Monitoring the Transcriptional Activity of Human Endogenous Retroviral HERV-W Family Using PNA Strand Invasion into Double-Stranded DNA.

In the presented assay, we elaborated a method for distinguishing sequences that are genetically closely related to each other. This is particularly important in a situation where a fine balance of the allele abundance is a point of research interest. We developed a peptide nucleic acid (PNA) strand invasion technique for the differentiation between multiple sclerosis-associated retrovirus (MSRV) and ERVWE1 sequences, both molecularly similar, belonging to the human endogenous retrovirus HERV-W family. We have found that this method may support the PCR technique in screening for minor alleles which, in certain conditions, may be undetected by the standard PCR technique. We performed the analysis of different ERVWE1 and MSRV template mixtures ranging from 0 to 100% of ERVWE1 in the studied samples, finding the linear correlation between template composition and signal intensity of final reaction products. Using the PNA strand invasion assay, we were able to estimate the relative ERVWE1 expression level in human specimens such as U-87 MG, normal human astrocytes cell lines and placental tissue. The results remained in concordance with those obtained by semi-quantitative or quantitative PCR.

[Effects of the cigarette smoking on the newborn clinical parametrs and the accumulation of cadmium and lead in the placenta of women from Upper Silesia]. / Wplyw palenia papierosów na parametry noworodka oraz na kumulacje kadmu i olowiu w lozysku kobiet z Górnego Slaska.

AIM: The aim of the study was to determine the content of cadmium and lead in different parts of the placenta and fetal membranes of women who were exposed to cigarette smoke. The correlation between the two chemical elements and the impact of the Cd and Pb accumulation on newborn parameters were established. MATERIALS AND METHODS: The study material was collected immediately after delivery from 40 patients of the Department of Obstetrics and Gynecology Katowice. The marginal and central parts of the placenta and fetal membranes (amnion) were taken. The women were divided into two groups: smokers and non-smokers. Metal concentration in placenta was determined by flame atomic absorption spectrometry (FAAS). Bioethical Commission approved of the study RESULTS: In both studied groups, smokers and non-smokers, the presence of cadmium and lead was detected. Smokers turned out to have accumulated more of the investigated heavy metals in the placenta and fetal membranes. In the analyzed groups of women of smokers and non-smokers, differences in the content of the studied metals were found, but they were not statistically significant. Differences in newborn parameters in the two groups of women occurred, but again they lacked statistical significance. The level of lead increases along with the increase in the amount of cadmium, which proves the existence of a statistically significant correlation between them (p = 0.000). CONCLUSIONS: The number of smoked cigarettes increases cadmium content in maternal placenta and fetal membranes. No significant differences in newborn parameters of either smoker or non-smokers were observed, which may indicate women's adaptation to the environment containing cigarette smoke. The placenta and fetal membranes are biomarkers of the level of toxic exposure for the developing baby.