Abstract The 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.
ABSTRACT Due to its economic worth, cotton (Gossypium hirsutum L.) is grown in almost 70 countries and provides income for more than 250 million people. Therefore, producing cotton with having some desired characteristics that includes extended biotic and abiotic stress tolerance, improved fiber quality, promoted nutritional content and increased yield is the main objective for cotton biotechnology. To achieve this goal, many tissue culture and gene transfer techniques are being developed and used throughout the years. As applications for the gene transfer, the Agrobacterium-mediated, particle bombardment and pollen tube pathway-mediated methods are most successfully used and in conjunction with this, meristematic shoot tips as explants are efficiently utilized in gene transfer methods. In this study, the main objective was to report an efficient protocol for a greater recovery of transgenic cotton plant using Agrobacterium tumefaciens-mediated transformation. For this, one of the cotton strains (Cukurova 1518) cultivated widely in Turkey was chosen and meristematic shoot tips as explant sources, and GFP and NPTII genes as reporter and marker genes were used, respectively. The effective post co-cultivation conditions were provided via using the selection regime in vitro. Finally, the current results showed highly reproducible protocol developed could be used to produce transgenic cotton plants expressing desired traits or can be utilized as a model system to study the expression of particular genes.
Since the commercialization of transgenic crops in 1996, the biotech crop planted area has continuously increased. The European consumers have particularly been sceptical about transgenes in food products and EU (European Union) has enacted very complex legislation. The area of food analytics requires continuous development and improvement of detection methods to track the legislative framework and respond to consumers requirements. In the last decade, real-time PCR (polymerase chain reaction) based methods have been the methods of choice for numerous laboratories, but for various reasons, end-point PCR based methods have still been used. In our research, 73 samples of food and feed were analysed for the presence of common elements of transgene construct – Cauliflower Mosaic Virus 35S promoter (P-35S) and Agrobacterium tumefaciens Nopaline Synthase Terminator (T-NOS), using end-point PCR based methods. These samples had been previously tested for the presence of the same elements using validated real-time PCR based methods. Comparison of the used methods sensitivity showed that real-time PCR based methods have undeniable advantage. More important factor is specificity, and the fact that the list of approved Genetically Modified Organisms (GMOs) is constantly increasing necessitates updating of validation methods procedures. Considering upward trend of approved GMOs, it is important to pay more attention to the improvement and specialization of GMO detection methods.
Abstract Although prostate cancer accounts for the highest number of newly diagnosed cases of cancer in men, it represents a specific diagnostic challenge in modern oncology. The standard diagnosis of prostatic carcinoma begins with the screening of serum concentrations of PSA (Prostate Specific Antigen). If the concentration of serum PSA levels is above 4 ng/mL, the patient is further referred to a digital rectal examination in order to determine an increase in prostate volume. In cases where enlargement of the prostate is observed, the next step is biopsy of prostate tissue. This physically painful and invasive approach to confirm the diagnosis is often unnecessary because, in many cases, the patohistologic analysis determines diagnosis of benign prostatic hyperplasia, and not a tumor. In this study, we investigated the possibilities of detection and measurement of the relative level of gene expression of the KLK3 (Kallikrein-related peptidase 3), PCA3 (Prostate Cancer Gene 3) and TEMPRSS: ERG (Transmembrane protease serine2 and in-ETS erythroblostosis virus E26 oncogene homolog) genes from the urine samples of patients with prostatic diseases and healthy controls. Urine was the sample of choice because it is taken in a non-invasive manner, and could potentially serve to make better selection to biopsy. One of the selected genes (KLK3) differed significantly in the samples of various pathological conditions of the prostate, and therefore we consider that its further investigation is reasonable.
Of the four species of the genus Satureja (Lamiaceae) that are recognized in Bosnia and Herzegovina, S. subspicata has the the widest distribution. It is taxonomically challenging species of geographically limited distribution and little data on its genetic diversity throughout its range is available. We sampled six geographically distinct populations from Bosnia and Herzegovina and applied nrDNA (ITS1, ITS2), chloroplast markers (matK and trnL) and AFLP to examine genetic diversity of S. subspicata in the center of its distribution range and to explore the possibility of establishing the species DNA barcode. AFLP analysis showed large genetic differentiation among populations as well as moderate correlation between genetic distance among populations and geographic distance among locations. MatK has not proven useful in distinguishing S. subspicata from sympatric species. However, nrDNA sequences provided necessary resolution power, with ITS2 being more informative. Estimates of evolutionary divergence between nrDNA sequences obtained in our research and homologous sequences of sympatric Satureja deposited in the GenBank reveal closer relationship between geographically proximate populations of different species and slight divergence within S. subspicata sequences pool. This outcome highlights the importance of considering overall genetic diversity across the distribution range of a species when assigning DNA barcode.
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’ age
Genetics, as a science of the future, is very important part of Biology teaching. Within educational system in Bosnia and Herzegovina it is studied in limited number of clasess with a minimum of practical excercises, in both elementary and secondary schools. Limited understanding of scientific facts result in prejudices or uninformed attitudes towards novel biotechnologies that are largely based on non-reviewed and non-scientific information often incorrectly or inadequately presented in mass-media. Purpose of this research was to estimate the level of knowledge about selected topics in genetics among high school students in three Mostar high school institutions and differences in the level of knowledge between students from different high school programs – grammar school and vocational medical school. The research was conducted in 2017 among students of fourth grade of three high school institutions: ''Grammar School Mostar'', ''Medical high school Mostar'' and Medical high school ''Sestre milosrdnice''. Notable differences in knowledge level between two vocational programs are observed as well as variance in learning outcomes of the same program presented in two formal languages in Bosnia and Herzegovina
Satureja subspicata and S. horvatii are endemic species of the Balkan Peninsula and often used in traditional medicine in Bosnia and Herzegovina to treat different health conditions. We aimed to analyze the unevaluated apoptotic, genotoxic and cytotoxic effects of two Satureja species, as well as their content of phenolics that are mainly responsible for the plant's biological activity. Apoptotic and geno/cytotoxic activities of S. subspicata and S. horvatii were investigated in vitro in human lymphocyte culture and in vivo in mice. The content of the main phenolics in plant extracts was determined by ultra-high pressure liquid chromatography-MS-MS (UHPLC–MS/MS). Genotoxic and cytotoxic activities of Satureja extracts were evaluated in vitro by applying a cytokinesis-block micronucleus cytome assay in human lymphocyte culture and in vivo applying a mice reticulocytes micronucleus assay. SALSA RT-MLPA R011-C1 apoptosis assay was used for measuring the relative expression of 44 genes associated with the regulation of the apoptotic pathways in human lymphocyte cultures treated with different concentrations of two Satureja extracts. The first analysis of phenolic compounds in S. horvatii and S. subspicata determined by an UHPLC-MS/MS method revealed high levels of rosmarinic and caffeic acids. Minor genotoxic potential was determined in relation to the tested concentrations while no cytostatic and cytotoxic effects were revealed in vitro. However, when applied in concentrations of 200 mg/kg per os, aqueous extracts of two Satureja species significantly decreased frequency of reticulocytes micronuclei in treated mice against controls. Extracts of S. subspicata and S. horvatii in concentrations of 0.2 mg/mL, regardless of solvent used, downregulated pro-apoptotic and upregulated anti-apoptotic genes, showing anti-apoptotic activity. Our results indicate that the registered anti-genotoxic and anti-apoptotic activity is most likely related to the high level of phenolic acids (particularly rosmarinic and caffeic) in the tested extracts.
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