The subject of this multidisciplinary forensic archaeological-anthropological research is a near-complete skeleton of a woman aged 40–49 years with possible perimortal cranial trauma, found within a known archaeological site from the 7th−3rd century calBCE. The skeleton (without any artefacts) was exhumed by speleologists from a depth of 14 metres, 2.8 m below the sedimentary deposit, in a deep and narrow karst abyss known as Studňa na Jame, which is located in the district of Liptovský Mikuláš in the Low Tatra Mountains (Liptov Region, north Slovakia). Enthesopathies rank her among strong, physically-working individuals living in a mountainous terrain. Analysis of δ13C and δ15N testify to a terrestrial diet with high animal protein. A Bayesian chronological model with two 14C dates and osteological prior information suggested her death occurred in the 19th−20th centuries calCE. Oral history research in the nearby village concluded that reportedly, after 1870, an adult woman of a known name had gone missing. Genealogical and archival research produced her date of birth and later corrected the earliest possible date of missing to 1891. Analysis of nuclear and mitochondrial aDNA extracted from her molar were used for identification of the skeletal remains. Her only found living female offspring could not be sampled for DNA due to ethical reasons. Therefore, profound archival genealogical research was conducted and two living distant matrilineal relatives were identified. They were chosen as probands and were DNA matched as relatives of the studied woman, whom we refer to as LM. Hence, we could add her date of birth as new prior information in the Bayesian chronological model and, eventually, estimate her date of death to 1891–1911 calCE. In Slovakia, this is the first forensic archaeological-anthropological case of successful identification of a missing person from skeletal remains using a strong, multidisciplinary, case-specific research toolkit rooted both in sciences and humanities.

In this case report, we describe an individual with Pearson syndrome, representing the first reported case in Slovakia. The patient was 1.5-year-old boy with pancytopenia including macrocytic anemia, neutropenia and thrombocytopenia, pancreatic insufficiency, hepatopathy, psychomotor development delay, short stature and failure to thrive. The patient also had atypical symptoms for Pearson syndrome, including atypical limb proportions and facial dysmorphism, which contributed to the delay in correct diagnosis. In the whole exome sequencing (WES) analysis, virtual panels targeting genes associated with inborn errors of immunity and anemia were selected based on the patient's clinical phenotype, however no pathogenic variant was identified within these panels. During the evaluation of secondary findings, a pathogenic deletion, m.10952_15371del, was detected in mitochondrial DNA in a heteroplasmic state (55.8% in peripheral blood), leading to the diagnosis. Subsequently, MLPA analysis confirmed this deletion in other patient tissues (urine, bone marrow aspirate, buccal swab) with the highest level of heteroplasmy (70%) detected in the urine sample. Our study emphasizes the importance of a comprehensive diagnostic approach, including the analysis of several tissues, especially in the diagnosis of clinically complex mitochondrial diseases.

X-linked severe combined immunodeficiency disease (X-SCID) is a form of inborn errors of immunity (IEI) associated with causal DNA variants of the IL2RG gene. Patients with X-SCID are characterized by a combination of cellular and humoral immunodeficiencies associated with increased susceptibility to infections. The presented cases constituted two unrelated male patients from the Slovak population. Proband A was primarily hospitalized at the age of three months because of recurrent fever, vomiting, and lethargy, and the atypical immunophenotype was determined to be T-B-NK-. For proband B, the first hospitalization occurred at the age of eight months because of generalized impetiginized dermatitis. Whole exome sequencing (WES) was performed via a comprehensive approach in patients with undefined IEI, and causal DNA variants were confirmed by Sanger sequencing. WES analysis in probands identified the currently undescribed hemizygous variants p.Asn84Thr and p.Val213Ala in the IL2RG gene. Segregation analysis of p.Asn84Thr indicated a de novo origin, and p.Val213Ala was detected only in the asymptomatic proband's mother. We comprehensively reconsidered and scored both variants based on biological and clinical aspects. Finally, taking all the information into account, we classified p.Asn84Thr as likely pathogenic and p.Val213Ala as likely pathogenic with mild penetrance based on the fulfilled ACMG (American College of Medical Genetics and Genomics) criteria for computational predictions, clinical correlations, localization at functional site, and de novo status. With the WES approach, we identified two novel, not yet reported, IL2RG variants in the Slovak population of X-SCID patients. These findings strengthen the fact that rapid and comprehensive molecular-genetic diagnostics of IEI is necessary for the early definition of precise diagnosis, which further enables appropriate treatment and patient management.

Objective: The aim of this study is to investigate the potential genetic etiology of cribra orbitalia noted on human skeletal remains. Materials: We obtained and analyzed ancient DNA of 43 individuals with cribra orbitalia. The analyzed set represented medieval individuals from two cemeteries in western Slovakia, Castle Devin (11th-12th century AD) and Cifer-P acute accent ac (8th-9th century AD).Methods: We performed a sequence analysis of 5 variants in 3 genes associated with anemia (HBB, G6PD, PO), which are the most common pathogenic variants in present day of European populations, and one variant MCM6: c.1917 + 326 C>T (rs4988235) associated with lactose intolerance.Results: DNA variants associated with anemia were not found in the samples. The allele frequency of MCM6: c.1917 + 326 C was 0.875. This frequency is higher but not statistically significant in individuals displaying cribra orbitalia compared to individuals without the lesion. Significance: This study seeks to expand our knowledge of the etiology of cribra orbitalia by exploring the po-tential association between the lesion and the presence of alleles linked to hereditary anemias and lactose intolerance.Limitations: A relatively small set of individuals were analyzed, so an unequivocal conclusion cannot be drawn. Hence, although it is unlikely, a genetic form of anemia caused by rare variants cannot be ruled out. Suggestions for Further Research: Genetic research based on larger sample sizes and in more diverse geographical regions.

Recent studies have shown that mitochondria are involved in the pathogenesis of Covid-19. Mitochondria play a role in production of reactive oxygen species and induction of an innate immune response, both important during infections. Common variability of mitochondrial DNA (mtDNA) can affect oxidative phosphorylation and the risk or lethality of cardiovascular, neurodegenerative diseases and sepsis. However, it is unclear whether susceptibility of severe Covid-19 might be affected by mtDNA variation. Thus, we have analyzed mtDNA in a sample of 446 Slovak patients hospitalized due to Covid-19 and a control population group consisting of 1874 individuals. MtDNA variants in the HVRI region have been analyzed and classified into haplogroups at various phylogenetic levels. Binary logistic regression was used to assess the risk of Covid-19. Haplogroups T1, H11, K and variants 16256C>T, 16265A>C, 16293A>G, 16311T>C and 16399A>G were associated with an increased Covid-19 risk. On contrary, Haplogroup J1, haplogroup clusters H+U5b and T2b+U5b, and the mtDNA variant 16189T>C were associated with decreased risk of Covid-19. Following the application of the Bonferroni correction, statistical significance was observed exclusively for the cluster of haplogroups H+U5b. Unsurprisingly, the most significant factor contributing to the mortality of patients with Covid-19 is the age of patients. Our findings suggest that mtDNA haplogroups can play a role in Covid-19 pathogenesis, thus potentially useful in identifying susceptibility to its severe form. To confirm these associations, further studies taking into account the nuclear genome or other non-biological influences are needed.

Background: During the last 20 years, X-chromosomal STR markers have become widely used in forensic genetics and paternity testing. Nevertheless, to exploit their full potential in any given population, a reliable reference dataset needs to be established. Since no relevant studies concerning these markers have been performed on the Slovak population so far, we decided to analyse several commonly used markers in this population. Aim: To create an informative set of Slovak population data concerning X-STR markers. Subjects and methods: We genotyped 378 individuals and analysed 12 loci (DXS10148, DX10135, DXS8378, DXS7132, DXS10079, DXS10074, DXS10103, HPRTB, DXS10101, DXS10146, DXS10134 and DXS742) localised in four distinct linkage groups. Results: Our analysis showed that the most informative marker is DXS10135 (PIC = 0,927) and the most informative linkage group (LG) is LG1 with 149 different haplotypes. This analysis also confirmed linkage disequilibrium for two pairs of markers (DX10101-DX10103 and DX10101-HPRTB) within LG3 in female samples. No statistically significant departure from HWE was observed for any locus. Moreover, the interpopulation comparison of 8 European populations based on haplotype frequencies showed no statistically significant FST values in any LG, except for LG2 in comparison with the German population. Conclusion: We created a haplotype database for forensic analyses and kinship testing in Slovakia, as well as the CE dataset which can be used to further increase the decision power in similar analyses in the future.

Orofacial clefts are a common developmental anomaly in living individuals; however, skulls with clefts are relatively rarely found in archaeological specimens. The presented research is dealing with the first and only historical skeletal case of cleft lip and palate from the territory of Slovakia. The skeletal remains belonged to a juvenile male impiously buried in a storage pit together with an adult male. The position of the skeletons strongly suggests that the deposition of the two bodies was a single event, dated to the mid-seventeenth century or in the late eighteenth century calAD. A juvenile male found in a squatting position at the bottom of the pit was affected by an incomplete unilateral cleft palate combined with a right-sided cleft lip. The defect was associated with dental anomalies and defects such as supernumerary teeth and enamel hypoplasia, as well as dysmorphology of the facial complex. Both the affected juvenile male and the adult male consumed meat, indicating their stable, if not higher, social status. That, coupled with the fact that the individual with cleft lived to the age of 16–18 years, suggests that at least this younger male must have been well cared for and lived in an economically and socially stable environment. The buried males were not biologically related and were probably of Western European origin. All these facts indicate that they were travelling foreigners from the higher class who died unexpectedly with a possibility of a violent death.

During rescue archeological excavation in 2018, the skeletal remains of a young male with an unusually large and rare lytic focus in the right iliac fossa, dating to the Middle Bronze Age, were found at the Mikulov site (Czech Republic). The presented communication considers various diseases that could be the cause of this pathological condition (tuberculosis, syphilis, mycosis, parasitic diseases, tumors, peritoneal abscess). The examination methods used in this case were macroscopic examination, radiological, histological, and genetic analyses. Differential diagnosis helps us to focus upon the most likely etiologies, and in our opinion, we feel this erosive lesion was probably caused by inflammation from a peritoneal abscess associated with chronic appendicitis.

BACKGROUND:Short tandem repeats (STRs) are genetic markers frequently used for human identification and paternity testing. They are highly mutable, which may occasionally lead to inconsistencies between the genotypes of parents and their children. As the mutation rates of individual STR markers can vary among populations, population-specific data are of high importance.AIM:To investigate the mutation rates of 16 STR markers in the Slovak population.SUBJECTS AND METHODS:In this study, we analysed the germline mutation rates of 16 STR markers (TH01, D3S1358, vWA, D21S11, D16S539, D1S1656, D19S433, SE33, D10S1248, D22S1045, D12S391, D8S1179, D2S1338, D2S441, D18S51 and FGA) in the Slovak population. At these loci, we analysed 42 096 allelic transfers and identified 61 mutation events.RESULTS:The loci with the highest overall mutation rates were SE33 and FGA, while no mutations were identified in TH01, D19S433 and D22S1045. The average paternal mutation rate was higher than the maternal mutation rate. All but one mutation consisted of gains or losses of a single repeat unit and the overall mutation rate was estimated to be 1.45 x10-3 per meiosis.CONCLUSION:This study provides data which can be used to further strengthen the correct paternity index calculations and reliability of paternity testing in Slovakia.