Identification of six Listeria species by real-time PCR assay.

UNLABELLED: The Listeria genus comprises 10 recognized species. Listeria monocytogenes causes listeriosis in humans and other animals primarily via contaminated food or animal feed. Listeria ivanovii causes listeriosis in animals and on rare occasions in humans. The identification of nonpathogenic species of Listeria in foods indicates that conditions exist that support the growth of pathogenic strains and is used to facilitate the implementation of control and prevention measures. This study shows the development and evaluation of a 5'exonuclease real-time PCR assay for the rapid identification of Listeria seeligeri, Listeria welshimeri, L. monocytogenes, L. ivanovii, Listeria grayi and Listeria innocua. The assay consists of two triplexes that were evaluated using 53 cultures of Gram-positive bacteria, including 49 Listeria spp. from human, animal, food or food-processing environments. The assay was rapid, specific and reproducible and could identify each of the six species from a mixture of strains. The developed assay proved to be a powerful means of rapidly identifying Listeria species and could be usefully implemented in busy specialist reference laboratories. SIGNIFICANCE AND IMPACT OF THE STUDY: The identification of species of Listeria from foods is important to monitor pathogenic strains and facilitates the implementation of control measures. This study shows the development and evaluation of a 5'exonuclease real-time PCR assay for the rapid identification of L. seeligeri, L. welshimeri, L. monocytogenes and L. ivanovii, L. grayi, L. innocua. The developed assay proved to be specific, rapid and reproducible and therefore could be implemented in busy specialist reference laboratories.

Influence of clay incorporation on the physical properties of polyethylene/Brazilian clay nanocomposites.

High density polyethylene/Brazilian clay nanocomposites were prepared by the melt intercalation technique. A montmorillonite sample from Boa Vista/PB, Northeast of Brazil, was organically modified with esthearildimethylammonium chloride (Praepagen WB) quaternary ammonium salt. The unmodified and modified clays with the quaternary ammonium salt were introduced in 1, 2, 3 and 5 wt% in a PE polymer matrix. The dispersion analysis and the interlayer distance of the clay particles were obtained by X-ray diffraction (XRD) and transmission electron microscopy (TEM). The mechanical properties of tensile and the flammability of the nanocomposites were studied. In general, the mechanical properties of the systems presented superior values compared to the matrix. The systems showed a reduction on the burning rate, indicating that the flammability resistance of nanocomposites was improved.

Urinary hypoxanthine and pseudouridine as indicators of tumor development in mesothelioma-transplanted nude mice.

A human mesothelioma was heterotransplanted to nude mice, and the urinary excretions of hypoxanthine, xanthine, pseudouridine, orotic acid, 7-methylguanine, and 1-methylhypoxanthine have been followed before and after the tumor transplantation. The compounds were measured by means of isotachophoresis, which has been found a rapid and precise method. The tumor reached maximum size within 30 days, and at this time a significantly increased excretion of pseudouridine and hypoxanthine was observed. Tumor growth was stopped by chemotherapy (vincristine, cyclophosphamide, prednisolone, and Adriamycin), and corresponding to this, a decrease occurred in both pseudouridine and hypoxanthine excretion to normal values.

Decrease in Le(x) expression in esophageal adenocarcinomas arising in Barrett's epithelium.

Fifty esophageal adenocarcinomas were investigated for their expression of Le(a), Le(x), and Le(a)-Le(x). Among the 50 adenocarcinomas, 17 cases developed in Barrett's epithelium. Those 17 differed from the other 33 cases by expressing much less Le(x). Fifty-nine percent of Barrett's adenocarcinomas were Le(x) negative compared with 24% of the non-Barrett's carcinomas. All Barrett's adenocarcinomas showed less than 50% Le(x) whereas 50% of non-Barrett's carcinomas showed between 50 and 100% expression. The statistical correlation coefficient for this association was P < 0.001. Normal gastric cardia epithelium showed the same Le(x) expression in both groups. In the Barrett group, Le(x) expression decreased from normal through intestinal metaplasia and dysplasia to adenocarcinoma. This progression was not seen in the non-Barrett group. Loss of Le(x) expression may prove useful in following patients with Barrett's epithelium in evaluating progression toward a malignant process. No difference in expression of Le(a) and Le(a)-Le(x) was found between Barrett's and non-Barrett's carcinomas.

Omeprazole in the long-term treatment of severe gastro-oesophageal reflux disease in patients with systemic sclerosis.

Twenty-five patients with systemic sclerosis and severe gastro-oesophageal reflux disease were treated with 20-80 mg omeprazole daily for up to 5 years. Efficacy of treatment was assessed by symptom score, by endoscopic and histopathological surveillance of the oesophageal and gastric mucosa, and by laboratory screening including serum gastrin concentration. Statistically significant relief of symptoms and healing of oesophagitis confirmed the efficacy of this treatment. However, complete healing of oesophagitis was not achieved in half of the patients due to residual gastro-oesophageal acid reflux. Repeated adjustments of the maintenance dose of omeprazole may be needed for this group of patients. From the safety point-of-view, nothing was observed to discourage the long-term use of omeprazole in this group of patients.

Major or local surgery for cure in early rectal and sigmoid carcinoma--a prospective evaluation.

The results of different types of treatment for early cancer in 158 out of 1,300 patients with rectal and sigmoid cancer were evaluated in a prospective study from 1979 to 1984. Radical surgery was performed in 121 patients (mean age 66 years) with cancer Dukes' A (median diameter 3 cm), while 37 patients (mean age 71) with smaller cancers (median 2 cm) had polypectomy or local excision. Post-operative complications were significantly more frequent in the first group. No patients in any of the two groups with carcinoma in pedunculated adenomas or within the upper half of the submucosa or above 9 cm from the anal verge had recurrent cancer. CEA-measurements had no prognostic value. No difference was found in crude or cancer-related death between the two groups. The overall results support the use of local treatment in elderly patients with complicating disease, having small cancers, not penetrating the tunica muscularis externa of the rectosigmoid wall.

Monitoring of omeprazole treatment in gastro-oesophageal reflux disease.

OBJECTIVE: To test our standard dosing regimen in omeprazole treatment of gastro-oesophageal reflux disease (GORD) and to determine whether 'non-responders' could be pinpointed. DESIGN: A reverse dose-response examination using increasing doses of omeprazole. The study was conducted as an open consecutive clinical study. Response was measured by 24-h pH-metry, symptoms, endoscopy and histopathology. SETTING: All patients had been referred to one of the partaking departments for evaluation of oesophageal reflux symptoms. PATIENTS: A total of 62 patients were included, 29 with systemic sclerosis and 33 consecutively included patients suffering from idiopathic oesophageal reflux. RESULTS: Approximately one-third of the patients required doses higher than 40 mg of omeprazole/day (up to 140 mg/day) to abolish GOR. No cases of tachyphylaxia or bile-induced oesophagitis were seen in this study. In all patients subjected to dose titration we were able to achieve healing of oesophagitis assessed by symptom scoring, endoscopy and histopathology. No prediction of final dose of omeprazole could be made. CONCLUSION: Four weeks after reaching a dose level of omeprazole that ensured the abolition of GOR, healing of oesophagitis according to endoscopic/histological evaluation was obtained in all patients. Persistent oesophagitis, i.e. bile induced, was not found.

Pain in arthritis and musculoskeletal disorders: the role of coping skills training and exercise interventions.

There is growing recognition of the limitations of conventional, biomedical approaches to the management of pain in individuals having arthritis and musculoskeletal disorders. This article provides an overview of newly developed biopsychosocial approaches to the management of pain in this population. The presentation is divided into three sections. In the first section, a biopsychosocial model of pain is presented. This model highlights the role that biological factors (eg., disease severity, comorbid conditions), cognitive-behavioral factors (eg., thoughts, emotions, and behaviors), and environmental factors (eg., spouse or family responses to pain behavior) can play in influencing the pain experience. In the second section, we provide an overview of two newly developed treatment protocols based on the biopsychosocial model of pain: a pain coping skills training protocol and an exercise training protocol. Practical aspects of implementing these protocols are illustrated by highlighting how they are applied in the management of patients having persistent osteoarthritic pain. In the final section of the article, we pinpoint several important future directions for research in this area. Future studies need to explore the utility of combining pain coping skills and exercise training protocols. In addition, there is a need to identify variables that predict patients' response to biopsychosocial treatments.

[Proctoscopic versus histologic diagnosis of rectal polyps]. / Rektoskopisk versus histologisk diagnose af rektalpolypper.

On the basis of the endoscopic appearance, 71 rectal polyps were assessed as adenomata or non-neoplastic polyps, after which the endoscopic diagnosis was compared with the results of histological examination. The diagnosis based on macroscopic examination of the polyps proved correct in only 62% of the cases. It is concluded that the diagnosis of adenoma cannot be established solely on the proctoscopic appearance of a polyp and, as adenomata are premalignant, removal of all polyps found at proctoscopy is recommended.

[Surgical treatment of cancer of the cardia and the esophagus]. / Kirurgisk behandling af cancer cardiae og cancer esophagi.

During the period 1977 to 1989, 379 patients with cancer cardiac and cancer esophagi were admitted. The ages ranged from 18-88 years with an average of 65 years. The treatment concept was basically unchanged during the study period. Resection as described by Ivor Lewis was employed as palliative or curative treatment when resection was considered possible. Intubation of the esophagus was employed when no other treatment was considered possible. Resection was employed in 251 patients, eight of these emergencies on account of perforation or haemorrhage. Bypass operations were employed in five patients, intubations in 63 and no surgical treatment was undertaken in 60 patients. The all over operative mortality for resections was 11.2% and for elective operations 9.9%. The operative mortality increased with the TNM stage of the tumour and was 3.2% for stages I + II, 8.2% for stage III and 24.4% for stage IV. The complication ration was 42%. 15% of the patients submitted to resection required reoperation. Anastomotic leaks were encountered in 18 patients and nine of these died. Pulmonary complications were the most frequent and resulted in ten early postoperative deaths. The mortality connected with intubations was 15%. The over all five-year survival rate after resections was 10.4%. In stages I + II this was 42% and 8% in stage III. None of the patients in stage IV survived for two years. The longest survival after intubation was less than one year.

[Liver transplantation in Denmark. First-year experiences]. / Levertransplantation i Danmark. Det første års erfaringer.

During the first 12 months of the Danish Liver Transplantation program, which began in October 1990, 21 transplantations were performed in 11 women, six men and three children. One patient required a retransplant. Fourteen operations were performed electively and six patients were transplanted for acute and subacute fulminant liver failure and coma, two patients had reduced size livers because of large donor liver. There were no peroperative deaths. One of the elective patients died after three weeks from multiorgan failure and sepsis. Two of the emergency patients died after 20 and 22 days. One from graft dysfunction due to stenosis of the celiac trunk and the other of exudative pericarditis. One patient died from chronic rejection and CMV-infection after seven months. Complications were relatively few and acute rejection occurred in 40% of the patients. Fifteen patients are discharged with normal liver function and 11 of these were back at work, school or previous functions in the home. It is concluded that these results are comparable to the best results from other centres but that 21 transplants in 12 months must be a minimum activity.

Gastrin and enteroglucagon cells in human antra, with special reference to intestinal metaplasia.

In a consecutive material consisting of 24 stomachs resected due to adenocarcinoma, intestinal metaplasia occurred in 21. Gastrin-producing cells (G-cells) were found to be distributed in a sporadic manner in antra with intestinal metaplasia. Not a single G-cell could be demonstrated in areas with metaplasia, while in the nonmetaplastic areas the distribution of the G-cells corresponded to that of the middle part of the mucosa. This means, that an error can occur when determining the quantity of G-cells, and can explain the previous controversial results regarding the density of G-cells. Enteroglucagon containing cells (GLI-cells) on the contrary were demonstrated in areas with intestinal metaplasia in antra of 19 of the stomachs showing intestinal metaplasia but never in the nonmetaplastic mucosa. This indicated that metaplasia also includes the endocrine cells. The identification of the G-cells and the GLI-cells was carried out by means of indirect immunoperoxidase technique combined with alcian blue pH 2,6-PAS staining.

Evaluation of the visual prostate symptom score in a male population with great language diversity and limited education: a study from Namibia.

BACKGROUND: A visual prostate symptom score (VPSS) using pictograms was developed to assess the force of the urinary stream, urinary frequency, nocturia and quality of life (QoL). OBJECTIVE: To compare the VPSS with the international prostate symptom score (IPSS) and maximum (Qmax) and average (Qave) urinary flow rates in men from diverse language groups with limited schooling. METHODS: Men with lower urinary tract symptoms admitted to the urology ward at Windhoek Central Hospital, Namibia, were evaluated. Patients who were unable to complete the questionnaires alone were assisted by a doctor or nurse. Local ethics committee approval was obtained. Statistical analysis was performed using Student's t-test and Spearman's rank correlation test. RESULTS: One hundred men (mean age 56.3 years, range 20.1 - 95.4) were evaluated over a period of one year. All the men understood one or more of 15 languages, and 30 were illiterate; 32 had <5 years of schooling, 34 had 5 - 9 years and 34 had >9 years. The VPSS took significantly less time to complete than the IPSS. There were statistically significant correlations between the total VPSS and IPSS scores, between the four VPSS questions and the corresponding IPSS questions, and between Qmax and Qave and the VPSS total and VPSS questions on the force of the urinary stream and QoL. CONCLUSION: The VPSS pictograms depicting the force of the urinary stream and QoL correlated significantly with Qmax and Qave, indicating that they can be used as single-item questions to rapidly assess bladder outflow obstruction in men with limited education.

Engineered high-affinity affibody molecules targeting platelet-derived growth factor receptor ß in vivo.

Platelet-derived growth factor receptor (PDGFR) ß is a marker of stromal pericytes and fibroblasts and represents an interesting target for both diagnosis and therapy of solid tumors. A receptor-specific imaging agent would be a useful tool for further understanding the prognostic role of this receptor in vivo. Affibody molecules constitute a class of very small binding proteins that are highly suited for in vivo imaging applications and that can be selected to specifically recognize a desired target protein. Here we describe the isolation of PDGFRß-specific Affibody molecules with subnanomolar affinity. First-generation Affibody molecules were generated from a large naive library using phage display selection. Subsequently, sequences from binders having a desired selectivity profile and competing with the natural ligand for binding were used in the design of an affinity maturation library, which was created using a single partially randomized oligonucleotide. From this second-generation library, Affibody molecules with a 10-fold improvement in affinity (K(d)=0.4-0.5 nM) for human PDGFRß and a 4-fold improvement in affinity (K(d)=6-7 nM) for murine PDGFRß were isolated and characterized. Complete reversible folding after heating to 90 °C, as demonstrated by circular dichroism analysis, supports tolerance to labeling conditions for molecular imaging. The binders were highly specific, as verified by dot blot showing staining reactivity only with human and murine PDGFRß, but not with human PDGFRα, or a panel of control proteins including 16 abundant human serum proteins. The final binder recognized the native conformation of PDGFRß expressed in murine NIH-3T3 fibroblasts and human AU565 cells, and inhibited ligand-induced receptor phosphorylation in PDGFRß-transfected porcine aortic endothelial cells. The PDGFRß-specific Affibody molecule also accumulated around tumoral blood vessels in a model of spontaneous insulinoma, confirming a potential for in vivo targeting.