Quantitative analysis of the dystrophin gene by real-time PCR

Duchenne and Becker muscular dystrophy (DMD/BMD) are severe X-linked neuromuscular disorders caused by mutations in the dystrophin gene. Our aim was to optimize a quantitative real-time PCR method based on SYBR® Green I chemistry for routine diagnostics of DMD/BMD deletion carriers. Twenty female relatives of DMD/BMD patients with previously detected partial gene deletions were studied. The relative quantity of the target exons was calculated by a comparative threshold cycle method (ΔΔCt). The carrier status of all subjects was successfully determined. The gene dosage ratio for non-carriers was 1.07±0.20, and for carriers 0.56±0.11. This assay proved to be simple, rapid, reliable and cost-effective

Anthropogenetic variability in the groups of homo- and heterosexually oriented individuals

This population-genetic study compares morphophysiological and genetic variability in a group of homosexually oriented individuals from Serbia (N=96) with control group of heterosexual individuals (N=96) using a test of determination of homozygously recessive characteristics in humans (HRC-test). Results of our study revealed a statistically significant difference in the mean values of genetic homozygosity (control group 5.0+0.2 ; homosexuals 3.4 +0.1 HRCs, out of 20 observed characteristics) the differences in the distribution type, as well as in the variances of presence of specific combinations of such traits. These results suggest a complex polygenic difference between two observed systems. There is no difference in the degree of genetic homozygosity between the genders in each tested group of individuals. However, both homosexually oriented females and males have significantly lower mean values of HRCs compared to female and male heterosexuals. [Projekat Ministarstva nauke Republike Srbije, br. 175093

Anthropogenetic variability in groups of children from regular and special schools from different localities in Serbia

This population-genetic study compares morpholophysiological and genetic variability in five control groups of individuals (children from five regular schools, N= 996) with children from that many special schools (N= 736) from Serbia, by using a test of determination of homozygously recessive characteristics in humans (HRC-test). Genetic homozygosity degree showed not only statistically significant difference between the mean values obtained for two groups of studied samples (control group 6.95± 0.07; children from special schools 8.63± 0.08 HRCs, out of 30 analyzed characteristics), but also differences in the type of distribution, as well as the presence of specific combinations of such traits. Results of comparisons done in different places (Kraljevo, Nis, Vranje, Leskovac, Pirot) showed the same tendency- the increase of genetic homozygosity and relative decrease of variability in samples of children from special schools. The number of HRCs among individuals from control groups varied from 2 to 15/30, and from 3 to 16/30 among children from special schools. It is possible that increased recessive homozygosity present in the group of children from special schools leads to increase of genetic loads, what may cause easier expression of some physiological and mental abilities that children from special schools have.A great individual variation in amount of genetic homozygosity that exists among human individuals may influence their potentials for different kinds of adaptation, including their mental abilities, physical capacities or resistance to different diseases. [Projekat Ministarstva nauke Republike Srbije, br. 175093

Morphogenetic Variability and Hypertension in Ischemic Stroke Patients—Preliminary Study

In this study, we evaluated and compared the morphogenetic variability and the degree of recessive homozygosity in patients with manifested ischemic stroke compared to healthy controls. We have evaluated 120 patients with manifested ischemic stroke, of which 64 did not have hypertension and 56 have hypertension. For comparison, we additionally tested 194 healthy individuals without manifested ischemic stroke (controls). For the estimation of the degree of recessive homozygosity, we have performed the homozygously recessive characteristics (HRC) test and tested 19 HRCs. There was a significant difference in the individual variations of 19 HRCs between the controls and patients with manifested ischemic stroke (∑χ2 = 60.162, p < 0.01). The mean values of the tested HRCs significantly differed between the controls and group with manifested ischemic stroke (Controls − 5.71 ± 1.61, Ischemic stroke group − 6.25 ± 1.54, p = 0.012). For the tested individuals with hypertension, the mean values of HRCs did not significantly differ between the controls and those that had manifested ischemic stroke (Controls − 5.28 ± 1.75, Ischemic stroke group − 5.64 ± 1.48, p = 0.435). We found a significant difference in the frequencies of HRCs between those with and without hypertension for controls (p < 0.003) and for those with manifested ischemic stroke (p < 0.001). There are increased degrees of recessive homozygosity along with decreased variability in patients with manifested ischemic stroke compared to controls

Anthropogenetic Variability in the Group of Individuals with Febrile Seizures: Population-Genetic Study

Febrile seizures (FS) are the most common neurological disorder in childhood and are a great stress for parents due to their dramatic clinical appearance. Using test for determination of homozygously recessive characteristics in humans (HRC test) we analyzed presence, distribution, and individual combination of 20 selected genetically controlled morphophysiological traits among FS patients (N=121) and control (N=121) to determine a possible deviation in the homozygosity level and genetic loads in the group of affected children and whether there is a predisposition to the occurrence of FS. The results of our study show a statistically significant difference in the mean values of the HRC tested (x¯HRC/20 CN = 3.2 ± 0.2; x¯HRC/20 FS = 4.6 ± 0.2, t= 5.74 , p< 0.0001), as well as in the distribution and variability of two studied samples (VC=55,3%, VFS= 39,6%), which indicates a complex polygenic difference among the tested groups of subjects. The differences in the degree of genetic homozygosity and variability are also present between the genders (t Cf/FSf = 4.12; t Cm/FSm = 3.98; p <0.0001) (VCf=56.9%, VFSf= 39.3%; VCm=54.1%, VFSm=40.1%). Obtained results indicate the enlargement of recessively homozygous genetic loads in the group of children with FS which may represent some kind of predisposition for expressivity of this type of seizures

Genetic and Environmental Dispositions for Cardiovascular Variability: A Pilot Study

Background: The aim of our study was to evaluate the degree of genetic homozygosity in the group of patients with coronary artery disease (CAD), as well as to evaluate morphogenetic variability in CAD patients regarding the presence of investigated risk factors (RF) compared to a control sample of individuals. Additionally, we aimed to evaluate the distribution of ABO blood type frequencies between tested samples of individuals. Methods: This study analyzed individual phenotype and morphogenetic variability of 17 homozygously-recessive characteristics (HRC), by using HRC test in a sample of 148 individuals in CAD patients group and 156 individuals in the control group. The following RF were analyzed: hypertension, diabetes mellitus, hyperlipidemia, and smoking. Results: The mean value of HRC in CAD patients is significantly higher, while variability decreases compared to the control sample (CAD patients: 4.24 ± 1.59, control sample: 3.75 ± 1.69; VCAD-patients = 37.50%, VC = 45.07%). There is a significant difference in individual variations of 17 HRC between control sample and CAD patients (χ2 = 169.144; p &lt; 0.01), which points out to different variability for tested genes. Mean values of HRC significantly differed in CAD patients in regard to the number of RF present. A blood type (OR = 1.75) is significant predictor for CAD, while O blood type (OR = 0.43) was significantly associated with controls. Conclusion: There is a higher degree of recessive homozygosity in CAD patients versus individuals in the control sample, and the presence of significant variations in the degree of recessive homozygosity as the number of tested RF increases