A PRELIMINARY STUDY OF THE PALEOLITHIC AND NEOLITHIC CONTRIBUTION THE EUROPEAN MTDNA FLOW IN SHAPING THE GENETIC STRUCTURE OF RECENT BOSNIAN POPULATION

The mitochondrial DNA (mtDNA) polymorphisms in Bosnian human population was analyzed by means of hypervariable segment I and II (HVSI and HVSII) sequencing and restriction fragment-length polymorphism analysis of the mtDNA coding region. The results suggest that shaping the genetic structure of recent Bosnian population likely to be affected by the expansion from the European glacial refuges area at the end of the Last Glacial Maximum (LGM), postglacial expansions from southwestern refuges of Europe, the Italian Peninsula and the dispersion in periods of more recent historical events, from the East European Plain. Especially interesting feature of the Neolithic expansion in this area is the ancient African/South Asian haplogroup N1a with the HVSI variant 16147G, which is almost absent in Europe. The haplotyps HVSI with variant 16147G suggest the colonization of the Northeast Bosnia region by Neolithic communities in the Early Neolithic period of expansion through Europe, as evidenced by the archaeological remains of the Starcevo culture

Molecular Variation of Rh, MN, Duffy, Kidd, Kell, and Lutheran Blood Groups in the Human Population of Bosnia and Herzegovina

Six blood groups (Rh, MN, Duffy, Kidd, Kell and Lutheran) were investigated among three major ethnic groups (Bosniaks, Bosnian Croats, and Bosnian Serbs) as well as ten regional subpopulations across Bosnia and Herzegovina (Krajina, Posavina, North-Eastern Bosnia, Eastern Bosnia, Middle Bosnia, Central Bosnia, Sarajevo Region, Eastern Herzegovina, Central Herzegovina, and Western Herzegovina). This is the first study that introduces the molecular genetic typing of five blood groups within the population of Bosnia and Herzegovina with the exception of RhD blood group. The sample consisted of 450 buccal swabs from unrelated individuals. Five blood group systems (RhD, RhC, RhE, Kidd, MN) were genotyped by the PCR-SSP method, while three blood group systems (Kell, Duffy, Lutheran) were genotyped by the PCR-RFLP method. Minor variation of genetic diversity was observed within the three major ethnic groups in B&H as well as within the ten subpopulations stratified according to geographical criteria. No genetic differentiation among ethnic groups was noticed. These results are in agreement with the results of previous studies based on different molecular genetics markers, which indicate that the three ethnic groups of Bosnia and Herzegovina belong to the same gene pool. A similar level of genetic variance was observed within regional subpopulations with no significant genetic differentiation among them. Comparison of intrapopulation genetic diversity of the Bosnian and Herzegovinian population with other European and non-European populations, based on the three loci (RHD, MN and KEL), clearly showed that the level of genetic diversity of the Bosnia and Herzegovina population is within the European range

Decrease in body mass index: Personal genotyping, individual diet, and exercise plan

Introduction: Single nucleotide polymorphisms (SNPs) have lately been used for prediction of metabolic processes that may be related to obesity. The aim of our study was to examine the association of SNPs of several genes with obesity and physical activity in 18 healthy volunteers. Methods: We used buccal swabs to collect and extract DNA from 18 volunteers. Pyrosequencing was used for molecular analysis of 13 polymorphisms in 10 genes (APOA2, MTHFR, MCM6, peroxisome proliferators-activated receptor gamma, FABP2, beta-2-adrenergic receptor (ADRB)2, ADRB3, A-actinin-3, angiotensin-converting enzyme, and FUT2). The volunteers’ personal data included body mass index (BMI), dietary practice and information on daily fitness and workout routine. Association between the 13 observed gene polymorphisms and individual BMI status (normal or overweight) was analyzed. Results of the DNA analysis were used for the expert evaluation by nutritionists and physiologists to obtain optimal regulation of nutrition and exercise. The volunteers had a dietary and fitness program for 12 months which they tracked by filling in a suitable study form. Results: 14 volunteers had a moderate genetic predisposition for abdominal adipose-tissue accumulation, while 4 of them had genotypes not associated with abdominal fat tissue accumulation. A statistically significant difference was found between the value of BMI before and after the implementation of personalized training and nutrition plan within the group of overweight volunteers (paired sample t=3.382; p = 0.006; exact p = 0.015). The single-locus F-test showed no association between the gene polymorphisms and BMI values. In addition, no correlation was detected between the gene polymorphisms and amount of BMI reduction prior and after the implementation of the personalized training and nutrition plan within the overweighed group of volunteers. Conclusion: Optimal nutrition and training plan are crucial for the BMI reduction as observed in the overweighed volunteers after the 12-month personalized training and individualized nutrition plan. However, the analyzed polymorphisms were not significantly associated with the obesity in this study

Identifying Pollen Donors and Success Rate of Individual Pollinizers in European Plum (Prunus domestica L.) Using Microsatellite Markers

European plum cultivars (Prunus domestica L.) are hexaploid and partially self-fertile or self-sterile requiring compatible pollinizers with overlapping bloom times. Therefore, inter-planting of different pollinizer cultivars is recommended. In order to identify successful pollinizers of the plum cultivars ‘Edda’, ‘Opal’ (self-fertile), ‘Jubileum’, ‘Reeves’, ‘Mallard’, ‘Avalon’, ‘Cacanska Lepotica’ (self-fertile), and ‘Valor’, 60 fruits per cultivar were collected from nine orchards in 2017 and 2018, all of which were located in Ullensvang, western Norway. DNA extraction was subsequently conducted from the obtained embryos, followed by genetic characterization using seven microsatellite markers. Tissue samples from all possible pollinizers were collected during the summer of 2017 and the same DNA approach was conducted. Results showed that ‘Opal’ was the most successful pollinizer among the investigated plum cultivars. The main exception was ‘Cacanska Lepotica’, which consistently displayed very high level of self-pollination. The most successful foreign pollinizer of ‘Opal’ was ‘Mallard’. However, in more than two thirds of embryos extracted from ‘Opal’ fruits self-fertilization was determined. ‘Reeves’ was identified as the most successful pollinizer among embryos collected from ‘Valor’. Among the five cultivars (‘Edda’, ‘Jubileum’, ‘Reeves’, ‘Mallard’, and ‘Avalon’) that did not display self-pollination, the pollinizer success rate of ‘Opal’, ranged from 36.5% (‘Mallard’) to 93.5% (‘Edda’) in 2017, while in 2018 this rate ranged from 43.5% (‘Jubileum’ and ‘Reeves’) up to 96.5% (‘Edda’). Overall, genotyping embryos using SSRs (simple sequence repeats) proved an effective method in determining the success rate of individual pollinizers among European plum cultivars.publishedVersio

Frequency of BRCA1 Mutations in B&H Breast and Ovarian Cancer Patients

Incidence of breast cancer ranges from 27 per 100,000 in Middle Africa and Eastern Asia to 92 per 100,000 in Northern America. It is the fifth most common cause of death from cancer in women, with an estimated 522,000 deaths per year (6.4% of the total). Autosomal dominant inheritance of these cancers is characterized by transmission of cancer predisposition from generation to generation, with around 5-10% of all breast cancers being associated with inherited mutations in BRCA1, BRCA2 and other genes.  Breast and ovarian cancers are strongly associated with BRCA1 and BRCA2 mutations. In this study, we genotyped BRCA1 gene for large genomic rearrangements in breast and ovarian cancer patients from Bosnia and Herzegovina, with aim to assess frequency of large BRCA1 mutations (exon deletions/duplications) in this group. We collected 59 breast cancer samples, as well as other data concerning patients’ histopathological parameters of tumor, like age at diagnosis, cancer type, TNM class, cancer grade, as well as estrogen, progesterone and Her2/neu expression. Following DNA extraction from breast cancer samples (tissue after biopsy), BRCA1 mutations were identified by Multiplex Ligase - Dependent Probe Amplification (MLPA) analysis. Biostatistical analyses were conducted using MedCalc v.9.2.0.0 software. In all statistical tests p<0.05 was considered significant. Mean age at diagnosis was 54±1.75 (range 17 – 80). BRCA1 genomic rearrangements were found in 22% of breast and ovarian cancer patients. Statistically significant associations and correlations were found between BRCA1 genomic rearrangements and cancer type, estrogen, progesterone and Her2/neu expression, but not cancer grade, size, invasiveness or patients’ ag

Pollinizer efficacy of several ‘Ingeborg’ pear pollinizers in Hardanger, Norway, examined using microsatellite markers

‘Ingeborg’ is currently the main commercial pear cultivar grown in Norway. However, fruit set and subsequent yields of this cultivar have proven to be variable and overall low averaging 10–20 t·ha−1. Pear seeds found in ‘Ingeborg’ fruits are often underdeveloped, suggesting that incomplete fertilization might be a major cause of poor fruit set. In some years, sporadically unfavorable environmental conditions during and immediately after pollination in Hardanger district, western Norway, have resulted in poor fruit set of ‘Ingeborg’. In this study, the pollinizer efficacy of several pollinizers, namely ‘Clara Frijs’, ‘Herzogin Elsa’, ‘Anna’, ‘Colorée de Juillet’, and ‘Belle lucrative’, from several orchards located in the Hardanger district was investigated using 12 microsatellite markers for two growing seasons (2014 and 2016). Pollinizer efficacy was estimated by genotyping ‘Ingeborg’, each individual pollinizer, as well as normally developed seeds from ‘Ingeborg’ fruit, and conducting gene assignment analyses to identify the pollen contribution from each of the pollinizer cultivars. In addition, S-allele genotyping was conducted, and only one pollinizer, ‘Anna’, was identified as being semicompatible with ‘Ingeborg’, whereas all other pollinizers were fully compatible. ‘Clara Frijs’ and ‘Belle lucrative’ were identified as the most efficient pollinizers probably because these cultivars were abundant compared with all other pollinizers within all, but one of the examined orchards. Higher yields could not be attributed to a particular pollinizer, and genetic effects associated with the triploid nature of ‘Ingeborg’ are most likely implicated as a cause behind the low and variable yield of this cultivar.publishedVersio

Pollinizer efficacy of several ‘Ingeborg’ pear pollinizers in Hardanger, Norway, examined using microsatellite markers

‘Ingeborg’ is currently the main commercial pear cultivar grown in Norway. However, fruit set and subsequent yields of this cultivar have proven to be variable and overall low averaging 10–20 t·ha−1. Pear seeds found in ‘Ingeborg’ fruits are often underdeveloped, suggesting that incomplete fertilization might be a major cause of poor fruit set. In some years, sporadically unfavorable environmental conditions during and immediately after pollination in Hardanger district, western Norway, have resulted in poor fruit set of ‘Ingeborg’. In this study, the pollinizer efficacy of several pollinizers, namely ‘Clara Frijs’, ‘Herzogin Elsa’, ‘Anna’, ‘Colorée de Juillet’, and ‘Belle lucrative’, from several orchards located in the Hardanger district was investigated using 12 microsatellite markers for two growing seasons (2014 and 2016). Pollinizer efficacy was estimated by genotyping ‘Ingeborg’, each individual pollinizer, as well as normally developed seeds from ‘Ingeborg’ fruit, and conducting gene assignment analyses to identify the pollen contribution from each of the pollinizer cultivars. In addition, S-allele genotyping was conducted, and only one pollinizer, ‘Anna’, was identified as being semicompatible with ‘Ingeborg’, whereas all other pollinizers were fully compatible. ‘Clara Frijs’ and ‘Belle lucrative’ were identified as the most efficient pollinizers probably because these cultivars were abundant compared with all other pollinizers within all, but one of the examined orchards. Higher yields could not be attributed to a particular pollinizer, and genetic effects associated with the triploid nature of ‘Ingeborg’ are most likely implicated as a cause behind the low and variable yield of this cultivar.publishedVersio

S-Genotyping and Seed Paternity Testing of the Pear Cultivar ‘Celina’

The diploid Celina/QTee® (‘Colorée de Juillet’ × ‘Williams’), one of the most promising pear cultivars developed by the Norwegian breeding program Graminor, was launched in 2010. In Norway, the flowering is medium to late, while the fruits ripen in the beginning of September. The fruits are attractive with an intense red blush (50%) on a green background. Although, ‘Celina’ is cultivated in the most climatically suitable regions for fruit cultivation, present in Norway, unfavorable environmental conditions for pear pollination can have a very negative effect on fruit set and consequent yield. The aim of this study was to determine the S-alleles of ‘Celina’, as well as its frequently used pollinizers, and, through paternity testing of ‘Celina’ seeds, give a recommendation regarding the most important pollinizers of this pear cultivar. In order to accomplish this, ‘Celina’ and its potential pollinizers were all S-genotyped. After harvest, seeds collected from ‘Celina’ fruit in 2017 and 2018 were genotyped using eleven microsatellite markers. Genomic DNA was also extracted from leaf material collected from ‘Celina’, as well as from five pear cultivars used as pollinizers in the three examined orchards, and analyzed using the same marker set. Subsequently a simple sequence repeat (SSR) database was constructed and used for gene assignment analyses with the aim of quantifying pollen donor contribution from individual pollinizers. The obtained results indicate that ‘Anna’, the only examined pollinizer that was fully cross-compatible with ‘Celina’, together with ‘Fritjof’, the genotype which had the highest flowering overlap with ‘Celina’, proved to be the most successful pollinizers across all seasons and orchards. Although both cultivars were ubiquitous in the examined orchards, either as planted trees or as branches introduced during the flowering period, they were the most abundant pollinizers in only one orchard each. It is therefore possible to conclude that pollinizer abundance has a secondary significance in pollinizer success within investigated ‘Celina’ orchards.publishedVersio

Inhibitory potential of phytochemicals on five SARS-CoV-2 proteins: in silico evaluation of endemic plants of Bosnia and Herzegovina

AbstractThe outbreak of the coronavirus disease 2019, caused by the SARS-CoV-2 virus, has prompted global health concerns. In response, researchers have been conducting investigations on active compounds in plants that may hold the potential to inhibit the proliferation of the virus. The aim of this study was to simulate and predict structural interactions of selected compounds isolated from 28 endemic plants of Bosnia and Herzegovina against the main protease (Mpro), papain-like protease (PLpro), RNA-dependent RNA polymerase (RdRp), spike glycoprotein and uridylate-specific endoribonuclease (NendoU) of SARS-CoV-2. The majority of compounds, especially hesperidin, showed great binding affinity to the target proteins. The highest affinity for Mpro was observed for genistein and hesperidin, while in terms of structural interactions, both compounds achieved interactions of interest. Hesperidin and luteolin were the compounds with the highest binding affinity for PLpro, but no significant interactions were observed. For RdRp, hesperidin and quercetin showed the highest binding affinity, where both compounds formed interactions of interest. Hesperidin and fisetin were the compounds with the highest binding affinity for spike glycoprotein, and both compounds achieved significant interactions. The highest affinity for NendoU was obtained for hesperidin and isorhamnetin, where both compounds formed interactions of interest. Although these findings appear encouraging, further research is needed, which includes in vitro and in vivo assessments, along with clinical trials, to provide evidence for the potential therapeutic uses of these plants