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Here, we present a case study of twin boys aged 2 and 7 years who both met the diagnostic criteria for autism spectrum disorders (ASDs) based on the standard diagnostic instruments ADOS-2 and ADI-R. The clinical indication for genetic diagnostics in the first boy was autism with high severity of symptoms, delayed speech development, and mild facial dysmorphia. The second boy’s indication was autism with moderate severity of symptoms, delayed speech development, mild facial features, slowed psychomotor development, and microcephaly. The microarray-based analysis of chromosome aberrations revealed a heterozygous 977,456 bp deletion of region 10q26.3 in both boys. The region includes 28 genes, some of these genes are important in the development of the central nervous and urogenital systems, and heterozygous deletions in this region have been associated with mental retardation, growth and development disorders, and craniofacial anomalies. The whole exome sequencing confirmed the presence of this deletion in both boys and, at the same time, led to the identification of a pathogenic SNV variant in the TRIO gene in the boy with microcephaly and delayed psychomotor development, which may explain the different phenotype of both boys. However, the segregation analysis of these variants in the family revealed that the microdeletion was inherited from the asymptomatic father, and the c.2149C > T variant in the TRIO gene was inherited from the asymptomatic mother, making the diagnostic finding uncertain. This case highlights that when pathogenic or likely pathogenic variants are inherited from unaffected parents, the clinical phenotype may result from a combined burden of multiple rare variants and polygenic risk, underscoring the importance of a comprehensive genomic analysis in complex cases. Thus, we emphasize the importance of utilizing available methods, such as whole exome sequencing besides microarray-based comparative genomic hybridization, in the genetic diagnosis of autism patients in Slovakia.

Molecular diagnostics of hereditary breast and/or ovarian cancer is mainly based on detection of BRCA1 and BRCA2 germline mutations in suspected families. The aim of the study was to determine the frequency, age and geographical distribution in 130 Slovak hereditary breast and ovarian cancer (HBOC) families diagnosed within the years 2000-2004. Mutation screening was performed by single-strand conformation polymorphism (SSCP), heteroduplex analysis (HDA) and sequencing of PCR products showing an abnormal migration pattern. Twenty of 130 (15.6%) HBOC suspected families were found to carry mutations in BRCA1 or BRCA2 genes. The glossary data from the National Cancer Registry of Slovakia (NCRS) were compared with the results from HBOC suspected kindreds. Age distribution of breast cancer onset in our study group showed the highest proportion of onset in HBC families within the 5 th decade of life, while NCRS reports at least a ten year later onset. These findings confirmed that cases of breast cancer under 50 years of age can be used as one of the principal criteria to assign a family as a hereditary breast and/or ovarian cancer kindred. In contrast with unselected ovarian cancer cases, about 75% of all HOC index cases were diagnosed between 40 and 49 years of age. To study the geographical distribution of hereditary breast and/or ovarian cancer, Slovakia was divided into three parts. The distribution of HBOC suspected families approximately follows this division, with an increasing number in the western area of the country.

Although the rabbit is a frequently used biological model, the phenotype of rabbit adipose-derived mesenchymal stem cells (rAT-MSCs) is not well characterized. One of the reasons is the absence of specific anti-rabbit antibodies. The study aimed to characterize rAT-MSCs using flow cytometry and PCR methods, especially digital droplet PCR, which confirmed the expression of selected markers at the mRNA level. A combination of these methods validated the expression of MSCs markers (CD29, CD44, CD73, CD90 and CD105). In addition, cells were also positive for CD49f, vimentin, desmin, α-SMA, ALDH and also for the pluripotent markers: NANOG, OCT4 and SOX2. Moreover, the present study proved the ability of rAT-MSCs to differentiate into a neurogenic lineage based on the confirmed expression of neuronal markers ENO2 and MAP2. Obtained results suggest that rAT-MSCs have, despite the slight differences in marker expression, the similar phenotype as human AT-MSCs and possess the neurodifferentiation ability. Accordingly, rAT-MSCs should be subjected to further studies with potential application in veterinary medicine but also, in case of their cryopreservation, as a source of genetic information of endangered species stored in the gene bank.

Endothelial progenitor cells (EPCs) have been broadly studied for several years due to their outstanding regenerative potential. Moreover, these cells might be a valuable source of genetic information for the preservation of endangered animal species. However, a controversy regarding their characterization still exists. The aim of this study was to isolate and compare the rabbit peripheral blood- and bone marrow-derived EPCs with human umbilical vein endothelial cells (HUVECs) in terms of their phenotype and morphology that could be affected by the passage number or cryopreservation as well as to assess their possible neuro-differentiation potential. Briefly, cells were isolated and cultured under standard endothelial conditions until passage 3. The morphological changes during the culture were monitored and each passage was analyzed for the typical phenotype using flow cytometry, quantitative real–time polymerase chain reaction (qPCR) and novel digital droplet PCR (ddPCR), and compared to HUVECs. The neurogenic differentiation was induced using a commercial kit. Rabbit cells were also cryopreserved for at least 3 months and then analyzed after thawing. According to the obtained results, both rabbit EPCs exhibit a spindle-shaped morphology and high proliferation rate. The both cell lines possess same stable phenotype: CD14−CD29+CD31−CD34−CD44+CD45−CD49f+CD73+CD90+CD105+CD133−CD146−CD166+VE-cadherin+VEGFR-2+SSEA-4+MSCA-1−vWF+eNOS+AcLDL+ALDH+vimentin+desmin+α-SMA+, slightly different from HUVECs. Moreover, both induced rabbit EPCs exhibit neuron-like morphological changes and expression of neuronal markers ENO2 and MAP2. In addition, cryopreserved rabbit cells maintained high viability (>85%) and endothelial phenotype after thawing. In conclusion, our findings suggest that cells expanded from the rabbit peripheral blood and bone marrow are of the endothelial origin with a stable marker expression and interesting proliferation and differentiation capacity.

DGAT1 encodes diacylglycerol O-acyltransferase (EC 2.3.1.20), a microsomal enzyme that catalyzes the final step of triglyceride synthesis. It was shown that the QTL variation is most likely caused by a nonconservative base substitution in the candidate gene DGAT1 changing lysine to alanine (K232A) in the enzyme diacylglycerol O-acyltransferase (DGAT). In particular, the allele encoding the lysine 232 variant proved to be more efficient with regard to milk fat synthesis. The objective of the present study was to determine the allele frequencies and to verify the effects of the two DGAT variants (K232A polymorphism, A and K alleles) on milk production traits in Slovak dairy cows. Samples of 196 dairy cows originating from 61 sires were genotyped for DGAT1 K232A polymorphism (A and K alleles) using the PCR-RFLP technique. The frequencies of DGAT1 alleles were 0.88 (A) and 0.12 (K). The performance data were collected during one season in order to minimize this effect. The overall milk yield (MILK), fat yield (FAT_Y), fat content (FAT_C), protein yield (PROT_Y), protein content (PROT_C) and age at first calving (AGE1) were studied. The effect of DGAT1 polymorphism on fat and protein content in milk was confirmed. Further study is needed for explanation of effect of DGAT1 on the age at first calving.