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U ovom radu opisan je sistem za detekciju pada, čiji je cilj pomoć starim i nemoćnim licima. U radu je dat detaljan uvid u sve delove realizovanog sistema za detekciju pada, kroz opis: hardverskih komponenti i prototipa uređaja, zatim izrade samog uređaja, programiranja mikrokontrolera za datu namenu, realizacije korisničkog interfejsa, formiranja baze podataka, kao i obuke modela mašinskog učenja zasnovanih na različitim algoritmima.

Introduction: The lives of most people around the world have changed since the onset of SARS-CoV-2 pandemic (SARS-CoV - severe acute respiratory syndrome corona virus). According to the WHO data, the first cases in Croatia were registered on February 26, in Bosnia and Herzegovina on March 5, and in Serbia on March 10, 2020. The assumption was that due to stress caused by imposed isolation, the alcohol consumption began to increase. Methods: In order to obtain information on alcohol consumption in the period before and during Covid-19 lockdown, an online anonymous survey was conducted, in which a total of 930 persons participated: 542 from Croatia, 219 from Bosnia and Herzegovina, and 169 from Serbia. Results: Among those who completed the survey were 659 women and 271 men, mostly between 20-45 years old, with higher or secondary education. Most were employed, living in urban areas with a larger population. The obtained results did not show the influence of lockdown on alcohol consumption. On the contrary, 20.21% of participants, who had previously consumed alcohol, answered that they stopped consuming during the lockdown. Only 0.43% of participants started consuming alcohol precisely because of isolation. Conclusions: This research sheds new light on the daily practice of people during Covid-19 pandemic in the Western Balkans, known as a region where alcohol consumption is quite widespread

Introduction: There is a growing amount of evidence showing the significant analytical bias of steroid hormone immunoassays, but large number of available immunoassays makes conduction of a single comprehensive study of this issue hardly feasible. Aim of this study was to assess the analytical bias of six heterogeneous immunoassays for serum aldosterone, cortisol, dehydroepiandrosterone sulphate (DHEAS), testosterone, 17-hydroxyprogesterone (OHP) and progesterone using the liquid chromatography coupled to the tandem mass spectrometry (LC-MS/MS). Materials and methods: This method comparison study included 49 serum samples. Testosterone, DHEAS, progesterone and cortisol immunoassays were performed on the Abbott Architect i2000SR or Alinity i analysers (Abbott Diagnostics, Chicago, USA). DiaSorin’s Liaison (DiaSorin, Saluggia, Italy) and DIAsource’s ETI-Max 3000 analysers (DIAsource ImmunoAssays, Louvain-La-Neuve, Belgium) were chosen for aldosterone and OHP immunoassay testing, respectively. All immunoassays were evaluated against the LC-MS/MS assay relying on the commercial kit (Chromsystems, Gräfelfing, Germany) and LCMS-8050 analyser (Shimadzu, Kyoto, Japan). Analytical biases were calculated and method comparison was conducted using weighted Deming regression analysis. Results: Depending on the analyte and specific immunoassay, mean relative biases ranged from -31 to + 137%. Except for the cortisol, immunoassays were positively biased. For none of the selected steroids slope and intercept 95% confidence intervals simultaneously contained 0 and 1, respectively. Conclusions: Evaluated immunoassays failed to satisfy requirements for methods’ comparability and produced significant analytical biases in respect to the LC-MS/MS assay, especially at low concentrations.

Background: Carbohydrate sulfotransferases (CHST) were shown to be involved in carcinogenesis. The aim of the study was to assess the diagnostic value of serum CHST7 concentration in differentiation between lung cancer and non-malignant pulmonary inflammations. Methods: Clinical case-control study involving 125 participants was conducted: the control group containing cases of pneumonia and chronic obstructive pulmonary disease was compared to the lung cancer group composed of primary and metastatic cancers. Serum concentrations of CHST7 and routinely used markers including carcinoembryonic antigen (CEA), cytokeratin fragment 21-1 (CYFRA 21-1) and neuron-specific enolase (NSE) were determined for each participant using immunochemical methods. Statistical association, receiver operating characteristic (ROC) analysis and cross-validation were used for the evaluation of CHST7 either as a standalone biomarker or as a part of a biomarker panel. Results: In comparison to the control group, serum CHST7 was elevated in lung cancer (p Conclusions: Serum CHST7 is a promising biomarker for the differentiation between lung cancer and non-malignant pulmonary inflammations.

To the Editor We appreciate authors thoughtful comments to our published manuscript.[1 2][1] Since hypotension and cerebral hypoperfusion have been defined as risk factors for cognitive decline in cardiac surgery in previous studies, we monitored blood pressure during the study period in order to

Aim To investigate a potential of the clinical use of the soluble fms-like tyrosine kinase 1 (sFLT-1) to placental growth factor (PlGF) ratio from the perspective of a small hospital centre. Methods Maternal serum samples were analysed at 241/7-28 0/7, and 281/7-320/7 weeks of gestation. The level of sFLT-1 and PIGF was determined by immunoassay platform and used to calculate the sFLT-1/PIGF ratio in 35 pregnant women, and divided into subgroups according to preeclampsia occurrence at the time of delivery: preterm (≤37 weeks) or term (37-42 weeks’), and matched a control group. Results Patients in the preterm delivery group had a significantly higher incidence of intrauterine growth restriction, lower gestational age at the time of delivery, and lower infant birth weight compared to the other two groups. There was a negative correlation between the sFLT-1/PlGF ratio and GA and between the sFLT-1/PlGF ratio and birth weight at the time of delivery. The value of the sFLT-1/PlGF ratio was significantly higher in the preterm delivery PE group. All the PE group pregnancies ended with caesarean delivery compared to 25% in the control group. However, none of the patients from the PE group had any of the possible complications of preeclampsia nor did they require additional therapy such as blood transfusion or additional non-standard hypertensive therapy. Conclusion The sFLT-1/PlGF ratio could be used as an indicator for the development and estimation of the severity of PE to provide objective evidence for the management of preeclampsia patients, and as a predictive marker of preeclampsia at low cost.

The objective was to determine the impact of VKORC1 polymorphisms on warfarin anticoagulant therapy (stable warfarin maintenance dose, time required to reach therapeutic dose and time spent in therapeutic range) and its adverse events (overanticoagulation and bleeding events, time to first overanticoagulation or bleeding event, and therapy for bleeding events) in Croatian patients. Blood samples were collected from 186 patients on stable warfarin therapy. VKORC1 1173C>T and VKORC1 – 1639G>A gene polymorphisms were analyzed using real-time PCR. Prothrombin time international normalized ratio (INR) values were determined and overanticoagulation as well as bleeding events were recorded. Both tested VKORC1 gene polymorphisms (VKORC1 1173C>T and VKORC1 –1639G>A) were in perfect linkage disequilibrium. Genotype analysis showed that 33.9% of patients were homozygous for wild- type, 46.8% were heterozygous and 19.4% were homozygous for the variant allele. We have found a statistically significant difference between variantallele carriers and wild-type patients in stable warfarin maintenance dose (p < 0.001) and incidence of bleeding events (p = 0.040). Patients homozygous for variant-allele were more likely to experience an overanticoagulation event in the first 30 days of therapy (p = 0.040). Our study showed that VKORC1 1173C>T and VKORC1 –1639G>A gene polymorphisms are associated with stable warfarin maintenance dose and adverse events of warfarin therapy.

Single nucleotide polymorphisms (SNPs) in the promotor regions of cytokine genes included in angiogenesis may influence prostate cancer (PCa) development via regulation of the pathways of tumor angiogenesis. The aim of the present study was to investigate the association of IL-1 female +3954 (rs1143634) and IL-10-1082 (rs1800896) polymorphisms with PCa risk and aggressiveness in eastern Croatian patients. One hundred twenty PCa patients and 120 benign prostatic hyperplasia (BPH) controls were genotyped using real-time PCR (LightCycler Instrument, Roche Diagnostics) and the melting curve analysis method. There was no significant difference in the frequency of genotypes for the two polymorphisms between PCa patients and controls (Χ2 = 0.857, p = 0.355 for IL-female 1; Χ2 = 0.026, p = 0.872 for IL-10). Carriers of the IL-10-1082AG variant were found to be associated with the Gleason score (GS)7 (AA versus GA+GG, OR = 3.47, 95% CI 1.11-10.88, p = 0.033). There was no significant difference in the frequency of genotypes for the two polymorphisms and the presence of metastatic disease in PCa patients. These results suggest that tested SNPs associated with differential production of IL-1 female and IL-10 are not risk factors for PCa and do not correlate with the presence of distant metastasis in eastern Croatians. We found that IL-10-1082 GA+/or GG carriers have a higher risk of developing PCa with GS7 in eastern Croatians.

Acute pyelonephritis is a severe disease which is sometimes difficult to recognize based on clinical symptoms and routinely available diagnostic tests, especially in young children. The aim of this study was to assess the diagnostic value of urinary neutrophil gelatinase-associated lipocalin (uNGAL) as a biomarker of acute pyelonephritis.In this case-control study we analyzed 134 children (median age 2.5 years) who were admitted to the Pediatric Clinic of University Hospital Centre Osijek, Croatia. Eighty of them had acute pyelonephritis, while 54 children had febrile state of different etiology including cystitis and they represented the control group. uNGAL, white blood cells, C-reactive protein, urinanalysis, urine culture, kidney ultrasound and a dimercaptosuccinic acid scintigraphic scan were done for each child. uNGAL was measured using chemiluminiscent microparticle immunoassay on ARHITECT i1000SR (Abbott Diagnostics, IL, USA).uNGAL values were significantly higher in children with acute pyelonephritis compared to the control groups (113.6 ng/mL vs. 10.2 ng/mL, puNGAL can be a useful diagnostic biomarker in acute pyelonephritis in children, but also in differentiating cystitis from febrile states other than UTI.

A single nuclear polymorphisms (SNPs) in the promoter region of the tumor necrosis factor-alpha (TNF-alpha) gene are involved in regulation of expression levels of TNF-alpha and therefore are associated with oncogenesis of several cancers. Aim of our study was to investigate the effect of G---A polymorphism at -308 position in the promoter region of the TNF-alpha gene on prostate cancer (CalphaP) susceptibility in a subset of patients from Eastern Croatia. Study population consisted of 240 patients (120 with CalphaP, 120 controls). They were genotyped for TNF-alpha G-308A polymorphism using real-time PCR (LightCycler Instrument, Roche Diagnostics) and melting curve analysis method. X(2) test was used to compare distribution of TNF-alpha polymorphism genotypes between patients and control group. Relative risk was estimated by the odds ratio (OR). There was no significant statistical difference (X(2)=0.000, DF=1, p=1, OR=1, 95%CI=0.5537-1.8059) between patients and control group. Besides, data of CalphaP patients were stratified according to pathohistological diagnosis (PHD) by Gleason score and groups were compared according to TNF-alpha genotypes. Also, all patients and CalphaP patients were grouped according to prostate volume (V) into three groups: V50 mL, V50-100 mL, V100 mL. These groups were also compared according to TNF-alpha genotypes. There were no significant statistical differences between any of groups. Our findings suggest that TNF-alpha -308 SNP is not associated with CalphaP in Eastern Croatia population.

Vitamin K epoxide reductase (VKOR) is a key enzyme in the y-carboxylation of proteins associated with important bodily functions (coagulation, bone metabolism, etc.). This feature is particularly used in warfarin therapy, which blocks the VKOR enzyme and leads to production of dysfunctional coagulation proteins. Genetic factors, particularly vitamin K epoxide reductase complex 1 (VKORC1) gene, have greatest influence on warfarin therapy. The aim of this study was to determine the distribution of VKORC1 1173CT and VKORC1 -1639GA polymorphisms, which are most important for warfarin therapy. We investigated 420 unrelated healthy individuals, mostly blood donors, from region of the Eastern Croatia. Both investigated polymorphisms were in perfect linkage disequilibrium (LD) and showed identical results. 151 patients (36%) were homozygous for the wild-type (C/C and G/G), 196 (47%) were heterozygous (C/T and G/A) and 73 (17%) were homozygous for the variant allele (T/T and A/A). Number of normal alleles among individuals was 498 (59.3%), and number of variant alleles was 342 (40.7%). The data obtained are in good agreement with the results of studies in other European populations.

Introduction It is well known that, beside age, race and familiar factor, dietary factors play big role in epidemiology of prostate cancer (CaP). The Western lifestyle, particularly the higher intake of fat, meat, and dairy products, is also associated with risk for CaP. Obesity is also linked to CaP, mainly with mortality rather than incidence. We wanted to see and show if there is difference in body mass index (BMI) in patients with biopsy diagnosed CaP and patients in control group. Patients and methods We have included 235 patients in our research was (age 42-84 yrs.). In all patients transrectal biopsy of prostate was done. In 117 patients out of 235 is diagnosed CaP, in rest of 118 patients patohistological finding did not show CaP (control group). BMI and PSA level were determinated to all patients. Student t-test was used to assign difference between BMI in our 2 groups. Results Mean BMI in group of patients with CaP was 27.625 kg/m2 (SD 5.680), mean BMI in control group was 27.692 kg/m2 (SD 3.987). There was no statistical significant difference in BMI in out two groups. When we grouped our patients according to their BMI (underweight 30 kg/m2), we noticed decrease of PSA level with increase of BMI. In our research there was no underweight patients, 64 of them were with normal BMI and mean PSA 69.48 ng/ml ; 120 of them were overweight with mean PSA 55.15 ng/ml and 51 of them were obese patients with mean PSA 33.16 ng/ml. Discussion General adiposity, as measured by BMI, is not in relationship with CaP incidention in our research, but obese is connected with lower PSA level with possible hemodilution in background. Question is – should we decrease the threshold of PSA in obese patients for prostate biopsy?

Aim Prostate cancer (PC) is most common cancer in men and benign prostate hyperplasia (BPH) is condition affecting majority of elder men with great impact on everyday life. Both genetic and environmental factors are involved in development of PC. Epidemiological studies have shown that inflammation, toxins, hormonal changes or trauma are risk factors of PC, suggesting that chronic inflammation can lead to PC. IL-6 is a cytokine expressed in inflammation processes. Polymorphisms in gene for IL-6 are responsible for different production of IL-6. Aim of our prospective study was to analyze correlation of inflammation and IL- 6 high producer (GG or GC genotype) with presentation and symptoms of PC and BPH. Methods 120 PC and 120 BPH patients, age matched groups, were included after giving informed consent. In all patients history was taken and urological exam was done before biopsy was performed. Genomic DNA was extracted from whole blood and genotype polymorphism was assessed by real-time PCR. Results PC in family history was more common in PC group (8, 3% vs. 5.8%, p=0.169), prostate were larger in BPH group (76 ccm vs. 51 ccm, p=0.0001) and inflammation on histology was more common in BPH group (91.3% vs. 49.1%, p=0.001). There was no difference in mean IPSS score between groups (14 vs. 13, p=0.488). High producer genotype (GG or GC) of IL-6 was present in 86.7% of BPH patients and 80.8% of PC patients (p=0.147) and there was uniform distribution of high producers between PC groups (by differentiation, p=0.601). There was no difference in high producer genotype in patients with or without inflammation on histology in both groups (p=0.209 and p=0.300), but high producers are more common in BPH then in PC patients with inflammation (87, 3% vs. 77.6%, p=0.09). High producer genotype is evenly distributed when prostate size was accounted (volume 81 ccm) in BPH group, but more high producers are noted in larger prostates of PC patients (73%, 79%, 85%). However, this difference is not significant when prostate of same size are compared between groups. Discussion We found no difference in IL-6 producers between PC and BPH patients in our population. Inflammation is more common in BPH and ratio of IL-6 high producers is much higher in BPH patients with inflammation, suggesting limited role in PC. Higher ratio of high producers in patients with larger prostate can be explained with higher growth promotion by mediators of inflammation. Although some studies have not found a positive role of high producer IL-6 in PC development, contradictory results of published results have lead to proposed explanation of ethnic differences in IL-6 producers and effect on population.

Gas chromatography coupled with mass spectrometry (GC-MS) is often used as a confirmatory technique for positive results obtained with immunoassay screening tests, but it can also be used as a screening method in clinical and forensic toxicology. Because of different reasons (amount of sample available, urgency, cost of analysis, etc.), laboratory is often in position to perform just one extraction and screening per sample. Influence of different extraction procedures on results is usually overlooked, but it can have great impact on obtained results, both quantitatively and qualitatively. We prepared four different levels of commercial urine toxicology control samples Liquichek (BIO-RAD), containing 12 drugs-ofabuse using three different extraction procedures: liquid– liquid extraction system TOXI-TUBES A (VARIAN), liquid–liquid extraction system Chem Elut (VARIAN) and solid phase extraction system based on Amberlite XAD2 polymeric adsorbent columns (Sigma–Aldrich). All extracts were analyzed with GC-MS (SHIMADZU GCMS-QP5050A). Obtained results are presented and influence of extraction procedure on GC-MS screening results is evaluated for every extraction procedure.

A fast method for qualitative analysis of methadone inhuman urine using gas chromatography- mass spectrometry (GC-MS) is described. Urine samples were extracted by liquid-liquid extraction using Toxi-tubes A, and were not derivatized. Limit of detection was 25 ng/ml. Method performance in terms of recovery, repeatability and intermediate precision was studied and found to be acceptable for clinical analysis of methadone.

Methadone is a synthetic opioid, often used in the treatment of narcotic addiction. The oral use of methadone can suppress withdrawal symptoms, but in contrast to heroin, does not give any euphoric effect. However, this sensation can be induced by combination with alcohol, benzodiazepines, tramadol, etc. Therefore, regular urine testing should be carried out. We analyzed urines of 25 patients on methadone therapy with 3 methods: enzyme multiplied immunoassay (EMIT), thin layer chromatography (TLC) and gas chromatography mass spectrometry (GC–MS). We assessed the efficacy of these methods in verification of tramadol abuse in patients on methadone substitution therapy.