Alternaria alternata NADP-dependent mannitol dehydrogenase is an important fungal allergen.

BACKGROUND: Alternaria alternata is one of the most important allergenic fungi worldwide. Mannitol dehydrogenase (MtDH) has previously been shown to be a major allergen of Cladosporium herbarum and cross-reactivity has been demonstrated for several fungal allergens. OBJECTIVE: The present study's objective was to clone the MtDH from an A. alternata cDNA library, express and purify the recombinant non-fusion protein and test its IgE-binding properties. Methods A cDNA library prepared from A. alternata hyphae and spores was screened for mannitol dehydrogenase by DNA hybridization with the radioactively labelled C. herbarum homologue as a probe. The resulting clone was sequenced and heterologously expressed in Escherichia coli as a recombinant non-fusion protein, which was purified to homogeneity and analysed for its IgE-binding capacity. RESULTS: The coding sequence of the full-length cDNA clone comprises 798 bp encoding a protein with a molecular mass of 28.6 kDa and a predicted pI of 5.88. Protein sequence analysis revealed an identity of 75% and a homology of 86% between the MtDHs of A. alternata and C. herbarum. The functional mannitol dehydrogenase was expressed in the E. coli strain BL21(DE3) transformed with the vector pMW172 and purified to homogeneity. The enzyme catalyses the NADPH-dependent conversion of d-fructose to d-mannitol. In IgE-ELISA and immunoblots, MtDH is recognized by 41% of A. alternata-allergic patients. In vivo immunoreactivity of the recombinant MtDH was verified by skin prick testing. Finally, inhibition-ELISA experiments confirmed cross-reactivity between the MtDHs of A. alternata and C. herbarum. CONCLUSION: Mannitol dehydrogenase (Alt a 8) represents an important new allergen of the ascomycete A. alternata that might be suitable for improving diagnostic and therapeutic procedures.     

Preconception Care for Improving Perinatal Outcomes: The Time to Act

Maternal and Child Health Journal -     

Endobronchial ultrasound: new insight for the diagnosis of sarcoidosis.

A diagnosis of sarcoidosis should be substantiated by pathological means in order to thoroughly exclude other diseases. The role of real-time endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the diagnosis of sarcoidosis has not been reported. The purpose of the present study is to evaluate the diagnostic yield of EBUS-TBNA in demonstrating the pathological features of sarcoidosis. In total, 65 patients with suspected sarcoidosis, with enlarged hilar or mediastinal lymph nodes on computed tomography, were included in the study. Patients with a suspected or known malignancy or previously established diagnosis of sarcoidosis were excluded. Convex probe endobronchial ultrasonography integrated with a separate working channel was used for EBUS-TBNA. Surgical methods were performed in those in whom no granulomas were detected by EBUS-TBNA. Patients were followed up clinically. EBUS-TBNA was performed on a total of 77 lymph node stations in 65 patients. A final diagnosis of sarcoidosis was made for 61 (93.8%) of the patients. The remaining four patients were diagnosed as having Wegener's granulomatosis (n=1) or indefinite (n=3). In patients with a final diagnosis of sarcoidosis, EBUS-TBNA demonstrated noncaseating epithelioid cell granulomas in 56 (91.8%) of the patients. No complications were reported. Endobronchial ultrasound-guided transbronchial needle aspiration proved to be a safe procedure with a high yield for the diagnoses of sarcoidosis.     

Embolisierende Thrombose bei Fehlbildung der V. cava inferior

Persistent left-sided inferior vena cava (VCI) is a rare venous anomaly, its prevalence being estimated at 0.2-0.5%. Thrombotic occlusion of a VCI has been reported in only a few of these cases. We report the case of a 24-year old woman who suffered an acute thrombosis in a left-sided VCI and recurrent pulmonary embolism. After thrombectomy the course was uneventful. The diagnostic approach and the treatment strategy are discussed with reference to the literature.     

Comparative assays of the rpoB gene for identification of Mycobacterium tuberculosis isolated from patients in Sudan.

OBJECTIVES: To characterise mycobacterial clinical isolates based on amplification of the rpoB gene. SETTING: One hundred and thirty-five mycobacterial isolates cultured from suspected pulmonary tuberculosis (TB) patients were identified phenotypically. Molecular characterisation of the isolates was performed based on amplification of the rpoB gene, using duplex polymerase chain reaction (DPCR), PCR-restriction fragment length polymorphism (RFLP) and nested PCR-based sequence analysis techniques. RESULTS: The DPCR assay identified 129 of 135 (95.5%) clinical isolates as Mycobacterium tuberculosis complex species. Restriction enzyme analysis of the rpoB PCR product using Hind II identified 134 of the 135 (99.3%) isolates as M. tuberculosis complex, while nested PCR sequence analysis of the rpoB gene identified 133/133 examined isolates (100%) as M. tuberculosis species. No mycobacteria other than M. tuberculosis (MOTT) were detected among the studied isolates. CONCLUSION: DPCR, PCR/RFLP Hind II and nested PCR sequence analysis of the rpoB gene techniques showed comparable efficiency in the characterisation of Mycobacterium isolates. Nested PCR sequence analysis of the rpoB gene was superior to PCR/RFLP for characterisation of suspected M. tuberculosis isolates, while the DPCR technique showed less sensitivity. As PCR-RFLP requires less sophisticated laboratory facilities than nested PCR sequence analysis, it would be more appropriate to be adopted for accurate characterisation of mycobacteria in countries with a weak infrastructure.     

Impact of mass media on knowledge about tuberculosis control among homemakers in Delhi.

SETTING: Homes in Delhi, India. OBJECTIVE: To study the reach of mass media campaigns and their impact on awareness about tuberculosis (TB) control among homemakers/housewives. DESIGN: A community-based cross-sectional survey among homemakers residing in Delhi for more than 6 months. RESULTS: Of a total of 920 women interviewed, about 74.2% had seen specific TB-related health messages in one or more of the mass media. The maximum number of subjects could recall having seen billboards or television campaigns. The percentage of respondents who had correct information about various aspects of the disease was higher among those who had seen TB campaigns on any of the mass media. The effectiveness of radio and newsprint in communicating TB messages was found to be more limited than that of television and billboards. CONCLUSION: The mass media can be effective in getting messages about TB across to the community of women who are homemakers, especially in developing countries. In view of our findings, it may be recommended that television and billboards be used as tools for reaching out to them with specific campaigns regarding TB control, and that the use of these media should be strengthened further.     

Mycobacterium caprae in cattle and humans in Croatia.

SETTING: This paper describes an outbreak of human and related bovine tuberculosis (TB) caused by Mycobacterium caprae in Croatia. A 13-year-old boy clinically presented enlargement of cervical lymph node with consecutive isolation of M. caprae. His 7-year-old sister, who had no clinical signs of disease, hyper-reacted to the purified protein derivative (PPD) test (>25 mm) and peribronchial infiltration was found by radiology. The children came from a family that ran a small-sized cattle dairy farm. DESIGN: All cattle on the farm were subjected to cutaneous TB testing: six of the 14 reacted positive, while three were suspicious. The entire herd was slaughtered, their carcasses examined and collected material subjected to pertinent diagnostic procedures. RESULTS: Gross examination findings consistent with TB were observed in the PPD-positive cows. Mycobacteria isolated from the boy and cattle were identified by classical and molecular methods, confirming M. caprae as the causative agent. CONCLUSION: Although not bacteriologically proven, consumption of raw milk or non-pasteurised milk products from infected dairy cattle was suspected as the source of infection in humans. Our findings confirm the domination of M. caprae among cattle in Croatia and represent the first evidence of M. caprae infection in humans in Croatia.     

Factors related to health system delays in the diagnosis of pulmonary tuberculosis in Estonia.

OBJECTIVE: To estimate health system delays (HSD) in the diagnosis of pulmonary tuberculosis (PTB) and its risk factors after major social changes in Estonia, and to assess the ability of a completely reformed health care system to diagnose patients with PTB. METHODS: All newly detected symptomatic culture-positive patients with PTB aged > or = 16 years from Southern Estonia during 2002-2003 (n = 185) were interviewed. HSD was defined as the interval from a patient's first contact with a medical provider to the date of TB diagnosis. RESULTS: The factors significantly associated with HSD greater than the median (19 days) and the 75th percentile (40 days) were smear negativity, absence of cough among symptoms, absence of chest X-ray during the first visit and age > 60 years. A significantly shorter HSD was determined in non-Estonians and unemployed patients. HSD was not associated with the specialty of the doctor first contacted by the patient. CONCLUSION: This study in Southern Estonia shows that the health care system is still managing the diagnosis of PTB without significant delays, even after substantial modifications in the health care system resulting from social reform in a post-socialist country, and that family physicians can manage PTB patients successfully.