Leukocytes are isolated by centrifugation after specific lysis of erythrocytes

Aim To investigate the prevalence of common genetic variants that can serve as markers of thrombophilia and warfarin pharmacogenetics in Bosnia and Herzegovina. Methods The study was performed between August and October 2017 on 130 healthy unrelated adult volunteers from Bosnian-Herzegovinian population sample. The prevalence of the following genetic variants was determined: F5 c.1601G>A (factor V Leiden), F2 c.*97G>A (factor II or prothrombin mutation), F13A1 (factor XIII) c.103G>T, MTHFR (methylenetetrahydrofolate reductase) c.665C>T and c.1286A>C, as well as PAI-1 (plasminogen activator inhibitor 1) c.-816A>G and c.-844G>A as markers of thrombophilia risk, and *2 and *3 alleles of CYP2C9 (cytochrome P450 2C9) and five variants of VKORC1 (vitamin K epoxide reductase complex subunit 1) as markers of warfarin pharmacogenetics. DNA was isolated from buccal swabs using salting out method, while genotyping was performed using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. Results Minor allele frequencies for two main thrombophilia risk factors, F5 c.1601G>A and F2 c.*97G>A were 0.023 and 0.008, respectively. Combined data for the markers of warfarin pharmacogenetics imply that 57.4% study participants can be expected to metabolize warfarin at an extensive, 40.3% at intermediate, and 2.3% at a poor rate. Conclusion This study reports the first extensive population genetic data for thrombophilia and warfarin pharmacogenetic markers in Bosnia and Herzegovina. Allele frequencies of genetic variants are within the general average for European populations, and their presence implies the necessity of introduction of personalized medicine in warfarin-mediated antithrombotic therapy.

Several association studies focusing on FTO gene polymorphisms have been published in the past years; however, the association between FTO -related conditions and FTO gene variants remains unexplained. Population genetics and association studies of different populations provide a valuable tool for further research. Thus, the aim of this systematic review is to summarize current knowledge on the FTO SNP rs9939609, and its association with presumably related conditions. The study included original research articles collected from PubMed and ResearchGate databases that were published in the period between 2007 and November 2017, and that provide information on rs9939609 mutant allele frequency and its probable association with any condition suspected of being related to the mutant allele. Genotype data was extracted and analyzed, and missing data was obtained from secondary sources. Short summaries of relevant studies from primary sources are organized in an overview table. The results of the systematic review suggest that mutant allele A is the most prevalent in European populations and least frequent on the Far East. In addition, it has been concluded that allele A is a good tool for the prediction of an increased risk of higher-than-normal BMI in a person carrying it, as well as that allele A should be further analyzed as a possible risk marker for type 2 diabetes mellitus and polycystic ovary syndrome development.

Novel oral anticoagulants (NOACs) are becoming a therapy of choice in everyday clinical practice after almost 50 years during which warfarin and related coumarin derivatives were used as the main anticoagulants. Advantages of NOACs over standard anticoagulants include their predictable pharmacodynamics and pharmacokinetics, stable plasma concentrations and less drug-drug and food-drug interactions. However, pharmacogenetics has its place in administration of NOACs, as considerable interindividual variations have been detected. In this review, previous findings in pharmacogenetics of dabigatran, rivaroxaban, apixaban and edoxaban are summarized, along with recommendations for studying genes encoding metabolically important enzymes for four selected NOACs. Future directions include identification of clinically relevant SNPs, and change in optimum dosage for patients who are carriers of significant variants.

The region of Western Balkans has been inhabited since the Paleolithic era and was the route of the spread of farming from the Middle East to Europe during the Neolithic era. In the present study, Y-STR data from European populations have been used to construct median-joining networks. The study was performed using Whit Athey’s Haplogroup Predictor, Y Utility and Network 4 software packages to predict Y haplogroups, construct networks, perform clustering of closely related Y chromosomes and calculate time estimates between individual nodes. The results of the study imply that geographically close populations cluster together at both Balkan and European levels. It was observed that an elevated number of study populations and individual haplogroups increases the possibility that individuals of different ethnic background cluster within the same or neighboring clades of network. Subsequent time estimates, performed based on the mutation frequency between the ancestral node and its descendant nodes, revealed that I2a haplogroup within the Western Balkan region has the most compact clustering (age, estimated at 3109 years), followed by Hg E1b1b which has the second most compact clustering (4896 years). The obtained results are nonetheless in accordance with previously published research investigating the frequency of Y haplogroups based on Y-SNP variant frequencies, indicating that Western Balkan countries are mainly represented by I2a subclade (average for six countries 32.3%), followed by E1b1b and R1a (average for six countries of 21.5% and 17%, respectively).

The region of Western Balkans has been inhabited since the Paleolithic era and was the route of farming from the Middle East to Europe during the Neolithic era (7th millennium BC). In the present study, Y-STR data from worldwide populations have been used to construct median-joining networks. The study was performed using Whit Athey’s Haplogroup Predictor, Y Utility and Network 4 software packages to construct networks, perform clustering of closely related Y chromosomes and calculate time estimates between individual nodes. The results of the study imply that geographically close populations cluster together on both worldwide and European level. It was observed that an elevated number of study populations and individual haplogroups increases the possibility that individuals of different racial and ethnic background cluster within the same or neighboring clades of network. An example is the case of the Nigerian population clustering closely with the populations from the Western Balkans. Subsequent time estimates performed based on the mutation frequency between the ancestral node and its descendant nodes revealed that I2a is the oldest haplogroup in the major area of the Balkan Peninsula (estimated separation time from its ancestral state: 4858 years), followed by haplogroups E1b1b (4088 years) and R1a (3910 years). This study is based on data collected from a single database and, therefore, gives approximations of the relative time distance between the nodes. Our results are nonetheless in accordance with previously published papers investigating the frequency of Y haplogroups based on Y-SNP variant frequencies, indicating that Western Balkan countries are mainly represented by I2a subclade (average for six countries 35.93%), followed by the other two haplogroups (average for six countries 23.16% and 10.62% regarding R1a and E1b1b, respectively).

β-Glucosidase was purified from Brassica oleracea by salting out with ammonium sulfate and hydrophobic interaction chromatography. Results demonstrated that the enzyme is a dimer (130 kD) made up of one major (80 kD) and one minor subunit (50 kD). The pH optimum is 6.0, with 50% of the enzyme's original activity remaining between pH 4.0 and pH 7.0. The temperature optimum is 35C, and activity did not decrease after two hours of exposure to this temperature. The activity of the enzyme was investigated on four substrates, 4-Nitrophenyl β-D-glucopyranoside (p-NPG), ortho-Nitrophenyl-β-D-glucopyranoside (o-NPG), para-Nitrophenyl-β-D-galactoside (p-NPGal) and ortho-Nitrophenyl-β-D-galactoside (o-NPGal), and km values were shown to be 0.755 mM, 0.174 mM, 0.988 mM and 0.213 mM, while Vmax values were 604 U/mg, 38 U/mg, 556 U/mg and 308 U/mg, respectively. The enzyme is completely inhibited by gluconolactone and glucose against p-NPG as substrate, with ki values of 0.038 mM and 0.64 mM, respectively. To our knowledge, this is the first study demonstrating purification and characterization of β-glucosidase from broccoli, thus providing a better understanding of its role in the plant, and establishing a basis for further research. Practical Applications To our knowledge, this is the first study demonstrating purification and characterization of β-glucosidase from broccoli, thus providing a better understanding of its role in the plant, and establishing a basis for further research. The results of this research highlight the potential of the enzyme isolated from broccoli for further research. Succeeding efforts would involve optimization of this procedure for increasing the enzyme yield, in order to make it a viable candidate for industrial application.

The relationship between single nucleotide polymorphisms (SNPs) and phenotypes is noisy and cryptic due to the abundance of genetic factors and the influence of environmental factors on complex traits, which makes the idea of applying artificial neural networks (ANNs) as universal approximates of complex functions promising. In this study, we compared different ANN architectures and input parameters to predict the adult length of Pacific lampreys, which is the primary indicator of their total migratory distance. Feedforward and simple recurrent network architectures with a different range of input parameters and different sizes of hidden layers were compared. Results indicate that the highest performing ANN had an accuracy of 67.5% in discriminating between long and short specimens. Sensitivity and specificity were 62.16% and 70.73%, respectively. Our results imply that feedforward ANN architecture with a single hidden neuron is enough to solve the problem of specimen classification. Nonetheless, while ANNs are useful at approximating functions with unknown relationships in the case of SNP data, additional work needs to be performed to ensure that the chosen SNP markers are related to functional regions related to the examined trait, as the use of non-specific markers will result in the introduction of noise into the dataset.

abstract In a study of the Bosnian-Herzegovinian (B&H) population, Y-chromosome marker frequencies for 100 individuals, generated using the PowerPlex Y23 kit, were used to perform Y-chromosome haplogroup assignment via Whit Athey's Haplogroup Predictor. This algorithm determines Y-chromosome haplogroups from Y-chromosome short tandem repeat (Y-STR) data using a Bayesian probability-based approach. The most frequent haplogroup appeared to be I2a, with a prevalence of 49%, followed by R1a and E1b1b, each accounting for 17% of all haplogroups within the population. Remaining haplogroups were J2a (5%), I1 (4%), R1b (4%), J2b (2%), G2a (1%), and N (1%). These results confirm previously published preliminary B&H population data published over 10 years ago, especially the prediction about the B&H population being a part of the Western Balkan area, which served as the Last Glacial Maximum refuge for the Paleolithic human European population. Furthermore, the results corroborate the hypothesis that this area was a significant stopping point on the “Middle East—Europe highway” during the Neolithic farmer migrations. Finally, since these results are almost completely in accordance with previously published data on B&H and neighboring populations generated by Y-chromosome single nucleotide polymorphism analysis, it can be concluded that in silico analysis of Y-STRs is a reliable method for approximation of the Y-chromosome haplogroup diversity of an examined population.

Forensic botany refers to the application of plant biology disciplines in crime investigations. Since plants are widespread in most habitats, they often provide valuable clues in criminal investigations. Hence, the primary role of forensic botany is to establish a connection between the victim or suspect and the crime scene. The forensic applications of forensic botany include many aspects including recognition of plant evidence at the crime scene, collection and preservation of plant evidence, maintenance of the chain of custody, and a testimony for the admissibility of this evidence in court. For the development of new efficient methods for plant identification and individualization, the initial step is optimization of DNA isolation from plant material. The aim of this work is to optimize the DNA isolation protocol of different leaf and non-leaf plant tissues, including pollen grains, using the salting out method from Taraxacum officinale, a species of the Asteraceae family. Since the species is very common in ecosystems with antropogenic influence, the need to optimize the DNA isolation protocol is of high importance. The efficiency of the isolation protocol was evaluated using spectrophotometry (concentration and purity) and gel electrophoresis. The results demonstrate that salting out can be used for the isolation of DNA from Taraxacum officinale. However, optimization is required, depending on the part of the plant from which DNA is extracted.