Development of a miniaturized DNA microarray for identification of 66 virulence genes of Legionella pneumophila.

INTRODUCTION: For the last five years, Legionella sp. infections and legionnaire's disease in Poland have been receiving a lot of attention, because of the new regulations concerning microbiological quality of drinking water. This was the inspiration to search for and develop a new assay to identify many virulence genes of Legionella pneumophila to better understand their distribution in environmental and clinical strains. The method might be an invaluable help in infection risk assessment and in epidemiological investigations. MATERIAL/METHODS: The microarray is based on Array Tube technology. It contains 3 positive and 1 negative control. Target genes encode structural elements of T4SS, effector proteins and factors not related to T4SS. Probes were designed using OligoWiz software and data analyzed using IconoClust software. To isolate environmental and clinical strains, BAL samples and samples of hot water from different and independent hot water distribution systems of public utility buildings were collected. RESULTS: We have developed a miniaturized DNA microarray for identification of 66 virulence genes of L. pneumophila. The assay is specific to L. pneumophila sg 1 with sensitivity sufficient to perform the assay using DNA isolated from a single L. pneumophila colony. Seven environmental strains were analyzed. Two exhibited a hybridization pattern distinct from the reference strain. DISCUSSION: The method is time- and cost-effective. Initial studies have shown that genes encoding effector proteins may vary among environmental strains. Further studies might help to identify set of genes increasing the risk of clinical disease and to determine the pathogenic potential of environmental strains.

How echinoccocosis affects potential cancer markers in plasma: galectin-3, sN-cadherin and sE-cadherin? A preliminary report.

BACKGROUND: An increasing number of publications are suggesting that galectin-3 (Gal-3) and soluble cadherin fragments, such as E-cadherin (sE-CAD) and N-cadherin (sN-CAD), may be considered as cancer markers. Despite the promising results of the studies, there are no data concerning their levels in the plasma of echinococcosis patients. In most cases, echinoccocosis affects the liver, and its symptoms and disease course are very similar to those of liver cancer. The aim of the present study was to observe whether echinococcosis affects the concentration of soluble sN-CAD, sE-CAD fragments and Gal-3 in plasma and to determine which of them could be considered reliable liver cancer markers for further research. METHODS: The concentrations of sN-CAD, sE-CAD and Gal-3 in the EDTA plasma of patients suffering from echinococcosis (N = 20), liver cancers (N = 10) and healthy subjects (N = 20) were measured using the ELISA method. RESULTS: The plasma concentration of sE-CAD was lower (p = 0.0381), and that of Gal-3 higher (p = 0.0288), in echinococcosis than in the healthy group. However, only the concentration of sE-CAD differed significantly among the three analysed groups. In echinococcosis there was a correlation between the sE-CAD and CRP levels (rs = 0.79; p = 0.0066) as well as a correlation between the sE-CAD level and the number of leukocytes (rs = 0.65; p = 0.0210) in the blood. CONCLUSIONS: Echinococcosis affects the concentration of soluble sE-CAD fragments and Gal-3 in plasma. sE-CAD can be considered as a marker for differentiation between liver cancer and echinoccocossis, a parasitic liver disease similar in symptoms. Further study is required to confirm these preliminary results. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/2115657402650448.

Tularaemia after tick exposure - typical presentation of rare disease misdiagnosed as atypical presentation of common diseases: a case report.

A 44-year-old female was admitted because of tender, enlarged inguinal lymph nodes with a history of tick bite five weeks earlier. In the place of a tick bite on the skin a small ulcer was present. The primary symptoms before admission suggested typical diseases related to tick bite such as Lyme borreliosis and tick-borne encephalitis, what corresponded with positive IgM ELISA test for Lyme borreliosis. The course of disease however clarified the diagnosis of tularaemia, which is a relatively rare disease in Poland (6 cases per 40 million population are reported annually). The ultimate diagnosis was confirmed by serological tests.

Association of ocular toxoplasmosis with type I Toxoplasma gondii strains: direct genotyping from peripheral blood samples.

Toxoplasma gondii strains were genotyped directly from blood samples of patients with ocular toxoplasmosis. Analysis of nontranscribed spacer 2 revealed that all detected strains belonged to type I, suggesting an association of ocular toxoplasmosis with this type. The method shows the usefulness of blood samples for genotyping in ocular toxoplasmosis.

Informed consent and the use of placebo in Poland: ethical and legal aspects.

The concept of informed consent was one of the most fruitful ideas that deeply changed the relationships between physicians and their patients from paternalism to respect for the personal autonomy of subjects needing professional medical care. The great progress in medicine, also involving the pharmaceutical industry, has created an increasing need to perform different clinical and experimental trials. The evolution of clinical research in the last decades has influenced strongly the design of these studies. One of the most important changes in this field has been the use of placebo groups in double-blind controlled studies. The controversies have involved not only the use of placebo when standard or proven treatment was available, but also some specific problems concerning the procedure of obtaining informed consent in such trials. This paper briefly presents the evolution of informed consent in Poland as well as different ethical and legal problems concerning informed consent and the use of placebo controls in clinical trials.