RESUMO
OBJECTIVES: Studies indicate the complex nature of the genetic structure of the European Roma which has been shaped by different effects of their demographic history, while preserving their ancestral Indian origin. The primary aims of this study were to present for the first time the paternal profiles of the Roma from Bosnia and Herzegovina based on the data from Y-chromosome STR loci, identify the components of non-Roma paternal gene flow into the Roma, and evaluate the genetic relationships with other European Roma populations. MATERIALS AND METHODS: In this study, 110 DNA samples of unrelated males from Roma populations residing in different regions of Bosnia and Herzegovina were genotyped using the 23 Y-STR loci included in the PowerPlex Y23 system. RESULTS: The analysis of the genetic structure of the Bosnian-Herzegovinian Roma revealed intra-country population substructuring and indicated differing genetic affinities between the Bosnian-Herzegovinian Roma and other European Roma populations. The paternal genetic structure of the Bosnian-Herzegovinian Roma has two components: an ancestral component represented by haplogroup H1a1a-M82, and European component presented by haplogroups I1-M253, I2a1a2b-L621, J2a1a-L26, J2a1a1a2b2a3~Z7671, J2b2a-M241, G2a2b2a1a1b-L497, and E1b1b-M215. CONCLUSION: Genetic relations between the Bosnian-Herzegovinian Roma and other European Roma are shaped by different influences on their demographic history. The data suggest that the paternal gene pool of the Roma from Bosnia and Herzegovina might be a consequence of an early separation of the proto-Roma population and the later gene flow as well as factors of the isolation that accompany the Roma populations in some Bosnian-Herzegovinian regions.
Assuntos
Cromossomos Humanos Y , Roma (Grupo Étnico) , Cromossomos Humanos Y/genética , Pai , Genética Populacional , Genótipo , Haplótipos , Humanos , Masculino , Repetições de Microssatélites , Roma (Grupo Étnico)/genéticaRESUMO
Background: Bosnia and Herzegovina is a multinational and multireligious country, located in the western part of the Balkan Peninsula. Migrations through history were a key factor in the genetic identity of the Bosnian-Herzegovinian population.Aim: To analyse genetic polymorphisms of 22 autosomal short tandem repeat (STR) loci in the population of Bosnia and Herzegovina and to compare STR allele frequencies for STR loci with the reference data for European populations.Subjects and methods: The study was conducted among 600 unrelated individuals from all regions of Bosnia and Herzegovina. Genotyping was performed using the PowerPlex® Fusion amplification kit. Allele frequencies and statistical parameters were calculated, as well as the genetic distance among analysed populations through the construction of a neighbor-joining dendrogram.Results: STR loci included in the PowerPlex® Fusion amplification kit showed high discriminatory power indicating their reliability for human identification and paternity testing. The neighbor-joining dendrogram based on the results of genetic distance analysis showed that the Bosnian and Herzegovinian population has the greatest genetic distance from Turkish and Hungarian populations and greatest similarity with Croatian, Slovenian, and Serbian populations.Conclusion: The results of this study strongly support the application of 22 autosomal genetic markers for paternity testing and personal identity testing and are in agreement with most previous human studies in the investigated human populations.
Assuntos
Cromossomos Humanos , Frequência do Gene , Repetições de Microssatélites , Polimorfismo Genético , Bósnia e Herzegóvina , Genética Forense , Marcadores Genéticos , HumanosRESUMO
BACKGROUND: Tuzla Canton is the most populated region in the ethnically mixed territory of Bosnia and Herzegovina, whose genetic analysis could provide an insight into past demographic events. AIM: Analysis of 23 Y-chromosome STR markers in the population of Tuzla Canton and investigation of the genetic relationship of the male population of the Tuzla Canton and that of the larger Bosnian and Herzegovinian population as well as neighbouring and other European populations. SUBJECTS AND METHODS: The study was conducted among 100 unrelated healthy adult males from Tuzla Canton that have been genotyped using 23 Y-STR loci included in the PowerPlex Y23 kit. Statistical parameters such as haplotype diversity and allele frequencies were calculated, as well as the Rst-based genetic distances between the new dataset and those from Bosnia and Herzegovina and elsewhere, which were then visualised through multi-dimensional scaling plot and neighbour-joining phylogenetic tree analyses. RESULTS: The PowerPlex Y23 kit has shown high discrimination capacity, as all 100 individuals have unique haplotypes. The newly incorporated loci seem to be highly informative. Population comparison reveals no statistically significant differences between the study population and the general Bosnian-Herzegovinian population, and between the study population and neighbouring populations. CONCLUSION: These results could be used as an additional investigation of the genetic relationship between the regional populations in Bosnia and Herzegovina and neighbouring human populations, as well as for further human population and forensic genetics studies.
Assuntos
Cromossomos Humanos Y/genética , Variação Genética , Genótipo , Repetições de Microssatélites/genética , Bósnia e Herzegóvina , Frequência do Gene , Humanos , MasculinoRESUMO
AIM: To determine newest the most accurate allele frequencies for 15 short tandem repeat (STR) loci in the Bosnian and Herzegovinian population, calculate statistical parameters, and compare them with the relevant data for seven neighboring populations. METHODS: Genomic DNA was obtained from buccal swabs of 1000 unrelated individuals from all regions of Bosnia and Herzegovina. Genotyping was performed using PowerPlex® 16 System to obtain allele frequencies for 15 polymorphic STR loci including D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, and FGA. The calculated allele frequencies were also compared with the data from neighboring populations. RESULTS: The highest detected value of polymorphism information content (PIC) was detected at the PentaE locus, whereas the lowest value was detected at the TPOX locus. The power of discrimination (PD) values had similar distribution, with Penta E showing the highest PD of 0.9788. While D18S51 had the highest value of power of exclusion (PE), the lowest PE value was detected at the TPOX locus. CONCLUSION: Upon comparison of Bosnian and Herzegovinian population data with those of seven neighboring populations, the highest allele frequency differentiation was noticed between Bosnian and Herzegovinian and Turkish population at 5 loci, the most informative of which was Penta E. The neighbor-joining dendrogram constructed on the basis of genetic distance showed grouping of Slovenian, Austrian, Hungarian, and Croatian populations. Bosnian and Herzegovinian population was between the mentioned cluster and Serbian population. To determine more accurate distribution of allelic frequencies and forensic parameters, our study included 1000 unrelated individuals from all regions of Bosnia and Herzegovina, and our findings demonstrated the applicability of these markers in both forensics and future population genetic studies.
Assuntos
Repetições de Microssatélites , Adulto , Áustria , Bósnia e Herzegóvina , DNA/genética , Impressões Digitais de DNA , Frequência do Gene , Genótipo , Humanos , Polimorfismo Genético , TurquiaRESUMO
Modern Bosnia and Herzegovina is a multinational and multi-religious country, situated in the western part of the Balkan Peninsula in South-eastern Europe. According to recent archaeological fi ndings, Bosnia and Herzegovina has been occupied by modern humans since the Palaeolithic period. The structure of Bosnia-Herzegovina's human populations is very complex and specifi c, due to which it is interesting for various population-genetic surveys. The population of Bosnia and Herzegovina has been the focus of bio-anthropological and population genetics studies since the 19th century. The fi rst known bio-anthropological analyses of Bosnia-Herzegovina population were primarily based on the observation of some phenotypic traits. Later examinations included cytogenetic and DNA based molecular markers. The results of all studies which have been done up to date showed no accented genetic difference among the populations (based on geographical regions) with quite high diversity within them. Human population of Bosnia and Herzegovina is closely related to other populations in the Balkans. However, there are still many interesting features hidden within the existing diversity of local human populations that are still waiting to be discovered and described.
Assuntos
Citogenética , Genética Populacional , Bósnia e Herzegóvina , Humanos , FenótipoRESUMO
AIM: To present the results obtained in the identification of human remains from World War II found in two mass graves in Ljubuski, Bosnia and Herzegovina. METHODS: Samples from 10 skeletal remains were collected. Teeth and femoral fragments were collected from 9 skeletons and only a femoral fragment from 1 skeleton. DNA was isolated from bone and teeth samples using an optimized phenol/chloroform DNA extraction procedure. All samples required a pre-extraction decalcification with EDTA and additional post-extraction DNA purification using filter columns. Additionally, DNA from 12 reference samples (buccal swabs from potential living relatives) was extracted using the Qiagen DNA extraction method. QuantifilerTM Human DNA Quantification Kit was used for DNA quantification. PowerPlex ESI kit was used to simultaneously amplify 15 autosomal short tandem repeat (STR) loci, and PowerPlex Y23 was used to amplify 23 Y chromosomal STR loci. Matching probabilities were estimated using a standard statistical approach. RESULTS: A total of 10 samples were processed, 9 teeth and 1 femoral fragment. Nine of 10 samples were profiled using autosomal STR loci, which resulted in useful DNA profiles for 9 skeletal remains. A comparison of established victims' profiles against a reference sample database yielded 6 positive identifications. CONCLUSION: DNA analysis may efficiently contribute to the identification of remains even seven decades after the end of the World War II. The significant percentage of positively identified remains (60%), even when the number of the examined possible living relatives was relatively small (only 12), proved the importance of cooperation with the members of the local community, who helped to identify the closest missing persons' relatives and collect referent samples from them.
Assuntos
Impressões Digitais de DNA/métodos , Antropologia Forense/métodos , II Guerra Mundial , Osso e Ossos , Bósnia e Herzegóvina , Fêmur , Humanos , Repetições de Microssatélites , Mucosa Bucal/citologia , DenteRESUMO
AIM: To detect polymorphisms of 23 Y-chromosomal short tandem repeat (STR) loci, including 6 new loci, in a reference database of male population of Bosnia and Herzegovina, as well as to assess the importance of increasing the number of Y-STR loci utilized in forensic DNA analysis. METHODS: The reference sample consisted of 100 healthy, unrelated men originating from Bosnia and Herzegovina. Sample collection using buccal swabs was performed in all geographical regions of Bosnia and Herzegovina in the period from 2010 to 2011. DNA samples were typed for 23 Y STR loci, including 6 new loci: DYS576, DYS481, DYS549, DYS533, DYS570, and DYS643, which are included in the new PowerPlex® Y 23 amplification kit. RESULTS: The absolute frequency of generated haplotypes was calculated and results showed that 98 samples had unique Y 23 haplotypes, and that only two samples shared the same haplotype. The most polymorphic locus was DYS418, with 14 detected alleles and the least polymorphic loci were DYS389I, DYS391, DYS437, and DYS393. CONCLUSION: This study showed that by increasing the number of highly polymorphic Y STR markers, to include those tested in our analysis, leads to a reduction of repeating haplotypes, which is very important in the application of forensic DNA analysis.
Assuntos
Cromossomos Humanos Y/genética , Genética Populacional , Haplótipos , Repetições de Microssatélites/genética , Polimorfismo de Nucleotídeo Único/genética , Alelos , Bósnia e Herzegóvina , DNA/genética , Genética Forense , Marcadores Genéticos , Genótipo , Humanos , Masculino , Reação em Cadeia da Polimerase , Grupos Populacionais/genéticaRESUMO
Sexual violence represents a widespread social problem associated with serious lifelong consequences. In many cases, an outcome of sexual violence is the victim's unwanted pregnancy, usually ended in an abortion. The objective of this paper is to report five rape cases, including rapes of a minor and young woman, two incest cases and a case of human trafficking for sexual exploitation, where every case resulted in the victim's pregnancy. In each case, pregnancy was terminated in the first trimester or at the beginning of the second trimester in the relevant medical center or clinic. Fresh fetal blood or aborted tissue samples were delivered to our laboratory in order to perform paternity testing for the purpose of proving the crime. DNA extraction using Qiagen Dneasy™ Tissue Kit was optimized according to the sample type. Amplification of autosomal STR (Short Tandem Repeat) markers was performed using the PowerPlex®16 System. In two cases, mixtures of maternal and fetal DNA in the aborted fetal material were found. Using the LRmix Studio v.2.1.5 Software for interpreting DNA mixtures based on a probabilistic model, the likelihoods of maternal contribution and presence of fetal allelic variants inherited from the alleged father/suspect were calculated. Based on these results, we confirmed the presence of assumed fetal fractions (determined before software analysis) in the mixtures. In all cases, positive paternity proved the crime (probabilities of paternity >99.9999%). This cases report once again pointed out the importance of DNA analysis in the process of clarifying and solving forensic cases and demonstrated that the LRmix Studio v.2.1.5 Software can deal with complex cases such as sexual assaults.
Assuntos
Delitos Sexuais , Gravidez , Feminino , Humanos , Medicina Legal , Paternidade , Feto Abortado , Repetições de Microssatélites , DNA , Impressões Digitais de DNARESUMO
Medieval cemeteries Klisa-Guca Gora, Alihodze and Glavica-Han Bila located in the Travnik area (Travnik, Bosnia and Herzegovina) were archaeologically examined in the period 2011-2014, revealing human skeletal remains of 11 individuals in total. Archaeological skeletal samples, previously deposited in Travnik Homeland Museum (Travnik, Bosnia and Herzegovina) were subjected to genetic analysis. The aim of this research was to test familiar relationship of 11 individuals excavated from three medieval cemeteries and to predict Y-haplogroup for male individuals. In order to perform molecular-genetic characterisation of collected human skeletal remains, two systems of genetic markers were analysed: autosomal and Y-STR loci. Complete or partial data obtained by autosomal STR typing of 11 individuals were subjected to kinship analysis. Male sex was determined in eight samples out of 11. Direct relatives of the "brother-brother" type were detected in one case with high kinship probability (KP) value of 99.99996 %. Complete or nearly complete and usable Y-STR profiles were obtained for six out of eight male individuals. The presence of identical haplotypes at Y-STR loci and results of Y-haplogroup prediction suggest that all male individuals share the same paternal lineage and belong to J2a haplogroup. Overall, this study emphasises the usefulness, efficiency and sensitivity of STR markers in the molecular-genetic characterisation of old skeletal remains as well as the importance of employing additional markers like Y-STRs in archaeogenetic studies, besides traditionally used autosomal STR markers, in order to get a comprehensive information about close and distant relatives, and ancestry.
Assuntos
Restos Mortais , Impressões Digitais de DNA , Humanos , Masculino , Cromossomos Humanos Y , Impressões Digitais de DNA/métodos , Família , Marcadores Genéticos , Haplótipos , Repetições de Microssatélites , História MedievalRESUMO
Many heavy metals (HMs) are essential micronutrients for the growth and development of plants. However, human activities such as mining, smelting, waste disposal, and industrial processes have led to toxic levels of HMs in soil. Fortunately, many plant species have developed incredible adaptive mechanisms to survive and thrive in such harsh environments. As a widespread and ruderal species, Geranium robertianum L. inhabits versatile soil types, both polluted and unpolluted. Considering the ubiquity of G. robertianum, the study aimed to determine whether geographically distant populations can tolerate HMs. We collected soil and plant samples from serpentine, an anthropogenic heavy metal contaminated, and a non-metalliferous site to study the physiological state of G. robertianum. HMs in soil and plants were determined using flame atomic absorption spectrometry. Spectrophotometric methods were used to measure the total content of chlorophylls a and b, total phenolics, phenolic acids, flavonoids, and proline. Principal component analysis (PCA) was used to investigate the potential correlation between HMs concentrations gathered from various soil types and plant samples and biochemical data acquired for plant material. A statistically significant difference was observed for all localities regarding secondary metabolite parameters. A positive correlation between Ni and Zn in soil and Ni and Zn in plant matter was observed (p<0.0005) indicating higher absorption. Regardless of high concentrations of heavy metals in investigated soils, G. robertianum displayed resilience and was capable of thriving. These results may be ascribed to several protective mechanisms that allow G. robertianum to express normal growth and development and act as a pioneer species.
Assuntos
Geranium , Metais Pesados , Poluentes do Solo , Humanos , Geranium/metabolismo , Solo/química , Poluentes do Solo/análise , Metais Pesados/análise , Plantas/metabolismo , Monitoramento AmbientalRESUMO
CONTEXT: Broccoli [Brassica oleracea L. var. italica Plenck. (Brassicaceae)] contains substantial quantities of bioactive compounds, which are good free radical scavengers and thus might have strong antitumor properties. Enhancing production of plant secondary metabolites could be obtained with phytohormones that have significant effects on the metabolism of secondary metabolites. In that manner, in vitro culture presents good model for manipulation with plant tissues in order to affect secondary metabolite production and thus enhance bioactive properties of plants. OBJECTIVE: Estimation of the antioxidative and antitumor properties of broccoli cultivated in different in vitro conditions. MATERIALS AND METHODS: In vitro germinated and cultivated broccoli seedlings, as well as spontaneously developed calli, were subjected to Soxhlet extraction. Antioxidative activity of the herbal extracts was determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH(â¢)) radical method. Antitumor properties of the extracts were determined using crown-gall tumor inhibition (potato disc) assay. RESULTS: Three, 10, 20, and 30 days old broccoli seedlings, cultivated in vitro on three different Murashige-Skoog media, two types of callus, and seedlings from sterile filter paper were used for extraction. In total, 15 aqueous extracts were tested for antioxidative and antitumor potential. Three day-old seedlings showed the highest antioxidative activity. Eleven out of 15 aqueous extracts demonstrated above 50% of crown-gall tumor inhibition in comparison with the control. Tumor inhibition was in association with types and concentrations of phytohormones presented in growing media. DISCUSSION AND CONCLUSIONS: It is demonstrated that phytohormones in plant-growing media could affect the bioactive properties of broccoli either through increasing or decreasing their antioxidative and antitumor potential.
Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Brassica/química , Extratos Vegetais/farmacologia , Antineoplásicos Fitogênicos/isolamento & purificação , Antioxidantes/isolamento & purificação , Compostos de Bifenilo , Meios de Cultura , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/farmacologia , Picratos , Tumores de Planta/microbiologia , Fatores de TempoRESUMO
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
In our previous population study, we have used twelve Y-chromosomal short tandem repeats loci incorporated in the PowerPlex Y System to determine Y-STR diversity in B&H human population. With intent to obtain additional verification of the previously obtained results as well as to establish specific reference for a local B&H population, we have decided to test DNA samples collected from 100 unrelated healthy male Canton Sarajevo residents (from Sarajevo region) for the same twelve Y-linked short tandem repeats loci. Qiagen DNeasy Tissue Kit (Qiagen, GmbH, Hilden, Germany) was used for DNA extraction from buccal swabs and PowerPlex Y System (Promega Corp., Madison, WI) has been used to simultaneously amplify Y-STR loci by PCR. PowerPlex Y System includes 12 STR loci: DYS19, DYS385a, DYS385b, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439. The total PCR reaction volume was 5 microL. PCR amplifications were carried out in PE GeneAmp PCR System Thermal Cycler (ABI). Electrophoresis of the amplification products was preformed on an ABI PRISM 310 genetic analyzer (ABI, Foster City, CA) according to the manufacturer's recommendations. The raw data were compiled and analyzed using the accessory software: ABI PRISM Data Collection Software and Genemapper version 3.2. In addition, we have compared the obtained "Sarajevo" dataset with the data previously generated for the entire Bosnian and Herzegovinian population, as well as with the available data on geographically close (neighboring) European populations. The results of this study will be used as guidelines in additional improving of research into genetic relationship among recent local B&H populations, both isolated and open, which is a long-term project in our country.
Assuntos
Cromossomos Humanos Y/genética , Genética Populacional , Sequências de Repetição em Tandem/genética , Alelos , Bósnia e Herzegóvina , Croácia , Variação Genética , Geografia , Humanos , Masculino , Valores de ReferênciaRESUMO
This study was conducted to confirm preliminary anthropological research indicating the specificity of isolated Selska Valley populations and implement clear mapping of genetic distances between neighboring populations and similar "inland island" populations from the region. The sample consisted of 86 unrelated individuals born in the Selska Valley from the lowland villages (Bukovica, Sevlje, Dolenja Vas, Selca, Zelezniki and Zali Log) and the mountain villages (Podlonk, Prtovc, Spodnje Danje, Zgornja Sorica and Spodnja Sorica). The for mentioned 15 STR loci (D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX and FGA) were analyzed and statistical analysis was applied to determine population-genetics and forensic parameters. The frequencies of 15 STR loci from isolated populations of Slovenian villages, Slovenia, Bosnian mountain villages and Bosnia and Herzegovina were analyzed to calculate genetic distances between them. Our results confirmed a similar genetic pattern between the Selska Valley mountain villages and Slovenian general population as well as Bosnian mountain villages and general Bosnian population. Even if the sample size was relatively small and examined populations were geographically isolated, observed genetic variation within the whole population was relatively high and comparable with neighboring populations. Additional analysis proved that the Selska Valley population is genetically closest to the Slovenian and Austrian populations. On the other hand, comparison with similarly patterned populations confirmed that this population could be recognized as "inland island" population in genetic terms.
Assuntos
Variação Genética/genética , Repetições de Microssatélites/genética , População Branca/genética , Bósnia e Herzegóvina , Frequência do Gene/genética , Genética Populacional , Humanos , População Rural , EslovêniaRESUMO
The floods in Bosnia and Herzegovina in May 2014 caused landslides all over the country. In the small village of Serici, near the town of Zenica, a landslide destroyed the local cemetery, relocated graves, and commingled skeletal remains. As the use of other physical methods of identification (facial recognition, fingerprint analysis, dental analysis, etc.) was not possible, DNA analysis was applied. DNA was isolated from 20 skeletal remains (bone and tooth samples) and six reference samples (blood from living relatives) and amplified using PowerPlex® Fusion and PowerPlex® Y23 kits. DNA profiles were generated for all reference samples and 17 skeletal remains. A statistical analysis (calculation of paternity, maternity, and sibling indexes and matching probabilities) resulted in 10 positive identifications. In this study, 5 individuals were identified based on one reference sample. This has once again demonstrated the significance of DNA analysis in resolving the most complicated cases, such as the identification of commingled human skeletal remains.
Assuntos
Restos Mortais , Impressões Digitais de DNA , DNA/isolamento & purificação , Osso e Ossos/química , Bósnia e Herzegóvina , Cemitérios , Cromossomos Humanos Y , Feminino , Inundações , Humanos , Masculino , Repetições de Microssatélites , Linhagem , Reação em Cadeia da Polimerase , Dente/químicaRESUMO
The goal of this study was to examine the effectiveness of 6 STR markers application (D21S1435, D21S11, D21S1270, D21S1411, D21S226 and IFNAR) in molecular genetic diagnostics of Down syndrome (DS) and to compare it with cytogenetic method. Testing was performed on 73 children, with the previously cytogenetically confirmed Down syndrome. DNA isolated from the buccal swab was used. Previously mentioned loci located on chromosome 21 were simultaneously amplified using quantitative fluorescence PCR (QF PCR). Using this method, 60 previously cytogenetically diagnosed DS with standard type of trisomy 21 were confirmed. Furthermore, six of eight children with mosaic type of DS were detected. Two false negative results for mosaic type of DS were obtained. Finally, five children with the translocation type of Down syndrome were also confirmed with this molecular test. In conclusion, molecular genetic analysis of STR loci is fast, cheap and simple method that could be used in detection of DS. Regarding possible false results detected for certain number of mosaic types, cytogenetic analysis should be used as a confirmatory test.
Assuntos
Cromossomos Humanos Par 21/genética , Análise Citogenética , Síndrome de Down/diagnóstico , Síndrome de Down/genética , Repetições de Microssatélites/genética , Técnicas de Diagnóstico Molecular , Criança , Estudos de Coortes , Feminino , Loci Gênicos/genética , Humanos , Masculino , Mosaicismo , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Translocação Genética/genéticaRESUMO
Despite their known toxic properties, various Helleborus species are used as medicaments in folk medicine to treat some diseases and health conditions. As the main mechanism of many cytostatic drugs is based on their cytotoxic activity, there is potential for the toxicity of hellebore to be used in anticancer therapy. This study tested the geno- and cytotoxic effects of extracts of three hellebore taxa (Helleborus odorus, Helleborus multifidus and Helleborus hercegovinus) on meristemic onion (Alliumcepa L.) cells and human lymphocytes. Treatments with Helleborus extracts induced cytotoxic and cytostatic effects in meristemic onion cells as well as in cultivated cytokinesis-blocked human lymphocytes. Cytokinesis-block micronucleus cytome assay indicated that treatments with hellebore extracts induce genotoxic effects in human lymphocytes, and that the significant mechanism of their antiproliferative activity is apoptosis induction.
Assuntos
Citotoxinas/isolamento & purificação , Citotoxinas/farmacologia , Helleborus/química , Apoptose/efeitos dos fármacos , Células Cultivadas/efeitos dos fármacos , Citocinese/efeitos dos fármacos , Dano ao DNA , Relação Dose-Resposta a Droga , Helleborus/genética , Humanos , Linfócitos/efeitos dos fármacos , Testes para MicronúcleosRESUMO
The objective of this study was to determine total phenolic and flavonoid contents, antioxidant and antimicrobial activities of methanolic extracts from the leaves and barks of three Alnus species. The phenolic and flavonoid contents of extracts were determined spectrophotometrically using Folin-Ciocalteau and aluminium chloride methods, respectively. In addition, antioxidant activity of the extracts was determined using 1,1-diphenyl-2-picrylhydrazyl radical scavenging method. The antimicrobial activity was performed by disc diffusion assay against six reference bacterial strains including Gram-negative and Gram-positive bacteria and two fungal strains. Extract of Alnus viridis bark contained the highest amounts of total phenolics (780 mg CAT/g), while extract of A. viridis leaves had the highest amount of flavonoids (30.01 mg RUT/g). All extracts showed antioxidant activity higher than thymol, which was used as a positive probe. The largest diameters of inhibition zone (25 mm) were recorded with Bacillus subtilis 168 M and Staphylococcus aureus ATCC 6538.
Assuntos
Alnus/química , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/farmacologia , Antioxidantes/isolamento & purificação , Antioxidantes/farmacologia , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Fenóis/isolamento & purificação , Fenóis/farmacologia , Antibacterianos/química , Anti-Infecciosos/química , Antioxidantes/análise , Antioxidantes/química , Compostos de Bifenilo/farmacologia , Flavonoides/análise , Flavonoides/química , Testes de Sensibilidade Microbiana , Fenóis/análise , Fenóis/química , Picratos/farmacologia , Casca de Planta/química , Extratos Vegetais , Folhas de Planta/química , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacosRESUMO
This study was undertaken in order to evaluate possible antioxidative and antiproliferative activities of three Helleborus taxa. The dry leaves and roots of three Helleborus taxa were extracted with ethanol and water. A phytochemical evaluation of the selected extracts was performed using spectrophotometric methods and a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity assay was used for measuring the antioxidative activity of extracts. The antiproliferative activity of the three Helleborus taxa was studied using Burkitt's lymphoma B cells (BJAB) cell lines. The phytochemical evaluation showed that the leaves contain high levels of total phenolic and flavonoid content. Results from the DPPH assay indicated that the activity of the ethanol and water extracts of the leaves was higher than that of positive control (thymol). Extracts from the roots of H. odorus also displayed higher antioxidant activity than the positive probe, while H. mulifidus and H. hercegovinus root extracts were less effective. A statistically significant correlation between total phenolic content and antioxidative properties indicates that these compounds contribute to the antioxidant activity. The highest percentage of cell growth inhibition was observed when testing the water root extracts of H. multifidus (50.14%) and H. hercegovinus (49.04%). In contrast, the water leaf extract of H. hercegovinus exhibited the lowest inhibition of cell growth (8.59%), although it showed strong antioxidant activity.