The Dinaric endemic plant species Moltkia petraea (Tratt.) Griseb. is often called a "living fossil" of ancient Tertiary flora, with great importance for Bosnia and Herzegovina’s biodiversity. Considering its narrow and limited distribution range, insufficient data on the molecular background of this species is given so far. Due to the presence of various secondary metabolites that interfere with the DNA, isolation of nucleic acids from plant cells is known to be challenging. Even in closely related species it is necessary to optimize DNA isolation protocol in order to obtain high quality PCR amplifiable DNA. We collected 91 samples from five populations in Herzegovina. Doyle and Doyle (1987) CTAB protocol was modified by adding vitamin C (ascorbic acid) to the cell lysis buffer to improve DNA yield and quality. trnL(UAA) intron and nrDNA (ITS1, ITS2) molecular markers were applied to demonstrate amplifiability of isolated DNA and elucidate the intra- and interpopulation genetic diversity. Our results suggest a successful PCR amplification for 81% of the analyzed samples. PCR-RFLP analysis of trnL(UAA) revealed that all individuals in five populations have the same haplotype based on the obtained enzymatic profile for three enzymes (TaqI, HinfI, HindII). Alignment and comparison of ITS sequences didn’t reveal any hypervariable portion that could be informative in elucidating the genetic diversity of M. petraea populations. Further studies with additional application of microsatellite loci, RAPD and AFLP methods are necessary in an attempt to get insights into the genetic diversity of M. petraea.

The focus of this study was microsatellite diversity of crossbred horses raised in Bosnia and Herzegovina. Genomic DNA was extracted from blood samples of 20 individuals (KBA group – 7 individuals, crosses between Bosnian and Herzegovinian mountain horse and Arabian horse; KBR group – 9 individuals, crosses between Bosnian and Herzegovinian mountain and Belgian horses, crosses between Bosnian and Herzegovinian mountain horses and Holstein, crosses between Bosnian and Herzegovinian mountain and Lipizzaner horses and KBN group – 4 individuals, crosses between Bosnian and Herzegovinian mountain horse with an unknown origin of the other parent). The samples were profiled using 17 microsatellite markers. This method consisted of multiplex PCR procedure and generated reasonable amplification across all the loci. All samples were genotyped successfully. Considering all the observed parameters, VHL20 locus showed the highest microsatellite diversity. Locus HMS7 was the least variable in KBR group, while HMS1 locus was the least diverse in KBN group. The highest microsatellite diversity in KBA group was found at AHT5 locus while HTG6 locus was the least diverse. Obtained results suggest that the investigated populations of crossbred horses from Bosnia and Herzegovina are not affected by substantial loss of genetic diversity, as indicated by the presence of reasonably high level of genetic variation. An increase in the inbreeding coefficient and sufficient heterogenity in KBN group indicate occurrence of consanguineous mating. The present research contributes to the knowledge of current status of genetic structure of the investigated crossbred horses.

Cervical cancer represents a serious health problem affecting women worldwide especially in developing countries due to low socioeconomic status, inadequate health-care infrastructure, weaknesses in education on this particular issue and lack of effective screening programmes. The primary aim of this study was to assess alternative screening method for the improvement of cervical cancer prevention in conditions of Bosnia and Herzegovina (B&H), which could be applicable in other developing countries as well. The study was conducted on 101 subjects who provided their self-sampled vaginal swabs and/or cervical specimens collected by their gynecologists. Universal Human Papilloma Virus (HPV) primer set optimized to detect a wide range of HPV types was used for HPV genotyping from obtained swab samples in multiplex PCR. Amplicons were analyzed in agarose gel and Agilent 2100 bioanalyzer – a platform based on microfluid technology. Inter-rater agreement kappa (MedCalc2) was used to assess concordance between results of cervical and vaginal sample analysis. Out of 39 subjects who provided their vaginal and cervical samples, results of HPV detection mismatched in 10% of the cases. Inter-rater agreement showed good strength of coincidence between the results of cervical and vaginal sample analysis (kappa=0,748, CI=95 %). We presented an alternative PCR method for the detection of HPV based on vaginal self sampling which is affordable, informative, simple and applicable with high coverage level of defined targeted population and potentially significant in the given cultural and socioeconomic context.

‘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. 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 inHardanger district, westernNorway, have resulted in poor fruit set of ‘Ingeborg’. In this study, the pollinizer efficacy of several pollinizers, namely ‘Clara Frijs’, ‘Herzogin Elsa’, ‘Anna’, ‘Color ee 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. BP 10273 pear (‘Conference’ · ‘Bonne Louise’) was bred at SLU Balsg ard (Swedish University of Agricultural Sciences), and after evaluation in Western Norway, it was named ‘Ingeborg’ in 1994 (Hjeltnes and Ystaas, 1993). This cultivar has become the most widely planted commercial pear variety grown in Norway, including the Hardanger district, western Norway. ‘Ingeborg’ is a triploid (3x) and is believed to be the result of fertilization of an unreduced diploid (2n) egg cell from ‘Conference’ with a haploid (n) pollen cell from ‘Bonne Louise’ (Sehic et al., 2012). Although ‘Ingeborg’ possesses good pomological traits that are highly desirable to Norwegians, fruit set and subsequent yields of this cultivar tend to be erratic and significantly lower than other pear varieties grown in Norway (Meland and Frøynes, 2014). Yields vary significantly between different orchards in the Hardanger region within any one growing season and parthenocarpy may play a role. Seeds extracted from ‘Ingeborg’ fruits are frequently underdeveloped. Triploids typically have low fertility due to a reproductive barrier whereby three sets of chromosomes cannot be divided evenly during meiosis yielding unbalanced segregation of chromosomes (Phillips et al., 2016). Triploids are typically highly infertile; however, limited fertility and seed production can result from the formation of apomictic embryos or through the union of aneuploid or unreduced gametes (Ramsey and Schemske, 1998). It should be noted that pears, which are auto-incompatible, may have seeds even if they are self-pollinated. Ny eki et al. (1998) found that even during self-pollination, pears can bear fruits, which are a) entirely seedless (parthenocarpic), or b) the seeds were empty or flat without any viable germination, or c) some viable seeds developed at a low rate (0.5% to 2%) in addition to empty seeds. Incompletely formed seeds, low seed number per fruit, or both have reduced sink strength (Weinbaum et al., 2001), which results in lower fruit weight and decreased yields. Self-fertilization in European pears (Pyrus communis L.), similar to other fruit species of the Rosaceae family, is prevented by gametophytic self-incompatibility (Crane and Lewis, 1942). Consequently, interplanting of suitable pollinizer genotypes in pear orchards is essential for fertilization of the ovules, which in itself is necessary for a successful set of an optimum crop load (Webster, 2002). Identifying cross-compatible pear cultivars is traditionally accomplished with testcrosses and more recently using polymerase chain reaction (PCR) based S-genotyping (Mota et al., 2007; Quinet et al., 2014; Sanzol, 2009). However, planting cross-compatible pollinizers, which have coincidental flowering time overlap with the main commercial cultivar, does not always guarantee consistently high yields. In addition, environmental variables, such as rainfall, temperature, and cloud cover, may also negatively affect pollinators and the effective pollination period (EPP) (Sanzol and Herrero, 2001). EPP is defined as the difference between the ovule longevity minus the time between pollination and fertilization (Williams, 1965). Because of generally unfavorable environmental conditions for pear pollination during the Nordic spring, ‘Ingeborg’ orchards have been established using multiple pollinizer cultivars. Despite this, yields are often low and erratic in some ‘Ingeborg’ orchards in Hardanger, Norway, and this requires further investigation to identifywhich of the pollinizers is the most effective, both in high and poor yielding orchards. Determining pollen compatibility of individual pollinizers may be accomplished by 1722 HORTSCIENCE VOL. 52(12) DECEMBER 2017 genotyping progeny plants produced by germinating seeds extracted from pear fruits of the main commercial cultivar and using the obtained molecular data to identify the male parent. However, the occurrence of aneuploid seedlings with poor viability, frequent among triploids (Zhang and Park, 2009) such as ‘Ingeborg’, makes the above procedure impractical. Consequently, genetic analyses should be performed on the pear seeds themselves instead of the progeny plants. The seedlings that are produced from seeds will be primarily aneuploids because of unbalanced chromosome segregation in meiosis (Brownfield and Kohler, 2011) with poor viability due to the triploid nature of ‘Ingeborg’. However, limited number of progeny frommaternal triploids could also be diploids (generational reversion) and tetraploids (fertilization from unreduced gametes from one or both parents) (Phillips et al., 2016). Microsatellite markers or simple sequence repeats (SSRs) have proven efficient in parent-offspring analyses on pear (Kimura et al., 2003). Although a comparative study has shown that the identification of a highly informative set of single-nucleotide polymorphism (SNP) from a large panel showed significantly more accurate individual genetic assignment compared with the combination of SSR loci (Glover et al., 2010), Moore et al. (2014) found that microsatellite markers are accurate genetic markers for genetic assignment, especially in combination with informative SNPs. In the case of plant parentage, pollination and dispersal analyses, and microsatellites with their various limitations remain an important genetic marker (Ashley, 2010). In addition, there are several readily available microsatellite markers at present, developed from either apple (Gianfranceschi et al., 1998; Liebhard et al., 2002) or pear (Fern andezFern andez et al., 2006), that can be used in the genetic analyses of European pear genotypes. In this study, pollination efficacy of several commonly used ‘Ingeborg’ pollinizers in the Hardanger region was investigated using microsatellite markers. To examine the causes of fertilization between Ingeborg vs. all pollinizer cultivars, molecular analyses of S alleles were performed. Materials and Methods The environmental conditions in Ullensvang, a municipality of Hardanger, Norway’s biggest fruit producing region, during flowering were conducive to pollination of ‘Ingeborg’ in 2014 and 2016 (Table 1). Because of the unfavorable climatic conditions in 2015 [low minimum temperatures (<7.3 C) and prolonged heavy rainfall during bloom] that contributed to the low fruit set and insufficient even for research sampling, this year was excluded. Dates of first bloom (BBCH 60), full bloom (80% of blossoms open), and petal fall (80%) (Jackson and Looney, 1999) for ‘Ingeborg’ in six commercial orchards as well as for five pollinizer cultivars during 2014 and 2016 are presented in Table 2, confirming that there was sufficient overlap between all pollinizers and ‘Ingeborg’ in both 2014 and 2016. At harvest, 50 randomly sampled ‘Ingeborg’ fruit were gathered from each of the six commercial orchards. Fruits were cut open and all pear seedswere extracted. Orchard size, yield, and age of the six different orchards are presented in Table 3. Pear producers did not provide beehives for pollination, but neighboring farms that are growing cherries and plums are renting beehives for pollination. The distance between some pear orchards and these beehives was 100 m and more. Molecular and phenology analyses. Tissue samples (leaves) for DNA analyses were collected in the Spring of 2014 from a single tree of the main commercial cultivar (Ingeborg) and from pollinizer genotypes (‘Clara Frijs’, ‘Herzogin Elsa’, ‘Anna’, ‘Color ee de Juillet’, and ‘Belle lucrative’) present in the analyzed orchards. The genomic DNA was isolated from 70 to 80 mg of leaf powder using the CTAB method (Cullings, 1992; Doyle and Doyle, 1987). Extraction and isolation of genomic DNA from pear seeds were conducted according to Padmalatha et al. (2008). As it was impossible to obtain enough high-quality DNA from a single seed for the genetic characterization, well developed seeds collected from each individual orchard were mixed and ground together to obtain a single homogenous sample. Twelve SSR primer pairs (Table 4) were chosen based on their polymorphism observed in a previous study on European pears (Gasi et al., 2013). All PCR reactions were carried out in accordance with the protocol described by Gasi et al. (2013

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.

This study presents genetic data for nine Native American populations from northern North America. Analyses of genetic variation focus on the Pacific Northwest (PNW). Using mitochondrial, Y chromosomal, and autosomal DNA variants, we aimed to more closely address the relationships of geography and language with present genetic diversity among the regional PNW Native American populations. Patterns of genetic diversity exhibited by the three genetic systems were consistent with our hypotheses: genetic variation was more strongly explained by geographic proximity than by linguistic structure. Our findings were corroborated through a variety on analytic approaches, with the unrooted trees for the three genetic systems consistently separating inland from coastal PNW populations. Furthermore, analyses of molecular variance support the trends exhibited by the unrooted trees, with geographic partitioning of PNW populations (FCT = 19.43%, p = 0.010 ± 0.009) accounting for over twice as much of the observed genetic variation as linguistic partitioning of the same populations (FCT = 9.15%, p = 0.193 ± 0.013). These findings demonstrate a consensus with previous PNW population studies examining the relationships of genome-wide variation, mitochondrial haplogroup frequencies, and skeletal morphology with geography and language.

Aim To determine newest the most accurate allele frequencies for 15 short tandem repeat (STR) loci in the Bosnian and Herzegovinian population, calculate statistical parameters, and compare them with the relevant data for seven neighboring populations. Methods Genomic DNA was obtained from buccal swabs of 1000 unrelated individuals from all regions of Bosnia and Herzegovina. Genotyping was performed using PowerPlex® 16 System to obtain allele frequencies for 15 polymorphic STR loci including D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, and FGA. The calculated allele frequencies were also compared with the data from neighboring populations. Results The highest detected value of polymorphism information content (PIC) was detected at the PentaE locus, whereas the lowest value was detected at the TPOX locus. The power of discrimination (PD) values had similar distribution, with Penta E showing the highest PD of 0.9788. While D18S51 had the highest value of power of exclusion (PE), the lowest PE value was detected at the TPOX locus. Conclusion Upon comparison of Bosnian and Herzegovinian population data with those of seven neighboring populations, the highest allele frequency differentiation was noticed between Bosnian and Herzegovinian and Turkish population at 5 loci, the most informative of which was Penta E. The neighbor-joining dendrogram constructed on the basis of genetic distance showed grouping of Slovenian, Austrian, Hungarian, and Croatian populations. Bosnian and Herzegovinian population was between the mentioned cluster and Serbian population. To determine more accurate distribution of allelic frequencies and forensic parameters, our study included 1000 unrelated individuals from all regions of Bosnia and Herzegovina, and our findings demonstrated the applicability of these markers in both forensics and future population genetic studies.

Raspberry cultivars are clonally propagated and therefore all plants belonging to a single cultivar represent the same genotype. Cultivar integrity of raspberry plantlings placed on the market in Bosnia and Herzegovina (B&H) is based on examining of morphological traits, which is not a reliable tool for genetic identification. In this study plantlings declared as cultivar ‘Polana’ were genotyped using seven microsatellites, in order to gain preliminary insight into the genetic integrity of raspberry plantlings marketed in B&H. Plant tissue (leaves) from 10 raspberry plants were randomly sampled from a batch of plantlings sold by major fruit nursery in Bosnia and Herzegovina. Along with these samples, four reference cultivars with confirmed identity (‘Polka’, ‘Autumn Bliss’, ‘Heritage’ and ‘Polana’) were also included in the study. Seven primer pairs amplified 31 alleles, or on average 4.4 alleles per locus. UPGMA cluster analysis, based on the Jaccard similarity coefficient, revealed that among the ten samples declared as ‘Polana’ plantlings only five were genetically identical to any of the other samples. The cluster analyses also exposed that none of the ten samples declared as ‘Polana’ seedlings were in fact identical or even closely related to the ‘Polana’ reference cultivar or any of the other reference cultivars. These findings clearly show that the genetic identity of primocane raspberry plantlings , currently sold in Bosnia and Herzegovina, needs to be tested using objective and reliable methods rather than simple morphologic observation.