Noninvasive determination of fetal RHD status by examination of cell-free DNA in maternal plasma.

BACKGROUND: Cell-free fetal DNA in maternal plasma opens the way for routine risk-free diagnosis of fetal D status of D- mothers. The focus was on accuracy of RHD typing and confirmation of fetal DNA in maternal plasma while RHD was not detected. STUDY DESIGN AND METHODS: Plasma DNA was extracted (by manual and/or automatic method) from 255 D- pregnant women and amplified in exons 7 and 10 and intron 4 of RHD gene with real-time polymerase chain reaction. The presence of fetal DNA was confirmed by testing SRY and, when negative, by one of 11 different polymorphisms found in the father but not in the mother. The results were compared with the D status of the newborns. RESULTS: After exclusion of 25 cases (10%) because of material shortage, in 230 cases (90%) available for complete study, the predictive value of the procedure of fetal RHD testing (RHD genotyping plus confirmation of fetal DNA) was 99.6 percent. SRY detection confirmed fetal DNA presence in maternal plasma in all boys, whereas the detection of various polymorphisms in all girls but one. CONCLUSIONS: Fetal RHD genotyping from maternal plasma may be used with confidence, although additional polymorphisms for confirmation of fetal DNA should be included for 100 percent predictive value (instead of 99.6%).

A symbiotic concept of autoimmunity and tumour immunity: lessons from vitiligo.

Vitiligo is a skin disease in which melanocytes (MCs) are eradicated from lesional epidermis, resulting in disfiguring loss of pigment. MCs are destroyed by MC-reactive T cells, as well as other non-immune and immune components. Similarities exist between the autoimmunity observed in vitiligo and the tumour immunity observed in melanoma immuno-surveillance. An analysis of these mechanisms might lead to the development of new therapies for both vitiligo and melanoma.

DRB1 alleles in relation to severity of liver disease in patients with chronic hepatitis C.

The aim of our study was analysis of relation between HLA class II antigens and the liver disease severity in chronic hepatitis C (CHC) patients. The subject of analysis was data obtained from 134 CHC patients with disease confirmed by histopathologic test (F/M: 62/72; age 16-74; average age 41.4 +/- 12.7 yrs), HCV RNA-positive, HbsAg- and HIV-negative with no coexistence of any other liver diseases. Liver biopsy specimens were estimated according to Ishak's criterions (grading 0-18; staging 0-6). HLA DRB1 alleles were determined by a commercial method INNOLiPA DRB (Innogenetics, Belgium). Statistical analysis considered alleles occurring with frequency higher than 10%. The necroinflammatory activity (average grading score) was compared in groups of patients with- and without particular allele. The frequency of each allele's occurrence was analyzed according to patients sex, age and staging score of liver fibrosis. In statistical analysis t-Student test and chi-squared test with or without Yates' correction were applied. Statistically significant correlation was found between occurrence of DRB1*13 and DRB1*07 alleles and necroinflammatory activity intensification, and between occurrence of DRB1*13 allele and progression of liver disease. Mild liver damage, instead, expresses statistically significant relation with DRB1*11 allele.

Local immune response in skin of generalized vitiligo patients. Destruction of melanocytes is associated with the prominent presence of CLA+ T cells at the perilesional site.

In situ immune infiltrates in lesional, perilesional, and nonlesional skin biopsies from patients with vitiligo were analyzed by immunohistochemistry and compared with immune infiltrates found in the skin of normal healthy donors and relevant disease controls. An increased influx of activated skin-homing T cells and macrophages were seen in the perilesional biopsies. The overall percentages of cutaneous leukocyte-associated antigen-positive (CLA+) T cells were similar to those found in normal healthy donors. This is compatible with the similar expression of E-selectin. Most strikingly, however, the CLA+ T cells in perilesional skin were mainly clustered in the vicinity of disappearing melanocytes, and 60% to 66% of these interacting T cells expressed perforin and granzyme-B. The perforin+/granzyme-B+ cells were not seen in locations different from that of disappearing melanocytes. Interestingly, the majority of the infiltrating T cells were HLA-DR/CD8+. Another hallmark of the present study is the focal expression of intercellular adhesion molecule (ICAM)-1 and HLA-DR in the epidermis at the site of interaction between the immune infiltrates and the disappearing melanocytes. The data presented in this study are consistent with a major role for skin-homing T cells in the death of melanocytes seen in vitiligo.

Follow-up studies of a representative sample of children with severe mental handicap in Poland.

A representative sample of 260 Polish children, aged 5-14 years, identified between 1964 and 1965 in an epidemiological study as severely mentally handicapped (IQs ranging from 0 to 51) were followed up twice: (I) after 10 years (in 1975-1976); and (II) after 23 years (in 1987-1988). At both follow-ups, all surviving subjects were contacted, and psychological and sociological data were gathered; the findings presented here pertain mostly to follow-up II. The great majority of subjects (85%) lived with their families, and the remainder resided in an institution. Only 10% of subjects (living with one exception in families) currently had a higher level of intellectual functioning than that of severe mental handicap. Among this group were individuals of relatively better health, without speech disorders, who had attended schools, had some vocational training, had been or were employed, and had families of their own, i.e. were self-dependent with a life-style similar to that of other people of their age and social background. Of the remainder, about 40% retained the same relative level of intellectual and social functioning, and about 50% deteriorated, particularly those in institutions. This latter group has remained in the role of permanent children, depending on others for care and maintenance.