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Daria Ler

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Background Serostudies are important resources when following pandemics and predicting their further spread, as well as determining the length of protection against reinfection and vaccine development. The aim of this study was to update data on the prevalence of seropositive individuals in Canton Sarajevo, Bosnia and Herzegovina (B&H) from September 2020 to May 2021. Methods Anti-SARS-CoV-2 antibodies were quantified using an electrochemiluminescence immunoassay. Results Compared to the period April–July 2020, when anti-SARS-CoV-2 antibodies were detected in 3.77% of samples, one year later (May 2021) the estimated percentage within the same population of the urban Canton Sarajevo was 29.9% (5,406/18,066). Of all anti-SARS-CoV-2 Ig-positive individuals, 53.27% were men, and 69.00% were of 50 years of age or younger. Also, the current update found the individuals 50 years of age or younger to be more frequently anti-SARS-CoV-2 Ig positive compared to older individuals. On the other hand, higher median anti-SARS-CoV-2 Ig levels were found in individuals > 50 years old than in younger individuals, as well as in men compared to women. Seropositivity gradually increased from September 2020 to May 2021, with the lowest frequency of positive cases (3.5%) observed in September 2020, and the highest frequency (77.7%) in January 2021. Conclusion Our results provided important seroprevalence data that could help in planning restrictive local public health measures to protect the population of Sarajevo Canton, especially considering that at the time of the study the vaccines were virtually inaccessible to the general population not belonging to any of the high-priority groups for vaccination.

Introduction Serological detection of SARS-CoV-2-specific immunoglobulins G (IgG) and M (IgM) antibodies is becoming increasingly important in the management of the COVID-19 pandemic. Methods We report the first results of COVID-19 serological testing in Bosnia and Herzegovina on 2841 samples collected and analysed in 2 medical institutions in Sarajevo. Antibody detection was performed using commercially available kits. Results In the first cohort, 43 IgM-positive/IgG-negative and 16 IgM-positive/IgG-positive individuals were detected, corresponding to 3.41% of participants having developed antibodies. In the second cohort, 4.28% participants were found to be IgM-negative/IgG-positive. Conclusions Our results suggest the need for population-wide serological surveying in Bosnia and Herzegovina.

Emina Huseinbegović, Daria Ler, M. Suljagić

Congenital cytomegalovirus (CMV) infection is the most common infectious cause of birth defects. It may cause both, immediate and long term health problems in infants. These include variety of symptoms, such as hearing loss, microcephaly, jaundice, hepatosplenomegaly and seizures. In more severe cases CMV infection can cause the death of an unborn baby and loss of pregnancy. Despite being one of the most extensively studied vertically transmitted infections recently, the adverse effects of vertically transmitted CMV infection are still not well presented to the general public, resulting in a low awareness among potential expectant mothers in Bosnia and Herzegovina. This study aims to elucidate the sensitivity of urine samples for CMV detection in infants as well as to reflect the importance of quantitative real-time PCR (qRT-PCR) in diagnostics of CMV infection in infants. qRT-PCR was used in this study as a technique for the screening of CMV DNA in a cohort of patients based in Sarajevo Canton. These results have shown that urine samples are sufficiently sensitive for the detection of CMV DNA in infants. Furthermore, the simultaneous analysis of several patients has shown a higher number of CMV DNA copies amplified in urine compared to blood samples, derived from the same patient, thus proposing urine as a reliable sample of choice for congenital CMV diagnostics. These findings may propose a need to classify qRT-PCR CMV test among one of the recommended first-trimester pregnancy screening tests, which could help in early detection of CMV infection in Sarajevo Canton.

Tea Bećirević, M. Suljagić, Daria Ler

Cervical cancer is one of the leading causes of cancer in women, worldwide. Infection with human papillomavirus (HPV) has been accepted as the primary cause for the development of invasive cervical cancer and its precursor lesions. Despite HPV infection has been proposed as an indispensable factor for cervical cancer development, only a subset of neoplastic lesions with HPV infection persist and progress to invasive cancer. This suggests us that other molecular events are also involved in cancer progression. Aim of this study was to extract mRNA from cytobrush-collected healthy and HPV infected cervical epithelial cells and investigate various RNA extraction and purification protocols for assessment of RNA yield and quality. Taking into consideration that cervical cancer screening is based on the cytology based Papanicolaou test (Pap test), main challenge is to investigate whether the samples obtained by regular Pap testing can be used for gene expression analysis. For this purpose, a total of 68 cervical specimens were previously tested for HPV infection. Following HPV testing, samples were submitted to RNA extraction and compared to the products after additional purification step involving DNase I. Products obtained after different RNA extraction and purification methods were visualized using 2% agarose gel electrophoresis. In conclusion, DNase I based RNA purification represents a necessary step for the assurance of a high-quality extracted RNA used for gene expression analysis studies. Reliance on commercial kits for RNA extraction only, without performing additional purification step can lead to errors in drawing final conclusions and/or to false negative gene expression profiling, affecting the overall diagnostic procedure. According to obtained results, the type of sampling used in this study was not suitable for the subsequent gene expression analysis.

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.

Jasminka Prguda-Mujic, K. Milde‐Langosch, V. Mueller, M. Suljagić, J. Ćorić, Daria Ler

BACKGROUND Metastasis-Associated in Colon Cancer-1(MACC1) was first identified as a transcriptional activator of the HGF/MET pathway. Deregulation of HGF/MET signaling is reported as a prognostic marker for tumorigenesis, early stage invasion, and metastasis which is associated with poor clinical outcome in breast cancer patients. The aim of the present study was to further investigate the prognostic or predictive value of MACC1 expression in breast cancer. MATERIALS AND METHODS We analyzed the MACC1 expression in 105 primary breast cancer samples by Western-Blot analysis and immunohistochemistry. RESULTS A significant correlation of high MACC1 expression with shorter disease-free survival was found within the group of lymph-node-negative patients. Additionally, an association of high MACC1 expression and shorter disease-free survival was observed within estrogen receptor positive tumors and patients without adjuvant chemotherapy. CONCLUSION Our results support a biologic role and potentially open the perspective for the use of MACC1 as a prognostic marker for treatment decision in breast cancer patients.

Background: The serum prostate specific antigen for the early detection and screening for prostate cancer are very common used among physicians as the best screening tool for prostate cancer. The result of prostate specific antigen levels discriminates whether or not a prostate biopsy should be performed. The lack of specificity is a limitation of PSA as tumor marker, increased PSA concentrations are found not only in patients with prostate cancer but also in patients with benign prostatic disease. The object of this study was to improve the specificity and sensitivity of prostatic cancer detection. We evaluated total PSA levels, free PSA levels and the prostate volume in asymptomatic patients which came for routine check without medical history of prostate cancer. Methods: We received medical record of 90 patients between 50-60 years. Total and free PSA in serum was measured with the analyzer Architeckt i2000 SR. Prostate volume was determined by transrectal ultrasound. Results : The ratio of total and free PSA levels to prostate volume was significantly (p < 0.001) between all three groups. It was observed that increased prostate volume correlates with increased level of total and free PSA in serum. Conclusion: Early studies have demonstrated the advantage of measuring prostate volume with PSA total and free levels in serum as a useful tool for early diagnosis of prostate cancer. Data from this study on 90 patients with total PSA in the range from 0,22-7,0 ng/ml confirmed the well known correlation. All three parameters total PSA, free PSA and prostate volume showed significant correlation and a useful tool in prediction of prostate cancer for Bosnia and Herzegovina men.

Tea Bećirević, M. Hukić, Daria Ler

Human papillomavirus (HPV) is one of the most common sexually transmitted disease (STD) worldwide. HPV is a small, double-stranded DNA virus that infects mucosal and coetaneous epithelia trough tiny cuts and abrasion, exposed by the cells of the basal layer. If diagnosed on time HPV can be successfully treated, however, in some cases it can lead to the development of tumor. Most of cervical tumors contain HPV DNA, and majority of them contain high-risk types HPV16 and HPV18. Different risk factors are associated with HPV infection, including behavioral and biological predispositions. Aim of this study is to genotype potentially infected patients on high-risk types HPV DNA and to correlate the results with patient’s different biological and lifestyle factors. For this purpose 20 gynecological smear samples were collected from women, previously subjected to the survey. Methodology included DNA extraction and real-time polymerase chain reaction (RT-PCR). Results showed that out of 20 patients five were positive for high risk HPV. Four of five positive patients were positive on HPV16 type of which one had HPV16 together with others high risk types. One of five positive patients was positive on HPV18 type and other high risk types not identified. Final outcome indicates the correlation of potentially endangered patients with specific sexual behavior and lifestyles, and furthermore represent the general consensus and awareness level this disease has on the

N. Bajram, Daria Ler

This paper introduced a general class of mathematical models, Markov chain models, which are appropriate for modeling of phenomena in the physical life, medicine, engineering and social sciences. Application of Markov chains are quite common and have become a standard tool of decision making. What matters in predicting the future of the system is its present state, and not the path by which the system got to its present state. Two methods are presented that exemplify the flexibility of this approach: the regular Markov chain and absorbing Markov chain. The long-term trend in absorbing Markov chains depends on the initial state. In addition, changing the initial state can change the final result. This property distinguishes absorbing Markov chains from regular Markov chains, where the final result is independent of the initial state.  The problems are formulated by using the Wolfram Mathematical Programming System.

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