Mucopolysaccharidosis III: Molecular basis and treatment.

Mucopolysaccharidoses (MPSs) are known as rare genetic diseases which are caused by mutation in the enzyme heparin sulfate, which normally leads to degradation and accumulation of glycosaminoglycans in the cells. There are 11 types of MPSs, whereby neuropathy may occur in seven of them (MPS I, II, IIIA, IIIB, IIIC, IIID and VII). Accumulation of degraded heparin sulfate in lysosomes causes cellular dysfunction and malfunction of several organs. However, the exact molecular mechanism how protein degradation and storage leads to cellular dysfunction is not understood, yet. Nonetheless, several genetic and biochemical methods for diagnosis of MPSs are available nowadays. Here we provide an overview on known molecular basis of MPS in general, including enzyme defects and symptoms of MPS; however, the main focus is on MPS type III together with potential and perspective therapy-options.

Association of biomarkers of inflammation and HLA-DRB1 gene locus with risk of developing rheumatoid arthritis in females.

Rheumatoid arthritis (RA) is an autoimmune disease causing chronic inflammation of the joints. Multiple factors, including HLA-DRB1 gene variants, influence the susceptibility to RA. The HLA-DRB1 gene is part of a family of genes called the human leukocyte antigen (HLA) complex. In this study, we compared the inflammatory biomarkers values, including erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP), between patients with RA and healthy control group of females of the Public Institution Health Centre of Sarajevo Canton. In addition, we estimated the frequencies of the HLA-DRB1 gene variants and their association with the risk for RA development in females. The haematological and biochemical tests were completed on automated analyzers. To assess the association between the HLA-DRB genes and the risk of RA in females, low-resolution genotyping of the HLA-DRB1, DRB3, DRB4, and DRB5 gene loci was performed by the sequence-specific polymerase chain reaction method (PCR-SSP). ESR and CRP were the most sensitive acute-phase reactants in females with RA and there was a correlation between ESR and CRP values in RA patients. There was significantly positive association between of the HLA-DRB1*03, *04, *08, *10, *11, and *14 variants and elevated values of ESR in RA patients, but negative between HLA-DRB1*03, *13 and *15 alleles and elevated CRP values. Furthermore, our results confirm genetic susceptibility to RA in a female population to the members of the HLA-DRB1*04 and *03 allelic groups, the DRB1*04/DRB1*04 and DRB1*03/DRB1*04 genotypes, and the DRB1*04-DRB4* or DRB1*03-DRB3* haplotypes, which, therefore, represent risk factors for the development of this disease. According to our results, the DRB1*01/DRB1*15 and DRB1*07/DRB1*16 genotypes and the HLA-DRB5 gene locus represent a protective factor for RA. The presence of specific HLA-DRB1 gene variants increases the risk of developing RA, while other variants provide protection against disease. Therefore, HLA typing could be helpful in the prediction of RA development and establishing and confirming a definitive diagnosis of autoimmune diseases in some subjects. A strong association with the higher levels of ESR and CRP could be used to establish definitive diagnosis and introduce of early treatment of RA to prevent the occurrence of RA symptoms.

The Frequency of the 677C>T and 1298A>C Polymorphisms in the Methylenetetrahydrofolate Reductase (MTHFR) Gene in the Population.

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.

The Genetic Variation of CYP2D6 Gene in the Bosnian Population.

INTRODUCTION: Genetic polymorphism is associated with individual responses to medication and susceptibility to diseases, and it represents the basis for individualized medical treatment and drug genomics studies. Genetic variation at CYP2D6 is high, both among populations and among individuals in the same population. AIM: The aim of the study was to investigate the CYP2D6 gene duplication and the non-synonymous single-nucleotide polymorphisms (SNP) 100C>T in the CYP2D6 gene in members of the Bosnian population. MATERIAL AND METHODS: The blood samples were collected from 151 unrelated and healthy donors from Bosnian populations consisted of 94 females and 57 males. Duplex long-range PCR was used to determine whether individuals carrying CYP2D6 gene duplication. The resulting PCR product of 5.1 kb, containing all nine CYP2D6 exons, was used as template for detection of the CYP2D6 100C>T allele by nested PCR. RESULTS: The CYP2D6 gene duplication frequency found in the Bosnian population (2.73%) was related to the frequencies of this allele in other Caucasians. The gene duplication is the result of inheritance of more than two copies of the fully functional CYP2D6 alleles. In contrast, the frequency of the CYP2D6 100C>T variant, with possibly damaging function, in the Bosnian population (15.56%) was significantly higher when compared with the other Caucasians but significantly lower when compared with Asians. CONCLUSION: CYP2D6 metabolizes many commonly prescribed drugs. Variations in the gene encoding this enzyme have been associated with individual differences in drug metabolism rates. The individuals with multiple CYP2D6 gene copies metabolize drugs more rapidly and therapeutic plasma levels will not be achieved at ordinary drug dosages. The non-synonymous coding SNP (100C>T) were predicted to have damaging effects on the protein function.

The reality of cancer treatment in a developing country: the effects of delayed TKI treatment on survival, cytogenetic and molecular responses in chronic myeloid leukaemia patients.

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.

Assessment of the genotoxicity and cytotoxicity in environmentally exposed human populations to heavy metals using the cytokinesis-block micronucleus cytome assay.

The cytokinesis-block micronucleus cytome (CBMN-Cyt) assay was developed as a system for evaluating DNA damage, cytostasis, and cytotoxicity. The aim of the present study was to estimate levels of micronuclei (MNi), nucleoplasmic bridges (NPBs), nuclear buds (NBUDs), cell death (apoptosis/necrosis), nuclear division index, and nuclear division cytotoxicity index values in the peripheral blood lymphocytes of environmentally exposed subjects to heavy metals from five Bosnian regions, characterized by different exposure to heavy metals. The study was performed using CBMN-Cyt assay, considering factors, such as age, gender and smoking habits and their possible effects on analyzed parameters. In total, 104 healthy subjects were selected (49.04% females and 50.96% males; average age, 35.41 years; 51.92% smokers and 48.08% nonsmokers). There was significant difference between the frequency of NBUDs in Tuzla as compared to the control group. Furthermore, there was observed a statistically significant difference for the frequency of NPBs between Zenica, Olovo, and Kakanj when compared with the controls. Males showed a significantly higher number of apoptotic cells than females in controls. There were significant differences between smokers and nonsmokers in the frequency of NPBs in controls (higher in nonsmokers) and necrotic cells in Olovo (higher in nonsmokers). The pack years of smoking significantly influenced the number of necrotic cells in controls and the frequency of NBUDs in the overall sample. The results of the present study provide evidence of significantly increased frequency of NPBs and NBUDs in exposed subjects, suggesting that these endpoints are highly sensitive markers for measuring genotoxicity.

The effect of age, sex, and lifestyle factors on micronucleus frequency in peripheral blood lymphocytes of the Bosnian population.

This study confirmed that the frequency of human lymphocyte biomarkers measured with the cytokinesis-block micronucleus cytome (CBMNcyt) assay, is associated with age, sex, and lifestyle factors. Cytogenetic analysis was carried out on samples from 100 healthy individuals living in Bosnia and Herzegovina. Cells were cytologically scored for viability status, defined by the proportion of necrotic and apoptotic cells; mitotic status, corresponding to the proportion and ratios of mono-, bi- and multinucleated cells; the nuclear division index and chromosomal damage, determined by the presence of micronuclei, nucleoplasmic bridges or nuclear buds of bi-nucleated cells. Ageing is positively associated with the frequency of cytogenetic biomarkers. The micronucleus frequency in females was significantly higher than the micronucleus frequency in males. The micronucleus frequency is positively associated with family history of cancer. Furthermore, it is positively correlated with smoking: the frequency is higher in male subjects with a smoking habit than in female smokers. However, alcohol is observed to decrease the frequency of apoptotic cells in human lymphocytes. The frequency of micronuclei was positively correlated with exposure to medication. Lastly, the frequency of nuclear buds and apoptotic and necrotic cells negatively correlated with exposure to radiation.

Chromosomal and nuclear alterations in root tip cells of allium cepa L. Induced by alprazolam.

INTRODUCTION: Alprazolam is a triazolobenzodiazepine used in panic disorders and other anxiety states. Target organ of Alprazolam is CNS, causing depression of respiration and consciousness. AIM: This study aimed to estimate the genotoxic potential of Alprazolam using Allium cepa test. METHODS: Allium cepa is one of the most suitable plants for detecting different types of xenobiotics. The test enables the assessment of different genetic endpoints making possible damage to the DNA of humans to be predicted. RESULTS: Alprazolam induced chromosomal (anaphase bridges, breaks, lagging and stickiness, abnormal spiralisation, multipolarity and polyploidy) and cytological aberrations, especially nuclear alterations (nuclear buds, fragmented nucleus and apoptotic bodies, cells without nucleus, binucleated and micronucleated cells), morphological alterations in shape and size of cells, spindle disturbance and polar deviation in root tip meristem cells of Allium cepa at all tested concentrations. Alprazolam also caused significant inhibition of mitotic index in these cells. CONCLUSION: These changes in cells are indicators of genotoxic potential of Alprazolam suggesting a need for further in vitro studies on animal and human lymphocytes as well as in vivo studies.

Comparative analysis of dermatoglyphic traits in Albanian and Turkish population living in Kosovo.

The aim of the study was to compare quantitative dermatoglyphic traits of two ethnic groups with different origin and customs, living on the same territory. The dermatoglyphic prints were collected from 800 inhabitants of the Dukagjin valley in southwest Kosovo, of Albanian (400) and Turkish (400) ethnic origin. The quantitative analysis comprised the number of ridges and triradii on the fingers, and the number of ridges in the interdigital areas on the palm (a-b, b-c, and c-d) as well as the size of the atd angle. The statistical analysis showed significant differences between the Albanian and the Turkish males for two fingers and pattern intensity index left, and on palms for a-b rc and c-d rc on both hands and b-c re on the left hand, and between females for six fingers and almost all palmar traits. Significant inter-population variations were observed for most palmar areas in both sexes but more pronouncedly in females. The differences found between the examined population groups show that admixture between Albanian and Turkish population in Kosovo has been small, and the Turks have retained their ethnic identity for several centuries.

[Radiation-induced chromosome aberrations]. / Hromosomske aberacije inducirane radijacijom.

Chromosomal aberrations in peripheral lymphocytes have been used as biomarkers of radiation exposure. The results obtained for this cytogentic study suggest that 86 persons who examined were with structural chromosome aberrations. The sample was included 100 persons who were exposed to radiation professionally. The most frequent persons were found with one, two, three and four chromosome aberrations, and 19 persons had six and more aberrations, while 341 total chromosome aberrations were registried. The most frequent radiation-induced chromosome aberrations in vivo were chromatid breaks, acentric fragments, acentric minutes and isochromatid breaks, while dicentrics, rings, quadriradials and triradials, translocations (chromosome rearrangements), marker chromosomes and chromosome repulsions were less frequent. These results suggest that are the most frequent chromosome single breaks, while more complex recombinations are less frequent. Obtained results when compared with results of standard showed that 32.56% of total number of persons with aberrations or 28% total number of analyzed persons found was different from standard. Chromosomal aberrations in lymphocytes suggest that these persons were probably exposed to ionizing radiation.

The genotoxicity of vitamin C in vitro.

The genotoxic effects of Vitamin C (ascorbic acid) on human lymphocytes in vitro were estimated by analyzing and identifying various chromosome abnormalities, in relation to the concentration of Vitamin C. Testing concentrations of Vitamin C induced different aberrations including the impairment of spindle function. The spindle disturbances can result in mitotic arrest, multipolar spindles and multipolar segregation, errors in chromosome segregation, formation of chromosome bridges and chromosome laggards. The most frequent irregularities were found in anaphase and telophase. A certain number of lymphocytes were arrested at anaphase or telophase (in colchicine-untreated cultures of human lymphocytes). Testing concentrations of ascorbic acid did not induce a significant increase in the number of aneuploid mitoses and were not clastogenic except at the highest concentration (1,000 microg/ml) in colchicine-treated cultures, and in colchicine-untreated cultures of human lymphocytes the pulverization of chromosome was observed. Vitamin C changed the mitotic index value of lymphocytes notably at the higher concentrations (250, 500 and 1,000 microg/ml).

[Unusual chromosome translocation 46, XX, t(1;16) (p13;p13) as a possible cause of retarded development of carrier]. / Neobicna hromosomska translokacija 46, XX, t(1;16) (p13;p13) kao moguci uzrok usporenog razvoja nosioca.

Unbalanced changes of chromosomes with loss or extra genetic material often cause changing phenotype. The main characteristics unbalanced karyotype involves mental retardation and numbers of anomalies, especially if the change involves autosoms. In this paper child with retard development and de novo translocation between chromosome 1 and chromosome 16 was described. Cytogenetics survey was carried out to establishing possible cause in relation with retard development and it involved standard cytogenetics method and method of differential staining (G-banding) that enables identification of certain structural rearrangement of chromosomes. Chromosome aberration was translocation of part of p arm of chromosome 1 on p arm of chromosome 16, that is, it presents abnormal karyotype 46, XX, t(1;16)(p13;p13) that might be one of the cause of retard development of child.