Corticosteroids regulate a number of physiological processes and are synthetic analogs of the natural steroid hormones produced by the adrenal cortex. As drugs, corticosteroids are non-inflammatory and are used for the treatment of plethora of conditions which include arthritis, kidney, skin, lungs or thyroid disorders, for the treatment and relief of symptoms of allergies and symptoms of some gastrointestinal disorders. In addition, glucocorticoids can regulate the effects of inflammatory disorders, including sepsis, autoimmune diseases, and allergies. These conditions are potentially fatal. Consequently, this drug class is among the most commonly prescribed globally. One representative of corticosteroid class of drugs is dexamethasone which is used to treat allergies, adrenal problems, arthritis, asthma, diseases of blood or bone marrow, inflammation, kidney diseases, different types of skin conditions, and episodes of multiple sclerosis. Virulence factors help bacteria colonize the host at the level of the cell. In their nature, these factors are secretory, associated with the membrane or present in the cytosol. Secretory factors allow bacterium to circumvent the host immune response, while membrane factors aid bacterium in adhesion to the host cell. Finally, cytosol factors help bacteria adapt metabolically, physiologically, and morphologically to their changing environment. One such factor is aspartyl proteinase, a protein that degrades other proteins and is a virulence factor in many pathogens playing a role in the host invasion process. Another important virulence factor is the ability to form biofilms, which can render bacteria resistant to antimicrobials. Despite the widespread use of corticosteroids, including dexamethasone, little is known about their possible influence on the expression of virulence factors such as aspartyl proteinase. If such a connection is to exist the use of corticosteroids could elicit pathogenesis in certain microbes. In the here-presented study we wanted to investigate the effects of dexamethasone on the growth, expression of aspartyl proteinase and biofilm formation in three E. coli strains that were previously isolated from patients suffering from urinary tract infection. To this aim, we amended the growth media with 0.5 mg/mL dexamethasone. Bacterial growth was measured over the period of 24 hours and the effect of dexamethasone was established at different time points. Administration of 0.5 mg/mL glucocorticoid drug dexamethasone did not significantly affect bacterial growth. However, it resulted in an increase in concentration of secreted E. coli virulence factor aspartyl proteinase, which increased up to 2.6-fold for some E. coli strains. In addition, we noted the increased biofilm formation in to three out of four studied strains. This study indicates dexamethasone as a possible trigger molecule for the expression of virulence factor aspartyl proteinase in E. coli.

Limited knowledge exists about the effects of commonly used diuretic medications on the human normal flora. Thus, we investigated potential stimulatory effects of diuretic drug furosemide on urogenital tract microbiota in women. Three strains of E. coli and C. albicans with different biofilm forming capacities were obtained from female patients diagnosed with urinary tract infections. All tested strains were treated with two different concentrations of furosemide drug, in comparison to non-treated strains as the negative control. At specific time intervals, samples were obtained from growing culture and analyzed for their proliferation rate, aspartyl proteinase excretion and biofilm formation ability. E. coli and C. albicans strains significantly increased their aspartyl proteinase excretion under furosemide treatment. This effect was frequently observed after 16 hours of incubation at 37oC. This drug has also increased the biofilm forming capacities of E. coli and C. albicans strains. Interestingly, both E. coli and C. albicans non-biofilm former strains, gained the capacity of biofilm formation when treated with furosemide at certain concentrations. E. coli control became a weak biofilm former after 48 hours of incubation, while non-biofilm former C. albicans strain became a weak biofilm former in dose-dependent fashion, after 48 hours incubation with furosemide in concentration of 0.1 mg/mL, and after 16 hours of incubation with furosemide in concentration of 0.5 mg/mL. Loop diuretic drug furosemide is able to increase the microbial virulence and turn commensal microbes into opportunistic pathogens. Additionally, the results suggest that enzyme aspartyl proteinase might act as a signal molecule for the biofilm formation, leading to the increased microbial pathogenicity.

Abstract Lavender and immortelle essential oils (EOs) are widely used to treat a spectrum of human conditions. The aim of this study was to investigate cyto/genotoxic effects of lavender and immortelle EOs using plant cells (Allium cepa) and human lymphocytes, as well as their antimicrobial potential using nine strains of bacteria and fungi. Our results for lavender and immortelle EOs showed that the frequency of chromosome aberrations (CAs) was increased in comparison with controls. For both oils, increased frequency of apoptosis for all concentrations, as well as the frequency of necrosis (0.10/0.30 µl/ml for lavender/immortelle, respectively) was demonstrated. In human lymphocytes, differences for minute fragments between immortelle oil (0.10 µl/ml) and controls were observed. Increased frequency of apoptosis was detected for immortelle oil (0.20 µl/ml), while both oils (0.20; 0.30 µl/ml lavender, and immortelle at all concentrations) induced higher frequency of necrosis in comparison with controls. Lavender EO was effective against all tested Gram-positive and Gram-negative bacteria, while immortelle EO inhibited only Gram-positive bacteria. Both oils exhibited antifungal effect. Our results demonstrated that lavender and immortelle EOs showed cyto/genotoxic effects in both, plant and human cells, as well as antimicrobial properties. Further studies are needed to strengthen these findings.

The identification of mutually exclusive somatic mutations shared among myeloproliferative neoplasm (MPN) subtypes has provided a powerful tool for studying disease evolution. Clinical features, gene mutations, and survival over 18 years were analyzed in MPN patients. One hundred thirty-eight MPN patients were subcategorized according to MPN subtypes: essential thrombocythemia (ET, n = 41), polycythemia vera (PV, n = 56), primary myelofibrosis (PMF, n = 10), and MPN unclassified (MPN-U, n = 31). Patient characteristics included clinical parameters, overall survival (OS), and mutational status of the Janus kinase 2 (JAK2), calreticulin (CALR), and myeloproliferative leukemia virus oncogene (MPL) genes. We compared hematologic and clinical features of JAK2V617F-ET vs. CALR-mutated ET vs. JAK2V617F-PV patients. JAK2V617F-patients had higher values of erythrocytes, hemoglobin, and hematocrit compared to CALR-mutated patients (p < 0.05). The mutant allele burden in JAK2V617F-PV and JAK2V617F-ET patients directly correlated with erythrocyte, hemoglobin, and hematocrit values, but it inversely correlated with platelet count. Thus, mutant allele burden was an indicator of the clinical phenotype in JAK2V617F-MPN patients. OS was not affected by the mutational status. In general, mutated JAK2, CALR, and MPL genes left specific hematological signatures.

Objectives This study aims to investigate the low-resolution human leukocyte antigen (HLA)-B locus polymorphisms between unrelated healthy individuals and patients with diagnosis of seronegative spondyloarthropathies and determine risky and protective allelic groups and genotypes. Patients and methods The study included 104 healthy control individuals (52 males, 52 females; median age 43 years; range 2 to 76 years) and 96 patients (43 males, 53 females; median age 28.5 years; range 2 to 67 years) diagnosed with: ankylosing spondylitis (AS) (n=19), reactive arthritis (n=19), psoriatic arthritis (n=28) and undifferentiated spondyloarthropathies (n=30). Genomic deoxyribonucleic acid was extracted from peripheral blood to detect allelic groups of HLA class I and II. Single-specific-primer polymerase chain reaction was used for HLA genotyping and visualization of products after their separation on 1.5% agarose gel for horizontal gel electrophoresis. Results Significantly increased frequency was found for HLA-A*02 and HLA-B*27 allelic variants in all groups of patients. The increased frequency of the HLA-B*35 allelic group in the control group represents the protective gene variant for the occurrence of AS. The predisposing genotype (HLA-B*27/B*44 and B*27/B*51) for the onset of disease was only found in AS patients. Conclusion This study shows the strong association of HLA-B*27 antigen with spondyloarthropathies, which is considered a risk variant of the gene for the onset of disease. Protective and risky allelic variants and genotypes are rare and their detection as well as increased frequency are possible if larger numbers of patients are involved.

Background: The gene for 5,10-methylenetetrahydrofolate reductase (NAD(P)H) or MTHFR gene encodes protein methylenetetrahydrofolate reductase (MTHFR), an enzyme important in folate metabolism. Aim: The aim of this study was to determine the frequencies of 677C>T and 1298A>C polymorphisms in the MTHFR gene of healthy subjects from the population. Material and methods: The blood samples were collected from 164 unrelated and healthy donors from population consisted of 98 females and 66 males. Both the MTHFR 677C>T and 1298A>C single nucleotide polymorphisms (SNPs) were analyzed by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Linkage disequilibrium (LD) between pair of SNPs was calculated through Haploview analysis. Results: The frequency of MTHFR 677T allele in the population (32.62%) was in agreement with the frequency of this allele in most other populations, however, the frequency of MTHFR 1298C allele (38.41%) was higher than that reported for most other populations in the world. Haploview analysis showed a relatively strong LD between 677C>T and 1298A>C SNPs with D′ values of 0.87. Conclusion: Regarding the two MTHFR polymorphisms, three of the nine combined genotypes were present in 87.2% of the population. 33.54% subjects were complex heterozygous (677CT/1298AC genotype), 34.15% subjects had 677CC/1298AC and 19.51% of 677CT/1298AA genotype. The subjects with 677TT genotype had a 1298AA or 1298AC genotype while subjects with 1298CC genotype had only 677CC genotype. The subjects with 677CC/1298AA genotype were only 3.05%. We were not found triple 677CT/1298CC and quadruple 677TT/1298CC mutation suggesting decreased viability of embryos with increased numbers of mutant alleles.

AIM: The research was conducted by genotyping two Human Leukocyte Antigen (HLA) gene classes. The main objective of this research was to investigate distribution and frequency of the allelic groups, genotypes and haplotypes in the gene loci of HLA class I (HLA-A*, -B*, -C*) and HLA class II (HLA-DRB1*, -DQB1*) in patients included in the program of cadaveric renal transplantation. MATERIAL AND METHODS: Our study covered 186 blood samples of patients who are registered on the list for cadaveric renal transplantation in Federation of Bosnia and Herzegovina and included 59 control, healthy unrelated individuals. For the HLA typing, we have used three different methods: micro lymphocyte cytotoxicity test (MLCT), Polymerase Chain Reaction (PCR) – Sequence Specific Primers (SSP) and PCR – Sequence-Specific Oligonucleotides (SSO) or Luminex technology. All patients and cadaveric donors were tested using the three methods because the system is polymorphic. RESULTS: Analysis of the results of genotyping HLA class I gene loci identified dominant HLA-A*02, HLA-B*35, HLA-C*07 allelic groups. Analysis of the HLA class II gene loci genotyping showed that HLA-DRB1*11 and HLA-DQB1*03 loci had the highest incidence in HLA class II. CONCLUSION: Based on our results and previous research, there were no observed differences between allelic frequencies and genotypes of healthy people and people with ESRD. Differences between allelic groups occurred, but they were not statistically significant, except HLA-C*01 (p = 0.020).

In this research, we examined the influence of low voltage (9 V) electric current frequencies (1.4 and 17 Hz), laser irradiation (648 and 532 nm) and combined treatment (one frequency and one laser beam) on the viability of baker’s yeast (Saccharomyces cerevisiae). Each treatment was conducted using modified methods and equipment in air-filter equipped working chamber. Staining was performed by a non-vital/vital staining technique that has shown an increase in viability of all samples. Counting of yeast cells in 1 ml of sample gave us several positive results in terms of different treatments, cell viability and increase in the number of healthy cells. Treatment with electric current at higher frequencies (4 and 17 Hz) showed increased cell death counts and, although compensating by an increase in viability, the 17 Hz frequency was considered more hazardous. The most adequate treatments (both increased viability and cell count) were the combined treatments (1 Hz/4 Hz + one of the two laser beams). Although, all electric treatments show certain increases in cell viability, combined treatments (1 or 4 Hz coupled with green or red laser beam) show the most promise in achieving both increased cell viability and increased cell counts. Acta Biol Szeged 60(2):139-144 (2016) KEy WoRdS cell viability electrical current laser irradiation Saccharomyces cerevisiae Submitted October 2, 2016; Accepted December 5, 2016 *Corresponding author. E-mail: alen_hamzic@hotmail.com

Introduction: Treatment of cancer has been subject of great interest. Researchers are continuously searching for new medicines. In this sense, ruthenium complexes have big potential. Some evidences suggest that ruthenium compounds possess anticancer activities. We synthesized two recently published ruthenium(III) complexes with bidentate O,N and tridentate O,O,N Schiff bases derived from 5-substituted salicylaldehyde and aminophenol or anilineare. These compounds showed affinity for binding to the DNA molecule, however, insufficient data are available regarding their possible toxic effects on biological systems.Methods: In the present study we evaluated genotoxic, cytotoxic, and cytostatic effects of Na[RuCl2(L1)2] and Na[Ru(L2)2], using the Allium cepa assay.Results: Different toxic effects were observed depending on the substance, tested concentration, and endpoint measured. In general, the tested compounds significantly lowered the root growth and mitotic index values as compared to the control group. Additionally, a wide range of abnormal mitotic stages, both clastogenic and non-clastogenic were observed in the treated cells. Na[RuCl2(L1)2] significantly increased the frequency of sticky metaphases, chromosome bridges, micronuclei, impaired chromosome segregation, as well as number of apoptotic and necrotic cells over the controls. In contrast, Na[Ru(L2)2] did not show significant evidence of genotoxicity with regard to chromosome aberrations and micronuclei, however, significant differences were detected in the number of apoptotic and necrotic cells when the highest concentration was applied.Conclusions: In this study we demonstrated antiproliferative effects of Na[RuCl2(L1)2] and Na[Ru(L2)2]. At clinical level, these results could be interesting for further studies on anticancer potential of the ruthenium(III) complexes using animal models.

Cancer patients in developing and low‐income countries have limited access to target therapies. For example, tyrosine kinase inhibitor (TKI) therapy for chronic myeloid leukaemia patients (CML) is often delayed. In Bosnia, 16% of patients received immediate TKI treatment (<3 months of diagnosis), while 66% of patients received therapy after a median 14‐month wait period. To assess the effect of delayed treatment on outcome, three patient groups were studied according to the time they received TKI treatment (0–5 months, 6–12 months and >13 months delay). The primary endpoints were complete cytogenetic (CCyR) and major molecular response (MMR) at 12 months. At 12 months of therapy, CCyR and MMR rates on imatinib decreased significantly: CCyR was achieved in 67% of patients in the immediate imatinib treatment group, 18% of patients in 6–12 months group and 15% of patients in >13 months wait group. MMR rates at 12 months occurred in 10% of patients with immediate treatment, 6% of those in 6–12 months group and 0% of patients in >13 months wait group. However, CCyR and MMR rates in patients on nilotinib were not associated with duration of treatment delay. Our data suggests that the deleterious effect of a prolonged TKI therapy delay may be ameliorated by the more active TKI nilotinib.

Objectives: To calculate the frequency of BCR-ABL1 splice variants (e14a2, e13a2 and e1a2) in a group of Bosnian patients with chronic myeloid leukemia (CML) and compare it with the data reported in other populations. Comparisons between cytogenetic and therapy outcomes in patients with BCR-ABL1 e13a2 and e14a2 transcripts was also aim of this study. Methods: Forty six (46) CML patients, hospitalized at the University Clinical Center Tuzla, were subjected to cytogenetic and RTPCR analysis. Results: Out of 46 patients, 33 (72%) patients expressed e14a2, followed by e13a2 seen in 10 (22%) patients. Two patients (4%) displayed both e14a2 and e1a2 forms of BCR-ABL1. One patient(2%) showed e1a2 transcript of BCR-ABL1. In a subgroup of 19 CML patients, treated with Imatinib, patients with e13a2 transcript had higher complete cytogenetic response (CCgR) compared to those with e14a2 transcript. Conclusion: The frequency of BCR-ABL1 e13a2 and e14a2 molecular isoform in patients with CML included in our research is in concordance with other researches. Continuation of the study with a larger number of patients is required to confirm these preliminary observations. Key words: BCR-ABL1 transcripts, CML, cytogenetic response, Philadelphia chromosome.