Luteolin and delphinidin are the flavonoids with known protective roles. They inhibit genotoxic effects induced by halogenated boroxine (HB) in vitro. Statistically significant decrease in the frequency of micronuclei and nuclear buds and suppression of the occurrence of aberrant cells were observed before, but mechanism of its anti-genotoxic activity is still not clear. In our experiment we aimed to quantify HB effects on the relative expression of CAT (catalase) gene and explore antioxidative effects of luteolin and delphinidin via restoration of CAT gene activity. Cell cultures from peripheral blood lymphocytes of five healthy donors were established and treated with independent and concomitant treatments of HB with luteolin or delphinidin. Total RNA was isolated from harvested cells and reverse-transcribed. SYBR based Real-Time PCR amplification method was used. Analysis of results included normalization of ratio of target (CAT) and housekeeping (GAPDH) gene and statistical analysis (REST®). Luteolin itself lead to downregulation of relative CAT gene expression as well as HB. But simultaneous treatment of HB and bioflavonoids lead to upregulation. Delphinidin as independent treatment and as simultaneous treatment caused upregulation of relative CAT gene expression. Obtained results may indicate protective role of delphinidin and luteolin to oxidative damage caused by HB, and also that new approaches to the treatment applications of HB should include bioflavonoids and monitoring corresponding antioxidant system. Our findings indicate that there is a quantifiable effect of luteolin and delphinidine on antioxidant genes which could be used in exact monitoring of oxidative stress related events.

Abstract Although prostate cancer accounts for the highest number of newly diagnosed cases of cancer in men, it represents a specific diagnostic challenge in modern oncology. The standard diagnosis of prostatic carcinoma begins with the screening of serum concentrations of PSA (Prostate Specific Antigen). If the concentration of serum PSA levels is above 4 ng/mL, the patient is further referred to a digital rectal examination in order to determine an increase in prostate volume. In cases where enlargement of the prostate is observed, the next step is biopsy of prostate tissue. This physically painful and invasive approach to confirm the diagnosis is often unnecessary because, in many cases, the patohistologic analysis determines diagnosis of benign prostatic hyperplasia, and not a tumor. In this study, we investigated the possibilities of detection and measurement of the relative level of gene expression of the KLK3 (Kallikrein-related peptidase 3), PCA3 (Prostate Cancer Gene 3) and TEMPRSS: ERG (Transmembrane protease serine2 and in-ETS erythroblostosis virus E26 oncogene homolog) genes from the urine samples of patients with prostatic diseases and healthy controls. Urine was the sample of choice because it is taken in a non-invasive manner, and could potentially serve to make better selection to biopsy. One of the selected genes (KLK3) differed significantly in the samples of various pathological conditions of the prostate, and therefore we consider that its further investigation is reasonable.

MODY (maturity-onset diabetes of the young) is an autosomal dominant form of diabetes that is usually manifested before the 25-year of life. This type of diabetes is caused by defects in the primary insulin secretion. There are several types of MODY, which are monogenic diseases, where mutations in a single gene are responsible for a particular type of MODY. Currently, there are eleven types of MODY, from which the most common types are MODY 2 and MODY 3 (with mutations on GCK and HNF1A genes, respectively). We identified very rare MODY 7 type of diabetes in three family members by MLPA analysis.

Abstract Plant bioflavonoids are widely present in the human diet and have various protective properties. In this study, we have demonstrated the capacity of delphinidin and luteolin to increase human telomerase reverse transcriptase (hTERT) expression level and act as protective agents against halogenated boroxine-induced genotoxic damage. Halogenated boroxine K2(B3O3F4OH) (HB), is a novel compound with potential for the treatment of both benign and malignant skin changes. In vivo and in vitro studies have confirmed the inhibitory effects of HB on carcinoma cell proliferation and cell cycle progression as well as enzyme inhibition. However, minor genotoxic effects of HB are registered in higher applied concentrations, but those can be suppressed by in vitro addition of delphinidin and luteolin in appropriate concentrations. Fresh peripheral blood samples were cultivated for 72 h followed by independent and concomitant treatments of HB with luteolin or delphinidin. We analyzed the differences in relative hTERT expression between series of treatments compared with controls, which were based on normalized ratios with housekeeping genes. The obtained results have shown that selected bioflavonoids induce upregulation of hTERT that may contribute to the repair of genotoxic damage in vitro.

Incidence of breast cancer ranges from 27 per 100,000 in Middle Africa and Eastern Asia to 92 per 100,000 in Northern America. It is the fifth most common cause of death from cancer in women, with an estimated 522,000 deaths per year (6.4% of the total). Autosomal dominant inheritance of these cancers is characterized by transmission of cancer predisposition from generation to generation, with around 5-10% of all breast cancers being associated with inherited mutations in BRCA1, BRCA2 and other genes.  Breast and ovarian cancers are strongly associated with BRCA1 and BRCA2 mutations. In this study, we genotyped BRCA1 gene for large genomic rearrangements in breast and ovarian cancer patients from Bosnia and Herzegovina, with aim to assess frequency of large BRCA1 mutations (exon deletions/duplications) in this group. We collected 59 breast cancer samples, as well as other data concerning patients’ histopathological parameters of tumor, like age at diagnosis, cancer type, TNM class, cancer grade, as well as estrogen, progesterone and Her2/neu expression. Following DNA extraction from breast cancer samples (tissue after biopsy), BRCA1 mutations were identified by Multiplex Ligase - Dependent Probe Amplification (MLPA) analysis. Biostatistical analyses were conducted using MedCalc v.9.2.0.0 software. In all statistical tests p<0.05 was considered significant. Mean age at diagnosis was 54±1.75 (range 17 – 80). BRCA1 genomic rearrangements were found in 22% of breast and ovarian cancer patients. Statistically significant associations and correlations were found between BRCA1 genomic rearrangements and cancer type, estrogen, progesterone and Her2/neu expression, but not cancer grade, size, invasiveness or patients’ age

Conventional screening and diagnostic procedures in prostate complaints rely on PSA (Prostate Specific Antigen) concentration which is not specific for prostate cancer and frequently leads to unnecessary invasive procedures in order to exclude malignant disease. It is estimated that approximately 50% of persons who underwent tissue biopsy did so based on false positive PSA value. Therefore a proper and timely differential diagnosis of malignant disease using non-invasive techniques remains one of the biggest challenges in medicine. Urine is the invaluable source of biological information contained in small molecules i.e. RNA that is easily accessible and detectable using molecular genetics techniques. We describe economical and fast method for relative expression analysis applicable to any target gene using urine as a sample. Efficient non-invasive method for identification of malignant or high risk cases prove useful in reduction of patient distress during the diagnostic procedure and significantly reduce healthcare costs.

Cervical cancer represents a serious health problem affecting women worldwide especially in developing countries due to low socioeconomic status, inadequate health-care infrastructure, weaknesses in education on this particular issue and lack of effective screening programmes. The primary aim of this study was to assess alternative screening method for the improvement of cervical cancer prevention in conditions of Bosnia and Herzegovina (B&H), which could be applicable in other developing countries as well. The study was conducted on 101 subjects who provided their self-sampled vaginal swabs and/or cervical specimens collected by their gynecologists. Universal Human Papilloma Virus (HPV) primer set optimized to detect a wide range of HPV types was used for HPV genotyping from obtained swab samples in multiplex PCR. Amplicons were analyzed in agarose gel and Agilent 2100 bioanalyzer – a platform based on microfluid technology. Inter-rater agreement kappa (MedCalc2) was used to assess concordance between results of cervical and vaginal sample analysis. Out of 39 subjects who provided their vaginal and cervical samples, results of HPV detection mismatched in 10% of the cases. Inter-rater agreement showed good strength of coincidence between the results of cervical and vaginal sample analysis (kappa=0,748, CI=95 %). We presented an alternative PCR method for the detection of HPV based on vaginal self sampling which is affordable, informative, simple and applicable with high coverage level of defined targeted population and potentially significant in the given cultural and socioeconomic context.

Introduction: Single nucleotide polymorphisms (SNPs) have lately been used for prediction of metabolic processes that may be related to obesity. The aim of our study was to examine the association of SNPs of several genes with obesity and physical activity in 18 healthy volunteers. Methods: We used buccal swabs to collect and extract DNA from 18 volunteers. Pyrosequencing was used for molecular analysis of 13 polymorphisms in 10 genes (APOA2, MTHFR, MCM6, peroxisome proliferators-activated receptor gamma, FABP2, beta-2-adrenergic receptor (ADRB)2, ADRB3, A-actinin-3, angiotensin-converting enzyme, and FUT2). The volunteers’ personal data included body mass index (BMI), dietary practice and information on daily fitness and workout routine. Association between the 13 observed gene polymorphisms and individual BMI status (normal or overweight) was analyzed. Results of the DNA analysis were used for the expert evaluation by nutritionists and physiologists to obtain optimal regulation of nutrition and exercise. The volunteers had a dietary and fitness program for 12 months which they tracked by filling in a suitable study form. Results: 14 volunteers had a moderate genetic predisposition for abdominal adipose-tissue accumulation, while 4 of them had genotypes not associated with abdominal fat tissue accumulation. A statistically significant difference was found between the value of BMI before and after the implementation of personalized training and nutrition plan within the group of overweight volunteers (paired sample t=3.382; p = 0.006; exact p = 0.015). The single-locus F-test showed no association between the gene polymorphisms and BMI values. In addition, no correlation was detected between the gene polymorphisms and amount of BMI reduction prior and after the implementation of the personalized training and nutrition plan within the overweighed group of volunteers. Conclusion: Optimal nutrition and training plan are crucial for the BMI reduction as observed in the overweighed volunteers after the 12-month personalized training and individualized nutrition plan. However, the analyzed polymorphisms were not significantly associated with the obesity in this study.