‘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.

Introduction: In spite of measures to avoid invasive mechanical ventilation, many preterm infants are still artificially ventilated. The need for intubation and positive pressure ventilation is associated with so-called ventilator-induced lung injury (VILI) and bronchopulmonary dysplasia (BPD). A lot of strategies are made in order to minimise VILI. One of these strategies is the use of permissive hypercapnia, in which clinicians use more gentle ventilatory strategies and accept higher than “normal” alveolar partial pressure of carbon dioxide (PaCO2) values. Although there are promising studies about the use of permissive hypercapnia in preterm infants, we are still not sure if and when this mode of treatment is safe.Aim: The aim of this study is to investigate conditions in which permissive hypercapnia is safe to prematurely born infants regarding their survival.Methods: The present study was conducted in a tertiary research and educational hospital, NICU, Pediatric Clinic, Clinical Center University of Sarajevo (Sarajevo, Bosnia and Herzegovina). All infants had chest X-ray at admission, and were treated for respiratory distress syndrome (RDS) with nasal continuous positive airway pressure (nCPAP), conventional mechanical ventilation (CMV), or high frequency oscillatory ventilation (HFOV). At admission we registered data regarding birth weight (BW), gestational age in weeks (GW), Apgar score and prenatally given steroids. Inclusion criteria were fulfilled by 200 infants. According to their mean PaCO2, patients were divided into hypercapnia and normocapnia groups. We analyzed the outcome (survival) of these two groups.Results: The two groups didn’t differ regarding GW, prenatally given steroids, RDS severity, surfactant use, 1- and 5-minute Apgar score, nor according to their CRIB score. Groups had also similar survival. After performing ROC analysis we have found that infants born ≤ 27 GW and ≤ 1,000 g treated with permissive hypercapnia, and infants with normocapnia born ≤ 26 GW and ≤ 980 g, have a prediction of negative outcome regarding survival, with a high level of accuracy.Conclusions: This study shows that ventilation with permissive hypercapnia of preterm infants with RDS is not safe, considering survival in children with GW ≤ 27 and BW ≤ 1,000 g.

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.

Helleborus multifidus Vis. is endemic Illyric-Adriatic species with distribution range in Italy, Slovenia, Croatia, Bosnia and Herzegovina, Montenegro and Albania. Although few studies reported different taxonomic categories for H. multifidus, this one is the first using molecular-genetic markers (trnL region and matK of chloroplast DNA and nuclear ITS1 and ITS2 region) for genetic characterization of H. multifidus presented at three localites in Bosnia and Herzegovina. The results revealed that PCRRFLP on trnL intron was not informative for testing interor intrapopulation diversity. Contrary, analysis of matK, ITS1 and ITS2 sequences showed differences between populations from Trebinje region and Kupreško polje, pointing to the need to include additional analyses in order to confirm these findings.

. Apple genetic resources in Norway are currently conserved within a number of local clonal archives. However, during establishment of these ex situ collections, primary focus was not on capturing as much of the diversity as possible, but instead on preserving cultivars of particular importance to specific fruit-growing areas. To identify redun- dancies within the collection as well as to assess the genetic diversity and structure of apple germplasm currently being conserved in Norway, eight microsatellites were used in genetic characterization of 181 apple accessions. Overall, 14 cases of synonym or possibly mislabeled accessions were identified, as well as several homonyms and duplicates within and among the analyzed collections. The information obtained should contribute to overall better management of the preserved germplasm. Bayesian analysis of genetic structure revealed two major clusters, one containing most of the foreign cultivars, while the other consisted mainly of traditional Scandinavian cultivars, but also some very winter-hardy genotypes such as ‘Charlamovsky’, ‘Gravenstein’, ‘Transparente Blanche’, and ‘Wealthy’. Analyses of molecular variance (AMOVA) detected a signifi- cant genetic differentiation among the clusters ( f CT = 0.077; P < 0.01).

UDK: 582.675.1:575(497.6) Helleborus multifidus Vis. is endemic Illyric-Adriatic species with distribution range in Italy, Slovenia, Croatia, Bosnia and Herzegovina, Montenegro and Albania. Although few studies reported different taxonomic categories for H. multifidus, this one is the first using molecular-genetic markers (trnL region and matK of chloroplast DNA and nuclear ITS1 and ITS2 region) for genetic characterization of H. multifidus presented at three localites in Bosnia and Herzegovina. The results revealed that PCR-RFLP on trnL intron was not informative for testing inter- or intrapopulation diversity. Contrary, analysis of matK, ITS1 and ITS2 sequences showed differences between populations from Trebinje region and Kupreško polje, pointing to the need to include additional analyses in order to confirm these findings.

The focus of this study was on genetic diversity of TB horse population raised in B&H. Genomic DNA was genotyped by using 17 microsatellite markers. A total of 103 alleles were detected. The average number of alleles per locus was 6.059 and effective number of alleles was 3.293. Means of observed and expected heterozygosity were calculated 0.645 and 0.696, respectively. The average PIC values was 0.649 and inbreeding coefficient was 0.090. Based on all observed parameters, ASB2 locus showed the highest genetic diversity while locus HMS2 was the least diverse. These results suggest that the population of TB horses from B&H is not affected by substantial loss of genetic diversity, indicating the presence of reasonably high level of genetic variability.

OBJECTIVES The population analysis of cardiovascular risk and non-risk genetic variation can help to identify adaptive or random demographic processes that shaped coronary incidence variation across geography. MATERIAL AND METHODS In this study, 114 single nucleotide polymorphisms and 17 tandem repeat polymorphisms from Nitric Oxide Synthases (NOS) regions were analyzed in 1686 individuals from 35 populations from Europe, North Africa, and the Middle East. NOS genes encode for key enzymes on nitric oxide availability, which is involved in several cardiovascular processes. These genetic variations were used to test for selection and to infer the population structure of NOS regions. Moreover, we tested whether the variation in the incidence of coronary events and in the levels of classical risk factors in 11 of these European populations could be explained by the population structure estimates. RESULTS Our results supported, first, the absence of clear signs of selection for NOS genetic variants associated with cardiovascular diseases, and second, the presence of a continuous genetic pattern of variation across European and North African populations without a Mediterranean barrier for gene flow. Finally, population structure estimates from NOS regions are closely correlated with coronary event rates and classical risk parameters (explaining 39-98%) among European populations. CONCLUSION Our results reinforce the hypothesis that genetic bases of cardiovascular diseases and associated complex phenotypes could be geographically shaped by random demographic processes.