin-treated murine macrophages (MΦ), or within immune-activated MΦ. Moreover, these results strongly correlated those of other
M. leprae
viability assays, including radiorespirometry-based and Live/Dead BacLight viability assays. The 16S rRNA/RLEP assay consistently identified the presence of
M. leprae
in eight multibacillary leprosy patient biopsy specimens prior to multidrug therapy (MDT) and demonstrated a decline in viability during the course of MDT. In contrast, the
sodA
/RLEP assay was able to detect the presence of
M. leprae
in only 25% of pretreatment biopsy specimens. In conclusion, new tools for
M. leprae
viability determination were developed. The 16S rRNA/RLEP RT-PCR
M. leprae
viability assay should be useful both for short-term experimental purposes and for predicting
M. leprae
viability in biopsy specimens to monitor treatment efficacy, whereas the
sodA
/RLEP RT-PCR
M. leprae
viability assay should be limited to short-term experimental research purposes.
Single Nucleotide Polymorphisms in the IS
900
Sequence of
Mycobacterium avium
subsp.
paratuberculosis
Are Strain Type Specific
Authors: Castellanos Elena , Aranaz Alicia , de Juan Lucia , Álvarez Julio , Rodríguez Sabrina , Romero Beatriz , Bezos Javier , Stevenson Karen , Mateos Ana , Domínguez Lucas 
Published: May 2009

ABSTRACT

Insertion sequence IS
900
is used as a target for the identification of
Mycobacterium avium
subsp.
paratuberculosis.
Previous reports have revealed single nucleotide polymorphisms within IS
900
. This study, which analyzed the IS
900
sequences of a panel of isolates representing
M. avium
subsp.
paratuberculosis
strain types I, II, and III, revealed conserved type-specific polymorphisms that could be utilized as a tool for diagnostic and epidemiological purposes.
Evaluation of CHROMagar
Acinetobacter
for Detection of Enteric Carriage of Multidrug-Resistant
Acinetobacter baumannii
in Samples from Critically Ill Patients
Authors: Gordon N. C. , Wareham D. W. 
Published: May 2009

ABSTRACT

CHROMagar
Acinetobacter
was used to screen stool and perineal swabs for enteric carriage of multidrug-resistant
Acinetobacter baumannii
in samples from critically ill patients. Results were compared with a molecular assay resulting in sensitivity and specificity of culture compared to PCR of 91.7% and 89.6%, respectively.
Rapid and Sensitive Detection of Shiga Toxin-Producing
Escherichia coli
from Nonenriched Stool Specimens by Real-Time PCR in Comparison to Enzyme Immunoassay and Culture
Authors: Grys Thomas E. , Sloan Lynne M. , Rosenblatt Jon E. , Patel Robin 
Published: May 2009

ABSTRACT

Shiga toxin (Stx)-producing
Escherichia coli
(STEC) bacteria are a frequent cause of food-borne gastroenteritis, hemorrhagic colitis, and hemolytic uremic syndrome. Because antimicrobial agents are generally contraindicated in patients infected with STEC, a sensitive and specific diagnostic test with rapid turnaround is essential. Current culture methods may fail to detect non-O157 STEC. We evaluated a Stx gene real-time PCR assay using hybridization probes and the LightCycler instrument with 204 prospectively collected stool specimens, which were also tested for Stx by enzyme immunoassay (EIA) (ProSpecT STEC; Remel, Lenexa, KS) and by culturing on chromogenic agar (Chromagar O157; BD BBL, Sparks, MD). In addition, 85 archived stool specimens previously tested for Stx (by EIA) and/or
E. coli
O157:H7 (by culture) were tested by PCR. Sample preparation for PCR included mixing the stool in sterile water and extraction of nucleic acid using the MagNA Pure LC instrument (Roche Diagnostics). The PCR assay had 100% sensitivity and specificity compared to EIA and culture for specimens collected prospectively (4 of 204 specimens were positive) and compared to culture and/or EIA for archival specimens (42 of 85 specimens were positive). Both the EIA and PCR produced positive results from a specimen containing an O103 serotype STEC in the prospective specimens, and the PCR test detected three positive specimens that contained nonviable STEC in the archived specimens. The PCR assay demonstrated 100% sensitivity and specificity compared to EIA and/or culture and more rapid turnaround than either EIA or culture.
Comparison of Chromogenic Media to BD GeneOhm Methicillin-Resistant
Staphylococcus aureus
(MRSA) PCR for Detection of MRSA in Nasal Swabs
Authors: Bischof Larry J. , Lapsley Linda , Fontecchio Karen , Jacosalem Dollie , Young Carol , Hankerd Rosemary , Newton Duane W. 
Published: May 2009

ABSTRACT

To select a method for detecting methicillin-resistant
Staphylococcus aureus
(MRSA) in nasal swabs, we compared BD GeneOhm MRSA PCR and various culture media (mannitol salt agar with cefoxitin, MRSA
Select
, CHROMagar MRSA, and Spectra MRSA). While PCR detection of MRSA was more rapid, MRSA
Select
and Spectra MRSA demonstrated performance equivalent to that of PCR with maximal detection at 24 h.