Associations Between Neuropsychophysiological and Dermatoglyphic Indicators in the Assessment of Human Health

The purpose of this study was to explore the relationship between psycho-physiological markers of human health and dermatoglyphic indicators in young people. Materials and Methods: The study included 920 healthy volunteers aged between 18 and 21 years. All volunteers underwent the following examinations: EEG, an assessment of the anxiety level according to the BAI, and dermatoglyphic scanning. Results: According to the data obtained, there was a statistically significant strong negative correlation between the stress load indicator and dermatoglyphic data, such as the summary delta index (DI) and summary ridge count. A strong positive correlation was found between the percentage of whorls and stress (r=0.88). The predominant increase in anxiety is characteristic of persons with total ridge count (TRC) on the thumb of the right hand in the range from 19 to 23. Conclusion: Results demonstrate the interrelationships (association) between psycho-physiological (anxiety level, stress load indicator) and dermatoglyphic markers (DI, TRC and whorl pattern type) in young healthy people

Alterations of the WNT7A Gene in Clear Cell Renal Cell Carcinomas

WNT7A (wingless-type MMTV integration site family, member 7A) is a known tumor suppressor gene of non-small cell lung carcinomas (NSCLC) and is frequently inactivated due to CpG-island hypermethylation in human cancers. The members of WNT family are involved in cell signaling and play crucial roles in cancer development. In the present work hypermethylation of the WNT7A gene was detected in 66% (29/44) of analyzed clear cell renal cell carcinomas (RCCs) using methyl-specific PCR (MSP). Moreover, bisulfite sequencing confirmed intensive hypermethylation of the 5-CpG island of the WNT7A gene. Methylation analysis revealed positive correlations between tumor stage, Fuhrman nuclear grade and WNT7A hypermethylation. Additionally, restoration of WNT7A gene expression in the A498 cell line by 5-aza-2-deoxycytidine treatment confirmed a direct contribution of hypermethylation in silencing of the WNT7A gene. High frequency of loss of heterozygosity (LOH) was demonstrated on chromosome 3p25 in regions surrounding the WNT7A gene. The frequent down-regulation of WNT7A gene expression was detected in 88% (15/17) of clear cell RCCs. We have also shown that the WNT7A gene possesses tumor suppression function by colony-formation and cell proliferation assays in RCC cell lines. In summary, the WNT7A gene is inactivated by genetic/epigenetic alterations in clear cell RCC and demonstrates tumor suppressor properties.Funding Agencies|State Fond of Fundamental Research|F46/457-2011|Swedish Cancer Society||Swedish Institute||Swedish Research Council||EACR||</p

Identification of the xenograft and its ascendant sphere-forming cell line as belonging to EBV-induced lymphoma, and characterization of the status of sphere-forming cells

Abstract Background We have characterized the human cell line arised from the Epstein–Barr virus (EBV) positive multiple myeloma aspirate subjected to the long-term cultivation. This cell line has acquired the ability to form free-floating spheres and to produce a xenograft upon transplantation into NOD/SCID mice. Methods Cells from both in vitro culture and developed xenografts were investigated with a number of analytical approaches, including pathomorphological analysis, FISH analysis, and analysis of the surface antigens and of the VDJ locus rearrangement. Results The obtained results, as well as the confirmed presence of EBV, testify that both biological systems are derived from B-cells, which, in turn, is a progeny of the EBV-transformed B-cellular clone that supplanted the primordial multiple myeloma cells. Next we assessed whether cells that (i) were constantly present in vitro in the investigated cell line, (ii) were among the sphere-forming cells, and (iii) were capable of internalizing a fluorescent TAMRA-labeled DNA probe (TAMRA+ cells) belonged to one of the three types of undifferentiated bone marrow cells of a multiple myeloma patient: CD34+ hematopoietic stem cells, CD90+ mesenchymal stem cells, and clonotypic multiple myeloma cell. Conclusion TAMRA+ cells were shown to constitute the fourth independent subpopulation of undifferentiated bone marrow cells of the multiple myeloma patient. We have demonstrated the formation of ectopic contacts between TAMRA+ cells and cells of other types in culture, in particular with CD90+ mesenchymal stem cells, followed by the transfer of some TAMRA+ cell material into the contacted cell

The LOH assays: status of the informative cases of clear cell RCC for the 3q25 region surrounding the <i>WNT7A</i> gene.

<p>D3S2403, D3S2385, D3S1252– microsatellite markers, white circles - homozygotes (non informative cases), grey circles – absence of the LOH, black circles – presence of the LOH, IC - informative cases, “+” - LOH positive sample, “−” - LOH negative sample.</p

Panel of <i>WNT7A</i> gene expression in clear cell RCC samples.

<p>R – values of expression in the form of the logarithmic ratio of tumor/normal tissue of the <i>WNT7A</i> gene relatively to the <i>TBP</i> gene.</p

Suppressive effect of <i>WNT7A</i> gene re-expression in RCC cell lines.

<p>Effect of <i>WNT7A</i> gene re-expression (A) on colony formation (B) for the A498, KRC/Y cell lines, and (C) cell proliferation assays for the A498 cell line; M – marker, 1 and 2– A498 cells were transfected by empty-pcDNA3.1 and WNT7A-pcDNA3.1 vectors, 3 and 4– KRC/Y cells were transfected by empty-pcDNA3.1 and WNT7A-pcDNA3.1 vectors, NC – negative control (H₂0). All experiments were performed in triplicate. Representative results are shown.</p

Clinical-pathological characteristics of clear cell RCC samples.

<p>Clinical-pathological characteristics of clear cell RCC samples.</p

Study of <i>WNT7A</i> gene methylation status in clear cell RCC.

<p>(A). Representative MSP analysis of the <i>WNT7A</i> gene by using methylated (M) and unmethylated (U) specific primers, PC – positive control, M.Sssi treated or untreated normal DNA, NC – negative control (H₂0), T4, T6, T10, T35, T37: tumor samples, N4, N6, N10, N35, N37: normal samples. (B). Sequencing of MSP products; white squares - 0–19% methylation at the CpG dinucleotide, grey squares - 20–59% methylation at the CpG dinucleotide, dark grey squares - 60–79% methylation at the CpG dinucleotide, black squares - 80–100% methylation at the CpG dinucleotide, T2, T4, T5, T6, T9, T10, T11: tumor samples. (C). Methylation status of the fifty two CpG dinucleotides of the <i>WNT7A</i> 5′-CpG island in tumor samples with a methylated <i>WNT7A</i> gene, where each CpG dinucleotide is shown by either a black square when methylated or a grey square when unmethylated; arrows indicate position of MSP primers, T4, T5, T6 are tumor samples. (D). Restoration of <i>WNT7A</i> expression by 5-aza-2′-deoxycytidine treatment of the A498 cell line, MSP-M - methylation analysis of the <i>WNT7A</i> gene by using methylated specific primers, NC – negative control (H₂0).</p