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The increasing prevalence of celiac disease (CD), especially in adults, its atypical clinical presentation, and the strict, lifelong adherence to gluten-free diet (GFD) as the only option for healthy state create an imperative need for noninvasive methods that can effectively diagnose CD and monitor GFD. Aim. Evaluation of anti-endomysium (EmA) and anti-tissue transglutaminase IgA (tTG-A) antibodies in CD diagnosis, GFD monitoring, and first degree relatives screening in CD adult patients. Methods. 70 newly diagnosed Greek adult patients, 70 controls, and 47 first degree relatives were tested for the presence of EmA and tTG-A. The CD patients were monitored during a 3-year period. Results. EmA predictive ability for CD diagnosis was slightly better compared to tTG-A (P=0.043). EmA could assess compliance with GFD already from the beginning of the diet, while both EmA and tTG-A had an equal ability to discriminate between strictly and partially compliant patients after the first semester and so on. Screening of first degree relatives resulted in the identification of 2 undiagnosed CD cases. Conclusions. Both EmA and tTG-A are suitable markers in the CD diagnosis, in the screening of CD among first degree relatives, having also an equal performance in the long term monitoring.

Reports of immunoglobulin E (IgE)-mediated allergic reactions to grapes and wine are limited in the literature. Nevertheless, grapes are widely grown and consumed in Mediterranean countries. The object of this prospective study was to present clinical features, in vivo and in vitro allergy testing, and human leukocyte antigen (HLA) serotyping in patients with recurring reactions to grapes and grape products. Eleven unrelated Greek patients, six men and five women (aged 16 - 44 years; mean, 26.9 years) were enrolled based on a documented history of IgE-mediated reactions to grapes, wine, or other grape products. Their evaluation included full history, reaction severity, clinical examination, skinprick tests with food allergens and molds, serum IgE, specific IgEs to the same allergen battery, and HLA typing. Patients reported 35 grape-induced anaphylaxis episodes ranging from moderate (more than one system involved but not prominent respiratory or cardiovascular symptoms; 45.5%) to severe (serious respiratory obstruction and/or hypotension and loss of consciousness; 54.5%). A causative agent was identified: wine, 10/35 (28.6%); red grapes, 9/35 (25.7%); stuffed vine leaves, 8/35 (22.9%); raisins, 3/35 (8.6%); white grapes, 2/35 (5.7%); wine vinegar, 2/35 (5.7%); and grape juice, 1/35 (2.9%). Other foods that induced anaphylaxis were apples (54.5%), cherries (18.6%), peaches (18.6%), and bananas (9.3%). Specific IgE values were in accordance with skin-prick tests reactivity. Concerning HLA typing, 9/11 possessed HLA-DR11(5) and -DQ7(3) and the remanning two possessed HLA-DR17(3) and -DQ2 antigens. Grapes, wine and other grape products might cause serious allergic reactions in sensitized individuals. The cosensitization and reaction incidence to other fruit allergens could be a basis for further investigation of panallergens of fruits. HLA class II antigens may contribute in genetic predisposition to these allergic reactions.

The need for confirmation or exclusion of biological father and / or biological mother is a social phenomenon, which is imposed by socio-economic and, sometimes, by moral-psychological factors. Modern science has significantly contributed to solving this problem, as many medical methods have been applied for this purpose. Biological markers that have been conducted for distinguishing between individuals were the human ABO blood groups, the Rh, MNS, Duffy, Kidd, and Kell systems, as well as the human leukocyte antigens (HLA) system. For a long time theHLA testing represented the standard testing in forensic genetics, but, due to the linkage disequilibrium and the predominance of certain HLA alleles and as the demand for parentage investigations is rapidly increasing during the recent years, this serological era has been replaced by molecular markers through the introduction of \"DNA profiling\", which is based on polymorphisms of short tandem repeats (STRs) loci. Nowadays, \"DNA profiling\" by analysis of STR loci is the method of choice for human identification and parentage investigations. This technique is the most informative, accurate, robust, rapid, cost-effective method of genotyping and has worldwide acceptance in the courts, as the probability of parentage will typically be greater than 99.99999%.

The aim of this study was to investigate the hypothesis that the expression of certain HLA antigens may constitute a risk marker for cardiovascular hypertrophy in subjects with arterial hypertension. We examined 158 subjects with newly diagnosed arterial hypertension. HLA class I (-A, -B, -Cw) and class II (-DR, -DQ) antigens were studied by two-step microlymphocytotoxic technique in peripheral T and B lymphocytes. Carotid intima-media thickness (IMT) was determined noninvasively by ultrasonography. The left ventricular mass was calculated according to the formula of Devereux and was normalized by the individual’s height (LVM/h). The individuals with DR13 and DR17 were characterized by higher values of IMT compared to those without these HLA (0.096 ± 0.018 cm v 0.085 ± 0.021 cm, P = .011, 0.100 ± 0.019 cm v 0.084 ± 0.021 cm, P = .012, respectively). The presence of HLA DQ7 was characterized by markedly higher values of IMT that just failed to reach statistical significance (0.091 ± 0.019 cm v 0.084 ± 0.022 cm, P = .045). Furthermore, subjects with HLA DQ7 and DR11 exhibited higher values of LVM/h in comparison to those without these HLA (191.3 ± 36.2 g/m v 166.9 ± 41.0 g/m, P = .029 and 194.6 ± 34.3 g/m v 166.6 ± 40.9 g/m, P = .034, respectively). Hypertensive subjects with HLA B51 tended to have lower LVM/h (166.6 ± 39.0 g/m with v 176.0 ± 41.7 g/m without HLA B51, P = .045). In conclusion, it can be postulated that certain HLA phenotypes exhibit an association with increased carotid IMT and left ventricular mass in hypertensive subjects. The determination of these antigens may help to identify subjects at high risk for cardiovascular events.

OBJECTIVE Nowadays, the application of DNA-typing in laboratory medicine is increasing rapidly for paternity/maternity disputes. The goal of this study was to evaluate the use of polymorphic microsatellite marker DNA analysis and to establish this analysis as the method of choice for parentage investigations. SUBJECTS AND METHODS Among 708 civil parentage tests addressed to our Laboratory previously examined for HLA class I (-A*,-B*,-Cw*), and class II (-DRB1*,-DQB1*,-DPB1*) alleles using PCR-SSOP and/or PCR-SSP methodologies, a cohort of 50 cases (137 individuals) of disputed parentage was selected. In these cases DNA-typing was generated from co-amplification of 15 autosomal STR DNA markers (D3S1358, HUMTH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, HUMCSF1PO, Penta D, HUMvWA, D8S1179, HUMTPOX, HUMFGA and the sex determining Amelogenin marker HUMAMEL), using fragment analysis methodology. RESULTS The evaluation of the results showed that 15 out of 50 cases, were sufficient for exclusion of fatherhood by both approaches (HLA and STRs). In all remaining 35 non-excluded cases, the PI value using HLA genotyping ranged from 76 to 6,452,794, whereas using aSTR genotyping ranged from 15,173 to 9.2 x 1010. In one non-excluded motherless case the alleged father showed one genetic discrepancy with the child at D21S11 locus, due to a mutation event. CONCLUSION The use of DNA-typing with 15 aSTR loci for parentage testing provides an accurate and high-sensitivity method which is simpler to perform and more rapid than an accepted standard technology, such as HLA genotyping. The analysis of aSTR loci offers a highly discriminating test suitable for trio paternity testing, increasing the W rate in comparison to HLA genotyping. Nevertheless, when a mutation event occurs in motherless cases, combination of HLA and STR polymorphisms offers high level of information, and also diminishes the possibility of false exclusion due to aSTRs mutations.

The angiotensin-converting enzyme (ACE) insertion/deletion polymorphism is an independent risk factor for cardiovascular disease. It has also been suggested that some HLA genes may contribute to the genetic susceptibility to essential hypertension. So far, an association between ACE polymorphism and HLA antigens in arterial hypertension has not been reported. We have studied 94 subjects with newly diagnosed essential hypertension, 49 men and 45 women (mean age, 52.3 ± 11.3 years), as well as 104 randomly selected, age- and gender-matched normotensive individuals (54 men and 50 women, mean age 48.7 ± 10.8 years). Both cohorts originated from the Greek population and lived in the greater Athens area. The ACE genotype was analyzed by polymerase chain reaction. HLA class I and II antigens were studied by serologic and molecular techniques. The prevalence of the ACE genotypes did not differ significantly between hypertensives and normal individuals. The casual blood pressure levels and the average ambulatory blood pressure levels were similar among the three ACE genotypes. Hypertensives with the ACE-DD genotype were characterized by an increased prevalence of the HLA-A2 antigen (50% v 31.4%, P < .005) and DR6 (16.7% v 11.4%, P < .01) in comparison to the normotensive subjects with the ACE-DD genotype. HLA-A24 was found more frequently among the hypertensives with the ACE-ID genotype than in the normal controls with the same genotype (35.5% v 26.4%, P < .05). ACE-DD genotype is associated with a high prevalence of specific HLA antigens. The coexistence of the ACE-DD genotype with certain HLA phenotypes could reveal a distinct hypertensive population with increased risk for cardiovascular events.

Alonso-Perez, E., Suarez-Gestal, M., Calaza, M., Witte, T., Papasteriades, C., Marchini, M., Migliaresi, S., Kovacs, A., Ordi-Ros, J., Bijl, M., Santos, M.J., Ruzickova, S., Pullmann, R., Carreira, P., Skopouli, F.N., D'Alfonso, S., Sebastiani, G.D., Suarez, A., Blanco, F.J., Gomez-Reino, J.J., Gonzalez, A.

The objective of this study was to determine the HLA class II associations of the anticardiolipin (aCL) and anti-b2GPI (ab2GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). A cohort of 577 European SLE patients was enrolled. aCL and ab2GPI were measured by ELISA methods. Molecular typing of HLA-DRB1, DRB3, DRB4, DRB5, DQA1 and DQB1 loci was performed by the polymerase chain reaction–sequence specific oligonucleotide probes (PCR–SSOP) method. aCL of IgG, IgM and IgA isotypes were detected in 22.8%, 14% and 13.9% of patients, respectively. IgG and IgM ab2GPI were detected in 20% of patients. aCL showed positive association with HLA DRB1*04, DRB1*0402, DRB1*0403, DRB1*07, DRB3*0301, DQA1*0201, DQA1*0301, DQB1*0302, and negative association with DQA1*0501, DRB3*0202. ab2GPI showed positive association with DRB1*0402, DRB1*0403, DQB1*0302. DRB1*0402 carried the highest relative risk for the presence of both aCL (RR = 8.1) and ab2GPI (RR = 4.6). Our results confirm the already described associations of aCL with HLA DR4 and DR7, but also demonstrate that, among the alleles at the DRB1*04 locus, the *0402 was most represented both in aCL and in ab2GPI positive patients. In addition, HLA class II associations of ab2GPI are for the first time extensively examined in a large cohort of European SLE patients.