Detection of virulence genes of Clostridium difficile by multiplex PCR

Antikainen J, Pasanen T, Mero S, Tarkka E, Kirveskari J, Kotila S, Mentula S, Könönen E, Virolainen‐Julkunen A‐R, Vaara M, Tissari P. Detection of virulence genes of Clostridium difficile by multiplex PCR. APMIS 2009; 117: 607–13. Clostridium difficile strains belonging to the PCR ribotype 027, pulse‐field gel electrophoresis (PFGE) type NAP1, toxinotype III and restriction endonuclease analysis group BI harbouring mutations in the tcdC gene and possessing binary toxin components A and B have been described to cause epidemics with increased morbidity and mortality. In the present study we developed a conventional multiplex PCR designed to detect selected virulence associated markers of the hypervirulent C. difficile PCR ribotype 027. The multiplex PCR assay detected the major toxins A and B, binary toxin components A and B as well as a possible deletion in the tcdC gene: a characteristic pattern of amplification products for the PCR ribotype 027 strains was detected. This rather simple method was specific for the screening of this hypervirulent C. difficile strain. The correlation between the multiplex PCR and PCR ribotyping methods was excellent. The sensitivity and specificity were 100% in our epidemiological situation. In conclusion, this multiplex PCR was found useful in the preliminary screening for the hypervirulent C. difficile PCR ribotype 027.     

OCCUPATIONAL ALLERGIC CONTACT DERMATITIS CAUSED BY 2,4,6-TRIS-(DIMETHYLAMINOMETHYL)PHENOL, AND REVIEW OF SENSITIZING EPOXY RESIN HARDENERS

Background. Epoxy resin compounds (ERG) include a large number of sensitizing chemicals such as epoxy resins (ER), hardeners (curing agents), and reactive diluents. Allergic contact dermatitis (ACD) caused by ERCS is often occupational. Materials and Methods. We report a patient, sensitized to a hardener of a two-component epoxy paint. Three conventional patch test sessions were performed to diagnose the causative chemical. We also review the literature on sensitizing epoxy-resin hardeners. Results. A 47-year-old nonatopic woman developed dermatitis from a two-component epoxy paint. Patch testing with epoxy resin was negative, but 2,4,6–tris(dimethylaminomethyl)phenol (tris-DMP), used in the paint hardener, induced an allergic patch test reaction. We also review briefly other epoxy hardeners that have caused allergic dermatitis, including: (1) aliphatic polyamines, e.g., ethylenediamine, diethylenetriamine, triethylenetetramine, 3-dimethylaminopropylamine, and trimethylhexamethylenediamine; (2) cycloaliphatic polyamines, e.g., isophoronediamine and 3,3′-dimethyl-4,4-diaminodicyclohexylmethane; (3) aromatic amines, such as 4,4′-diaminodiphenylmethane, m-phenylene diamine, and 1,3-xylylene diamine; (4) dicyanodiamide; (5) triglycidyl isocyanurate, an epoxy compound that may be used as an epoxy-resin hardener; and (6) additives in epoxy accelerators, such as hexavalent chromate. Conclusions. No one chemical can be used to screen for sensitization to the many different epoxy hardeners. Extensive patch testing may be required to reveal the hardener that has caused the allergy. The hardener, 2,4,6-tris-dimethylaminomethyDphenol (tris-DMP), is a new sensitizer. To verify ACD caused by tris-DMP, patch-testing at 1% in petrolatum is suggested.     

Variable expression of tenascin‐C,osteopontin and fibronectin in inflammatory myofibroblastic tumour of the lung

Kaarteenaho R, Sormunen R, Pääkkö P. Variable expression of tenascin‐C, osteopontin and fibronectin in inflammatory myofibroblastic tumour of the lung. APMIS 2010; 118: 91–100. The aim of this study was to analyse the expression of tenascin‐C, osteopontin and fibronectin in inflammatory myofibroblastic tumour of the lung, which is a rare tumour of unknown aetiology. Nine patients with an inflammatory myofibroblastic tumour of lung were studied by immunohistochemistry for the presence of tenascin‐C, osteopontin, fibronectin and alpha‐smooth muscle actin, which is a common marker for myofibroblasts. The ultrastructure of myofibroblasts was confirmed by transmission electron microscopy. The expression of tenascin‐C, osteopontin, fibronectin and alpha‐smooth muscle actin was also studied by immunoelectron microscopy. All cases displayed all of the studied extracellular matrix proteins and also alpha‐smooth muscle actin‐positive spindle‐shaped fibroblastic cells that were undoubtedly myofibroblasts. The immunoelectron microscopic studies demonstrated labelling for alpha‐smooth muscle actin in intracellular filament bundles within myofibroblasts, for fibronectin in the extracellular filaments of the fibronexus and for tenascin‐C extracellularly often adjacent to myofibroblasts. Labels for osteopontin were observed within myofibroblasts and plasma cells. These results demonstrate that tenascin‐C, osteopontin and fibronectin were expressed in all three kinds of subtypes of inflammatory myofibroblastic tumours of the lung and further, variable amounts of myofibroblasts could be observed by light and transmission electron microscopy as well as by immunoelectron microscopic techniques.     

HUMAN GRANULOSA CELLS CONTAIN INSULIN-LIKE GROWTH FACTOR-BINDING PROTEIN (IGF BP-1) mRNA

Previous studies have demonstrated expression of insulin-like growth factor-binding protein (IGF BP-1) in secretory and decidualized endometrium, in adult and fetal liver, and in HepG2 liver cancer cells. We have studied the expression of IGF BP-1 in various types of ovarian neoplasias, normal ovary, and granulosa cells from hyperstimulated human ovarian follicles by RNA blot hybridization. A single 1.6 kb mRNA species, similar to that present in human decidua, was identified in poly(A)RNA-containing preparations of granulosa cells and of a borderline malignant ovarian cystadenoma. This finding verifies the postulated production of IGF BP-1 by the human ovary.     

Pain and activity disturbance after paediatric day case adenoidectomy

Over the past two decades outpatient surgery has become standard practice in paediatric surgery. Adenoidectomy is a common surgical procedure in children. In this prospective survey pain and pain-related outcomes such as sleep and activity disturbance were evaluated in 167 children aged 1–7 years who had undergone adenoidectomy as a day case in Kuopio University Hospital. The survey questionnaire consisted of 76 structured questions about pain, pain medication, adverse effects and daily activities during first week after the operation. Eighty-three per cent of children had pain at home and 17% of them had moderate or severe pain on a four point verbal rating scale. Eighty per cent of children used pain medication at home. Pain medication did not cause any major adverse effects. Over 90% of children were back to normal daily activities during the first three postoperative days and nearly all were able to drink during the whole postoperative period. We conclude that pain is a common problem after adenoidectomy in children but most of the children return to normal activities within three days.     

Peripheral Arterial Disease—Natural Outcome

ABSTRACT Three hundred and twelve patients with peripheral arterial disease were followed up for 8 3/4 years or more (maximum 11 3/4 years) to assess the natural history of the disease and factors determining its outcome. Of the 312 patients, 188 (69%) died during the follow-up, 68% of the deaths having cardiovascular causes. The 10-year relative cumulative survival rate was 0.61 for males and 0.48 for females. The role of smoking as a risk factor could not be analysed without bias. In addition to known risk factors diabetes mellitus, cerebrovascular disease and coronary heart disease, the degree of peripheral arterial disease itself also proved to be a risk factor among men. The expected life lost for men with intermittent claudication was 20%, but 44.3% for men with advanced lower limb ischaemia (p<0.01). This difference could not be explained by the well-known association of advanced ischaemia and diabetes mellitus. The present results therefore suggest that the state of advanced ischaemia indicates larger involvement of the whole of the arterial tree and predicts fatal cardiovascular events among these patients.     

Occupational allergic rhinitis from guar gum

Three cases of allergic rhinitis from a vegetable gum, guar gum, have been detected. Two subjects were exposed to fine guar gum powder (Emco Gum 563, Meyhall Chemical AG, Switzerland), an insulator in rubber cables, when opening cables in a power cable laboratory. After 1–2 years' exposure the patients developed rhinitis. Scratch-chamber tests, nasal provocation tests, nasal eosinophilia and a RAST test proved their allergy. A third subject developed allergic rhinitis from another guar gum product (Meyproid 5306, Meyhall Chemical AG) after 2 years' exposure in a paper factory. A positive skin test and nasal provocation test confirmed the diagnosis. A fourth case of possible allergy to guar gum after exposure to Meyproid 5306 in a paper factory is also presented. No final diagnosis was reached in this case (in 1974). The present subjects, only one of whom was atopic, developed allergy within 2 years, although their exposure to guar gum was not especially heavy. Therefore, when handling guar, adequate ventilation facilities should be provided and protective clothing, including a respiratory mask, should be worn.