Identification of fetal DNA and cells in skin lesions from women with systemic sclerosis

BACKGROUND: Systemic sclerosis is a disease of unknown origin which often occurs in women after their childbearing years. It has many clinical and histopathological similarities to chronic graft-versus-host disease. Recent studies indicate that fetal stem cells can survive in the maternal circulation for many years post partum. This finding suggests that fetal cells persisting in the maternal circulation or tissues could be involved in the pathogenesis of systemic sclerosis by initiating a graft-versus-host reaction. METHODS: We used the polymerase chain reaction (PCR) to identify Y-chromosome sequences in DNA extracted from peripheral-blood cells and skin lesions from women with systemic sclerosis of recent onset. To confirm the PCR findings, we used fluorescence in situ hybridization of peripheral-blood cells and cells within chronic inflammatory-cell infiltrates in biopsy specimens of affected skin. RESULTS: Y-chromosome sequences were found in DNA from peripheral-blood cells in 32 of 69 women with systemic sclerosis (46 percent), as compared with 1 of 25 normal women (4 percent, P\u3c0.001), and in T lymphocytes from 3 women with systemic sclerosis who had male offspring. Furthermore, Y-chromosome sequences were identified in skin-biopsy specimens from 11 of 19 women with systemic sclerosis (58 percent); 9 of the 11 were known to have carried male fetuses. Nucleated cells containing Y chromosomes were detected by fluorescence in situ hybridization in paraffin-embedded sections of skin lesions from all seven women we tested whose skin-biopsy specimens contained Y-chromosome sequences. CONCLUSIONS: Fetal antimaternal graft-versus-host reactions may be involved in the pathogenesis of systemic sclerosis in some women

Improving the sensitivity of the heat-transfer method (HTM) for cancer cell detection with optimized sensor chips

© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. In this article, we increased the sensitivity of the heat-transfer method (HTM) for the detection of breast cancer cells (ZR-75-1 cells, see figure) in phosphate buffered saline (PBS). The effect of small technological changes on the limit of detection (LoD) of the methodology was examined. To this extent, polished aluminum substrates with a mirror finish were used, replacing the unpolished chips used in previous studies. These chips were coated with a polyurethane layer and imprinted for the target cell type, creating a so-called surface imprinted-polymer (SIP). Binding of target cells to the SIP resulted in an increase of the thermal resistance at the solid-liquid interface under study. Background thermal resistance measurements were performed with polished and unpolished aluminum substrates. In addition, the effect of using silver paste as thermal coupling between the aluminum chip and the copper heat provider was analyzed. The results of these experiments reveal that optimal thermal contact is achieved when directly coupling the copper heat provider to the polished side of the aluminum substrate as evidenced by a decrease in the baseline thermal resistance. In addition, noise levels on the heat-transfer resistance (Rth) signal decreased by a factor in the optimal configuration. Dose-response curves were obtained using the optimized methodology and were compared with results obtained with the original substrates. These quantitative experiments demonstrated an improvement of the LoD by approximately thirty percent. ZR-75-1 cells applied onto a home-made rubber stamp.status: publishe