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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.

Introduction: The aim was to evaluate the BD Barricor tubes by comparison with the BD Rapid Serum Tubes (RST) through measuring 25 analytes and monitoring sample stability after 24 hours and 7 days. Materials and methods: Samples of 52 patients from different hospital departments were examined. Blood was collected in BD RST and BD Barricor tubes (Becton, Dickinson and Company, Franklin Lakes, USA). Analytes were measured by Beckman Coulter AU 480 (Beckman Coulter, Brea, USA), Dimension EXL (Siemens Healthcare Diagnostics, Newark, USA) and ARCHITECT i2000SR (Abbott Diagnostics, Lake Forest, USA). Between-tube comparison for each analyte was performed, along with testing analyte stability after storing samples at 4 °C. Results: BD Barricor tubes showed unacceptable bias compared to BD RST tubes for potassium (K) (- 4.5%) and total protein (4.4%). Analyte stability after 24 hours was acceptable in both tested tubes for most of analytes, except for glucose, aspartate aminotransferase (AST) and lactate dehydrogenase (LD) in BD Barricor and free triiodothyronine in BD RST sample tubes. Analyte stability after 7 days was unacceptable for sodium, K, calcium, creatine kinase isoenzyme MB, AST, LD and troponin I in both samples; additionally for glucose, alkaline phosphatase and albumin in BD Barricor. Conclusion: All analytes, except K and total protein, can be measured interchangeably in BD RST and BD Barricor tubes, applying the same reference intervals. For most of the analytes, sample re-analysis can be performed in both tubes after 24 hours and 7 days, although BD RST tubes show better 7-day analytes stability over BD Barricor tubes.

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.

In this study, we present a case of falsely elevated oestradiol (E2) concentration, determined by two immunoassays, in a breast cancer patient receiving exemestane therapy. The positive bias of immunochemical measurements was revealed using liquid chromatography tandem mass spectrometry which showed undetectable E2 concentration. The discrepancy is expected to be a consequence of the structural resemblance of E2 and exemestane sharing the same steroidal backbone. Inaccurate laboratory findings in therapy monitoring, as in this case, may lead to unnecessary changes of 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.

Single nuclear 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-1β+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-β1; χ2=0.026, p=0.872 for IL-10). Carriers of the IL-10-1082A>G 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-1β 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 GS>7 in eastern Croatians.

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.

Cytochrome P450 1A1 (CYP1A1) is an enzyme participating in the bioactivation of various endogenous and environmental reactive compounds that can bind to DNA and thus induce cancerogenesis. Gene encoding the enzyme is expressed in the prostate tissue and is polymorphic. CYP1A1*2A gene polymorphism is associated with elevated enzyme activity and/or inducibility which can lead to accumulation of genotoxic compounds and consequently to cancerogenesis. We examined the association of this polymorphism with prostate cancer (PCa) risk and aggressiveness. The case-control study consisted of 120 PCa patients and 120 benign prostatic hyperplasia (BPH) controls, in Croatian population. Regarding aggressiveness, PCa patients were grouped according to the Gleason score (GS), tumor stage (T) and existence of distant metastasis (M). The polymorphism was analyzed using real-time polymerase chain reaction (PCR). We did not observe association of mutated allele with PCa risk, neither with PCa aggressiveness. Furthermore, frequency of polymorphic genotype was slightly higher in BPH group (16.6% vs. 14.2%, respectively) and also in less aggressive form of PCa (20.4% vs. 9.6% for GS

Vitamin K epoxide reductase (VKOR) is a key enzyme in the g-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 1173C>T and VKORC1 -1639G>A 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.

Svjedoci smo da je u školama sve više djece s kombiniranim teškoćama u učenju i razvoju. Poteškoće se očituju u lošijoj fi noj i gruboj motorici, držanju tijela, slabijoj koncentraciji, a česti su i poremećaji pažnje, pamćenja, hiperaktivnosti, disleksije i disgrafi je. Unatoč velikoj angažiranosti svih sudionika odgojno-obrazovnog procesa, željeni rezultati većinom izostaju. U ovom radu predstavljena je metoda ,,gimnastika za mozak“ (eng. Brain Gym) kojom se aktivira mozak za učenje pomoću integrirajućih pokreta. Naglasak je posebice na primjeni njezinih elemenata pri radu s djecom s poteškoćama u učenju. Njezina primjena i pozitivni rezultati poznati su u mnogim državama svijeta, a u Hrvatskoj se primjenjuje u nekoliko škola i vrtića te predstavlja način pomoći djeci s teškoćama u učenju. Određivanje ciljanih elemenata i vježbi gimnastike za mozak za svakog pojedinog učenika temelji se na određivanju dominantnog profi la učenika. U radu je predstavljen postupak određivanja dominantnog profi la učenika drugog razreda osnovne škole, primjena elemenata gimnastike za mozak; uključenost učenika, učiteljice, roditelja i stručne suradnice-defektologinje-logopedice te rezultati primjene te metode, posebice pri radu s djecom s teškoćama u učenju., We have been witnessing the fact that our schools have been occupied in increasing number of children with combined learning and developmental diffi culties. The diffi culties are refl ected in poor fi ne and rough motor skills, body posture, lesser concentration, attention and memory disorders, hyperactivity, dyslexia and dysgraphia. Despite huge involvement of all participants in the educational process, the desired results are largely absent. Brain Gym (gymnastics for the brain) is a method of activating the brain for learning through integrating movement, presented in this paper. The particular emphasis is on application of its elements while working with children with learning diffi culties. Its implementation and positive results are known in many countries but in Croatia it is applied in several schools and kindergartens and represents an aspect of helping children with learning diffi culties. The defi nition of target elements and Brain Gym exercises for each pupil is based on the determination of pupils’ dominant profi les. The paper has presented the process of dominant profi ling of primary school second-graders; the use of Brain Gym elements; the involvement of pupils, teachers, parents and a professional assistant – i.e. a special-education teacher-speech therapist; and the results of Brain Gym application, particularly when working with children with learning diffi culties., L’aumento del numero dei bambini con varie diffi coltà nello sviluppo e nell’apprendimento è sotto gli occhi di tutti. Le diffi coltà si manifestano mediante capacità motorie non all’altezza, nella postura, nelle diffi coltà nella oncentrazione; sono frequenti anche i problemi legati al livello di attenzione, di memoria, all’iperattività, dislessia, disgrafi a. Nonostante il grande impegno di tutti i soggetti coinvolti nel processo educativo, i risultati desiderati non vengono raggiunti. Nel presente lavoro viene presentato il metodo „ginnastica cerebrale” (brain gym) che attiva il cervello attraverso attività integrate. L’accento viene posto sull’applicazione dei metodi del „brain gym” nel lavoro con gli alunni con le diffi coltà nell’apprendimento. La sua applicazione e i risultati positivi raggiunti sono ormai noti in tutto il mondo mentre in Croazia viene usata in alcune scuole e in alcuni asili nido. Essa rappresenta un valido aiuto per i ragazzi con le diffi coltà nell’apprendimento. La defi nizione degli elementi mirati e degli esercizi di ginnastica cerebrale adatti ad ogni singolo alunno si basa sulla defi nizione del profi lo dominante tra gli alunni. In questa sede viene esposto il processo di defi nizione del profi lo dominante tra gli alunni delle classi seconde delle scuole elementari, l’uso della ginnastica cerebrale, la partecipazione degli insegnanti, degli alunni, genitori nonché della fi gura di diffetologo-logopedista. Vengono altresì esposti i risultati dell’applicazione del metodo in questione, in particolar modo nel lavoro con i bambini che hanno delle diffi coltà nell’apprendimento.

Diagnostic value of available tumor markers, such as cancer antigen CA 15-3 and carcinoembryonic antigen (CEA) in breast cancer is limited. There is an ongoing search for additional, potentially better diagnostic blood markers with improved clinical utility. The aim of this study is to evaluate performance of the approach based on routine blood tests accompanied by a statistical learning tool to the diagnosis of breast cancer.Blood was collected from total of 104 subjects which were divided into two groups: breast cancer patients and a control group that consisted of asymptomatic volunteers and patients who had benign breast lesions at the time of blood collection. Random forest statistical learning method and the external method validation have been applied to evaluate diagnostic performance of 31 routine blood tests.The applied statistical learning approach assigned the highest diagnostic importance to the anemia panel among all analyzed blood tests that also included CA 15-3. External validation has shown utility of selected statistical approach – we were able to select tests that provide a diagnostic accuracy comparable to some diagnostic tools described in literature and based on more demanding laboratory techniques, such as gene expression microarrays.Inclusion of tests for anemia significantly improves diagnostic accuracy for the breast cancer in comparison to the diagnostic accuracy of the CA 15-3 alone. Application of the random forests also enables the reduction of number of laboratory tests needed for the establishment of diagnosis. Differences in relevant test values between the cancer and control group are small but application of multiparametric statistical learning ensured diagnostic accuracy of 72.0% associated by a sensitivity of 64.7% and specificity of 84.9%.

The objective of the present study was to compare estradiol/progesterone ratios of different age categories of red deer hinds and use it as a predictor of estrus synchronization success and consequently conception rate. To accomplish this we used 38 red deer hinds to establish serum progesterone and estradiol levels in young (21 animals), mature (10 animals) and old (7 animals) hinds during the estrus synchronization procedure (transvaginal/cervical AI). The following estrus synchronization was used: at the start of the experiment each hind received a controlled intravaginal drug-releasing device (CIDR, Pharmacia&Upjohn, New Zealand) containing 0.3 g of progesterone. The device was removed on day 11, simultaneously with an application of 250 IU of Pregnant Mare Serum Gonadotropin (PMSG, Folligon® Intervet International, Boxmeer, Holland). Transvaginal/ cervical AI (artificial insemination) was performed 48 hours after CIDR withdrawal (day 13). Blood samples were obtained from the jugular vein using a Venoject® vacutainer without an anticoagulant for hormonal tests on the same experimental day (0, 11th and 13th day). A statistically (p lt 0.01) higher progesterone level was found in young hinds on the 11th day after controlled intravaginal drug-releasing device insertion. A significantly higher (p lt 0.01) estrogen level was observed in the young in regard to mature and old hinds on the expected day of estrus (13th day). Estradiol/progesterone ratios showed a statistically significant difference (p lt 0.01) on insemination day (13th day) between old and young hinds (98.67 : 46.59) and between old and mature hinds (98.67 : 51.79). Out of a total of 38 hinds only 9 had their offspring, 6 of the young and 3 of the mature hinds. Farmski držani jeleni podvrgavaju se različitim tehnikama asistirane reprodukcije, kako bi se dobile genetski vredne životinje. Između ostalog ove tehnike uključuju i sinhronizovanje estrusa i veštačko osemenjavanje. Malo se zna o odnosu između estradiola i progesterona kod košuta. Cilj ovoga rada je bio da se uporede odnosi estradiola i progesterona različitih starosnih kategorija košuta crvenog jelena i da se koristite za predviđanje uspeha sinhronizovanja estrusa i koncepcije. Da bi smo ovo postigli proveli smo postupak sinhronizacije estrusa (0. dan aplikacija intravaginalnih uložaka progesterona (CIDR, Pharmacia&Upjohn, New Zealand), 11. dan uklanjanje intravaginalnih uložaka i aplikacija 250 IU seruma ždrebnih kobila (PMSG, Folligon® Intervet International, Boxmeer, Holand). Košute su umjetno osjemenjene 48 sati nakon aplikacije (13. dan) duboko smrznutim sjemenom uvezenim iz Novog Zelanda. Uzorci krvi za analizu hormona uzeti su iz jugularne vene pomoću Venoject® vacutainera bez antikoagulansa u iste dane tretmana (0., 11. and 13. dan) i pri tome smo određivali nivo progesterona i estradiola u serumu mladih, zrelih i starih košuta. Koncentracija hormona odeređena je imunoenzimskom kemiluminiscencijom 'ECLIA' (Roche Elecsys 2010, Roche Diagnostics, Mannheim, Germany). Statistički (p lt 0.01) veći nivo progesterona primećen je kod mladih košuta 11 dana posle aplikacije progesteronskog intravaginalnog implantata. Statistički vrlo značajno viši (p lt 0.01) nivo estrogena primećen je kod mladih košuta u odnosu na druge. Trinaestog dana eksperimenta, odnosno na dan estrusa, odnos estrogen/progesteron se vrlo značajno razlikovao (p lt 0.01) između starih (98,67) i mladih (46,59) košuta, kao i između starih i zrelih košuta (98,67:51,79). Od ukupno 38 obrađenih košuta, 9 je imalo potomstvo, od čega je 6 iz kategorije mladih košuta i 3 iz kategorije zrelih.

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.