Characteristics of high- and low-risk individuals in the PRIORITY study: urinary proteomics and mineralocorticoid receptor antagonism for prevention of diabetic nephropathy in Type 2 diabetes.

AIM: To compare clinical baseline data in individuals with Type 2 diabetes and normoalbuminuria, who are at high or low risk of diabetic kidney disease based on the urinary proteomics classifier CKD273. METHODS: We conducted a prospective, randomized, double-blind, placebo-controlled international multicentre clinical trial and observational study in participants with Type 2 diabetes and normoalbuminuria, stratified into high- or low-risk groups based on CKD273 score. Clinical baseline data for the whole cohort and stratified by risk groups are reported. The associations between CKD273 and traditional risk factors for diabetic kidney disease were evaluated using univariate and logistic regression analysis. RESULTS: A total of 1777 participants from 15 centres were included, with 12.3% of these having a high-risk proteomic pattern. Participants in the high-risk group (n=218), were more likely to be men, were older, had longer diabetes duration, a lower estimated GFR and a higher urinary albumin:creatinine ratio than those in the low-risk group (n=1559, P<0.02). Numerical differences were small and univariate regression analyses showed weak associations (R2 < 0.04) of CKD273 with each baseline variable. In a logistic regression model including clinical variables known to be associated with diabetic kidney disease, estimated GFR, gender, log urinary albumin:creatinine ratio and use of renin-angiotensin system-blocking agents remained significant determinants of the CKD273 high-risk group: area under the curve 0.72 (95% CI 0.68-0.75; P<0.01). CONCLUSIONS: In this population of individuals with Type 2 diabetes and normoalbuminuria, traditional diabetic kidney disease risk factors differed slightly between participants at high risk and those at low risk of diabetic kidney disease, based on CKD273. These data suggest that CKD273 may provide additional prognostic information over and above the variables routinely available in the clinic. Testing the added value will be subject to our ongoing study. (European Union Clinical Trials Register: EudraCT 2012-000452-34 and Clinicaltrials.gov: NCT02040441).

Low-level 45,X/46,XX mosaicism is not associated with congenital heart disease and thoracic aorta dilatation:prospective magnetic resonance imaging and ultrasound study.

OBJECTIVE: To establish the prevalence of risk factors for aortic dissection, such as bicuspid aortic valve, aortic coarctation and ascending aorta dilatation, in women with low-level 45,X/46,XX mosaicism undergoing an in-vitro fertilization (IVF) procedure. METHODS: The study group comprised 25 women with low-level 45,X/46,XX mosaicism (ranging from 3.3% to 10.0%) who were referred to two reproductive medicine units between 2009 and 2013 because of infertility and who underwent subsequent karyotyping. In accordance with the recommendation of the Practice Committee of the American Society for Reproductive Medicine for patients with Turner syndrome (TS), prior to the IVF procedure, all women underwent careful cardiovascular screening for congenital heart disease and thoracic aorta dilatation, including standard cardiac examination, echocardiography and non-contrast cardiac magnetic resonance imaging. Aortic size index (ASI, diameter of the ascending aorta normalized to body surface area) and the prevalence of coarctation of the aorta and of bicuspid aortic valve were compared with findings previously reported in women with TS and the general population. RESULTS: Bicuspid aortic valve without any stenosis or regurgitation was found in one woman in the study group with low-level 45,X/46,XX mosaicism, a statistically significantly lower prevalence of bicuspid aortic valve than that reported in women with TS. Aortic coarctation was not identified in any individual. The ASI was below the 95th percentile in all cases and the mean value was significantly lower than the mean reference values for both the general population and women with TS. CONCLUSION: Compared with the general population, the prevalence of risk factors for aortic dissection was not found to be higher in women with low-level 45,X/46,XX mosaicism without any noticeable features except infertility.

[Factors influencing success rate of FISH in pre-implantation genetic diagnostics]. / Faktory ovlivnující úspesnost FISH pri preimplantacní genetické diagnostice.

OBJECTIVE: Information on preimplantation diagnostics (PGD) by means of the method of fluorescent in situ hybridization (FISH) and some factors, which principally influence the results obtained with this technique from the point of view of a molecular cytogeneticist. Critical steps of work with one cell in PGD-FISH are summarized based on data from literature as well as from the authors, own experimental experience. The individual steps are analyzed and possibilities of execution are presented. TYPE OF STUDY: A review. SETTING: Institute of Medical Genetics and Fetal Medicine of Faculty Hospital and Medical Faculty of Palacky University, Center of Assisted Reproduction at Obstetric-Gynecological Clinic, Faculty Hospital, Olomouc. SUBJECT AND METHODS OF STUDY: Introduction into the PGD problem. A survey and analysis of factors decisive for the success of the FISH method for PGD. - Selection of embrya for the experimental phase of PGD-FISH. - Selection of fixation method and pretreatment, application of suitable probes. - Localization of the fixed cell. - Interpretation of the results obtained. Recommendation of the mode of their successful solution. CONCLUSION: The paper presents a survey of the PGD-FISH method. Some critical aspects in the analysis of unicellular preparation are pointed out and evaluated and possibilities and procedures of the optimal solution are suggested.

[Importance of genetic testing in couples with reproductive disorders]. / Význam genetického vysetrení u páru s poruchami reprodukce.

OBJECTIVE: To determine the prevalence of chromosomal aberrations in infertile couples undergoing in vitro fertilization (IVF). DESIGN: Cytogenetic analysis of peripheral blood lymphocytes in the group of patients undergoing IVF. Detection of chromosomal aberrations in the fetuses after IVF. SETTING: Department of Medical Genetics and Fetal Medicine, Medical Faculty, Palacký University and the University Hospital, Olomouc. METHODS: Cultivation of peripheral blood lymphocytes or fibroblasts of amniotic fluid. Using fluorescent in situ hybridization in cases of mosaicism. RESULTS: Out of 638 patients undergoing treatment for male or female infertility, 595 had normal karyotype and 43 (6.8%) had abnormal karyotype. There were detected 9 (1.4%) cases of balanced chromosomal rearrangements, 2 (0.31%) cases of deletion of Y chromosome, 2 (0.31%) cases of inversion, 2 (0.31%) cases of marker chromosome, 5 (0.78%) cases of gonosomal aneuploidy (47,XXY) and 23 (3.65%) cases of gonosomal mosaicism--out of the 22 (3.5%) cases of low-level mosaicism. In the small group of pregnant patients after IVF investigated for the risk of genetic disorders included in our study (n = 60) the frequency of chromosomal abnormalities was 9 (15%). CONCLUSIONS: Our data show that a high number of infertile couples is affected by chromosomal aberrations which occur more frequently in females than in males. It is caused by high frequency of low-level gonosomal mosaicism in the group of infertile women. Chromosomal analyses are highly recommended before each IVF procedure.

[Analysis of specific sequences in female patients with Turner syndrome--initial study]. / Analýza y specifických sekvencí u pacientek s turnerovým syndromem--úvodní studie.

BACKGROUND: DNA sequences from chromosome Y can cause gonadoblastoma development in patients with Turner syndrome (TS). Estimated risk is about 30%. The aim of the study is detection of Y-sequences of DNA level, calculation of mosaicism and its cytogenetic location. Clinical result of the study is the recommendation to gonadectomy of proved positive patients. METHODS AND RESULTS: Samples from 110 patients were collected. The PCR method and analysis of products on agarose gel was compared with analysis of DNA fragments from quantitative fluorescent (QF) PCR on capillary electrophoresis. The loci DYZ3, AMGX/Y and SRY were used for detection. The method QF PCR was effected for DYZ3 and AMGX/Y loci. The positive cases were examined by FISH method. Five (4.5%) and 3 (2.7%) positive cases were detected in DYZ3 and SRY resp. loci by electrophoresis on agarose gel. Seventeen (15.5%) and 7 (6.4%) positive cases were detected in DYZ3 and AMGX/Y resp. by capillary electrophoresis. The estimated mosaicism ranged from 1:5 to 1:100,000. CONCLUSIONS: QG PCR is the most sensitive method for diagnostics of Y-sequences. Simultaneously the incidence of Y-positive cells can be estimated. The positive cases with marker in karyotype were confirmed by FISH.

[Early fetal karyotyping and its role in prenatal diagnosis]. / Vcasná karyotypizace fetu a její prínos pro prenatální diagnostiku.

OBJECTIVE: Shift of indicated invasive examination in prenatal diagnostics towards the earlier phases of pregnancy with preservation of quality of cytogenetic detection. DESIGN: Cytogenetic and molecular-cytogenetic analysis of the chorionic villi after long term culture. SETTING: Department of Medical Genetics and Foetal Medicine, Faculty of Medicine, Palacky University Olomouc, Faculty Hospital in Olomouc. METHODS: Cultivation of fibroblasts developing from chorionic villi after enzymatic or mechanical disintegration and their karyotyping. Using fluorescent in situ hybridisation to identify the most common chromosomal aneuploidies and to determine gonosomes in indicated cases. RESULTS: Testing and optimisation of long term culture method and its routine use. Method was utilised so far in 12 patients and successfulness was 83%. Additional fluorescent in situ hybridisation was performed in 6 cases. CONCLUSION: Using long term culture method of chorionic villi as reliable and routine tool in prenatal diagnostics.

[Ca 125 during normal and pathologic pregnancy]. / Ca 125 v prubehu fyziologického a patologického tehotenství.

In 44 women in the first trimester of pregnancy a single examination of the Ca 125 serum level was made. In 10 women of the control group after mini-interruptions of pregnancy almost in all instances serum concentrations within the normal range were recorded. In 23 women with the diagnosis of spontaneous abortion in the first trimester a mean concentration of 58 U/ml of this protein was found. In a group of 11 women with the diagnosis of imminent abortion with a favourable prognosis, on the other hand, the mean serum concentration was only 42 U/ml, which is close to the mean concentration recorded in the control group of healthy pregnant women (46 U/ml). The difference between the two groups of pathological pregnancies is, however, not statistically significant.