Differential Detection of Five Mouse-Infecting Helicobacter Species by Multiplex PCR

Several species of helicobacter have been isolated from laboratory mice, including H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius, which appear to be the most common. The most widely used published method for molecular detection of these agents is PCR amplification of a conserved region of 16S rRNA, but differential speciation requires restriction enzyme digestion of the amplicons. This study was undertaken to determine PCR conditions that would simultaneously and specifically identify each of the five common species without restriction enzyme analyses. First, we designed novel and specific PCR primers for H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius, using sequences from the heterologous regions of 16S rRNA. Because of comigration of amplified products, we next identified P17, an H. bilis-specific protein; P25, an H. hepaticus-specific protein; and P30, an H. muridarum-specific protein by screening genomic DNA expression libraries of each species. Primers were designed from these three genes, plus newly designed, species-specific 16S rRNA primers for H. rodentium and H. typhlonius that could be utilized for a five-plex PCR. The sizes of the amplicons from H. bilis, H. hepaticus, H. muridarum, H. rodentium, and H. typhlonius were 435, 705, 807, 191, and 122 bp, respectively, allowing simultaneous detection and effective discrimination among species.     

The survival and recognition of Mycoplasma meleagridis grown at 37 degrees C and then maintained at room temperature

This study was made because of the difficulty of resuscitating Mycoplasma meleagridis from agar cultures received from the field. The type strain of Mm was found viable following culture at 37 degrees C for as long as 10 days and maintenance at room temperature for a further 14 days but only when few colonies were present in the original culture. It is suggested that for the satisfactory subculture of this organism the original culture on mycoplasma agar should not exceed 1 week, or no more than a few days should the colonies appear after this time, and that the period at room temperature should also not exceed 1 week. Non-viable colonies, if small, may however be recognised by immunofluorescence.     

Recombinant Helicobacter bilis Protein P167 for Mouse Serodiagnosis in a Multiplex Microbead Assay

Infection of mice with Helicobacter bilis is widespread in research and commercial mouse colonies. Therefore, sensitive, specific, and high-throughput assays are needed for rapid and accurate testing of mice in large numbers. This report describes a novel multiplex assay, based on fluorescent microbeads, for serodetection of H. bilis infection. The assay requires only a few microliters of serum to perform and is amenable to a high-throughput format. Individual microbead sets were conjugated to purified, H. bilis-specific, recombinant proteins P167C and P167D and bacterial membrane extracts from H. bilis and Helicobacter hepaticus. For detecting H. bilis infection in the microbead multiplex assay, P167C and P167D provided significantly higher sensitivities (94 and 100%, respectively) and specificities (100 and 95%, respectively) than membrane extract (78% sensitivity and 65% specificity). Microbead multiplex assay results were validated by enzyme-linked immunosorbent assay. Purified recombinant proteins showed low batch-to-batch variation; this feature allows for ease of quality control, assay robustness, and affordability. Thus, recombinant antigens are highly suitable in the multiplex microbead assay format for serodetection of H. bilis infection.     

Cross-cultural adaptation,validity and reliability of Turkish version of Oxford Ankle Foot Questionnaire for children with congenital talipes equinovarus

BackgroundThe purpose of the study was to evaluate the reliability and validity of the Turkish version of the Oxford Ankle Foot Questionnaire (OxAFQ) to provide cultural adaptation.MethodsThis study involved translation, back translation, and cross-cultural adaptation. Forty-nine patients with congenital talipes equinovarus were evaluated using the Turkish version of OxAFQ. Turkish version of the Childhood Health Assessment Questionnaire (CHAQ) was used as a gold standard to validate the Turkish version of the OxAFQ. The validation was assessed with Spearman correlation analysis by using CHAQ. The reliability of the questionnaire was assessed with Cronbach alpha (internal consistency) and exploratory factor analysis.ResultsHigh validity was found between OxAFQ and CHAQ (r = -0.422?0.292) (p < 0.01). Reliability analysis showed that OxAFQ had a high level of Cronbach alpha (α = 0.88?0.96) and internal consistency (ICC = 0.90?0.96).ConclusionThe Turkish version of OxAFQ is a valid, reliable and useful quality of life questionnaire in patients with congenital talipes equinovarus and it is proper for use by health professionals and researchers.     

CYP2D6 polymorphism in systemic lupus erythematosus patients

Objectives: To determine whether patients with idiopathic systemic lupus erythematosus (SLE) are associated with impaired CYP2D6 activity and to gain insight into whether there is an association between particular CYP2D6 genotypes and susceptibility to SLE, and whether CYP2D6 polymorphism is linked to any specific clinical features of SLE. Methods: Debrisoquine sulfate (10 mg p.o.) was given to 159 healthy volunteers and 39 idiopathic SLE patients. Genotypic assay was carried out in 80 healthy volunteers and 32 patients. A 10-ml blood sample was drawn for genotypic assay. Debrisoquine and 4-hydroxydebrisoquine were determined in 8-h urine samples. Blood samples were analysed for the presence of mutations in the CYP2D6 gene, by using polymerase chain reaction (PCR) specific for CYP2D6*3 and CYP2D6*4 alleles. Results: The metabolic ratio of debrisoquine to 4-hydroxydebrisoquine ranged from 0.01 to 86.98 in healthy subjects and from 0.02 to 96 in SLE patients. We observed the poor metabolizer(PM) debrisoquine phenotype in three of 39 patients with idiopathic SLE (7.6%) and five of 159 healthy subjects (3.1%). There was no significant difference in the frequency of PM phenotypes between idiopathic SLE and healthy subjects (Fisher's exact test, P = 0.19). No significant difference in the distribution of overall genotypes and allele frequencies were observed between the two groups. No significant relationships were found between specific clinical features and the overall genotype. Conclusion: The results of this study confirm that CYP2D6 activity is not impaired in SLE and that there is no association between SLE and phenotypic CYP2D6 status. The results also showed that there was no difference in the frequency of CYP2D6A and CYP2D6B alleles between controls and patients with SLE. Received: 14 May 1998 / Accepted in revised form: 19 October 1998     

Lidocaine 10% in the endotracheal tube cuff: blood concentrations, haemodynamic and clinical effects

The purpose of this study was to evaluate the effects (common haemodynamic variables, peak cuff pressures, the incidence of reaction ('bucking') during extubation and the incidence of sore throat after operation) of lidocaine 10% instilled into the endotracheal tube cuff in intubated patients. Plasma concentrations of lidocaine were assayed. Seventy ASA class I-II patients scheduled for plastic surgery were studied. Patients were randomly divided in two groups: the cuff of the endotracheal tube was inflated with either lidocaine 10% (group L) or with saline (group S) immediately after endotracheal intubation. In group L patients, the haemodynamic changes were less (P < 0.05), and the peak cuff pressure was lower (P < 0.01) than for group S. At extubation, more patients reacted ('bucked') in group S (70.5% vs. 19.4%, P < 0.01). The incidence and severity of sore throat were significantly lower in group L 1 and 24-h after extubation. Plasma lidocaine concentrations did not reach toxic values. Lidocaine 10%, compared with saline, in the endotracheal tube cuff was associated with less disturbance of haemodynamic responses and less incidence of bucking during tracheal extubation. Lidocaine was also effective in reducing of incidence and severity of sore throat after operation.     

Surgical treatment of pulmonary hydatid cysts in children

OBJECTIVE: Hydatid disease is a parasitosis and endemic in many sheepraising areas; it is still an important health problem in Turkey. We report our experience with childhood hydatid cyst and discuss the concepts of treatment. METHODS: The clinical courses of 128 children with thoracic and liver hydatid cyst operated on from 1994 to 2000 were reviewed. The group consisted of 71 boys and 57 girls aged from 8 months to 16 years. Intact cysts were found in 144 patients and ruptured cysts in 68. RESULTS: In the postoperative course we have encountered 20 perioperative complications in 16 patients. The most common complication was residual pleural space and delayed air leakage, which occurred in 9 patients. There was no early death. CONCLUSION: Surgery is the treatment of choice for most patients with pulmonary hydatid disease. The aim of surgery is evacuation of the cyst, removal of the endocyst, and management of the residual cavity. Conservative surgical methods that preserve lung parenchyma should be preferred.     

Collar-type osteophyte of the femur in young adults: is it a harbinger of intra-articular osteoid osteoma?

Variable clinical and radiological findings for intra-articular osteoid osteoma (OO) of the hip joint make its diagnosis difficult. Because radiographs commonly do not identify the nidus, MR imaging becomes the second line of study. However, because the appearance varies, findings on MR images can be confusing. We found “collar type osteophyte” of the femur i.e. an osteophyte rim around the femoral neck, to be a conspicuous finding of intra-articular OO. Here, this feature will be emphasized and intra-articular OOs will be discussed, with a review of the literature.