Novel Therapy Approaches in β-Thalassemia Syndromes — A Role of Genetic Modifiers

The β-thalassemia syndromes are heterogeneous autosomal recessive hereditary disorders, caused by alterations in the HBB gene and characterized by absent or reduced β-globin chain synthesis. The β-thalassemia phenotypes are variable, ranging from severe, transfusion-dependent thalassemia major to mild, asymptomatic thalassemia trait. This interpatient clinical variability has swayed researchers toward identifying genetic modifiers for these disorders. Primary modifiers refer to type of alterations affecting β-globin gene. Secondary modifiers include variations in genes affecting α/β-globin chain equilibrium, such as genes involved in the γ-globin gene expression and genes affecting the amount and stability of α-globin chains. Tertiary modifiers are gene variations affecting the phenotype with regard to the complications caused by β-thalassemia syndromes. A role of secondary genetic modifiers in ameliorating the clinical phenotype has been observed. Secondary genetic modifiers are the most common targets for modern therapy and could be located within α- and γ-globin genes or outside globin gene cluster. The most potent secondary modifier genes are γ-globin genes. Production of fetal hemoglobin (HbF) trough adulthood ameliorates the severity of β-thalassemia phenotype. Large family and genome-wide association studies have shown that regions outside of the β-globin gene cluster are also implicated in γ-globin gene expression regulation. HBS1-MYB intragenic region and BCL11A gene have been particularly studied. Variants within these loci, along with γ-globin gene variants, account for approximately 50% of the HbF level variation, suggesting that additional factors are involved (transcription regulators (KLF1), regulators of α-globin chain stability (AHSP), epigenetic regulators (FoP)). Until recently a definitive cure for β-thalassemia could be achieved with bone marrow transplantation. However, it is available for less than 30% of the patients and bears a significant risk of morbidity and mortality. Alternative strategies, such as gene therapy and development of induced pluripotent stem cells (iPSCs) have been explored. The targets for gene therapy are hematopoietic stem cells, which are harvested from patient bone marrow or peripheral blood, purified by immunoselection, transduced by “therapeutic gene” aimed at correcting the effect of defective β-globin gene, and returned to the patient. Various types of vectors have been considered for gene transfer, including non viral (tRNK and ribozymes) and viral (retroviral and lentiviral vectors). In the past few years, iPSCs emerged as an interesting candidate for gene transfer. The feature that makes these cells appealing in the field of gene therapy is their susceptibility to gene correction by homologous recombination. Therapy protocols based on molecular basis of β-thalassemia are the best example of novel approaches in disease treatment

THE INFLUENCE OF DIFFERENT CONCENTRATIONS OF PLANT ESSENTIAL OILS ON GROWTH AND REPRODUCTION OF Salmonella enteritidis

Plant essential oils have been reported to possess antimicrobial properties and therefore have potential usage as natural antimicrobials of food. The aim of the study was to examined the antimicrobial effect of sweet basil and thyme essential oils against growth and reproduction of Salmonella enteritidis reference strain ATCC 13076 (S. enteritidis RS) and Salmonella enteritidis epidemical strain (S. enteritidis ES) cultivated on plate. Therefore, the samples were prepared as a dip application from different concentrations of sweet basil and thyme essential oils (1; 2.5 and 5%) with initial concentration of bacteria from 109 CFU/mL and were cultivated on plate. The control samples were prepared as dip application of bacteria without added essential oils. All samples were exposed at 37°C and 46°C. The growth of S. enteritidis RS and S. enteritidis ES was observed only in the control samples without added sweet basil and thyme essential oils. There wasn’t any growth of both Salmonella еnteritidis strains in the samples dipped in the 1; 2.5 and 5% sweet basil and thyme essential oils. The results from the ANOVA indicate that the utilized essential oils in combination with temperature regime was significantly (p <0.001) reduced the CFU number of the both strains of Salmonella enteritidis. These results support the possibility of using sweet basil and thyme essential oil as natural preservatives in food to contribute in the reduction of Salmonella enteritidis at acceptable levels in view to prevent the risk for consumers

Konvektivno sušenje parčića ploda batata (Ipomoea batatas (L.) Lam), zavisnost od predtretmana

The dried of pieces of sweet potatoes (Ipomoea batatas (L.) Lam) was carried in experimental convective cabinet drier with loess. This vegetable was rich in beta carotene and was not sufficiently in our countries. The dried process was carried at 55oC, and weight changes were measured every 15 minutes. Were performed three pretreatments: dried with pretreatment, blanched with the 1% vitamin C and 5% NaCl and without pretreatment. Blanched was carried in water at 60oC, for 3 minutes. The moisture content of raw sweet potato was 76.40%, but in dried material value from 16.90 to 19.59%, respectively. Rehydration values were measure of the quality of the dried process. The best results were obtained to dried small pieces of sweet potato without pretreatment, than rehydration was 83.37%.Sušenje parčića ploda batata (Ipomoea batatas (L.) Lam) ili slatkog krompira vršeno je u eksperimentalnoj konvektivnoj sušari sa lesama. Ova povrtarska biljka bogata je beta karotenom, a nedovoljno se koristi u našoj zemlji. Proces sušenja odvijao se na 55oC, a merena je promena mase na svakih 15 minuta. Rađena su tri predtretmana blanširanjem: bez dodataka (predtretman I), sa dodatkom 1% vitamina C (predtretman II), sa dodatkom 5% NaCl (predtretman III) i sušenje bez predtretmana (kontrola). Blanširanje je vršeno u destilovanoj vodi temperature 60oC za 3 minuta, sa i bez dodataka. Sadržaj vlage u sirovom plodu batatu bio je 76,40%, a u osušenom materijalu kretao se od 16,90 do 19,59 %. Stepen rehidratacije korišćen je kao mera kvaliteta procesa sušenja. Najbolji rezultati dobijeni su kod sušenja parčića ploda batata bez predtretmana, čiji je stepen rehidratacije bio 83,37%

Antibacterial activities of rosemary essential oils and their components against pathogenic bacteria

There are many known essential oils, of which only 300 are commercially important for different types of industry. Antimicrobials are used in food for two main reasons: to control natural spoilage processes and to prevent/control growth of micro-organisms, including pathogenic microorganisms. The aim of this review was to investigate antibacterial effects of rosemary essential oils (EOs) on some Gram-positive and Gram-negative bacteria: Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa. Listeria monocytogenes, Salmonella enteritidis, Salmonella typhimurium. Rosmarinus officinalis essential oil is very important for its medicinal uses and its powerful antibacterial, cytotoxic, anti-mutagenic, antioxidant, anti-phlogistic and chemopreventive properties. The antimicrobial and other biological activities of rosemary essential oil are directly correlated with the presence of bioactive volatile components. To this end, one possibility has been the use of rosemary essential oils (EOs) and the compounds found there in as alternative antimicrobial food preservatives. The future will see investigation of food applications of the naturally occurring antimicrobials, especially the effectiveness of rosemary EOs, individually and in combination with other parts of plant extract, other effective EOs and other food-processing techniques. Keywords: antibacterial effect, essential oil, rosemary, pathogenic bacteri

Quantitative traits in wheat (Triticum aestivum L. cv. 'Novosadska rana 5') grown on pseudogley soil depending on lime rates

The effect of the use of different lime rates on the pH values and subsequently on the quantitative traits in wheat (Triticum aestivum L. cv. 'Novosadska rana 5') was investigated on the pseudogley type of soil. Of the quantitative traits in wheat, spike length, number of spikes per m2, grain mass per spike, number of grains per spike and 1000-grain weight, were studied as the most significant indicators of its yield. The studies were carried out during 2010 and 2011. The trial was set up following a randomized block design with four liming variants along with three replications, in which the experimental field was 70 m2 in area. In both study years, along with the previously determined experimental conditions, the variant of CaCO3, used with an amount of 2 t/ha, was found to have the highest effect on enhancing all the parameters of wheat. Based on the results obtained throughout the current research work, it could be inferred that, despite falling under a lower quality class, pseudogley soil could promote usable optimal biological potential of the wheat variety ('Novosadska rana 5'), if adequately treated with lime along with the application of cropping practices.Key words: Pseudogley, lime rates, wheat, grown

The influence of different concentrations of plant essential oils on growth and reproduction of Salmonella enteritidis

Plant essential oils have been reported to possess antimicrobial properties and therefore have potential usage as natural antimicrobials of food. The aim of the study was to examined the antimicrobial effect of sweet basil and thyme essential oils against growth and reproduction of Salmonella enteritidis reference strain ATCC 13076 (S. enteritidis RS) and Salmonella enteritidis epidemical strain (S. enteritidis ES) cultivated on plate. Therefore, the samples were prepared as a dip application from different concentrations of sweet basil and thyme essential oils (1%; 2.5% and 5%) with initial concentration of bacteria from 109 CFU/mL and were cultivated on plate. The control samples were prepared as dip application of bacteria without added essential oils. All samples were exposed at 37°C and 46°C. The growth of S. enteritidis RS and S. enteritidis ES was observed only in the control samples without added sweet basil and thyme essential oils. There was not any growth of both Salmonella еnteritidis strains in the samples dipped in the 1%; 2.5% and 5% sweet basil and thyme essential oils. The results from the ANOVA indicate that the utilized essential oils in combination with temperature regime was significantly (p <0.001) reduced the CFU number of the both strains of Salmonella enteritidis. These results support the possibility of using sweet basil and thyme essential oil as natural preservatives in food to contribute in the reduction of Salmonella enteritidis at acceptable levels in view to prevent the risk for consumers. Key words: Salmonella, sweet basil, thym

Short communication: In vitro antimicrobial properties of basil and thyme essential oils against Salmonella Spp.

Food diseases are caused by consuming foods that have been contaminated by an infectious agent or a toxin produced by it. According to the WHO, 30% of people in industrialized countries suffer from foodborne diseases. Salmonella enteritidis even at 95.9% was the main etiological factor for salmonellosis in humans (De Knegt et al., 2015) and have established a trend of increasing the level of contamination of food products with Salmonella spp. in the Republic of North Macedonia. In order to protect food from contamination with pathogens and other harmful microorganisms, many scientists have examined the antifungal, antibacterial and antioxidative properties of essential oils (EOs) and their application in food technology. They are considered a safe and environmentally friendly alternative to the control of bacteria present in the food and food industry, but also to the control of other pathogenic microorganisms, especially those that are drug resistant (Yap et al., 2014). The aim of the study was to determine the antimicrobial effect of different concentrations of basil and thyme EOs on the growth of Salmonella enteritidis in laboratory conditions, inoculated in dough for making pasta with eggs and inoculums with flour and chicken egg

Antimicrobial effect of thyme essential oil against Salmonella enteritidis inoculated in flour and homogenized eggs

The aim of this study was to determine the antimicrobial effect of thyme essential oil against two strains of Salmonella enteritidis group D: reference strain ATCC 13076 and epidemical strain. The both strains of bacteria were inoculated separately up to concentration of 109 CFU/mL in the micellar solutions which were prepared as application of thyme essential oil in physiological solution up to final concentration of 1; 2.5 and 5%. That stock micellar solutions were used for preparing the mixture with white flour and homogenized eggs, separately. The prepared samples were cultivated on plate and exposed at a temperature of 37°C for 18 hours and at a temperature of 46°C for 9 hours (ISO 6579). GLM multivariate statistical model showed that the number of both strains of Salmonella enteritidis was significantly influenced by the thyme essential oil concentration, type of media and their interaction.The thyme essential oils might be useful in the control of Salmonella spp. in commercially produced food systems as an alternative of the chemical preservatives. Keywords: antimicrobial effect, thyme essential oil, Salmonella, flour, eggs

Antimicrobial effect of sweet basil essential oil against Salmonella enteritidis growth in vitro

The aim of the study was to examine the antimicrobial activity of sweet basil essential oil against two strains of Salmonella enteritidis group D: reference strain ATCC 13076 and epidemical strain. The both strains of bacteria were inoculated separately up to concentration of 109 CFU/mL in the micellar solutions which were prepared as application of sweet basil essential oil in physiological solution up to final concentration of 1; 2.5 and 5%. That stock micellar solutions were used for preparing the mixture with white flour and homogenized eggs, separately. In all samples was added 90 ml Salenit F broth and were exposed at a temperature of 37°C for 18 hours and at a temperature of 46°C for 9 hours. The samples were cultivated on plate for enumeration according ISO 6579. The results from the GLM multivariate statistical model indicate that the number of both strains of Salmonella enteritidis cultivated on plate was high significantly influenced by the concentration of the sweet basil essential oil, medium and also their interaction. These results support the possibility of using sweet basil essential oil as natural preservative in food like pasta, prepared from white flour and eggs. Keywords: antimicrobial effect, sweet basil essential oil, Salmonella, in vitro

Systematic documentation and analysis of human genetic variation in hemoglobinopathies using the microattribution approach

We developed a series of interrelated locus-specific databases to store all published and unpublished genetic variation related to hemoglobinopathies and thalassemia and implemented microattribution to encourage submission of unpublished observations of genetic variation to these public repositories. A total of 1,941 unique genetic variants in 37 genes, encoding globins and other erythroid proteins, are currently documented in these databases, with reciprocal attribution of microcitations to data contributors. Our project provides the first example of implementing microattribution to incentivise submission of all known genetic variation in a defined system. It has demonstrably increased the reporting of human variants, leading to a comprehensive online resource for systematically describing human genetic variation in the globin genes and other genes contributing to hemoglobinopathies and thalassemias. The principles established here will serve as a model for other systems and for the analysis of other common and/or complex human genetic diseases