RESUMO
Halogenated boroxine K2[B3O3F4OH] (HB), an inorganic derivative of cyclic anhydride of boronic acid, is patented as a boron-containing compound with potential for the treatment of both benign and malignant skin changes. HB has effectively inhibited the growth of several carcinoma cell lines. Because of the growing interest in autophagy induction as a therapeutic approach in bladder carcinoma (BC), we aimed to assess the effects of HB on metabolic phenotype and autophagy levels in 5637 human bladder carcinoma cells (BC). Cytotoxicity was evaluated using the alamar blue assay, and the degree of autophagy was determined microscopically. Mitochondrial respiration and glycolysis were measured simultaneously. The relative expression of autophagy-related genes BECN1, P62, BCL-2, and DRAM1 was determined by real-time PCR. HB affected cell growth, while starvation significantly increased the level of autophagy in the positive control compared to the basal level of autophagy in the untreated negative control. In HB-treated cultures, the degree of autophagy was higher compared to the basal level, and metabolic phenotypes were altered; both glycolysis and oxidative phosphorylation (OXPHOS) were decreased by HB at 0.2 and 0.4 mg/mL. Gene expression was deregulated towards autophagy induction and expansion. In conclusion, HB disrupted the bioenergetic metabolism and reduced the intracellular survival potential of BC cells. Further molecular studies are needed to confirm these findings and investigate their applicative potential.
Assuntos
Autofagia , Neoplasias da Bexiga Urinária , Humanos , Autofagia/efeitos dos fármacos , Linhagem Celular Tumoral , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/tratamento farmacológico , Neoplasias da Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/genética , Proliferação de Células/efeitos dos fármacos , Glicólise/efeitos dos fármacos , Fenótipo , Fosforilação Oxidativa/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Mitocôndrias/metabolismo , Mitocôndrias/efeitos dos fármacos , Antineoplásicos/farmacologia , Antineoplásicos/química , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , HalogenaçãoRESUMO
Air pollution, recognized as a human carcinogen, is a significant cause of death in industrial and developing countries, and Bosnia and Herzegovina (B&H) is one of the leading countries for air pollution-caused death rate and has the poorest urban air quality in Europe. Despite a population decrease, urban air pollution in B&H has increased due to traffic pollution and still intensive use of solid fuel for heating and cooking. Human biomonitoring studies, regarding the described air pollution, have not been conducted before, and particularly have not been conducted in the region of Sarajevo. Good health, well-being, and environmental protection are part of the 17 defined Sustainable Development Global Goals. Accordingly, this study aimed to determine baseline levels of DNA damage in a group of Sarajevo citizens and to compare seasonal variations in DNA damage in relation to the reported levels of air pollution. From 33 individuals included in the study, samples were collected in the summer and winter seasons. The buccal micronucleus cytome (BMCyt) assay and comet assay in leucocytes isolated from saliva were performed. Mean values and standard deviations of log-transformed tail intensity (%), tail length (µm), and tail moment results in winter were 1.14 ± 0.23, 2.20 ± 0.14, and 1.03 ± 0.29, respectively, while in the summer season those values were 1.19 ± 0.19, 2.25 ± 0.17, and 1.07 ± 0.25, respectively. No significant differences were found for the comet assay parameters. Nevertheless, BMCyt results showed significant increases in micronuclei (P = .008), binuclear cells (P = .04), karyolysis (P = .0003), condensed chromatin (P = .03), and pyknosis (P = .002) in winter. Although the results of comet and BMCyt assays are not in accordance, this study contributes to the human air pollution biomonitoring in Sarajevo, B&H, and based on the genotoxic effects of air pollution evidenced by the BMCyt biomarker further studies of this kind are necessary.
Assuntos
Poluição do Ar , Monitoramento Biológico , Humanos , Bósnia e Herzegóvina , Mucosa Bucal , Dano ao DNA , Poluição do Ar/efeitos adversos , Ensaio Cometa , Testes para Micronúcleos/métodosRESUMO
Bosnia and Herzegovina (B&H) is among the European countries with the highest rate of air pollution-related death cases and the poorest air quality. The main causes are solid fuel consumption, traffic, and the poorly developed or implemented air pollution reduction policies. In addition, the city of Sarajevo, the capital of B&H, suffers temperature inversion episodes in autumn/winter months, which sustain air pollution. Human biomonitoring studies may be confounded by the lifestyle of subjects or possible metabolic alterations. Therefore, this study aimed to evaluate Ligustrum vulgare L. as a model for air pollution monitoring by measuring DNA damage at one rural and two urban sites. DNA damage was measured as tail intensity (TI) in L. vulgare leaves, considering seasonal, sampling period, leaf position and staging, and spatial (urban versus rural) variation. Effects of COVID-19 lockdown on TI were assessed by periodical monitoring at one of the selected sites, while in-house grown L. vulgare plants were used to test differences between outdoor and indoor air pollution effects for the same sampling period. Significantly higher TI was generally observed in leaves collected in Campus in December 2020 and 2021 compared with March (P < 0.0001). Outer and adult leaves showed higher TI values, except for the rural site where no differences for these categories were found. Leaves collected in the proximity of the intensive traffic showed significantly higher TI values (P < 0.001), regardless of the sampling period and the stage of growth. In regards to the COVID-19 lockdown, higher TI (P < 0.001) was registered in December 2020, after the lockdown period, than in periods before COVID-19 outbreak or immediately after the lockdown in 2020. This also reflects mild air pollution conditions in summer. TI values for the in-house grown leaves were significantly lower compared to those in situ. Results showed that L. vulgare may present a consistent model for the air pollution biomonitoring but further studies are needed to establish the best association between L. vulgare physiology, air quality data, and air pollution effects.
Assuntos
Poluentes Atmosféricos , Poluição do Ar , COVID-19 , Adulto , Humanos , Poluentes Atmosféricos/efeitos adversos , Ensaio Cometa , Bósnia e Herzegóvina , Controle de Doenças Transmissíveis , Poluição do Ar/efeitos adversos , Monitoramento Ambiental/métodos , Material ParticuladoRESUMO
Apoptosis induction is a promising approach in targeting tumor cells. As halogenated boroxine (HB) shows antitumor activity, but its mechanism of action in hematological tumors remains unclear, in this study, we aimed to analyze apoptosis triggering in normal and UT-7 leukemia cells by HB. Methods for assessing cell viability and cytotoxicity, apoptosis detection, relative expression of 84 apoptosis-associated genes and BCL-2, and functional analysis were applied. Pronounced HB activities in inhibition of cell viability, cytotoxicity, and apoptosis induction with measurable differences between tumor and normal cells were found. HB modulated the expression of 21 genes, predominantly downregulated the antiapoptotic genes in leukemia. The functional association revealed HB's impact on inhibition of NF-κB signaling pathway. BCL-2 expression decreasing was found only in UT-7 leukemia. This study identified HB as an apoptosis inducer affecting leukemia but not normal cells considering mechanisms of selective activity that may be a great advantage of HB applications.
Assuntos
Boro , Leucemia , Apoptose , Linhagem Celular Tumoral , Humanos , Leucemia/tratamento farmacológico , Leucemia/patologia , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genéticaRESUMO
Imiquimod (IMQ) induced human-like psoriasis in mice has been shown to be effective in testing and development of novel treatments. The IMQ psoriasis model has become widely used animal model, however, it is not completely characterized in different rat strains. We aimed to evaluate IMQ and betamethasone treatment for induction and reversal of psoriatic lesions on macroscopic, histological, genetic as well as cytokines and chemokines activation levels. Wistar rats were treated topically with IMQ. Adopted Psoriasis Area Severity Index (PASI) was calculated at the baseline, after the IMQ-symptoms induction and after betamethasone-symptoms reversal. Systematic effects were studied on cytokines and chemokines levels in plasma. Skin biopsy was taken to assess histological symptoms and selected inflammatory cytokines and receptors genes expression levels. Reversal of skin lesions, after betamethasone treatment, was significant (p = 0.03). Histological differences between untreated and IMQ-treated skin were significant for some markers (p < 0.05) though not significantly decreased by betamethasone treatment. Fourteen genes were significantly up-regulated after the IMQ and four genes were down-regulated after skin lesions reversal by betamethasone. This work provides new insights on biological effects of imiquimod induced psoriasis and its reversal by betamethasone treatment in Wistar rats. It also contributes to general knowledge of the rat model usage for testing of novel anti-psoriasis drugs.
Assuntos
Betametasona/administração & dosagem , Citocinas/sangue , Imiquimode/efeitos adversos , Psoríase/induzido quimicamente , Psoríase/tratamento farmacológico , Transdução de Sinais/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Administração Cutânea , Animais , Modelos Animais de Doenças , Regulação para Baixo/efeitos dos fármacos , Imiquimode/administração & dosagem , Masculino , Pomadas , Psoríase/sangue , Psoríase/genética , Ratos , Ratos Wistar , Índice de Gravidade de Doença , Pele/metabolismo , Pele/patologia , Resultado do Tratamento , Regulação para Cima/efeitos dos fármacosRESUMO
Genotoxic and cytotoxic effects of curcumin and sunset yellow were tested by the chromosome aberration analysis and cytokinesis-block micronucleus cytome assay in human lymphocyte culture. Water solutions of food dyes, in concentrations of 1, 2, 4 and 8 mM, were added to the cultures at the beginning of the cultivation period. Concentrations of 4 and 8 mM of sunset yellow induced significant increase in frequencies of cells with chromosome aberrations. Tested concentrations of sunset yellow significantly associated with frequencies of structural aberrations, chromatid-type aberrations, total aberrant cells and micronuclei showing considerable dose dependent clastogenic activity. In higher analyzed concentrations, curcumin significantly increased only nuclear buds frequency, suggesting its potential genotoxicity, while sunset yellow showed dose-dependent genotoxic potential. Obtained results point toward favorization of natural coloring agents in food consumption and emphasize the need of controlled use of food colorants.
Assuntos
Compostos Azo/toxicidade , Curcumina/toxicidade , Corantes de Alimentos/toxicidade , Linfócitos/efeitos dos fármacos , Mutagênicos/toxicidade , Células Cultivadas , Aberrações Cromossômicas/induzido quimicamente , Humanos , Linfócitos/metabolismo , Linfócitos/patologiaRESUMO
Recently it was found that dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH), is a potent and highly specific inhibitor of precancerous cell processes. We conducted gene expression profiling of human melanoma cells before and after treatment with two concentrations (0.1 and 1 mM) of this boron inorganic derivative in order to assess its effects on deregulation of genes associated with tumor pathways. Parallel trypan blue exclusion assay was performed to assess the cytotoxicity effects of this chemical. Treatment with K2(B3O3F4OH) induced a significant decrease of cell viability in melanoma cellline at both tested concentrations. Furthermore, these treatments caused deregulation of more than 30 genes known as common anti-tumor drug targets. IGF-1 and hTERT were found to be significantly downregulated and this result may imply potential use of K2(B3O3F4OH) as an inhibitor or human telomerase and insulin-like growth factor 1, both of which are associated with various tumor pathways.
Assuntos
Antineoplásicos/farmacologia , Compostos de Boro/farmacologia , Regulação para Baixo/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/genética , Melanoma/tratamento farmacológico , Melanoma/genética , Telomerase/genética , Antineoplásicos/síntese química , Antineoplásicos/química , Compostos de Boro/síntese química , Compostos de Boro/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Fator de Crescimento Insulin-Like I/metabolismo , Melanoma/metabolismo , Melanoma/patologia , Estrutura Molecular , Relação Estrutura-Atividade , Telomerase/metabolismoRESUMO
Anti-proliferative effects of halogenated boroxine - K2(B3O3F4OH) (HB) - have been confirmed in multiple cancer cell lines, including melanoma, but the exact mechanism of action is still unknown. This study aimed to determine its cytotoxic effects on human Caucasian melanoma (GR-M) cell growth in vitro as well as on the expression of cell death-related genes BCL-2, BECN1, DRAM1, and SQSTM1. GR-M and peripheral blood mononuclear (PBM) cells were treated with different HB concentrations and their growth inhibition and relative gene expression profiles were determined using the Alamar blue assay and real-time PCR. HB significantly inhibited cell growth of both GR-M and PBM cells but was even more effective in GR-M melanoma cells, as significant inhibition occurred at a lower HB concentration of 0.2 mg/mL. GR-M BCL-2 expression was significantly downregulated (P=0.001) at HB concentration of 0.4 mg/mL, which suggests that HB is a potent tumour growth inhibitor. At the same time, it upregulated BCL-2 expression in normal (PBM) cells, probably by activating protective mechanisms against induced cytotoxicity. In addition, all but the lowest HB concentrations significantly upregulated SQSTM1 (P=0.001) in GR-M cells. Upregulated BECN1 expression suggests early activation of autophagy at the lowest HB concentration in SQSTM1 cells and at all HB concentrations in PBM cells. Our findings clearly show HB-associated cell death and, along with previous cytotoxicity studies, reveal its promising anti-tumour potential.
Assuntos
Leucócitos Mononucleares , Melanoma , Humanos , Leucócitos Mononucleares/patologia , Proteína Sequestossoma-1 , Morte Celular , Proteínas Proto-Oncogênicas c-bcl-2/farmacologia , Melanoma/genética , Melanoma/patologia , Apoptose , Linhagem Celular TumoralRESUMO
Background: Waterpipe, also known as a hookah or narghile, is a type of tobacco products consumption device. Recently it has been increasingly popular in Bosnia and Herzegovina and the region. Waterpipe consumers are predominantly adolescents and young adults. Many of them believe in slighter harmful effects of waterpipes, compared to cigarettes. We aimed to determine the DNA damage in oral leukocytes and buccal cells of young individuals who have smoked a waterpipe for more than one year. Methods: The study group consisted of 40 cigarette non-smokers who regularly smoked a waterpipe on average of once per week. As a control, 40 non-smoking individuals were selected to match smokers for age. All participants in the study were healthy male and female adults from Bosnia and Herzegovina, 18-30 years of age. Before sampling, detailed survey and informed consent have been provided by each participant. Comet assay in oral leukocytes and buccal micronucleus cytome (BMCyt) assay in exfoliated buccal cells were applied. Results: Almost half of waterpipe smokers (WPS) tasted waterpipe at 15-16 years of age. Comet assay analysis showed increased tail intensity, tail length, and tail moment values among WPS compared to non-smokers (NS) (p = 0.0001, p = 0.0067, and p = 0.0001, respectively). Frequencies of the micronucleated (p = 0.0004), binucleated (p = 0.01), karyorrhectic, (p = 0.0036), and pycnotic cells (p = 0.03) were significantly higher in WPS compared to NS group. Conclusions: Genotoxicity and DNA damage biomarkers were increased in oral leukocytes and exfoliated buccal cells of young waterpipe smokers from Bosnia and Herzegovina, compared to NS group.
RESUMO
Aim Chromosome translocations are considered as one of the most severe forms of genome defects. Because of the clinical significance of chromosome translocations and scarce data on the incidence of sporadic translocations in population of Bosnia and Herzegovina, we aimed to report sporadic translocation frequencies in samples karyotyped in our laboratory. Methods The study group consisted of 108 samples. Whole blood was cultivated in complete medium for 72 hours with the thymidine application at 48th hour to synchronize the cell culture. Metaphases were arrested by colcemid 60 minutes before harvesting. Following hypotonic treatment, cells were fixed and cell suspension was dropped on coded slides. Dried slides were subjected to conventional GTG (G-banding with trypsin-Giemsa) banding and analyzed under 1000x magnification in the accordance with ISCN (International System for Human Cytogenetic Nomenclature) and E.C.A. Cytogenetic Guidelines and Quality Assurance. Results The incidence of all detected sporadic translocations was 27.81 x 10-4 per metaphase. The incidence of sporadic translocations involving chromosomes 7 and 14, being considered as the most frequent sporadic translocations of the human karyotype in phytohaemagglutinin (PHA) stimulated lymphocytes, was 15.89 x 10-4 per metaphase. The most frequent breakpoints were 7p21, 14q11 and 14q21. Other detected sporadic translocation breakpoints were: 1q25, 3p22, 7p13, 7q11.22, 7q33, 14q23 and 19q13.4. Conclusion Higher incidence of sporadic translocations compared to the similar studies was registered. Since potential explanations for this issue are smaller sample size and higher exposure of examined population to genotoxic agents, further monitoring of sporadic translocation incidences is recommended.
RESUMO
Medical radiation exposures have been reduced significantly with modern equipment and protection measures. Biomonitoring of medical personnel can provide information concerning possible effects of radiation exposure. However, chromosome aberration (CA) analysis is now recommended only when the estimated effective dose is 200 mSv or higher. In this retrospective study in Bosnia and Herzegovina, we have measured the cytogenetic status of medical workers and healthy volunteers (controls). Peripheral blood samples from 66 medical workers exposed to low-dose ionising radiation and 89 non-exposed volunteers were collected for chromosome aberrations (CA) analysis and the cytokinesis-block micronucleus (CBMN) assay. Higher rates of chromatid and chromosome breaks, acentric fragments, double minutes, micronuclei, and micronucleated binuclear cells were observed in the control group, while the rate of nucleoplasmic bridges was higher in the medical workers group.
Assuntos
Linfócitos , Exposição Ocupacional , Bósnia e Herzegóvina , Aberrações Cromossômicas , Análise Citogenética , Humanos , Linfócitos/efeitos da radiação , Testes para Micronúcleos , Exposição Ocupacional/efeitos adversos , Radiação Ionizante , Estudos RetrospectivosRESUMO
Clinopodium alpinum subsp. orontium (K.Malý) Govaerts and Thymus bracteosus Vis. ex Benth. are endemic Lamiaceae species in Bosnia and Herzegovina with rather limited data about their cytotoxic and genotoxic effects. This study aimed to analyse phenolic compounds composition of C. alpinum subsp. orontium and T. bracteosus aqueous and dimethyl sulphoxide (DMSO) extracts and their cytotoxic and genotoxic potential in human peripheral blood lymphocyte cultures. Among 33 analytes, 17 were identified and quantified in the examined extracts with the rosmarinic and chlorogenic acids as main constituents. Genotoxic effects of extracts from both species are proven at the highest applied dose. T. bracteosus extracts and DMSO as a solvent exhibited stronger genotoxic potential that should be further investigated in tumour cell lines. Nevertheless, non-endemic species with similar phenolic composition and bioactivity should be the first choice for medicinal purposes.
Assuntos
Lamiaceae , Bósnia e Herzegóvina , Dano ao DNA , Dimetil Sulfóxido , Humanos , Fenóis/análise , Extratos Vegetais/toxicidadeRESUMO
We have examined antiproliferative, cytotoxic, and genotoxic potential of a halogenated boroxine dipotassium trioxohydroxytetrafluorotriborate (K2[B3O3F4OH]). The impact on cell growth was evaluated by alamarBlue assay in basal cell carcinoma culture. Cytostatic, cytotoxic, and genotoxic potential were evaluated in lymphocytes culture, applying cytokinesis-block micronucleus cytome assay and chromosome aberrations analysis. Tested concentrations (0.05, 0.1, 0.2, and 0.4 mg/mL) were correlated with inhibition of cell growth in basal cell carcinoma culture and with the lymphocytes proliferation. Clastogenic activity has been confirmed, without evidences of aneugenic activity, in human lymphocytes.
Assuntos
Antineoplásicos/farmacologia , Compostos de Boro/farmacologia , Proliferação de Células/efeitos dos fármacos , Aberrações Cromossômicas , Antineoplásicos/efeitos adversos , Antineoplásicos/química , Compostos de Boro/efeitos adversos , Compostos de Boro/química , Sobrevivência Celular/efeitos dos fármacos , Aberrações Cromossômicas/induzido quimicamente , Relação Dose-Resposta a Droga , Humanos , Linfócitos/efeitos dos fármacos , Linfócitos/patologia , Estrutura Molecular , Testes de Mutagenicidade , Células Tumorais CultivadasRESUMO
Cytogenetic biomarkers, such as micronuclei in peripheral blood or oral mucosa, are widely used for evaluation of exposure to genotoxins or carcinogens. Tobacco is one of the strongest carcinogens, responsible for development of different types of cancers. The aim of this study was to assess the genotoxicity of cigarette consumption in young smokers and to correlate results of cytogenetic analysis in peripheral blood lymphocytes and exfoliated buccal cells. The study was conducted on samples taken from 43 smokers and 44 non-smokers, young individuals from Bosnia and Herzegovina. Significantly higher frequency of micronuclei in peripheral blood lymphocytes was observed in smokers (p < 0.05). No significant correlations were found for age, duration and intensity of smoking, and frequency of micronuclei in lymphocytes. Significantly higher frequency of degenerated (apoptotic) buccal cells was also revealed in smokers (p < 0.05). The frequency of apoptotic cells in smokers was significantly influenced by the age of participants (F = 8.649; p < 0.01) and duration of smoking (F = 5.389; p < 0.05). Results of cytogenetic analysis conducted in peripheral blood and exfoliated buccal cells are in significant positive correlation, indicating complementarities of those analyses.
Assuntos
Carcinógenos/toxicidade , Micronúcleos com Defeito Cromossômico , Mucosa Bucal/efeitos dos fármacos , Mutagênicos/toxicidade , Fumar/efeitos adversos , Adulto , Bochecha , Análise Citogenética , Exposição Ambiental/análise , Feminino , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Masculino , Testes para Micronúcleos , Mucosa Bucal/ultraestrutura , Adulto JovemRESUMO
BACKGROUND: Boron and boron containing compounds are known for their biological and protective roles being non-toxic and non-mutagenic in low concentrations. Male rats were exposed to halogenated boroxine (HB), dipotassium-trioxohydroxytetrafluorotriborate K2[B3O3F4OH], a potential new boron-containing therapeutic, aiming to determine concentrations with no adverse effects on selected serum biochemical parameters and histomorphological features. METHODS: HB was prepared by reacting potassium hydrofluoride (KHF2) with boric acid in molar ratios 2:3 at room temperature and its primary structure contains 4 fluorine atoms substituted in 6-membered ring. In concentrations of 10, 25, 35 and 45â¯mg/kg, HB was administered intraperitoneally as a single dose. Biochemical parameters were observed 24 and 96â¯h following the treatment. Effects of HB on biochemical blood parameters were also observed 24â¯h following continuous nine days application in concentrations of 10â¯mg/kg intraperitoneally and 50â¯mg/kg per os. Histomorphological observation of kidneys, liver, spleen, lungs and heart was performed for all treated animals. RESULTS: Administration of single high dose of HB (35â¯mg/kg-45â¯mg/kg) effected high levels of urea and creatinine, which indicated renal injury that appeared to be temporary. Possible cause of concern is pancreatic injury indicated by elevated levels of serum amylase in the groups of animals that received the highest dosages of the substance. Histopathological examination of selected tissues revealed mild to moderate lesions in the kidneys and livers associated with administration of HB. CONCLUSION: Observation of biochemical serum parameters or histopathology of examined tissues revealed no adverse effects of HB either after the administration of single dose lower than 35â¯mg/kg or following repeated administration at 10â¯mg/kg. These dosages should be further considered for potential therapeutic applications.
Assuntos
Compostos de Boro/efeitos adversos , Animais , Creatinina/metabolismo , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Ratos , Ratos Wistar , Ureia/metabolismoRESUMO
AIM: To report on the use of STR, Y-STRs, and miniSTRs typing methods in the identification of victims of revolutionary violence and crimes against humanity committed by the Communist Armed Forces during and after World War II in which bodies were exhumed from mass and individual graves in Slovenia. METHODS: Bone fragments and teeth were removed from human remains found in several small and closely located hidden mass graves in the Skofja Loka area (Lovrenska Grapa and Zolsce) and 2 individual graves in the Ljubljana area (Podlipoglav), Slovenia. DNA was isolated using the Qiagen DNA extraction procedure optimized for bone and teeth. Some DNA extracts required additional purification, such as N-buthanol treatment. The QuantifilerTM Human DNA Quantification Kit was used for DNA quantification. Initially, PowerPlex 16 kit was used to simultaneously analyze 15 short tandem repeat (STR) loci. The PowerPlex S5 miniSTR kit and AmpF/STR MiniFiler PCR Amplification Kit was used for additional analysis if preliminary analysis yielded weak partial or no profiles at all. In 2 cases, when the PowerPlex 16 profiles indicated possible relatedness of the remains with reference samples, but there were insufficient probabilities to call the match to possible male paternal relatives, we resorted to an additional analysis of Y-STR markers. PowerPlex Y System was used to simultaneously amplify 12 Y-STR loci. Fragment analysis was performed on an ABI PRISM 310 genetic analyzer. Matching probabilities were estimated using the DNA-View software. RESULTS: Following the Y-STR analysis, 1 of the "weak matches" previously obtained based on autosomal loci, was confirmed while the other 1 was not. Combined standard STR and miniSTR approach applied to bone samples from 2 individual graves resulted in positive identifications. Finally, using the same approach on 11 bone samples from hidden mass grave Zolosce, we were able to obtain 6 useful DNA profiles. CONCLUSION: The results of this study, in combination with previously obtained results, demonstrate that Y-chromosome testing and mini-STR methodology can contribute to the identification of human remains of victims of revolutionary violence from World War II.
Assuntos
Osso e Ossos , Cromossomos Humanos Y , Vítimas de Crime , Antropologia Forense/métodos , Repetições de Microssatélites/genética , Militares , II Guerra Mundial , Humanos , Masculino , EslovêniaRESUMO
Twelve previously synthesized, biologically active 2,6,7-trihydroxyxanthen-3-one derivatives were evaluated in vitro for antiproliferative activity. Compounds were screened against HeLa, SW620, HepG2 and A549 tumor cell lines. Compound with the trifluormethyl group on C-4' position of the phenyl ring showed the best inhibitory activity towards HeLa and A549 tumor cells with IC50 of 0.7 and 4.1 µmol L-1, resp. Compound with chlorine and fluorine substituents on aryl ring showed the best antiproliferative activity against SW620 with IC50 of 4.1 µmol L-1 and against HepG2 tumor cell line with IC50 of 4.2 µmol L-1. Analyses of cytotoxic and genotoxic potential of the trifluormethyl derivative were performed with cytokinesis-block micronucleus cytome assay in human lymphocyte culture and revealed no genotoxic and cytotoxic effects. The most potent compounds were subjected to molecular docking simulations in order to analyse bindings to molecular targets and, at the same time, further support the results of experimental cytotoxic tests. Docking studies showed sites of importance in forming hydrogen bonds of the most potent compounds with targets of interest.
Assuntos
Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Mutagênicos/farmacologia , Células 3T3 , Células A549 , Animais , Linhagem Celular , Linhagem Celular Tumoral , Citotoxinas/farmacologia , Dano ao DNA/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Células HeLa , Células Hep G2 , Humanos , Camundongos , Simulação de Acoplamento Molecular/métodos , Relação Estrutura-AtividadeRESUMO
Modern Bosnia and Herzegovina is a multi-ethnic and multi-religion country, with a very stormy history. Certain archaeological findings indicate continuous population of its territory since the Paleolithic. In time, vast number of different factors jointly influenced fascinating diversity of local human populations. A great number of small, more or less isolated, indigenous populations, make this area quite attractive for population-genetic surveys of different levels and approaches. Austro-Hungarian military physicians conducted the very first known bio-anthropological analyses of Bosnia-Herzegovina population at the end of the 19th century. Thus, the first step towards resolving the genetic structures of local B&H human populations was made. The studies that followed (conducted throughout most of the 20th century) were primarily based on the observation of various phenotypic traits. This stage was followed by the examination of various cytogenetic and fundamental DNA based molecular markers. The efforts undertaken over the last three centuries revealed "human genetic treasure" in Bosnia and Herzegovina. However, even now, after all the studies that were conducted, many interesting features remain to be discovered and described within the existing local human populations.
Assuntos
Genética Populacional , População/genética , Antropologia , Bósnia e Herzegóvina , Citogenética , DNA , Marcadores Genéticos , Humanos , FenótipoRESUMO
Standard cytogenetic assays used in genotoxicology usually include chromosome aberrations analysis and micronucleus cytokinesis-block assay. Both tests originate on standard protocol for lymphocyte culture and can be used as complement or substitute to each other. Aim of this study was to evaluate complementarities between results of chromosome aberration analysis assay and results of micronucleus cytokinesis-block assay in representative sample of inhabitants from Bosnia and Herzegovina. The aim was achieved by calculating Pearson correlation coefficient and simple linear regression.
Assuntos
Aberrações Cromossômicas , Análise Citogenética/métodos , Micronúcleos com Defeito Cromossômico , Adulto , Células Cultivadas , Citocinese/fisiologia , Feminino , Humanos , Modelos Lineares , Linfócitos/patologia , Linfócitos/fisiologia , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos TestesRESUMO
Abstract Biological activity of boron-containing compounds (BCCs) has been well-known. Growing interest and numerous applications for BCCs have been reported. Boron and boron-containing acids show low acute toxicity in mammals but data on halogenated boroxine (HB) - dipotassium-trioxohydroxytetrafluorotriborate, K2(B3O3F4OH) acute toxicity have not been reported before. This compound, characterized as a potential therapeutic for skin changes, exhibits no observable genotoxicity in doses lower that 0.1 mg/ml in vitro and 55 mg/kg in vivo. It has also been confirmed as an antitumour agent both in vitro and in vivo as well as an inhibitor of enzymes involved in antioxidant mechanisms. The aim of this study was to assess the acute toxicity of HB and to determine the maximum tolerated dose as well as a dose free of any signs of toxicity in different test organisms. Acute toxicity of HB was tested in Sprague-Dawley and Wistar rats and BALB/c mice after single parenteral application of different doses. We determined doses free of any sign of toxicity and LD50 after single dose administration. LD50 of HB ranges from 63 to 75 mg/kg in different test models, meaning that HB shows moderate toxicity