Genetic patterns in Pinus nigra from the central Balkans inferred from plastid and mitochondrial data

Pinus nigra J.F. Arnold, European black pine, is a typical component of Mediterranean and sub-Mediterranean coniferous forests with highly fragmentary distribution. Western Mediterranean populations of this species have been studied genetically to date, while eastern populations from the central Balkans, which are larger and more abundant, are still genetically understudied. We analyzed seven populations of P. nigra representing all infraspecific taxa recognized within the central Balkans (subspecies nigra with varieties nigra and gocensis Dordevic; and subspecies pallasiana (Lamb.) Holmboe with varieties pallasiana and banatica (Endl.) Georgescu et Ionescu), with three chloroplast microsatellites (cpDNA SSRs) and one mitochondrial (mtDNA) locus. Although our molecular data failed to support circumscription of studied infraspecific taxa, we found that genetic patterns at both genomes are in accordance with those found previously in westward populations of this species, that is - exceptionally high levels of genetic diversity (H-T = 0.949) and low genetic differentiation (G(ST) = 0.024) at the cpDNA level, and moderate levels of genetic diversity (H-T = 0.357) and genetic differentiation (G(ST) = 0.358) at the mtDNA level. Based on genealogical relations of mtDNA types currently present in Balkans' and Iberian/African populations, we inferred that the ancestral gene pool of P. nigra already harbored polymorphism at position 328 prior to the divergence to two lineages currently present in westward and eastward parts of the species range distribution. Subsequent occurrence of three mutations, which distinguish these two lineages, suggests their long-term isolation

Cross-species amplification of nuclear est-microsatellites developed for other pinus species in pinus nigra

Due to the current lack of nuclear microsatellites (simple sequence repeats SSRs) specifically developed for Pinus nigra, an important European coniferous species, we cross-species amplified 12 EST-SSRs (expressed sequence tagged SSRs) developed for other Pinus species in P. nigra in order to delineate loci which can be used for assessing levels of genetic diversity and genetic structuring in this species. We amplified these loci in individuals from seven populations from the central Balkans representing four recognized infraspecific taxa of P. nigra (ssp. nigra, var. gocensis, ssp. pallasiana, and var. banatica). Contrary to expectations on high transferability of EST-SSRs into related species, only three out of 12 tested loci were successfully amplified in P. nigra, but they displayed lack/low levels of polymorphism or generated multilocus amplification products. Thus, our estimates on levels of genetic diversity (HE = 0.183) and genetic differentiation (F-ST = 0.007) were based on variability of a single locus harboring four alleles only and they should be taken with cautions. Our study highlights the need for the development of high-resolution molecular markers, such as co-dominant genic or genomic SSRs or predominantly biallelic SNPs, or utilization of anonymous dominant markers, such as AFLPs, for genotyping in P. nigra

THE ASSOCIATION BETWEEN MATERNAL ANTHROPOMETRY AND BLOOD PRESSURE IN PREGNANCY – RESULTS FROM THE CROATIAN ISLANDS’ BIRTH COHORT STUDY (CRIBS)

Elevated blood pressure (BP) in pregnancy, especially gestational hypertension and preeclampsia, can lead to serious pregnancy complications and adverse birth outcomes. A large body of literature already reported the effect of baseline body mass index (BMI) on changes in blood pressure during pregnancy. The aim of this study was therefore to define trajectory of systolic (SBP) and diastolic (DBP) blood pressure in 308 pregnant participants from the CRIBS study (146 from the mainland and 162 from the islands of Brač and Hvar) and to analyze the association of blood pressure with maternal BMI prior to pregnancy and maternal anthropometry during pregnancy. Pregnant women included in the CRIBS study had no history of chronic diseases. The BP of CRIBS participants was measured at least once in each trimester, and maternal pre-pregnancy weight was self-reported. All analyses were performed using SPSS 10.0. Results showed that pre-pregnancy BMI was the strongest predictor of pregnancy blood pressure. This association was evident for pre-pregnancy BMI independently (p<0.001), and it also persisted after adjusting for maternal age, education, income, parity, smoking and physical activity (p<0.05). The association between maternal anthropometry during pregnancy and blood pressure was not as strong and was therefore less informative. The study reinforces the role of BMI on SBP and DBP and highlights its importance during prenatal care monitoring. Significant association also emerged between blood pressure in pregnancy and place of residence (mainland vs. island). Women on Dalmatian islands have lower educational level, higher pre-pregnancy BMI and different levels of blood pressure than women from the mainland (namely, higher SBP and lower DBP). Such comparisons between mainland and island populations are valuable, because they can, in the long term, lead to better maternal health care on the islands

Branched-chain amino acid database integrated in MEDIPAD software as a tool for nutritional investigation of mediterranean populations

Branched-chained amino acids (BCAA) are essential dietary components for humans and can act as potential biomarkers for diabetes development. To efficiently estimate dietary intake, we developed a BCAA database for 1331 food items found in the French Centre d'Information sur la Qualité des Aliments (CIQUAL) food table by compiling BCAA content from international tables, published measurements, or by food similarity as well as by calculating 267 items from Greek, Turkish, Romanian, and Moroccan mixed dishes. The database embedded in MEDIPAD software capable of registering 24 h of dietary recalls (24HDR) with clinical and genetic data was evaluated based on archived 24HDR of the Saint Pierre Institute (France) from 2957 subjects, which indicated a BCAA content up to 4.2 g/100 g of food and differences among normal weight and obese subjects across BCAA quartiles. We also evaluated the database of 119 interviews of Romanians, Turkish and Albanians in Greece (27⁻65 years) during the MEDIGENE program, which indicated mean BCAA intake of 13.84 and 12.91 g/day in males and females, respectively, comparable to other studies. The MEDIPAD is user-friendly, multilingual, and secure software and with the BCAA database is suitable for conducting nutritional assessment in the Mediterranean area with particular facilities for food administration

CROSS-SPECIES AMPLIFICATION OF NUCLEAR EST-MICROSATELLITES DEVELOPED FOR OTHER Pinus SPECIES IN Pinus nigra

Due to the current lack of nuclear microsatellites (simple sequence repeats SSRs) specifically developed for Pinus nigra, an important European coniferous species, we cross-species amplified 12 EST-SSRs (expressed sequence tagged SSRs) developed for other Pinus species in P. nigra in order to delineate loci which can be used for assessing levels of genetic diversity and genetic structuring in this species. We amplified these loci in individuals from seven populations from the central Balkans representing four recognized infraspecific taxa of P. nigra (ssp. nigra, var. gocensis, ssp. pallasiana, and var. banatica). Contrary to expectations on high transferability of EST-SSRs into related species, only three out of 12 tested loci were successfully amplified in P. nigra, but they displayed lack/low levels of polymorphism or generated multilocus amplification products. Thus, our estimates on levels of genetic diversity (HE = 0.183) and genetic differentiation (F-ST = 0.007) were based on variability of a single locus harboring four alleles only and they should be taken with cautions. Our study highlights the need for the development of high-resolution molecular markers, such as co-dominant genic or genomic SSRs or predominantly biallelic SNPs, or utilization of anonymous dominant markers, such as AFLPs, for genotyping in P. nigra.Ministry of Education, Science, and Technology of the Republic of Serbia {[}173029, 173030

Biological activity of Pinus nigra terpenes-Evaluation of FtsZ inhibition by selected compounds as contribution to their antimicrobial activity

In the current work, in vitro antioxidant, antibacterial, and antifungal activites of the needle terpenes of three taxa of Pinus nigra from Serbia (ssp. nigra, ssp. pallasiana, and var. banatica) were analyzed. The black pine essential oils showed generally weak antioxidative properties tested by two methods (DPPH and ABTS scavenging assays), where the highest activity was identified in P. nigra var. banatica (IC50=25.08 mg/mL and VitC=0.67 mg (vitamin C)/g when tested with the DPPH and ABTS reagents, respectively). In the antimicrobial assays, one fungal (Aspergilus niger) and two bacterial strains (Staphylococcus aureus and Bacillus cereus) showed sensitivity against essential oils of all three P. nigra taxa. The tested oils have been shown to possess inhibitory action in the range from 20.00 to 0.62 mg/mL, where var. banatica exhibited the highest and ssp. nigra the lowest antimicrobial action. In order to determine potential compounds that are responsible for alternative mode of action, molecular docking simulations inside FtsZ (a prokaryotic homolog of tubulin) were performed. Tested compounds were the most abundant terpenoid (germacrene D-4-ol) and its structurally similar terpene (germacrene D), both present in all three essential oils. It was determined that the oxygenated form of the molecule creates stable bonds with investigated enzyme FtsZ, and that this compound, through this mechanism of action participates in the antimicrobial activity. (C) 2014 Elsevier Ltd. All rights reserved.Ministry of Education and Science of the Republic of Serbia {[}173029

Biological activity of Pinus nigra terpenes-Evaluation of FtsZ inhibition by selected compounds as contribution to their antimicrobial activity

In the current work, in vitro antioxidant, antibacterial, and antifungal activites of the needle terpenes of three taxa of Pinus nigra from Serbia (ssp. nigra, ssp. pallasiana, and var. banatica) were analyzed. The black pine essential oils showed generally weak antioxidative properties tested by two methods (DPPH and ABTS scavenging assays), where the highest activity was identified in P. nigra var. banatica (IC50=25.08 mg/mL and VitC=0.67 mg (vitamin C)/g when tested with the DPPH and ABTS reagents, respectively). In the antimicrobial assays, one fungal (Aspergilus niger) and two bacterial strains (Staphylococcus aureus and Bacillus cereus) showed sensitivity against essential oils of all three P. nigra taxa. The tested oils have been shown to possess inhibitory action in the range from 20.00 to 0.62 mg/mL, where var. banatica exhibited the highest and ssp. nigra the lowest antimicrobial action. In order to determine potential compounds that are responsible for alternative mode of action, molecular docking simulations inside FtsZ (a prokaryotic homolog of tubulin) were performed. Tested compounds were the most abundant terpenoid (germacrene D-4-ol) and its structurally similar terpene (germacrene D), both present in all three essential oils. It was determined that the oxygenated form of the molecule creates stable bonds with investigated enzyme FtsZ, and that this compound, through this mechanism of action participates in the antimicrobial activity. (C) 2014 Elsevier Ltd. All rights reserved.Ministry of Education and Science of the Republic of Serbia [173029