Bosa Mirjanic-Azaric

Background/Objectives: Low-density lipoprotein cholesterol (LDL-C) is a primary therapeutic target in cardiovascular risk assessment. While direct assays are available, LDL-C is frequently estimated using various formulas, whose performance can vary by population and lipid concentration. This study evaluates the analytical agreement and clinical classification performance of four formulas (Spasić–Kotur–Vujović, Friedewald, De Long, and Martin–Hopkins) against directly measured LDL-C. Methods: Lipid profiles from 3935 fasting adult patients were analyzed. LDL-C was directly measured via a homogeneous assay and compared with calculated values using linear regression and Bland–Altman analyses. Analytical bias and classification accuracy were assessed across stratified lipid levels. Results: Among the evaluated equations, the Spasić–Kotur–Vujović formula showed the closest agreement with directly measured LDL-C. It had the lowest mean absolute bias and mean percentage differences across most lipid strata, as well as narrower limits of agreement compared with Friedewald, De Long, and Martin–Hopkins formulas. Although all formulas showed strong correlations with directly measured LDL-C (Pearson’s r correlation coefficient = 0.956–0.958; intraclass correlation coefficient > 0.974), systematic underestimation of LDL-C was observed for all equations and became more pronounced with increasing total cholesterol, LDL-C, triglycerides, and non-high-density lipoprotein cholesterol concentrations. The Spasić–Kotur–Vujović formula achieved the highest overall classification accuracy (70%), followed by De Long (68%), Martin–Hopkins (61%), and Friedewald (58%). Conclusions: Despite strong correlations, clinically relevant differences in bias and classification exist among LDL-C formulas. The Spasić–Kotur–Vujović formula demonstrated superior agreement and accuracy in this population, suggesting it is a more reliable tool for routine laboratory practice. It should be noted that these findings reflect agreement with the specific homogeneous LDL-C assay used on the Alinity c platform (Abbott Laboratories,, Chicago, IL, USA) and may not be directly generalizable to other analytical systems.

Background/Aim: Rheumatoid arthritis (RA) is an autoimmune inflammatory disease, characterized by the production of numerous pro-inflammatory cytokines, such as tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), which lead to pathophysiological changes in innate and acquired immunity. The existing evidence shows that pro-inflammatory cytokines in rheumatoid arthritis impact monoaminergic neurotransmission, neurotropic factors, and synaptic activity, which may lead to the development of depression. Materials and Methods: In our study, we explored the association between TNF-α and IL-6, disease activity, and the degree of depression in patients with RA. The association between TNF-α and IL-6 and the Beck and Hamilton depression scales was analyzed in a group of 116 RA patients with depression. We investigated the same correlation in 45 patients with primary depression who represented the control group. Results: A Spearman test showed that IL-6 levels had a positive association with the Beck and Hamilton scales (p < 0.05) and that TNF-α had a positive association with the Hamilton scale (p < 0.05). Also, the Hamilton depression scale was the more sensitive scale in the detection of depressive symptoms. Conclusions: Our study indicates that elevated values of pro-inflammatory cytokines are associated with the degree of depression in patients with RA. Future preclinical and clinical studies will contribute to a better understanding of the pathophysiological mechanism of depression in patients with RA and may serve as the basis for new treatment modalities. By detecting depression promptly, with the help of the HAM-D as the more sensitive scale, we could influence the future modality of treatment, and with a multidisciplinary approach, we could ensure an improvement in the quality of life of patients with RA.

Numerous studies point to the significance of cathepsin B (CTSB) in the development of carcinoma. Therefore, the aim of this pilot study was to investigate the levels of cathepsin B (CTSB) and the expression of CTSB mRNA in the plasma of non-Hodgkin lymphoma (NHL) patients. Methods: The study included 44 newly diagnosed NHL patients and 35 healthy volunteers comprising the control group. CTSB in the plasma samples were detected using the enzyme-linked immunosorbent assay (ELISA). Results: The level of CTSB was significantly higher in NHL patients compared to control subjects: 15.28 (11.68–17.23) versus 11.57 (10.12–13.41), p = 0.003. In addition, a positive correlation between plasma CTSB mRNA and CTSB after therapy was observed (rho = 0.591, p = 0.026). Regarding redox parameters, we found a negative correlation between CTSB and the total antioxidant status (TAS) (rho = −0.499, p = 0.035), as well as a positive correlation with the total oxidant status (TOS) (rho = 0.576, p = 0.012). Conclusions: Targeting CTSB might have significant clinical relevance in the diagnostics of NHL.

Background: Recent findings point to the key role of cathepsin S (CTSS) in the survival of malignant cells, as well as the significance of the anti-apoptotic properties of high-density lipoprotein (HDL) that contribute to enhanced cell survival. The purpose of this study is to analyse CTSS as a potential biomarker in lymphoma. Also, in order to better understand the role of CTSS in the origin and development of lymphoma, its association with cystatin C (Cys C), lipids, and inflammatory markers was analysed. Methods: The study included 90 subjects: 11 Hodgkin (HL) and 44 B-cell non-Hodgkin lymphoma (NHL) patients, as well as 35 healthy subjects. CTSS was determined using the Invitrogen ELISA kit (Thermo Fisher Scientific, Inc., Waltham, MA, USA). The non-denaturing 3%-31% polyacrylamide gradient gel electrophoresis method was used to separate plasma HDL particles. Results: The level of CTSS was significantly higher in NHL patients than in control subjects: 12.20 (9.75-14.57) vs 9.97 (8.44-10.99), P<0.001. In NHL patients, there was a positive correlation between CTSS and the proportions of HDL3a, HDL3b, and the sum of the HDL3 subclasses (r=0.506, P<0.001; r=0.411, P=0.006, r=0.335, P=0.026, respectively). In addition, the area under the receiver operating characteristic curve (AUC curve) of CTSS was 0.766 (CI: 0.655-0.856) for NHL patients. There was no significant difference in CTSS values between the control group and patients with HL, nor significant correlations between CTSS and HDL subclasses in the HL group. Conclusions: CTSS is significantly elevated in patients with NHL and has the potential to be a new diagnostic bio - marker for the detection of NHL. Also, this study was the first to unveil the association between serum CTSS levels and the proportions of anti-apoptotic HDL3a and HDL3b subclasses in NHL patients.

Newer research points to alterations in the plasma redox status and the HDL subclass distributions in cancer. We aimed to assess the redox status and the HDL subclass distributions, lipids, and inflammatory markers in lymphoma patients in order to determine whether they were correlated with changes in FDG-PET/CT scans. At the beginning of this study, redox status, HDL subclasses, lipids, and inflammation biomarkers were determined in 58 patients with lymphoma (Hodgkin lymphoma, n=11 and non-Hodgkin lymphoma, n=47), and these same measurements were reassessed during their ensuing treatment (in 25 patients). Initially, the total oxidation status (TOS), the prooxidant–antioxidant balance (PAB), the OS index (OSI), the total protein sulfhydryl groups (SH-groups), and the advanced oxidation protein products (AOPP) were significantly higher in lymphoma patients as compared to healthy subjects, but the total antioxidant status (TAS) was significantly reduced. The PAB had a strong correlation with the CRP and interleukin-6 (rho=0.726, p<0.001; rho=0.386, p=0.003). The correlations between these parameters and the maximum standardized uptake values (SUVmax) were: PAB, rho=0.335 and p=0.010; SH-groups, rho=0.265 and p=0.044; CRP, rho=0.391 and p=0.002; HDL3b, rho=0.283 and p=0.031; HDL2b, rho= -0.294 and p=0.025; and HDL size, rho= -0.295 and p=0.024. The reductions in SUVmax between two follow-up points were associated with increases in the OSI, TOS, and SH-groups, as well as a reduction in the PAB and TAS. In conclusion, the redox parameters in patients with lymphoma were consistent with FDG-PET/CT findings. Targeting the redox status parameters and the HDL subclasses could be potential strategies in the molecular fight against lymphoma.

Current data suggest that aristolochic acid (AA) exposure is a putative cause of Balkan endemic nephropathy (BEN), a chronic kidney disease strongly associated with upper tract urothelial carcinoma. The cellular metabolism of AA is associated with the production of reactive oxygen species, resulting in oxidative distress. Purpose: Therefore, the aim of this study was to analyze individual, combined and cumulative effect of antioxidant gene polymorphisms (Nrf2 rs6721961, KEAP1 rs1048290, GSTP1AB rs1695, GSTP1CD rs1138272, GPX3 rs8177412 and MDR1 rs1045642), as well as GSTP1ABCD haplotypes with the risk for BEN development and associated urothelial cell carcinoma in 209 BEN patients and 140 controls from endemic areas. Experimental method: Genotyping was performed using polymerase chain reaction (PCR) and PCR with confronting two-pair primers (PCR-CTTP) methods. Results: We found that female patients carrying both variant GPX3 rs8177412 and MDR1 rs1045642 genotypes in combination exhibited significant risk towards BEN (OR 1 = 3.34, 95% CI = 1.16–9.60, p = 0.025; OR 2 = 3.79, 95% CI = 1.27–11.24, p = 0.016). Moreover, significant association was determined between GPX3rs8174412 polymorphism and risk for urothelial carcinoma. Carriers of variant GPX3*TC + CC genotype were at eight-fold increased risk of BEN-associated urothelial tumors development. There was no individual or combined impact on BEN development and BEN-associated tumors among all examined polymorphisms. The haplotype consisting of variant alleles for both polymorphisms G and T was associated with 1.6-fold increased risk although statistically insignificant (OR = 1.64; 95% CI = 0.75–3.58; p = 0.21). Conclusions: Regarding GPX3 rs8177412 polymorphism, the gene variant that confers lower expression is associated with significant increase in upper urothelial carcinoma risk. Therefore, BEN patients carrying variant GPX3 genotype should be more frequently monitored for possible upper tract urothelial carcinoma development.

Background: The pandemic of severe acute respiratory syndrome by coronavirus 2 (SARS-CoV-2) is a multi-system disease caused by a diffuse systemic process involving a complex interaction of the inflammatory, immunological and coagulative cascades. This study aims to identify the most effective biomarkers to predict poor outcome in intensive care unit (ICU) patients with severe COVID-19 disease. Methods: A single-centre retrospective observational study enrolled 69 deceased and 20 recovered patients treated in the ICU of the General Hospital Gradiska in the period from March 1, 2021. until April 1, 2022. We evaluated the leukocytes (WBC), lymphocytes (LYM), neutrophils (NEU), platelets (PLT), haemoglobin, neutrophil-lymphocyte ratio (NLR), platelet lymphocyte ratio (PLR), and systemic immune-inflammation index (SII). In addition, we evaluated the IL-6, ferritin, CRP, D-dimer, magnesium, bilirubin and lactate dehydrogenase. Results: Between deceased and recovered patients on admission to the ICU, there was a significant difference between the following parameters: WBC x109/L 11.50 (8.86-14.75) vs. 9.40 (5.90-11.90), p =0.026; NEU x109/L 10.15 (7.81-12.74) vs. 8.60 (4.80-10.30) p=0.022; NLR 15.01 (10.60-24.33) vs. 9.45 (5.10-14.90) p=0.02; SII 3712 (2240-6543) vs. 1949 (993-3720) p=0.003. The magnesium level increased significantly over time in the patients who died, while the haemoglobin level and platelet count decreased. ROC analysis showed the following AUC values: WBC 0.774; NEU 0.781; NLR 0.786; SII 0.776; D-dimer 0.741, and bilirubin 0.713. Conclusion: In this retrospective study WBC, NEU, NLR, SII, D-dimer, and bilirubin determined at hospital admission had a high value in predicting death among patients with severe COVID-19.

Background: The aim of this study was to determine the reference intervals (RIs) for thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3) and FT3/FT4 ratio using indirect methods. Methods: We analyzed 1256 results TSH, FT4 and FT3 collected from a laboratory information system between 2017 and 2021. All measurements were performed on a Siemens ADVIA Centaur XP analyzer using the chemiluminescent immunoassay. We calculated the values of the 2.5th and 97.5th percentiles as recommended by the IFCC (CLSI C28-A3). Results: The RIs derived for TSH, FT4, FT3 and FT3/FT4 ratio were 0.34-4.10 mIU/L, 11.3-20.6 pmol/L, 3.5-6.32 pmol/L and 0.21-0.47, respectively. We found a significant difference between calculated RIs for the TSH and FT4 and those recommended by the manufacturer. Also, FT3 values were significantly higher in the group younger than 30 years relative to the fourth decade (5.26 vs. 5.02, p=0.005), the fifth decade (5.26 vs. 4.94, p=0.001), the sixth decade (5.26 vs. 4.87, p<0.001), the seventh decade (5.26 vs. 4.79, p<0.001) and the group older than 70 years old (5.26 vs. 4.55, p<0.001). Likewise, we found for TSH values and FT3/FT4 ratio a significant difference (p <0.001) between different age groups. Conclusions: The establishing RIs for the population of the Republic of Srpska were significantly differed from the recommended RIs by the manufacturer for TSH and FT4. Our results encourage other laboratories to develop their own RIs for thyroid parameters by applying CLSI recommendations.

Background: An accurate measurement of serum thyroid stimulating hormone (TSH) is crucial for a thyroid disorder diagnosis and management. We compared the values of TSH between three automated immunoassays with aims to provide insights into variations in TSH levels that are important for a clinical decision: 2.50 mIU/L, 4.00 mIU/L and 10.00 mIU/L. Methods: We measured TSH with three different fully automated immunoassays: Abbott (Architect ci8200), Siemens (ADVIA Centaur XP) and Roche (Cobas e411). Serum was collected from 110 patients between August 2018 and January 2019. The results were compared using the Passing–Bablok regression method. Additionally, linear regression coefficients were calculated after logarithmic transformation of data. Results: Although all three regression coefficients were high (r² > 0.98), the slopes from Passing–Bablok plots for the correlation of Abbott with either Roche or Siemens were merely 0.66 and 0.73, respectively. The slope for the correlation of Roche and Siemens was 1.11. Consecutively, the results obtained by the Roche and Siemens methods were proportionally higher than those obtained by the Abbott method (38% and 52%, respectively) at all measured ranges. Conclusions: Although immunoassays correlated among themselves, they cannot achieve the same values for clinical decisions for hypothyroidism (clinical requirements). Clinicians should be conscious of these limitations. A harmonisation of the methods is needed to meet clinical requirements and to enable appropriate clinical decisions in cases of hypothyroidism. Likewise, we suggest the introduction of borderline and high risk values of TSH for hypothyroidism depending on immunoassays to avoid misdiagnosis.

Summary: Background: Cathepsin S (CTSS) is a cysteine protease involved in atherogenesis. We compared the plasma CTSS as well as other biomarkers of atherosclerosis in patients with abdominal aortic aneurysms (AAA) and aortoiliac occlusive disease (AOD), aiming to identify the underlying pathogenic mechanisms of the disease development. Also, we hypothesised that the level of plasma CTSS simultaneously increases with a decrease of plasma high-density lipoprotein cholesterol (HDL-C) values.Methods: 33 patients with AAA and 34 patients with AOD were included in this study.Results: There was no difference in the level of plasma CTSS between the two analysed groups (p=0.833). In the patients with AAA, the plasma CTSS was correlated with HDL-C (r = -0.377, p = 0.034) and total bilirubin (r =0.500, p = 0.003) while, unexpectedly, it was not correlated with cystatin C (Cys C) (r =0.083, p = 0.652). In the patients with AOD, the plasma CTSS correlated with triglycerides (r = 0.597, p< 0.001), only. When the patients were divided according to HDL-C (with HDL-C ≤0.90 and HDL-C >0.90 mmol/L), the plasma CTSS values differed among these groups (31.27vs.25.61 μg/L, respectively, p<0.001).Conclusions: These results provide the first evidence that CTSS negatively correlated with HDL-C and bilirubin in patients with AAA. It is possible that differences in the association of the CTSS and other markers of atherosclerosis can determine whether atherosclerotic aorta will develop dilatation or stenosis.