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T. Dujic

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Aim To assess the association of single nucleotide polymorphisms (SNPs) in the ACE2 and TMPRSS2 genes with COVID-19 severity and key biomarkers. Methods The study involved 750 COVID-19 patients from Bosnia and Herzegovina, divided into three groups: mild, moderate, and severe cases. Genetic variations within the ACE2 (rs2285666) and TMPRSS2 (rs2070788) genes were examined with real-time polymerase chain reaction. Biochemical markers were determined with standard procedures. Results There was a significant difference in the rs2070788 genotype distribution between patients with mild and moderate symptoms, but not between other groups. For the rs2285666 polymorphism, no significant difference in genotype distribution was found. In patients with mild symptoms, carriers of the GG genotype of rs2070788 had significantly higher total bilirubin levels than carriers of the AA genotype. Similarly, carriers of the TT genotype of rs2285666 had significantly higher activated partial thromboplastin time and international normalized ratio, and lower lactate dehydrogenase levels compared with the CC genotype. Among patients with severe symptoms, carriers of the GG genotype showed significantly higher potassium levels than carriers of the AA genotype, while carriers of the TT genotype showed significantly higher erythrocyte count as well as hemoglobin and hematocrit levels compared with the CC genotype. Conclusion This study highlights the role of genetic factors, particularly SNPs in the ACE2 and TMPRSS2 genes, in determining COVID-19 severity, aiding patient risk assessment and prognosis.

A. Šljivo, A. Bostandzija, A. Arnaout, A. Blekić, T. Dujic, M. Aljičević, Z. Tafi, A. Skopljak, B. Kudić et al.

Background A pilot study conducted in academic 2017/18 among undergraduates of the University of Sarajevo showed energy drinks to be most frequently consumed during academic activity, less frequently mixed with alcohol in leisure, and rarely in the sports activity. The aim of this study was to assess the impact of the COVID-19 pandemic on energy drinks consumption among undergraduates of the same University, with a focus on their consumption during exams. Study design A cross-sectional study was conducted by an online questionnaire. Methods The questionnaire, mainly based on the Consortium Nomisma-Areté questionnaire, was customized to compare energy drinks consumption before and during the COVID-19 pandemic, and distributed among students between July 26th, 2020 and April 3rd, 2021. Results Out of 1,045 students who chose to participate in the study (participation rate of 14.7%), 653 students, mostly women, attending the lower study years, reported energy drinks consumption. Both be-fore and during pandemic, overall energy drinks consumption was most frequently reported as rare [281 (43.9%) before, 326 (51.2%) during the pandemic], and exams-related energy drinks consumption as once or twice a week [156 (43.8%) before, 130 (42.1%) during pandemic]. The pandemic increased the number of frequent consumers (consumption of 4-5 energy drinks per week) in both overall [35 (5.5%) before, 46 (7.2%) during pandemic] and exams-related energy drinks consumption [42 (11.8%) before, 48 (15.5%) during pandemic]. Study year (OR=0.842; 95% CI 0.77-0.921; p<0.001), being single and living alone [OR=0.512; 95% CI 0.296-0.883; p=0.016), or living with a partner and children [OR=0.377; 95% CI 0.168-0.847; p=0.018) were identified as negative independent predictors for exams-related energy drinks consumption, while being a regular smoker (OR=0.429; 95% CI 0.223-0.875; p=0.011) appeared its new negative independent predictor during pandemic. Conclusions The pandemic seemed to decrease both the overall and exam-related energy drinks consumption among undergraduates of the University of Sarajevo with the exception of a portion of already frequent energy drinks consumers.

M. Malenica, A. Klisić, N. Meseldžić, T. Dujic, T. Bego, J. Kotur-Stevuljević

Background The aim of the study was to explore the mutual relationship between oxidative stress, inflammation and metabolic biomarkers in subjects with prediabetes (PRE), newly diagnosed type 2 diabetes patients (NT2D) and overt type 2 diabetes (T2D) using principal component analysis (PCA) as a thorough statistical approach. Methods Glycated hemoglobin, lipid parameters, inflammation (IL-6, CRP and fibrinogen) and oxidative stress markers pro-oxidants (AOPP, PAB, TOS) and antioxidants (PON1, tSHG, TAS) were measured. PCA was applied to explore the factors that the most strongly influenced glucoregulation. Results A total of 278 subjects were (i.e., 37 PRE, 42 NT2D and 99 T2D) were compared with 100 healthy subjects as a control group (CG). PCA emphasized 4 different factors explaining 49% of the variance of the tested parameters: oxidative stress-dyslipidemia related factor (with positive loading of TG and tSHG, and with negative loading of HDL-c and TAS), dyslipidaemia related factor (i.e., total cholesterol and LDL-c, both with positive loading), Anthropometric related factor (i.e., waist and hip circumference, both with positive loading) and oxidative stressInflammation related factor (i.e., PAB, fibrinogen, and CRP all with positive loading). Out of these 4 factors, only oxidative stress - dyslipidaemia related factor showed a significant predictive capability towards poor glucoregulation. An increase in this factor by one unit showed a 1.6 times higher probability for poor glucoregulation. Conclusions Redox imbalance (determined with lower TAS and higher tSHG), in addition to higher TG and lower HDLc was associated with poor glucoregulation.

Amela Bostandzic, V. Beslic, I. Šurković, Sefkija Balic, T. Dujic, Z. Ašimi, Azra Bureković

Background: IDegLira( fixed combination of GLP 1 receptor agonist and insulin) has been shown to be effective in improving the glucoregulation in patients previously treated with oral therapy as well as individual components, GLP-1 receptor agonist or basal insulin. Objective: The aim of this study is to examine the parameters of metabolic control in patients treated with IDegLira who were previously treated with premix insulin in several daily doses and to compare them with patients whose premix insulin dose was increased. Methods: The study included 100 patients who had been previously treated with two or three daily doses of premix insulin. Half of the patients were switched to IdegLira( group I), and half (group II) had their insulin dose increased according to the clinical assessment of the physician. Fasting glucose, 2h postprandial glucose, HbA1c, BMI and insulin dose were determined at baseline and at follow-up after 6 months. Results: Patients treated with IDegLira compared to patients whose insulin dose was increased achieved significantly lower fasting glucose (p <0.001), postprandial glucose (p <0.001), HbA1c (p <0.001), BMI (p <0.001) with a significantly lower insulin dose (p <0.001). Comparison of the same parameters within the groups of patients at the beginning and after 6 months showed that patients who were switched from insulin premix to IDegLira achieved significantly lower fasting blood glucose (p <0.001), postprandial glucose (p <0.001), HbA1c (p < 0.001), BMI (p <0.001) with significantly lower insulin dose within the fixed combination (p <0.001). Patients with gradually increased insulin dose achieved significant reduction in fasting glucose (p = 0.021) and postprandial glucose (p = 0.036),but with a significantly higher insulin dose (p = 0.005). There was also a slight increase in BMI that was not statistically significant (p = 0.267). Conclusion: The obtained data suggest that switching patients from a complex insulin regimen to a fixed combination of basal insulin and GLP 1 receptor agonist in comparison to increases in insulin dose results in a significant improvement in fasting glucose, postprandial glucose, HbA1c, and BMI. The results were achieved with a significantly lower daily insulin dose.

B. Prnjavorac, A. Mujaković, Lejla Prnjavorac, T. Bego, E. Jusufovic, E. Begić, Vildana Torlak-Arnaut, Meliha Mutapčić, H. Škiljo et al.

Aim To analyse the resolution of chest X-ray findings in relation to laboratory parameters in patients infected with acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in a two- month followup. Analysis of chest X-ray findings in the first few months after the disease is the main goal of our work. Methods Out of the total of 343 patients chest X-ray findings were followed in 269 patients. Patients were divided into groups according to the severity of findings. D-dimer, inflammatory markers, blood cell count, neutrophil lymphocyte ratio (NLR) were analysed. Chest X-ray was analysed during the hospitalization on the day of admission, on the third, the seventh and the fourteenth day (scoring method was used). After discharge chest X-ray was performed in a two-week follow-up, then after one and two months, and after three months if necessary. Results Incomplete chest X-ray resolution was identified in 24 (39.34%) patients with severe, 27 (22.31 %) patients with moderate and in three (3.91%) patients with mild findings. Statistical significance was established in overall score by comparison between all groups (p<0.001), and in the moderate compared to the mild group (p=0.0051). The difference of NLR in the severe compared to the moderate group was observed (p=0.0021) and in the severe group compared to the mild group (p=0.00013). Conclusion Chest X-ray findings persisted mostly in the severe group followed by the moderate and mild ones. Long-term followup is necessary for the appropriate treatment and prevention of fibrosis, and reduction of symptoms.

The antidiabetic drug gliclazide is partly metabolized by CYP2C19, the main enzyme involved in omeprazole metabolism. The aim of the study was to explore the interaction between omeprazole and gliclazide in relation to CYP2C19 phenotype using physiologically based pharmacokinetic (PBPK) modeling approach. Developed PBPK models were verified using in vivo pharmacokinetic profiles obtained from a clinical trial on omeprazole-gliclazide interaction in healthy volunteers, CYP2C19 normal/rapid/ultrarapid metabolizers (NM/RM/UM). In addition, the association of omeprazole cotreatment with gliclazide-induced hypoglycemia was explored in 267 patients with type 2 diabetes (T2D) from the GoDARTS cohort, Scotland. The PBPK simulations predicted 1.4–1.6-fold higher gliclazide area under the curve (AUC) after 5-day treatment with 20 mg omeprazole in all CYP2C19 phenotype groups except in poor metabolizers. The predicted gliclazide AUC increased 2.1 and 2.5-fold in intermediate metabolizers, and 2.6- and 3.8-fold in NM/RM/UM group, after simulated 20-day dosing with 40 mg omeprazole once and twice daily, respectively. The predicted results were corroborated by findings in patients with T2D which demonstrated 3.3-fold higher odds of severe gliclazide-induced hypoglycemia in NM/RM/UM patients concomitantly treated with omeprazole. Our results indicate that omeprazole may increase exposure to gliclazide and thus increase the risk of gliclazide-associated hypoglycemia in the majority of patients.

ABSTRACT Type 2 diabetes (T2D) has a continuously rising prevalence worldwide. Pharmacogenetics has been recognized as a promising concept for pharmacological treatment of T2D, as antidiabetic drugs are not equally effective and safe for all patients, and the costs of diabetes treatment are increasing. The latest published guidelines on T2D treatment firmly endorse the use of newer antidiabetic drugs, sodium-glucose cotransporter-2 inhibitors (SGLT2i), dipeptidyl peptidase-4 inhibitors (DPP-IVi), and glucagon-like peptide-1 receptor agonists (GLP-1RA), considering their satisfactory pharmacological effect and good safety profile. Furthermore, SGLT2i and GLP-1RA show protective effects in patients with established atherosclerotic cardiovascular disease and chronic kidney disease. However, there has been growing evidence that the effectiveness and safety of these drug classes could depend on genetic variability. Here, we summarized the results of the published studies on the pharmacogenetic biomarkers for the three drug classes. A number of genetic variations have been investigated so far. The explored candidate genes mostly encode drug targets, drug-metabolizing enzymes, and genes linked to T2D risk. Although many of the results are promising, it is still necessary to obtain more information from larger controlled studies to confirm their clinical significance. This approach may lead towards more personalized treatment for patients with T2D.

Background Altered levels of many hematological parameters have been directly associated with diabetes in adults, while studies on children with type 1 diabetes mellitus are lacking. The aim of this study was to determine hematological indices in diabetic Bosnian children in comparison to healthy controls as well as to correlate their levels to blood glucose and hemoglobin A1c. Methods 100 healthy and 100 children with type 1 diabetes mellitus (age 1-18) were included in this study. Complete blood count, hemoglobin A1c, and glucose were tested. Results were analysed by IBM SPSS Statistics version 23. Results Significant differences (p<0.05) between healthy and diabetic children were found in relation to HbA1c, glucose, mean platelet volume, the number of white blood cells and erythrocytes, hematocrit, hemoglobin and MCH values. No gender differences or significant age differences were seen for hemoglobin, hematocrit, and MCV, while platelets, MPV, and MCH differed by age only in healthy children. When diabetic children were classified according to HbA1c levels, significant differences were seen for erythrocyte count and hematocrit value (p=0.013 and 0.019, respectively). The number of erythrocytes and white blood cells correlated significantly with HbA1c (p=0.037 and 0.027, respectively). Conclusions Lower levels of erythrocytes, hematocrit, and hemoglobin in diabetic compared to healthy children indicate possible development of anemia, while higher MCV, MCH, and MPV values indicate an alteration in erythrocyte morphology. Hematological indices could be a useful inexpensive tool in the diagnosis and follow up of type 1 diabetes in children.

BACKGROUND Energy drinks (EDs) are non-alcoholic beverages that contain caffeine and other ingredients, marketed for their actual or perceived effects as stimulants, energizers and performance enhancers. The aim of this pilot study was to evaluate patterns of EDs consumption in leisure, sports, and academic activities over the last year among a group of pregraduate students of the University of Sarajevo, Bosnia and Herzegovina. STUDY DESIGN A cross-sectional study conducted by an online questionnaire-based survey. METHODS An anonymous questionnaire was mainly based on a Consortium Nomisma-Areté questionnaire [background information and consumer profile, general EDs consumption practices and reasons; alcohol mixed with EDs (AmEDs) consumption, EDs consumption in sports, consumption of other caffeinated beverages], and an additional part to evaluate EDs consumption during academic activities. RESULTS Out of 812 respondents from 22 faculties (participation rate of 23%), mean age 21.37 ± 1.98 years, 498 (61.7%) reported EDs consumption over the last year. Three main reasons for EDs consumption were to stay awake (58.2%), to enjoy the taste (46.8%), and to boost energy (38.0%). Energy drinks were mainly consumed less than once a month (70.5%), most frequently during academic activity (50.4%), less frequently mixed with alcohol for relaxation (21.5%), and only rarely in association with sports or other physical activity (10%). Drinking coffee (OR = 2.022; 95% CI 1.416-2.830; p < 0.001) and being a higher year student (OR = 0.723; 95% CI 0.639-0.819; p < 0.001) were independent predictors for EDs consumption; being single and living with parents (OR = 17.138; 95% CI 1.328-221.528; p = 0.030) for consumption of AmEDs; and being a man (OR = 2.251; 95% CI 1.493-3.392; p < 0.001) and living in urban environment (OR = 1.193; 95% CI 1.125-3.251; p = 0.017) for consuming EDs in association with sports or other physical activity. CONCLUSION Based on these preliminary data and taking low participation rate into account, EDs consumption seems not to be alarming among university students in our region. EDs are most frequently consumed during academic activity, less frequently mixed with alcohol for relaxation, and only rarely in association with sports or other physical activity. However, as EDs are increasingly aggressively promoted and easily accessible, the larger study is warranted to provide more reliable and up to date conclusions, and if necessary, to inform measures preventing health risks associated with EDs consumption.

The response to metformin, the most commonly used drug for the treatment of type 2 diabetes (T2D), is highly variable. The common variant rs7903146 C>T within the transcription factor 7 like 2 gene (TCF7L2) is the strongest genetic risk factor associated with T2D to date. In this study we explored the effects of TCF7L2 rs7903146 genotype on metformin response in T2D. The study included 86 newly diagnosed patients with T2D, incident users of metformin. Levels of fasting glucose, insulin, HbA1c, total cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides, and anthropometric parameters were measured prior to metformin therapy, and 6 and 12 months after the treatment. Genotyping of TCF7L2 rs7903146 was performed by the Sequenom MassARRAY® iPLEX® platform. At baseline, the diabetes risk allele (T) showed an association with lower triglyceride levels (p = 0.037). After 12 months of metformin treatment, the T allele was associated with 25.9% lower fasting insulin levels (95% CI 10.9-38.3%, p = 0.002) and 29.1% lower HOMA-IR index (95% CI 10.1-44.1%, p = 0.005), after adjustment for baseline values. Moreover, the T allele was associated with 6.7% lower fasting glucose levels (95% CI 1.1-12.0%, p = 0.021), adjusted for baseline glucose and baseline HOMA-%B levels, after 6 months of metformin treatment. This effect was more pronounced in TT carriers who had 16.8% lower fasting glucose levels (95% CI 7.0-25.6%, p = 0.002) compared to the patients with CC genotype. Our results suggest that TCF7L2 rs7903146 variant affects markers of insulin resistance and glycemic response to metformin in newly diagnosed patients with T2D within the first year of metformin treatment.

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