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Rijad Konjhodžić

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Objective: This comprehensive research aimed to thoroughly examine the effectiveness of a diode laser (445 nm) in combination with non-surgical treatment in patients with chronic periodontitis (CP) by evaluating a wide range of clinical and microbiological parameters. Materials and methods: Thirty-one subjects diagnosed with CP were included in this study. The total number of treated periodontal pockets was 862. The subjects were randomly assigned to group 1, which underwent scaling and root planing and laser therapy (SRP+L), and group 2, which underwent scaling and root planing (SRP) only. All respondents underwent a periodontal diagnostic protocol. The parameters plaque index (PI), gingival index (GI), bleeding on probing index (BOP), probing depth (PD), clinical attachment level (CAL), and tooth mobility (TM) were registered. Clinical periodontal measurements were performed at baseline and one and three months after therapy. Microbiological analysis was conducted on Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Tannerella forsythia (Tf) using real-time polymerase chain reaction (PCR). For microbiological analysis, samples were taken at baseline, immediately after therapy, and after three months. Laser irradiation was performed immediately after SRP. Results: All clinical parameters improved statistically from baseline to three months after therapy. For all examined clinical parameters, better results were achieved in group 1 than in group 2. This study showed a more significant reduction in Pg and Tf from baseline to three months in group 1 compared to group 2. Conclusion: These results showed that the diode laser wavelength 445 nm was also usable in treating periodontal diseases as an additional method to SRP.

Human mitochondrial genes MT-ATP6 and MT-ATP8 encode the subunits 6 and 8, respectively, of ATP synthase, a vital protein Complex V intricately involved in oxidative phosphorylation and ATP metabolism. This enzyme produces ATP from ADP in the mitochondrial matrix utilizing energy provided by the proton electrochemical gradient. Pathogenic mutations within these genes have been linked to various syndromes such as NARP syndrome, Leigh syndrome, mitochondrial myopathy with reversible cytochrome C oxidase deficiency, and progressive spastic paraparesis, among others. In our investigation, we sequenced 24 complete human mitochondrial genomes of healthy adult individuals from Bosnia and Herzegovina, each representing unique maternal lineage. Employing the Illumina MiSeq NGS platform and the Nextera XT DNA library preparation protocol, we obtained raw NGS reads. Subsequent analysis utilizing SAMtools enabled the identification of genetic variants within the MT-ATP6 and MT-ATP8 genes. We identified a total of 11 SNPs, including three in MT-ATP8 and eight in MT-ATP6, with none of them being associated with any mitochondrial diseases or conditions. Our results align well with previously reported genome variation data for European populations and set the groundwork for future mtDNA analysis for clinical purposes in Bosnia and Herzegovina.

Introduction: COVID-19 has been a major focus of scientific research since early 2020. Due to its societal, economic, and clinical impact worldwide, research efforts aimed, among other questions, to address the effect of host genetics in susceptibility and severity of COVID-19. Methods: We, therefore, performed next-generation sequencing of coding and regulatory regions of 16 human genes, involved in maintenance of the immune system or encoding receptors for viral entry into the host cells, in a subset of 60 COVID-19 patients from the General Hospital Tešanj, Bosnia and Herzegovina, classified into three groups of clinical conditions of different severity (“mild,” “moderate,” and “severe”). Results: We confirmed that the male sex and older age are risk factors for severe clinical picture and identified 13 variants on seven genes (CD55, IL1B, IL4, IRF7, DDX58, TMPRSS2, and ACE2) with potential functional significance, either as genetic markers of modulated susceptibility to SARS-CoV-2 infection or modifiers of the infection severity. Our results include variants reported for the first time as potentially associated with COVID-19, but further research and larger patient cohorts are required to confirm their effect. Discussion: Such studies, focused on candidate genes and/or variants, have a potential to answer the questions regarding the effect of human genetic makeup on the expected infection outcome. In addition, loci we identified here were previously reported to have clinical significance in other diseases and viral infections, thus confirming a general, broader significance of COVID-19-related research results following the end of the pandemic period.

Background: All viral genomes, including the SARS-CoV-2 virus, mutate over time, and some of these mutations can affect the characteristics of the virus, such as the ease of spread, the severity of the patient’s clinical picture, or the effect of vaccines, therapeutic drugs, diagnostic tools or other measures of public health and social protection. Because of all the above, it is imperative to carry out continuous sequencing of this pathogen. Objective: The main goal of this research was to obtain the highest quality genomic sequences of the SARS-CoV-2 virus, to compare the obtained sequences with the reference Wuhan-Hu-1 sequence and to obtain a high-quality genomic alignment in order to reconstruct the appropriate phylogenetic tree. Methods: For the purposes of this research, a next-generation semiconductor sequencing method was chosen. In this research, a total of 47 samples of nasopharyngeal and oropharyngeal swabs from patients from the human population of Bosnia and Herzegovina with a clinical diagnosis of COVID-19 were collected. Results: In the processed 47 samples, there are several monophyletic groups on the constructed phylogenetic tree, of which one sample belongs to the same monophyletic group as the Wuhan-Hu-1 reference sequence. Conclusion: The greater number of samples is needed for a more comprehensive approach. Therefore, the results of this research can act as a guideline for the design of effective measures and strategies in order to solve problems regarding future pandemics as efficiently as possible.

Nejla Muhovic - Pasic, Mirela Kahvic, Selma Karup, D. Pećar, E. Kandić, L. Salihefendić, Rijad Konjhodžić

Background: Human papillomavirus is a sexually transmitted infection and it is estimated that 75% of all women have been exposed to HPV infection in a certain period of life. High-risk types of HPV are considered to be one of the major causes of cervical cancer and its precursor intraepithelial neoplasia. Objective: The aim of this study was to investigate the degree of HPV infections and to provide more data on HPV genotype distribution among women in Bosnia and Herzegovina (B&H). Methods: Number of 375 samples were collected from different polyclinics in Sarajevo and were analyzed by Alea Genetic Center using Genomed f-HPV typing™ multiplex Fluorescent PCR kit for human papillomavirus genotyping. DNA required for this method is extracted from cervical swabs and amplified using a multiplex PCR reaction containing a set of 16 fluorescently labeled primers that recognize 16 HPV types. 14 HPV types are classified as high-risk (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68) and two are low-risk (6 and 11) HPV types. Results: Results showed that in the years 2018, 2019, and 2021, HPV type 16 is predominant causing the high-risk factor for CIN1, CIN2, CIN3, and cervical cancer development. HPV 18 infection rates decreased during the last four years of study. HPV 6 infection rates increased during that period of time. Conclusion: HPV 16 and HPV 18 are almost completely preventable by vaccination implying that the number of diagnosed cervical cancers in B&H could be much lower in the next decades if the HPV vaccination routine immunization program starts soon.

D. Primorac, V. Škaro, Petar Projić, S. Missoni, Ivana Horjan Zanki, Siniša Merkaš, J. Šarac, Natalija Novokmet, A. Ledić et al.

Aim To analyze an additional set of Y-chromosome genetic markers to acquire a more detailed insight into the diversity of the Croatian population. Methods A total of 518 Yfiler Plus profiles were genotyped. Allele frequencies, haplotype frequencies, and haplotype diversity were calculated by using the STRAF software v. 2.0.4. Genetic distances were quantified by Rst with AMOVA online tool from the YHRD. The evolutionary history was inferred with the neighbor-joining method of phylogenetic tree construction in the MEGAX software. Whit Athey's Haplogroup Predictor v. 5 was used for additional comparison with regional and other European populations. Results A total of 507 haplotypes were used for genetic STR analysis. An interpopulation study on 17 Y-STR markers showed the lowest genetic diversity between the Croatian and Bosnian-Herzegovinian populations and the highest between the Croatian and Irish populations. Additional interpopulation comparison with the original 27 Y-STR markers (for the population with available data) was also performed. A total of 518 haplotypes were used in the determination of haplogroup diversity. Haplogroup I with its sublineage I2a expressed the highest prevalence. The second most prevalent haplogroup was R, with its major sublineage R1a, except for the subpopulation of Hvar, where E1b1b was the second most prevalent haplogroup. Rare haplogroups also confirmed in this study were L, T, and Q. G1 was detected for the first time in the Croatian population. Conclusion We obtained a new insight into the differences between examined subpopulations of Croatia and their possible (dis)similarities with neighboring and distant populations.

Background: SARS-CoV-2 is a coronavirus that causes a respiratory disease, COVID-19. For COVID-19 testing, real-time PCR is considered gold standard and therefore many commercial SARS-Cov-2 detection kits are available. Objective: The aim of the study is to determine diagnostic values of 10 different commercially available SARS-CoV-2 detection kits, based on their Ct value. Methods: For this study thirty clinical nasopharyngeal samples were collected in ALEA Genetic Center. Twenty four of them were positive, while six were negative and used as a negative control. Positive samples were selected based on the day when first symptoms appeared. RNA was extracted using the same extraction method for all samples. For amplification and comparison of detection kits, the same RT- PCR instrument was used. Results: Accuracy, sensitivity, specificity and Cohen’s kappa coefficient were estimated to evaluate diagnostic values of the tested kits. This study showed that all kits showed 100% specificity. Accuracy, sensitivity and kappa coefficient varied among examined assays. Based on clinical features, LabGunTM COVID-19 Assay by LabGenomics proved to be the most sensitive, the most accurate and most specific. Therefore this assay was used as a reference kit. Conclusion: If things from practice are taken into account, accuracy and reliability of the tested commercial kits can vary compared to those obtained in this study where results were based on ideal functioning of the kits. When choosing the convenient commercial SARS-CoV-2 detection kit using RT-PCR method, many parameters need to be considered.

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.

D. Primorac, V. Škaro, Petar Projić, S. Missoni, Ivana Horjan Zanki, Siniša Merkaš, J. Šarac, Natalija Novokmet, A. Ledić et al.

Aim To analyze additional set of Y-Chromosome genetic markers to acquire a more detailed insight into the diversity of the Croatian population. Methods The total number of 518 Yfiler™ Plus profiles was genotyped. Allele, haplotype frequencies and haplotype diversity, were calculated using the STRAF software package v2.0.4. Genetic distances were quantified by Rst using AMOVA online tool from the YHRD. The evolutionary history was inferred using the neighbor-joining method of phylogenetic tree construction in MEGAX software. Whit Athey’s Haplogroup Predictor v5 was used for additional comparison with selected European populations. Results The total of 507 haplotypes were used for genetic STR analysis. The interpopulation comparison with the original 27 Y-STR markers shows the lowest genetic diversity between Croatian and Serbian population, and the highest between Croatian and Spanish population. Interpopulation study on 17 Y-STR markers shows the lowest genetic diversity between Croatian and Bosnian-Herzegovinian population, and the highest between Croatian and Irish population. Total of 518 haplotypes were used in the determination of haplogroup diversity. Haplogroup I with its sublineage I2a expressed the highest prevalence. Haplogroup R, with its major sublineage R1a, is the second most abundant in the studied Croatian population, except for the subpopulation of Hvar, where E1b1b is the second most abundant haplogroup. Rare haplogroups also confirmed in this study are L, T and Q. G1 is detected for the very first time in Croatian population. Conclusion New insight into differences between examined subpopulations of Croatia and their possible (dis)similarities with neighboring abroad populations was notified.

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