PCSK9 and Lipid Metabolism: Genetic Variants, Current Therapies, and Cardiovascular Outcomes.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) plays a crucial role in the modulation of lipid metabolism as a critical negative regulator of hepatic low-density lipoprotein receptor (LDLR) levels and circulating low-density lipoprotein (LDL) clearance. Numerous gain-of-function (GOF) mutations in PCSK9 have been identified as causing familial hypercholesterolemia (FH) by reducing LDLR levels, and loss-of-function (LOF) mutations associated with a hypercholesterolemia phenotype protective against atherosclerosis. PCSK9 represents an example of successful translational research resulting in the identification of PCSK9 as a major drug target for a lipid-lowering therapy. To explore the genetic constitution of PCSK9 and its biologic role, in this review, we summarize the current evidence of clinically significant PCSK9 genetic variants involved in lipid metabolism as well as emphasize the importance of PCSK9 inhibition for the improvement of cardiovascular outcomes by conducting a meta-analysis of the available data on the incidence of cardiovascular disease events.

Evaluation of COLIA1 gene rs1107946 polymorphism in relation to bone mineral density and fracture risk in Slovak postmenopausal women.

OBJECTIVES: The aim of the study was the evaluation of the rs1107946 polymorphism of the COLIA1 gene impact on bone mineral density and fracture risk in Slovak postmenopausal women. METHODS: One hundred and twenty-seven postmenopausal Slovak women with a diagnosis of osteopenia/osteoporosis were genotyped for rs1107946 polymorphism of the COLIA1 gene. Clinical and anthropometric data were obtained. DNA isolation was performed using a standard protocol. Genetic analyses of the rs1107946 polymorphism of the COLIA1 gene were performed by the TaqMan SNP genotyping assays. RESULTS: The study confirmed a statistically significant relationship using an association analysis between the rs1107946 polymorphism of the COLIA1 gene genotypes and body weight of the Slovak postmenopausal women with osteopenia/osteoporosis (p = 0.03). The study revealed a significant association of the risk T allele of the rs1107946 polymorphism of the COLIA1 gene with osteoporotic fractures (p = 0.038). The odds ratio confirmed 2.060 times higher risk of osteoporotic fractures in Slovak postmenopausal women with the presence of risk T allele of the rs1107946 COLIA1 gene polymorphism (OR = 2.060; 95% CI: 1.024-4.144). CONCLUSION: The results of this study revealed an association of T allele of the rs1107946 COLIA1 gene polymorphism with osteoporotic fractures in Slovak postmenopausal women with osteopenia/osteoporosis and suggest that the rs1107946 polymorphism of the COLIA1 gene may be a molecular biomarker usable in the management of osteoporosis.

Oral health status of 6- and 12-year-old children of Roma origin from Eastern Slovakia: a pilot study.

OBJECTIVE: The purpose of this study was to determine the prevalence and clinical effect of untreated dental caries in Roma children from eastern Slovakia using dmft/DMFT index and SiC index, association between dental caries development and oral hygiene, dietary habits and preventive dental care. METHODS: Dental caries were assessed by recording the dmft index (for primary dentition) and the DMFT index (for permanent dentition) that are used to assess the state of teeth, which expresses the current state of teeth or its development in an individual or the entire population. The SiC index was calculated as the mean dmft of one-third of the population with highest caries scores. The normality of data distribution was tested by Shapiro-Wilk test. P-value < 0.05 was considered statistically significant. Chi-square test was used to compare proportions (oral hygiene, dietary habits and preventive dental visit). Data were analysed using ordered logistic regression and t-test. The study includes questionnaire containing 5 questions about dietary habits, oral hygiene and preventive dental visit. RESULTS: The results of presented study confirmed higher average values of DMFT (3.24) in the population of 12-year-old Roma children and lower average values of dmft (2.5) in the second group (p < 0.05). The value of SiC index represented 6.10 in the group of six-year-old and 7.66 in twelve-year-old children. Logistic regression was performed to test the magnitude of the association between dental caries and related factors. There was statistically significant association between average value dmft/DMFT and dietary habits, oral hygiene, and preventive dental visit in both study groups. CONCLUSION: The study revealed insufficient oral hygiene of the Roma children population. Systematic implementation of preventive examinations for oral hygiene and health programmes are needed to promote oral health. The study represents a pilot study of the SiC index values in Roma minority population from eastern Slovakia.

Finding the candidate sequence variants for diagnosis of hypertrophic cardiomyopathy in East Slovak patients.

BACKGROUND: Hypertrophic cardiomyopathy is a heterogeneous myocardial disease. Mutations appearing in several genes might be a potential cause of the disease. The aim of the study was to analyze selected exons of the sarcomeric and non-sarcomeric genes, with the purpose to identify potential candidate genetic variants and to understand etiopathogenetic mechanisms of hypertrophic cardiomyopathy in East Slovak patients. METHODS: This study recruited 23 unrelated patients with hypertrophic cardiomyopathy, namely, 13 men and 10 women (mean age of 58.09±15.82 years) and 25 healthy controls in order to determine the candidate sequence variants, in the selected exons of six cardiomyopathy genes (MYBPC3, MYH7, NEBL, SCN5A, TNNI3, TNNT2), by conventional capillary-based Sanger sequencing method and standard protocols. RESULTS: Molecular genetic results confirmed the presence of 43 sequence variants in the selected exons of six cardiomyopathy genes, 58.14% of detected variants were novel. The majority of detected sequence variants were confirmed within exon 23 of MYH7 gene. Only 11 genetic alterations were predicted to be potentially pathogenic. CONCLUSIONS: In our study, we identified known and novel sequence variants in 23 unrelated patients with hypertrophic cardiomyopathy, but we did not observe any strong mutation hotspot. The results of our study assumed that exon 23 of MYH7 gene can be in potential affinity to hypertrophic cardiomyopathy in our cohort of patients. The sequence variants identified in this study may be further investigated in order to determine their functions in disease pathogenesis and improve management, diagnosis, and treatment in Slovak patients.

Analysis of selected genes associated with cardiomyopathy by next-generation sequencing.

BACKGROUND: As the leading cause of congestive heart failure, cardiomyopathy represents a heterogenous group of heart muscle disorders. Despite considerable progress being made in the genetic diagnosis of cardiomyopathy by detection of the mutations in the most prevalent cardiomyopathy genes, the cause remains unsolved in many patients. High-throughput mutation screening in the disease genes for cardiomyopathy is now possible because of using target enrichment followed by next-generation sequencing. The aim of the study was to analyze a panel of genes associated with dilated or hypertrophic cardiomyopathy based on previously published results in order to identify the subjects at risk. METHODS: The method of next-generation sequencing by IlluminaHiSeq 2500 platform was used to detect sequence variants in 16 individuals diagnosed with dilated or hypertrophic cardiomyopathy. Detected variants were filtered and the functional impact of amino acid changes was predicted by computational programs. RESULTS: DNA samples of the 16 patients were analyzed by whole exome sequencing. We identified six nonsynonymous variants that were shown to be pathogenic in all used prediction softwares: rs3744998 (EPG5), rs11551768 (MGME1), rs148374985 (MURC), rs78461695 (PLEC), rs17158558 (RET) and rs2295190 (SYNE1). Two of the analyzed sequence variants had minor allele frequency (MAF)<0.01: rs148374985 (MURC), rs34580776 (MYBPC3). CONCLUSION: Our data support the potential role of the detected variants in pathogenesis of dilated or hypertrophic cardiomyopathy; however, the possibility that these variants might not be true disease-causing variants but are susceptibility alleles that require additional mutations or injury to cause the clinical phenotype of disease must be considered.

Relationship between A163G osteoprotegerin gene polymorphism and other osteoporosis parameters in Roma and non-Roma postmenopausal women in eastern Slovakia.

BACKGROUND: The study was focused on evaluating the possible correlation between biochemical, anthropometric, and genetic indicators of osteoporosis in postmenopausal women. The frequency of genotypes and differences in measured parameters were evaluated within two ethnically different groups of women in Slovakia. METHODS: The study included 310 postmenopausal women divided into non-Roma and Roma groups. Based on results of densitometry, they were divided into control groups and women with osteoporosis and osteopenia. In all women, a genetic analysis of polymorphism of osteoprotegerin gene promotor region (A163G) was provided along with measurement of indicators of bone tissue metabolism. RESULTS: There is a particularly low incidence of osteoporosis in Roma women. We found a correlation between bone mineral density (BMD), body mass index, and waist and hip circumference in women with osteoporosis and in Roma women with osteopenia. The frequency of the AG genotype was higher in non-Roma women with osteoporosis, but reached only 10.7% in Roma women with osteopenia. While the presence of the G allele in the non-Roma population was accompanied by higher BMD and markers of osteoformation, it was accompanied by significantly higher concentrations of parathyroid hormone in the Roma population. CONCLUSION: The presence of the AG genotype has a different effect on bone metabolism in two ethnically diverse populations of women in Slovakia. In the general population, the presence of the G allele exhibited protective effects consistent with other studies, but in Roma population this appears to be the allele A. However, this requires a further study for confirmation and more detailed characterization of the differences between populations that have this work indicated.

Analysis of SCN5A Gene Variants in East Slovak Patients with Cardiomyopathy.

OBJECTIVE: Mutations in ion channels genes are potential cause of cardiomyopathy. The SCN5A gene (sodium channel, voltage gated, type V alpha subunit gene; 3p21) belongs to the family of cardiac sodium channel genes. Mutations in SCN5A gene lead to decreased Na+ current and ion unbalance. The SCN5A gene mutations are found in approximately 2% of patients with dilated cardiomyopathy (DCM), and they may be potential phenotype modifiers in hypertrophic cardiomyopathy (HCM). The role of SCN5A gene mutations in cardiomyopathy is not fully elucidated. METHODS: Three selected exons (12, 20, and 21) of the SCN5A gene in the cohort of 58 East Slovak patients with dilated and HCM were analyzed by the Sanger sequencing method in order to detect etiopathogenic mutations associated with dilated and HCM. RESULTS: The mutation screening of three selected exons of SCN5A gene in the cohort of 27 DCM, 12 HCM patients, and 16 controls identified 10 missense genetic variants. Three of them (T1247I, A1260D, and G1262S), all in exon 21 of the SCN5A gene, were potentially damaging and disease-causing variants. CONCLUSION: Data from this study demonstrate that SCN5A gene variants have important role in the etiopathogenesis of dilated and HCM.

Innovative health tracker that provides advanced functionalities to support and guide users in modifying their lifestyle: a Straussian ground theory approach.

Objective: Health can be described as the state of homeostasis and optimal functioning across various bio-psycho-social dimensions and processes, allowing an individual to adapt and respond effectively to extrinsic and intrinsic challenges. Our thoughts, choices, behaviors, experiences, and feelings shape our existence. By transitioning from unconscious reactions to conscious responses, we can establish novel habits and behaviors, actively embracing positive shifts in our lifestyle. Subjects and methods: The presented examination focuses on the smartwatch (SW), analyzing the incorporation of potentially progressive attributes that could enrich our lifestyle pursuits. The objective is not the health disorders themselves but the employment of wearable devices to create a strong sense of coherence in the Straussian grounded theory approach. The study had no subjects. Results: The potential of the SW has been partially explored in lifestyle intervention, modification, research, and practice. Conclusion: Based on our examination, creating an innovative SW capable of aiding individuals in better comprehending their behaviors and motivating them toward comprehensive changes in their lifestyle is a challenging yet attainable endeavor. Our ambition is to bring into existence SW capable of comprehensively measuring and evaluating interoception, circadian rhythm (CR), selected lifestyle pillars, and their associated components, and seamlessly integrating them into current SW features. It focuses on boosting motivation, maintenance, and amelioration regarding one's lifestyle. The novel approach strives to boost both immediate and underlying factors that actively contribute to improving one's metacognition.

Comparison of BMI and % BF determined by bioimpedance analysis for obesity screening in young women.

The body mass index (BMI) is used as the most common and simplest measure to predict obesity. The aim of the study was to compare the BMI method and % BF (body fat) determined by bioimpedance analysis for obesity screening and to evaluate the correlation between these methods. 200 women aged 18-25 years were included in the study. BMI ≥ 25/ ≥ 30 kg/m2 was the criterion for assessment of overweight/obesity. Body fat (BF) in % and kg was determined by bioimpedance analysis. For assessment of obesity by % BF, the criterion was ≥ 30% fat. Based on BMI, 4.5% of women were obese, 14.5% were overweight. According to % BF, up to 30% of women were obese. The largest differences between BMI and % BF categorization were found in the underweight and overweight groups. According to the BMI 43% of women in the underweight category and 34.5% in the overweight category had a % BF in the normal range. By correlation analysis, we found a strong positive statistically significant correlation in women between BMI and BF (%) (rs = 0.782, p < 0.01) and also between BMI and BF (kg) (rs = 0.880, p < 0.01). Both of these methods assess body composition in their own specific way, complement each other, and other combining them, we achieve more accurate results for determining the overall health status of an individual.

Anthropological analysis of skeletal remains from the site Vysná Mysla - Koscelek.

The archaeological site Vysná Mysla - Koscelek is located in the southeast of the Slovak Republic, and it is peculiar by its location in the middle of the woods near a thermal spring. Burials could be dated to the 13th to 14th centuries based on the presence of the ruins of the church, albeit the funeral practices could last until the 18th century. A total of 53 individuals were examined, in whom the age at death, sex and stature were morphoscopically and metric estimated by standard anthropological methods, and the presence of non-metric traits and pathological conditions was also recorded. The analysed group consisted of 40 adults and 4 adolescents, of which 16 were women and 2 possible females, 19 men and one probable male individual, three individuals of inconclusive sex and six individuals of unknown sex because of bad preservation of the human remains. The group of nonadults consisted of 6 children. Pathological conditions were present and recorded in 32% of 53 evaluated individuals. Due to the lack of historical written sources, this analysis represents an important study for the reconstruction of the life of historical population living in this area.

The frequency of factor V Leiden and prothrombin G20210A mutations in Slovak and Roma (Gypsy) ethnic group of Eastern Slovakia.

Factor V Leiden and prothrombin G20210A are the two most prevalent causes of inherited thrombophilia. The prevalence of these mutations varies widely in healthy Caucasian population. The aim of our study was to determine the frequency of factor V Leiden and prothrombin G20210A mutations in Slovak and Roma ethnic group from Eastern Slovakia. We analyzed 540 asymptomatic individuals (269 individuals of Slovak ethnicity and 271 individuals of Roma ethnicity) by real-time PCR method. The detected allele frequencies were 2.97 versus 6.64 % for factor V Leiden (p = 0.0049), and 0.74 versus 0.92 % for prothrombin mutation (p = 0.7463) in Slovak and Roma population, respectively. The Roma ethnic group had significantly higher prevalence of factor V Leiden mutation when compared to Slovak ethnic group. The allele frequency of factor V Leiden in ethnic Romanies from Eastern Slovakia was one of the highest in Europe. Our results confirm an uneven geographical and ethnic distribution of factor V Leiden.

Association of the FTO rs9939609 polymorphism with obesity in Roma/Gypsy population.

The rs9939609 SNP located in the first intron of the fat mass and obesity associated gene (FTO) has been found to be associated with common obesity mainly in populations of European descent. The Roma/Gypsy population as an ethnic minority of Asian Indian origin is well known for its adverse health status with a high prevalence of obesity. The main aim of this study was to examine the contribution of the rs9939609 FTO polymorphism to the high prevalence of obesity in the Roma/Gypsy population. Following a number of anthropometric measurements, the FTO rs9939609 polymorphism was genotyped in 312 Roma/Gypsy individuals. We observed significant differences in body mass index (BMI), waist circumference, and waist-to-hip ratio between different genotypes (P = 0.003, P = 0.012, and P = 0.03, respectively). The waist circumference in the subjects with AA genotype was about 7.1 cm larger than in those with TT genotypes (P = 0.005). However, the strongest association of minor allele A of the rs9939609 FTO polymorphism was found with BMI (odds ratio, 1.55; 95% confidence interval, 1.129-2.128; P = 0.007), even after adjusting for age, sex, and smoking status. This study provides the first report of allele and genotype frequencies for the rs9939609 polymorphism and also the first evidence of the association of the FTO variant with obesity in the Roma/Gypsy population.

Association between vitamin D receptor gene polymorphisms (Fok I, Cdx-2) and bone mineral density in Slovak postmenopausal women.

Osteoporosis is a skeletal disorder characterized by low bone mass and microarchitectural deterioration of bone tissue with consequent increase in bone fragility and fracture risk. Bone mineral density (BMD), the major determinant of osteoporotic fracture risk, has a particular genetic background. Vitamin D receptor (VDR) is implicated in the regulation of bone mineral density. The present study evaluates the association between Vitamin D receptor gene polymorphisms Fok I (rs2228570), Cdx-2 (rs11568820), bone mineral density and fracture risk in Slovak postmenopausal women. A total of 403 unrelated Slovak postmenopausal women aged 43-86 years were genotyped using TaqMan®SNP Genotyping Assays. Lumbar spine, femoral neck and total hip BMD/T-score were detected by dual energy X-ray absorptiometry (DEXA). We found the Fok I and Cdx-2 polymorphism in the VDR gene to be associated with osteoporotic fractures (non-vertebral fractures: Fok I p = 0.001; Cdx-2 p = 0.0000; all fractures: Fok I p = 0.0001; Cdx-2 p = 0.0000) (Fok I: OR = 0.50, 95% CI = 0.35-0.71; Cdx-2: OR = 0.25, 95% CI = 0.17-0.37). The present data suggest that VDR gene Fok I and Cdx-2 polymorphisms contribute to the determination of BMD in Slovak postmenopausal women and can probably be used with other genetic markers together to identify individuals at high risk of osteoporosis.

Differences in body fat distribution in women depending on the second to fourth digit ratio value.

Second to fourth digit ratio (2D:4D) is sexually dimorphic, however, the hypothesis claiming that masculine digit ratio in women is connected with masculine phenotype is not fully confirmed. The aim of the present study was to investigate differences in anthropometric parameters in women in relation to the digit ratio value. Cross-sectional research was conducted among 730 Polish female university students, aged 19-25 years. Anthropometric measurements were performed and data about socioeconomic status were collected. The results showed no statistically significant relationships between both right- or left-hand 2D:4D digit ratio and body height and BMI either in univariate linear regressions or in multivariate regression when SES was included in the models. The correlation between right-hand 2D:4D digit ratio and WHR also was not statistically significant. There were significant linear negative relationships between left 2D:4D and WHR. The significant linear negative relationships between both right and left 2D:4D and waist circumference and WHtR were found. Relationships between digit ratio and WC and WHtR were also tested by multiple regression analyses with SES included in the models. The results were statistically significant. Additionally, statistically significant differences in both right- and left digit ratios depending of the categories of waist circumference and WHtR were found. The lowest values of digit ratio were presented by the women with abdominal obesity. The results suggest that low values of digit ratio in women are associated with higher abdominal fat accumulation.

High frequency of SLC22A12 variants causing renal hypouricemia 1 in the Czech and Slovak Roma population; simple and rapid detection method by allele-specific polymerase chain reaction.

Renal hypouricemia is a rare heterogeneous inherited disorder characterized by impaired tubular uric acid transport with severe complications, such as acute kidney injury. Type 1 and 2 are caused by loss-of-function mutations in the SLC22A12 and SLC2A9 gene, respectively. A cohort of 881 randomly chosen ethnic Roma from two regions in Eastern Slovakia and two regions in the Czech Republic participated. Genomic DNA was isolated from buccal swabs and/or from blood samples. The c.1245_1253del and c.1400C>T genotypes were determined using polymerase chain reaction with allele-specific primers in a multiplex arrangement and/or direct sequencing of exon 7 and 9. Allele frequencies and genotypes were tested for Hardy-Weinberg equilibrium using the Chi-square test. 25 subjects were heterozygous and three were homozygous for the c.1245_1253del, while 92 subjects were heterozygous and two were homozygous for the c.1400C>T. Moreover, two participants were compound heterozygotes. Frequencies of the c.1245_1253del and c.1400C>T variants were 1.87 and 5.56 %, respectively. Our finding confirms an uneven geographical and ethnic distribution of SLC22A12 mutant variants. We found that the c.1245_1253del and c.1400C>T variants were present in the Czech and Slovak Roma population at unexpectedly high frequencies. Renal hypouricemia should be kept in mind during differential diagnostic on Roma patients with low serum uric acid concentrations.

Ethnic differences in the association of thrombophilic polymorphisms with obstetric complications in Slovak and Roma (Gypsy) populations.

AIMS: Hereditary as well as acquired thrombophilia is associated with a higher incidence of severe obstetric complications such as preeclampsia, spontaneous pregnancy loss, placental abruption, and fetal growth retardation. The aim of our study was to examine the association of selected thrombophilic polymorphisms (factor V Leiden, MTHFR C677T, and MTHFR A1298C) with pregnancy complications in the Slovak majority population and the Roma (Gypsy) ethnic population. The study included 354 women; 120 patients and 105 controls from the Slovak majority population, 50 patients and 79 controls from the Slovak Roma population. Genotyping was performed by the real-time polymerase chain reaction method using TaqMan(®) MGB probes. RESULTS: A statistically significant higher frequency of factor V Leiden (p=0.001, odds ratio [OR]=5.9) and MTHFR C677T polymorphism (p=0.011, OR=1.7) was observed in the Slovak majority patient group compared to the control group. The incidence of MTHFR A1298C polymorphism between patients and controls did not differ significantly. None of the three polymorphisms studied was in association with pregnancy complications in the group of Roma women. CONCLUSIONS: Our study has confirmed the variable distribution of selected thrombophilic polymorphisms in different ethnic groups as well as their various effects on the clinical phenotype.

TNFRSF11B gene polymorphisms, bone mineral density, and fractures in Slovak postmenopausal women.

Osteoporosis is a common disease that is characterized by low bone mineral density (BMD), deterioration in bone microarchitecture, and increased fracture risk. Due to its important role in bone biology, the TNFRSF11B gene, coding for OPG, has been considered as a candidate gene for osteoporosis. In this study, single nucleotide polymorphisms (SNPs) A163G, T245G, and G1181C (rs3102735, rs3134069, and rs2073618, respectively) within the TNFRSF11B gene were studied for association with BMD and fracture incidence in a cohort of 327 postmenopausal Slovak women. Genomic DNA was extracted and purified from peripheral blood leukocytes by the commercial kit JetQuick (Genomed GmbH, Germany) using a standard protocol. Genotyping was performed using the Custom TaqMan® SNP Genotyping Assays. The lumbar L1-L4 spine BMD (g/cm(2)) and T-score in the subgroup of Slovak postmenopausal women with osteoporotic fractures were significantly lower than those in the subgroup of women without fracture (p = 0.0025; p = 0.0009). We identified the T245G (rs3134069) polymorphism in the TNFRSF11B gene associated with osteoporotic fractures (vertebral fractures: p = 0.0320; non-vertebral fractures: p = 0.0005; all fractures: 0.0000). The polymorphism T245G (rs3134069) in the TNFRSF11B gene could be used together with other genetic markers to identify individuals at high risk of osteoporotic fractures. The results from the present study provided more evidence to reveal the role of TNFRSF11B gene polymorphisms in BMD and the risk of osteoporotic fractures.

Y-SNP analysis versus Y-haplogroup predictor in the Slovak population.

Human Y-chromosome haplogroups are important markers used mainly in population genetic studies. The haplogroups are defined by several SNPs according to the phylogeny and international nomenclature. The alternative method to estimate the Y-chromosome haplogroups is to predict Y-chromosome haplotypes from a set of Y-STR markers using software for Y-haplogroup prediction. The purpose of this study was to compare the accuracy of three types of Y-haplogroup prediction software and to determine the structure of Slovak population revealed by the Y-chromosome haplogroups. We used a sample of 166 Slovak males in which 12 Y-STR markers were genotyped in our previous study. These results were analyzed by three different software products that predict Y-haplogroups. To estimate the accuracy of these prediction software, Y-haplogroups were determined in the same sample by genotyping Y-chromosome SNPs. Haplogroups were correctly predicted in 98.80% (Whit Athey's Haplogroup Predictor), 97.59% (Jim Cullen's Haplogroup Predictor) and 98.19% (YPredictor by Vadim Urasin 1.5.0) of individuals. The occurrence of errors in Y-chromosome haplogroup prediction suggests that the validation using SNP analysis is appropriate when high accuracy is required. The results of SNP based haplotype determination indicate that 39.15% of the Slovak population belongs to R1a-M198 lineage, which is one of the main European lineages.

Founder mutations in NDRG1 and HK1 genes are common causes of inherited neuropathies among Roma/Gypsies in Slovakia.

Autosomal recessive forms of Charcot-Marie-Tooth disease (CMT) account for less than 10 % of all CMT cases, but are more frequent in the populations with a high rate of consanguinity. Roma (Gypsies) are a transnational minority with an estimated population of 10 to 14 million, in which a high degree of consanguineous marriages is a generally known fact. Similar to the other genetically isolated founder populations, the Roma harbour a number of unique or rare autosomal recessive disorders, caused by "private" founder mutations. There are three subtypes of autosomal recessive CMT with mutations private to the Roma population: CMT4C, CMT4D and CMT4G. We report on the molecular examination of four families of Roma origin in Slovakia with early-onset demyelinating neuropathy and autosomal recessive inheritance. We detected mutation p.R148X (g.631C>T) in the NDRG1 (NM_006096.3) gene in two families and mutation g.9712G>C in the HK1 (NM_033498) gene in the other two families. These mutations cause CMT4D and CMT4G, respectively. The success of molecular genetic analysis in all families confirms that autosomal recessive forms of CMT caused by mutations on the NDRG1 and HK1 genes are common causes of inherited neuropathies among Slovak Roma. Providing genetic analysis of these genes for patients with Roma origin as a common part of diagnostic procedure would contribute to a better rate of diagnosed cases of demyelinating neuropathy in Slovakia and in other countries with a Roma minority.

Unique frequencies of HFE gene variants in Roma/Gypsies.

The aim of this study was to assess the frequencies of three hemochromatosis gene (HFE) mutations in ethnic Roma/Gypsies in Slovakia. A cohort of 367 individuals representing general population and not preselected for health status was genotyped by TaqMan real-time PCR assay for C282Y, H63D and S65C mutations in HFE gene. A unique genetic profile was revealed: C282Y is found in the highest frequency of all Central European countries (4.90%), while the frequency of H63D mutation (4.09%) is lower than any reported in Europe so far. S65C mutation was not present in the cohort. These mutation frequencies can be explained rather by gene influx and genetic isolation than by genetic inheritance from a former Roma/Gypsy homeland.