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U ovom istraživanju proveli smo genetičku studiju povezanosti različitih tipova prirođenih anomalija s polimorfizmima gena uključenih u razvoj kao čimbenika rizika razvojnih poremećaja. Analizirali smo 140 uzoraka DNA izolirane iz tkiva parafinskih kocki od fetalno ili neonatalno umrle djece s prirođenim anomalijama a naspram kontrolne skupine zdrave djece kod četiri različita polimorfizma gena rs12329305 OSR1, rs10951154 HOXA1, rs9936833 u blizini FOXF1 i rs326119 MTRR. Spomenuti polimorfizmi analizirani su metodom real-time PCR. Analiza povezanosti napravljena je na razini alelske i genotipske distribucije između dvije ispitivane skupine. Značajnu alelsku (p=7×10−4) i genotipsku (p=0,0013) povezanost s prirođenim malformacijama našli smo u polimorfizmu rs12329305 gena OSR1. Dodatne analize povezanosti u istog polimorfizma pokazale su njegovu povezanost s podskupinom izoliranih anomalija (p=1.25×10−5) i to sa razvojnim anomalijama srca (p=5,12x10-8) i bubrega (p=4,18 x 10-5). Polimorfizam rs9936833 u blizini gena FOXF1 pokazao je genotipsku povezanost s prirođenim malformacijama (p=0,034), dok ostala dva polimorfizma nisu pokazala. Analizom distribucije genotipova ispitivanih polimorfizama uz uključen spol utvrdili smo da je OR za pojavnost prirođenih malformacija u skupini muškog spola za 1,8 puta veći nego u skupini ženskog spola (p=0,038). Rezultati ovog istraživanja po prvi put su pokazali da su polimorfizmi rs12329305 gena OSR1 i rs9936833 gena FOXF1, kao i muški spol, čimbenici rizika za nastanak različitih tipova prirođenih malformacija u mrtvorođene/neonatalno umrle djece., We tested the association of four development-related gene polymorphisms with congenital anomalies. We assumed that those polymorphic variants may be a risk factor for developmental disorders. We analyzed 140 DNA samples isolated from archived paraffin tissue of deceased patients in whom fetal/neonatal autopsy examination had shown congenital malformations versus a control group of healthy children for four different polymorphisms: OSR1 rs12329305, rs9936833 near FOXF1, HOXA1 rs10951154 and MTRR rs326119. These polymorphisms were genotyped using the TaqMan allelic discrimination assay. Association analysis was performed on the allelic and genotypic distribution between the two tested groups. Significant allelic and genotypic association with stillborn/neonatal death was observed for rs12329305 (p=7×10-4, p=0.0013, respectively). In addition, association analysis for the same polymorphism was shown in the subgroup with isolated anomalies (p=1.25×10−5), particularly in the subgroup of cases with kidney and heart anomalies (p=4.18 x 10-5, p=5.12×10-8, respectively). Polymorphism rs9936833 near the FOXF1 gene showed only genotypic association with congenital malformations (p=0.034), when we compared minor allele heterozygotes and homozygotes versus major allele homozygotes, while the other two polymorphisms were not revealed. We also found that the OR for congenital malformations occurrence was 1.8 times higher in males than in females (p = 0.038), when we analysed the genotypic distribution of four examined polymorphisms along with sex. This is the first study, as far as we know, that showed the OSR1 gene polymorphism rs12329305 and rs9936833 near FOXF1, as well as male sex, are risk factors for different types of congenital malformations in cases of stillborn/neonatal death.

In the initial diagnostics of arterial hypertension (AH) laboratory medicine is a cornerstone, along with a blood pressure (BP) measurement and an electrocardiogram. It mainly refers to routine blood and urine tests for diagnosis and monitoring primary hypertension and its associated conditions such as asymptomatic hypertension-mediated organ damage, chronic kidney disease and hypertensive disorders of pregnancy. In addition, long term non-fatal and fatal risks for cardiovascular (CV) events in hypertension are assessed based on clinical and laboratory data. Furthermore, laboratory medicine is involved in the management of hypertension, especially in monitoring the disease progression. However, antihypertensive drugs may interfere with laboratory test results. Diuretics, especially thiazides, can affect blood and urine sodium concentrations, or angiotensin- converting enzyme inhibitors and angiotensin receptor blockers can affect the blood biomarkers of the renin- angiotensin-aldosterone system (RAAS). It’s dysfunction plays a critical role in primary aldosteronism (PA), the most common endocrine disorder in secondary hypertension, which accounts for only small proportion of AH in relative terms but substantial proportion of hypertensives in absolute terms, affecting younger population and carrying a higher risk of CV mortality and morbidity. When screening for PA, aldosterone-to- renin ratio still contributes massively to the increased incidence of the disease, despite certain limits. In conclusion, laboratory medicine is involved in the screening, diagnosis, monitoring and prognosis of hypertension. It is of great importance to understand the preanalytical and analytical factors influencing final laboratory result.

Cilj istraživanja: istražiti da li je moguće uz pomoć analiziranih laboratorijskih parametara predvidjeti tijek i ishod bolesti COVID 19 kod starije populacije. Ispitanici i metode: Ispitanici su 32 osobe hospitalizirane u jedinici intezivne njege KBC Split radi COVID-19 infekcije preko 60 godina starosti od kojih je 8 preživljelih i 8 preminulih muškaraca te 8 preživljelih i 8 preminulih žena. Venepunkcija uzoraka seruma izvršila se u okviru standardnog uzorkovanja krvi u skladu s Nacionalnim preporukama za uzorkovanje venske krvi (Hrvatsko društvo za medicinsku biokemiju i laboratorijsku medicinu (HDMBLM) iz ožujka 2014.godine). Razine vitamina D analizirane su imunokemijskom elektrokemoluminiscentnom metodom proizvođača Roche na analizatoru Cobas e801. Razine kalcija, magnezija i fosfora analizirane su spektrofotometrijskom metodom proizvođača Roche na analizatoru Cobas c702. Broj leukocita, limfocita, neutrofilnih granulocita te trombocita su određeni na multiparametarskom hematološkom analizatoru, Advia 2120, za dijagnostiku in vitro u kliničkim laboratorijima. Pozitivan rezultat na virus SARS-Cov-2 je dokazan molekularnom PCR metodom. Rezultati: Napravljena je usporedba razine vitamina D u skupini osoba koje su preživjele u odnosu na skupinu umrlih osoba od posljedica COVID-19 bolesti te nije uočena statistička značajnost, preživjeli vs umrli (44.0 ± 29.7 vs 36.1 ± 18.3), p=0.376. Ono što se primjećuje je kako obje skupine imaju sniženu razinu vitamina D u serumu. Svim uključenim pacijentima je napravljena krvna slika, razina kalcija, magnezija i fosfora u serumu. Uočavamo najveću statističku značajnost kod omjera neutrofila i limfocita p=0.002, te da dob osobe utječe na ishod bolesti p=0.032 kao i ukupni broj leukocita p=0.028 i razina kalcija p=0.028. Izmjerene razine vitamina D u korelaciji s analiziranim laboratorijskim parametrima nisu pokazale međuzavisnost odnosno nema poveznice između razine vitamina D i broja leukocita, neutrofilnih granulocita, limfocita, trombocita, neutrofilno-limfocitnog omjera, trombocitno-limfocitnog omjera, kalcija, fosfora i magnezija. Multivarijabilna logistička regresijska analiza pokazala je da je samo broj leukocita statistički značajan prediktor preživljenja COVID-19 bolesti (OR 1.570, 95% CI 1.016 - 2.427; p = 0.041) kada se stavi u model zajedno s osnovnim karakteristikama poput dobi i spola te s drugim osnovnim laboratorijskim parametrima (OR 1.570, 95% CI 1.016 - 2.427; p = 0.041). Zaključak: Ishod bolesti COVID-19 kod istraživane populacije je lošiji što je dob starija te je još veći rizik uz visoki ukupan broj leukocita. Razina vitamina D je niska u proučavanoj skupini ali ne utječe na ishod bolesti i nema prognostički značaj. Neutrofilno-limfocitni omjer je prediktor tijeka COVID-19 bolesti te bi ga trebalo uzeti u obzir kod primjene terapije u liječenju. Objectives: Investigate is it possible to predict the course and outcome of COVID-19 disease in the elderly population with the help of analyzed laboratory parameters Subjects and methods: Respondents were 32 people hospitalized in the intensive care unit of KBC Split due to COVID-19 infection over 60 years of age, of which 8 survivors and 8 deceased men and 8 survivors and 8 deceased women. Venepuncture of serum samples was performed as part of standard blood sampling in accordance with the National Recommendations for Venous Blood Sampling (Croatian Society for Medical Biochemistry and Laboratory Medicine (HDMBLM) from March 2014). Vitamin D levels were analyzed by Roche immunochemical electrochemoluminescent method on a Cobas e801 analyzer. Calcium, magnesium and phosphorus levels were analyzed by spectrophotometric method from Roche on a Cobas c702 analyzer. A positive result for SARS-Cov-2 virus was proved by molecular PCR method. Results: A comparison of vitamin D levels in the group of survivors compared to the group of deaths due to COVID-19 disease was made and no statistical significance was observed, survivors vs deaths (44.0 ± 29.7 vs 36.1 ± 18.3), p = 0.376. What is noticeable is that both groups have lowered serum vitamin D levels. A blood count, calcium, magnesium and serum phosphorus levels were performed on all patients involved. We observe the greatest statistical significance in the ratio of neutrophils and lymphocytes p = 0.002, and that the age of the person affects the outcome of the disease p = 0.032 as well as the total number of leukocytes p = 0.028 and calcium levels p = 0.028. Measured vitamin D levels in correlation with the analyzed laboratory parameters did not show interdependence, there is no connection between the level of vitamin D and the number of leukocytes, neutrophilic granulocytes, lymphocytes, platelets, neutrophil-lymphocyte ratio, platelet-cell lymphocyte and lymphocyte lymphocytes. Multivariable logistic regression analysis showed that only leukocyte count was a statistically significant predictor of COVID-19 disease survival (OR 1,570, 95% CI 1,016 - 2,427; p = 0.041) when placed in the model along with basic characteristics such as age and sex and others basic laboratory parameters (OR 1,570, 95% CI 1,016 - 2,427; p = 0.041). Conclusion: The outcome of COVID-19 disease in the elderly population is worse in the older age and the higher is the risk with a high total leukocyte count. Vitamin D levels are low in the study group but do not affect disease outcome and have no prognostic significance. The neutrophil-lymphocyte ratio is a predictor of the course of COVID-19 disease and should be considered in the treatment of therapy.

Oral squamous cell carcinoma (OSCC) is frequently diagnosed in the advanced stages. The purpose of this paper is to determine the salivary values of SCCA1, SCCA2 and TROP2 in patients with T1N0M0 OSCC and to compare them with the values obtained from healthy individuals. Unstimulated (UWS) and stimulated (SWS) saliva was sampled from 29 patients with T1N0M0 OSCC and 29 sex- and age-matched healthy individuals. Statistical difference was observed in SCCA1 and SCCA2 levels both in UWS and SWS samples. TROP2 was not measurable in most of the salivary samples. Both SCCA1 and SCCA2 could represent potential biomarkers for the early-stage OSCC. Research on a larger sample and biomarker validation is needed to assess the clinical potential of SCCA1 and SCCA2 in the OSCC early diagnostics.

CILJ ISTRAŽIVANJA: Usporediti serumske koncentracije visoko osjetljivog troponina I i vrijednosti upalnog parametra hsCRP u STEMI i NSTEMI pacijenata. Utvrditi postoji li statistički značajna razlika u broju leukocita, diferencijalnoj krvnoj slici, aktivnosti serumskog enzima AST-a i glukoze u obje ispitivane skupine. ISPITANICI I METODE: Ovo istraživanje uključilo je 60 ispitanika sa simptomima srčanog zatajenja, a bili su liječeni na Klinici za bolesti srca i krvnih žila KBC-a Split u periodu siječnja 2018. do veljače 2019. godine. Uzorci su obrađeni i analizirani na Zavodu za medicinsko laboratorijsku dijagnostiku KBC Split. Prikupljeni su klinički podaci o dijagnozi ispitanika te su napravljena različita mjerenja i biokemijska ispitivanja. REZULTATI: Pacijenti su podijeljeni na STEMI (50%) i NSTEMI akutni infarkt miokarda (50%). Svim pacijentima izmjeren je sistolički i dijastolički tlak, kao i indeks tjelesne mase koji je pokazao pretilost kod 13,3% uključenih u istraživanje. Kod 78,3% pacijenata već je izvedena PCI. Lijekovi korišteni tijekom terapije su statini, varfarin, aspirin, a najčešće korišteni su ACE-inhibitori (50%) i beta blokatori (33,3%). Uspoređuju se komorbiditeti kao što su: šećerna bolest, pušenje, pretilost, a najčešće prisutna je arterijska hipertenzija (60%). Mjerenjem aktivnosti enzima AST-a u krvi uočene su značajne razlike kod STEMI (90,1 ± 108,9 IU/L) i NSTEMI (41,4 ± 28,3 IU/L), kao i kod broja neutrofila: STEMI (8,4 ± 2,9 x10^9/L) i NSTEMI(6,4 ± 2,3 x10^9/L). Značajne razlike uočene su i kod broja limfocita i leukocita. Ostali biokemijski parametri nisu ukazivali na značajne statističke razlike: hsCRP (STEMI (11,2 ± 17,0 mg/L) i NSTEMI (16,1 ± 26,8 mg/L), protrombinsko vrijeme, glukoza, natrij, eritrociti. Jedan od parametara bez značajne razlike je i visoko osjetljivi srčani troponin I: STEMI (417,8 ± 617,6 ng/L) i NSTEMI (314,9 ± 406,6 ng/L), iako je vidljiva njegova viša koncentracija kod STEMI ispitivane skupine. ZAKLJUČAK: Bez obzira na rezultate laboratorijskih pretraga, dijagnoza se mora postaviti zajedno s kliničkom prezentacijom pacijenta i zajedno s ostalim dijagnostičkim testovima. OBJECTIVES: To compare the serum concentrations of highly sensitive troponin I and the values of the inflammatory parameter hsCRP in STEMI and NSTEMI patients. To determine whether there was a statistically significant difference in leukocyte count, differential blood count, serum AST enzyme activity, and glucose in both study groups. PATIENTS AND METHODS: This study included 60 subjects with symptoms of heart failure, and were treated at the Clinic for Cardiovascular Diseases of the Clinical Hospital Center Split in the period from January 2018 to February 2019. The samples were processed and analyzed at the Department of Medical Laboratory Diagnostics, University Hospital Center Split. Clinical data on the diagnosis of the subjects were collected and various measurements and biochemical tests were performed. RESULTS: Patients were divided into STEMI (50%) and NSTEMI acute myocardial infarction (50%). All patients had systolic and diastolic blood pressure measured, as well as body mass index, which showed obesity in 13.3% of those included in the study. PCI has already been performed in 78.3% of patients. Medicaments used during therapy are statins, warfarin, aspirin, and the most commonly used are ACE inhibitors (50%) and beta blockers (33.3%). Comorbidities such as diabetes, smoking, obesity are compared, and arterial hypertension is most common (60%). By measuring the activity of AST in the blood, significant differences were observed in STEMI (90.1 ± 108.9 IU / L) and NSTEMI (41.4 ± 28.3 IU / L), as well as by measuring the number of neutrophils: STEMI (8, 4 ± 2.9 x10 ^ 9 / L) and NSTEMI (6.4 ± 2.3 x10^9/ L). Significant differences were also observed in the number of lymphocytes and leukocytes. Other biochemical parameters did not indicate significant statistical differences: hsCRP (STEMI (11,2 ± 17,0 mg/L) i NSTEMI (16,1 ± 26,8 mg/L), prothrombin time, glucose, sodium, erythrocytes. One of the parameters without significant difference is the highly sensitive cardiac troponin I: STEMI (417.8 ± 617.6 ng / L) and NSTEMI (314.9 ± 406.6 ng / L), although its higher concentration is visible in STEMI study groups. CONCLUSION: Regardless of the results of laboratory tests, the diagnosis should be established with the clinical presentation of the patient and other diagnostic tests.

Antinuclear antibodies (ANA) represent a family of autoantibodies targeting ubiquitous cellular constituents and are a hallmark of systemic inflammatory autoimmune rheumatic diseases named connective tissue diseases (CTD). The gold standard method for ANA determination is indirect immunofluorescence (IIF) on the human laryngeal epidermoid carcinoma cell line type 2 substrate (HEp-2), but with increasing demand for ANA testing, novel methods eased for automation emerged, which allows testing by staff less experienced in this specific field of laboratory diagnostic. In 2016 The working group (WG) for laboratory diagnostics of autoimmune diseases as part of the Committee for the Scientific Professional Development of the Croatian Society of Medical Biochemistry and Laboratory Medicine (CSMBLM) published the data of a survey regarding general practice in laboratory diagnostics of autoimmune diseases in Croatia. Results indicated high diversity in the performance of autoantibody testing as well as reporting of the results and indicated the need of creating recommendations for the assessment of ANA that would help harmonize diagnostics of systemic autoimmune rheumatic diseases in Croatia. This document encompasses twenty-seven recommendations for ANA testing created concerning indications for ANA testing, preanalytical, analytical, and postanalytical issues, including rational algorithm and quality control assurance. These recommendations are based on the relevant international recommendations and guidelines for the assessment of ANA testing and relevant literature search and should help to harmonize the approach in ANA testing and clarify differences in interpretation of the results obtained using different methods of determination.

Background:The aim of our study was to perform verification of serum indices on three clinical chemistry platforms.Methods:This study was done on three analyzers: Abbott Architect c8000, Beckman Coulter AU5800 (BC) and Roche Cobas 6000 c501. The following analytical specifications were verified: precision (two patient samples), accuracy (sample with the highest concentration of interferent was serially diluted and measured values compared to theoretical values), comparability (120 patients samples) and cross reactivity (samples with increasing concentrations of interferent were divided in two aliquots and remaining interferents were added in each aliquot. Measurements were done before and after adding interferents).Results:Best results for precision were obtained for the H index (0.72%–2.08%). Accuracy for the H index was acceptable for Cobas and BC, while on Architect, deviations in the high concentration range were observed (y=0.02 [0.01–0.07]+1.07 [1.06–1.08]x). All three analyzers showed acceptable results in evaluating accuracy of L index and unacceptable results for I index. The H index was comparable between BC and both, Architect (Cohen’s κ [95% CI]=0.795 [0.692–0.898]) and Roche (Cohen’s κ [95% CI]=0.825 [0.729–0.922]), while Roche and Architect were not comparable. The I index was not comparable between all analyzer combinations, while the L index was only comparable between Abbott and BC. Cross reactivity analysis mostly showed that serum indices measurement is affected when a combination of interferences is present.Conclusions:There is heterogeneity between analyzers in the hemolysis, icteria, lipemia (HIL) quality performance. Verification of serum indices in routine work is necessary to establish analytical specifications.

All key biological macromolecules are susceptible to carbonylation – an irreparable oxidative damage with deleterious biological consequences. Carbonyls in proteins, lipids and DNA from cell extracts have been used as a biomarker of oxidative stress and aging, but formation of insoluble aggregates by carbonylated proteins sometimes precludes their quantification. Since carbonylated proteins can correlate with morbidity and mortality, we have developed an in situ detection of total proteins, DNA, RNA, lipids and carbonyl groups at the level of the whole organism of the nematode C. elegans. The method enabled us to show that after UV irradiation, carbonylation colocalizes mainly with proteins, and was applied to improve the knowledge of mechanisms of E. coli cell death and C elegans aging. There is a growing body of evidence that carbonylation could be the root cause of cell death and aging. To explore this possibility, we have monitored in E. coli DNA damage and protein carbonylation after UVC irradiation. A method that detects in real time all emerging mutations, allows the exclusion of mutations as the leading cause of UVC-induced cell death. The recovery process of DNA is nearly complete within two hours after irradiation, while UVC-induced protein carbonylation remains doubled. Our new method reveals that UVC induces also an increase in lipid levels and changes in lipid pattern which could indicate membrane permeabilization. Since cell death by membrane disintegration follows the increase in protein carbonylation, we have concluded that cell function (proteome activity) is the primary target in cell death. This is not surprising given the fact that all cellular structures and functions (e.g., DNA and membrane integrity) depend on proteome activity. Moreover, we applied this method to explore the effects of chronic ionizing radiation of C. elegans on oxidative damage to its proteins. To follow the lifespan of rapidly reproducing animals after irradiation we used a conditional sterile glp-1 mutant. Our results confirmed that: (i) increase in protein carbonylation with C. elegans’ age establishes carbonylation as a good biomarker of aging and (ii) low-dose chronic ionizing radiation reduces the lifespan of sterile C. elegans whatever the dose/dose rate tested. However, there is also a rather unexpected result of reduced carbonylation after irradiation which could be explained by increased activity of proteasome in the germline ablated glp-1 worms. In addition, radiations induced a decrease in lipid levels, whatever the dose or the dose rate. Fat composition and fat content together could induce a beneficial or detrimental effect on longevity. All these findings are of particular importance. These parameters could become predictive biomarkers of biological consequences of radiation and aging. In conclusion, our new procedure enables us to monitor carbonylation in E. coli during death and in the nematode C. elegans during its life cycle, including the egg stage, through all the stress and aging. That opens up a possibility that the method might be applied for ex vivo diagnostic detection of oxidative carbonylation in the samples of human biopsies., Biološke makromolekule su osjetljive na oksidaciju, na primjer karbonilaciju proteina koja je nepopravljivo oštećenje s pogubnim biološkim posljedicama. Budući da korelira s oksidativnim stresom i starenjem, upotrebljava se kao biomarker. Karbonilacija se događa i na ostalim biološkim makromolekulama, a osim toga karbonilirani proteini stvaraju unutarstanične agregate, što pridonosi otežanoj kvantifikaciji karboniliranih proteina. Također, svi postupci koji uključuju ekstrakciju proteina mogu prouzročiti gubitak karboniliranih proteina. Budući da postoji jasna povezanost između karboniliranih proteina, staničnog morbiditeta i smrti te da su karbonilirani proteini mogući uzrok navedenog, pojavila se potreba za vjernom kvantifikacijom i lokalizacijom na nivou stanica i tkiva. Zato smo, koristeći odgovarajuće fluorescentne molekule, razvili metodu in situ detekcije ukupnih proteina, DNK, RNK, lipida i karbonilnih skupina na razini cijelog organizma nematode C. elegans. Pokazali smo da nakon UV zračenja karbonilacija kolokalizira uglavnom s proteinima, a ova metoda je također primijenjena za poboljšanje znanja o mehanizmu smrti kod bakterije E. coli i starenju nematode C. elegans. Postoji sve veći broj dokaza da bi karbonilacija tokođer mogla biti i uzrok stanične smrti. Da bismo istražili ovu mogućnost, pratili smo oštećenje DNK i karbonilaciju proteina kod bakterije E. coli nakon UV zračenja. Metoda koja u realnom vremenu otkriva sve nove mutacije i oštećenja DNK, omogućuje isključivanje mutacija kao vodećeg uzroka stanične smrti izazvane UV zračenjem. Proces oporavka DNK je završen unutar dva sata nakon zračenja, dok se razina karbonilacije proteina inducirana zračenjem udvostruči. Naša nova metoda kod ozračenih bakterija također otkriva povećanje razine lipida i promjene u njihovom uzorku koje bi mogle ukazivati na povećanu propusnost membrane. Budući da stanična smrt prati porast karbonilacije, zaključujemo da je stanična funkcija (aktivnost proteoma) primarna meta smrti stanica. Ova spoznaja ne iznenađuje budući da stanične strukture i funkcije (kao što su npr. integritet genoma i membrana) ovise isključivo o aktivnosti proteina. Naša metoda je također korištena za istraživanje učinaka kroničnog ionizirajućeg zračenja na životni vijek nematode C. elegans. Za praćenje životnog vijeka nematoda poslije zračenja smo koristili uvjetni sterilni glp-1 mutant. Naši rezultati potvrđuju sljedeće: (i) karbonilacija je opravdan biomarker starenja budući da stare nematode pokazuju višu razinu karbonilacije od mladih, što je očekivano i sukladno brojnim ranijim istraživanjima starenja, te (ii) kronično ionizirajuće zračenje niskim dozama smanjuje životni vijek sterilnih nematoda bez obzira na ispitivanu dozu/stopu doze. Međutim, kronično zračenje pokazalo je neočekivano smanjenje razine karbonilacije koje se može objasniti povećanom aktivnošću djelovanja proteasoma kod sterilnih nematoda. Zračenje je također izazvalo povećanje razine lipida kao odgovor na starenje te lipidni katabolizam kao odgovor na zračenje (također bez obzira na dozu). Konačno, sadržaj lipida i njihov udio mogu izazvati blagotvorno ili štetno djelovanje na dugovječnost. Ovi parametri mogu postati prediktivni biomarkeri bioloških posljedica zračenja i starenja. U konačnici, ova metoda omogućuje praćenje karbonilacije u pojedinačnim stanicama te prilikom stresa, starenja i bolesti tijekom cijelog životnog ciklusa nematode C. elegans, uključujući i stadij jajeta. Navedeno otvara put primjeni metode na ex vivo modelu odnosno na mogućnosti dijagnostike oksidativne karbonilacije na uzorcima ljudskih tkiva.

Uvod: Cilj ovog istraživanja bio je odrediti učinak hiperbaričnog tretmana (HB) na leukocitne proupalne biljege CD15s i CD11b. Dodatno, istraživanje opisuje promjene u izražaju CD77 i CD34 biljega na stanicama bubrežnog, plućnog i srčanog tkiva nakon hiperbaričnog tretmana. Materijali i metode: Izražaj CD11b i CD15s na leukocitima, kao i CD77 i CD34 biljega na suspenziji stanica bubrežnog, plućnog i srčanog tkiva u životinja nakon hiperbaričnog tretmana i u kontrolnim životinjama, određen je protočnom citometrijom. Rezultati: Hiperbarični tretman je statistički značajno povećao postotak leukocitnih CD15s+CD11b- stanica (od 1.71 ± 1.11 do 23.42 ± 2.85, P < 0.05). Hiperbarični tretman je statistički značajno smanjio postotak CD77+CD34- i CD77+CD34+ stanica u bubrežnom tkivu (od 16.35 ± 5.5 do 4.48 ± 1.28, P < 0.05). Hiperbarični tretman je statistički značajno povećao postotak CD34+ stanica u plućnom tkivu (od 3.27 ± 2.01 do 11.92 ± 6.22, P < 0.05). Naše istraživanje je prvo koje je pokazalo utjecaj hiperbaričnog tretmana na CD34+ stanice u srčanom tkivu kod štakora. Zaključak: Naši rezultati povećanog postotka endotelnog liganda CD15s+ na leukocitima, nakon hiperbaričnog tretmana, upućuju na njegovu ulogu u procesima oštećenja endotela. Pronaši smo također značajni porast postotka CD34+ stanica u srčanom tkivu kao i postotka CD34+ stanica u plućnom tkivu nakon hiperbaričnog tretmana, što može biti posljedica mehanizama oporavka oštećenog endotela usljed hiperoksije., Objective(s): The aim of this study was to estimate effects of hyperbaric (HB) treatment by determination of CD15s and CD11b leukocyte proinflammatory markers expression. In addition, this study describes changes in CD77 and CD34 expression on rat endothelial cells in renal, pulmonary and cardiac tissue following exposure to hyperbaric pressure. Materials and Methods: Expression of CD11b and CD15s on leukocytes, as well as CD77 and CD34 expression on endothelial cells in cell suspensions of renal, pulmonary and cardiac tissue in rats after hyperbaric treatment and in control rats were determined by flow cytometry. Results: Hyperbaric treatment significantly increased percentage of leukocytes expressing CD15s+CD11b- (from 1.71 ± 1.11 to 23.42 ± 2.85, P < 0.05). Hyperbaric treatment significantly decreased sum percentage of CD77+CD34- and CD77+CD34+ renal cells (from 16.35 ± 5.5 to 4.48 ± 1.28, P < 0.05). Hyperbaric treatment significantly increased percentage of CD34+ pulmonary cells (from 3.27 ± 2.01 to 11.92 ± 6.22, P < 0.05). Our study is the first reporting the hyperbaric environment influence on CD34+ heart cells in rats. Conclusion: The current findings of increased percentage of leukocytes expressing endothelial selectin ligand CD15s after hyperbaric treatment, point its role in endothelial damage. We found out a significantly increase in percentage of CD34+ cardiac cells as well as CD34+ pulmonary cells in rats after HB treatment which could be a part of repair mechanism of injured endothelium caused by hyperoxia.

CILJ: Utvrditi razine leptina i adiponektina u krvi pretile djece i adolescenata te saznati utječe li pojedini polimorfizam gena za sintezu leptinskog receptora na rezistenciju samog receptora na leptin i razinu leptina. Procijeniti utjecaj polimorfizama gena za adiponektin na njegovu razinu u krvi. MATERIJALI I METODE: Presječno istraživanje slučajeva i kontrola usporedbom ispitivane grupe od 30 pretile djece i adolescenata (dob 13.2±2.6 godina) sa kontrolnom skupinom od 30 djece normalne težinu usklađene dobi (dob 12.7±2.9 godina). U obje skupine izmjeren je indeks tjelesne mase (ITM) i opseg struka i bokova te sistolički i dijastolički krvni tlak. Izmjereni su i standardni metabolički parametri (GUK natašte, ukupni kolesterol i njegove frakcije, serumski trigliceridi). Osjetljivost na inzulin je procijenjena korištenjem inzulinemije natašte i HOMA-indeksa. Razine adiponektina i leptina određene su korištenjem ELISA metode. SNP-ovi su locirani PCR-RFLP metodom. REZULTATI: Serumska razina leptina bila je značajno veća (34.0±20.4 ng/mL u usporedbi s 9.1±6.4 ng/mL, p, OBJECTIVES: To determine serum levels of leptin and adiponectin of obese children and adolescents and to identify the influence of the polymorphisms of leptin receptor gene on leptin resistance and leptin levels. Furthermore, to examine the association between the polymorphisms of adiponectin gene and adiponectin levels. PATIENTS AND METHODS: A case-control study comparing a study group of 30 obese children and adolescents (age 13.2±2.6 years) to a normal weight age matched (age 12.7±2.9 years) control group of 30 children. In both groups body mass index (BMI) and waist and hip circumference, systolic and diastolic blood pressure were mesured. Also, the classical metabolic parameters (fasting glycemia, total cholesterol and its fractions, serum triglycerides) were measured. Insulin sensitivity was evaluated using fasting insulinemia and HOMA-index. Adiponectin and leptin levels were determined using ELISA method. PCR-RFLP based assay was utilized to genotype SNPs. RESULTS: Serum level of leptin was significantly higher (34.0±20.4 ng/mL versus 9.1±6.4 ng/mL, p

Introduction: Cobas 6000 (Roche, Germany) is biochemistry analyzer for spectrophotometric, immu-noturbidimetric and ion-selective determination of biochemical analytes. Hereby we present analytical validation with emphasis on method performance judgment for routine operation. Materials and methods: Validation was made for 30 analytes (metabolites, enzymes, trace ele-ments, specific proteins and electrolytes). Research included determination of within-run (N = 20) and between-run imprecision (N = 30), inaccuracy (N = 30) and method comparison with routine analyzer (Beckman Coulter AU640) (N = 50). For validation of complete analytical process we calculated total error (TE). Results were judged according to quality specification criteria given by European Working Group. Results: Within-run imprecision CVs were all below 5% except for cholesterol, triglycerides, IgA and IgM. Between-run CVs for all analytes were below 10%. Analytes that did not meet the required speci-fications for imprecision were: total protein, albumin, calcium, sodium, chloride, immunoglobulins and HDL cholesterol. Analytes that did not fulfill requirements for inaccuracy were: total protein, calcium, sodium and chloride. Analytes that deviated from quality specifications for total error were: total pro-tein, albumin, calcium, sodium, chloride and IgM. Passing-Bablok regression analysis provided linear equation and 95% confidence interval for intercept and slope. Complete accordance with routine ana-lyzer Beckman Coulter AU640 showed small number of analytes. Other analytes showed small pro-portional and/or small constant difference and therefore need to be adjusted for routine operation. Conclusions: Regarding low CV values, tested analyzer has satisfactory accuracy and precision and is extremely stable. Except for analytes that are coherent on both analyzers, some analytes require adjustments of slope and intercept for complete accordance.

Sedimentacija eritrocita (SE) i C reaktivni protein (CRP) su jednostavne, brze i neinvazivne laboratorijske pretrage, primjenjuju se za otkrivanje ili praćenje bolesnika sa sumnjom na upalnu bolest. Odabir SE ili CRP-a u procesu obrade i/ili liječenja bolesnika ovisi o dijagnostičkoj vrijednosti pretrage za određeno stanje ili bolest. SE je jedna od najstarijih laboratorijskih pretraga, kod koje brzina sedimentiranja ovisi o obliku i broju eritrocita te sastavu plazme. SE je povišena u nekim fiziološkim stanjima kao u trudnoći, a u brojnim patološkim stanjima povezana je najčešće s upalom, anemijom, paraproteinemijom, povećanom koncentracijom fibrinogena, prisustvom hladnih aglutinina. Referentni raspon za SE varira ovisano o dobi i spolu a kreće se od  15 mm/h do  49 mm/h. Eritrociti se u normalnim uvjetima međusobno odbijaju. Sijalinska kiselina se nalazi na membrani eritrocita i štiti stanice od ubrzane agregacije. U upalnim stanjima se pojačano sintetiziraju proteini akutnog odgovora organizma, fibrinogen ili imunoglobulini. Te bjelančevine inhibiraju zaštitno djelovanje sijalinske kiseline na površini eritrocita, pa oni brže sedimentiraju. Za određivanje brzine sedimentacije eritrocita Internacionalni odbor za standardizaciju u hematologiji (engl. International Council for Standardisation in Haematology, ICSH) preporučujeje metodu po Westergreenu, kojom se mjeri brzina taloženja eritrocita u standardiziranoj graduiranoj pipeti u određenom vremenu (1 sat) na sobnoj temperaturi. Posljednjih su se godina razvile nove, automatizirane metode od kojih su neke modifikacija klasične Westergreenove metode. Većina metoda za automatizirano određivanje sedimentacije koristi analizatore koji primjenjuju mikrofotometar sa infracrvenom zrakom za određivanje optičke gustoće uzorka. Metode su jednostavne i brze, ali nisu standardizirane. CRP je najznačajniji biljeg akutnoga upalnoga odgovora, osjetljiviji je od SE, u dobroj je korelaciji s težinom i opsegom upale. Unatoč mišljenju da promjene vrijednosti SE i CRP u podlozi imaju sličnu patofiziološku osnovu, poznate su razlike za istu bolest u vrijednostima među ova dva analita. Među proteinima akutne faze koji se u laboratoriju mogu jednostavno mjeriti CRP-je jedan od osjetljivijih testova sa brzim odgovorom. SE nije specifična za odgovor akutne faze upale i na nju utječe hematokrit, oblik i veličina eritrocita ... Preporučene metode za određivanje CRP-a su neobilježene ili izravne tehnike, npr.imunonefelometrija, imunoturbidimetrija i radijalna imunodifuzija U laboratorijskoj dijagnostici poznavanje značajki metoda za određivanje SE i CRP podrazumijeva , odabir prikladnog uzorka i metode, poznavanje ograničenja metode. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) are simple, fast and non-invasive laboratory tests, and are used for the detection or monitoring of patients with suspected inflammatory disease. Selection of ESR or CRP in the processing and / or treatment of a patient depends on the diagnostic value of the test for a given condition or disease. ESR is one of the oldest laboratory tests, in which the speed of the sedimentation depends on the shape and the number of red blood cells and plasma composition. ESR is elevated in some physiological states as in pregnancy, and in a number of pathological conditions it is associated with inflammation, anemia, paraproteinemia, increased concentration of fibrinogen, presence of cold agglutinins. The reference range for ESR varies dependent on age and sex and ranges from • 15 mm / h to • 49 mm / h. The red blood cells in normal conditions repel one another. Sialic acid is on the erythrocyte membrane and protects the cells of accelerated aggregation. In inflammatory conditions proteins of acute response are increasingly synthesized, fibrinogen or immunoglobulins. These proteins inhibit the protective effect of sialic acid on the surface of red blood cells, so they quickly settle down. To determine the erythrocyte sedimentation speed International Council for Standardization in Haematology (ICSH) recommends a method by Westergren, which measures the speed of sedimentation of erythrocytes in a standardized graduated pipette in a given time (1 hour) at room temperature. In recent years new, automated methods were developed of which some are modifications of classical Westergren method. Most methods for automated determination of sedimentation uses analyzers to apply microphotometer with infrared ray to determine the optical density of the sample. The methods are simple and fast, but they are not standardized. CRP is the most important marker of acute inflammatory response, it is more sensitive than ESR, and it is in good correlation with the severity and extent of inflammation. Despite the opinion that the value changes of ESR and CRP have similar pathophysiological basis in the background, there are differences in values between the two analytes for the same disease Among the acute phase proteins that can easily be measured in the laboratory, CRP is one of the more sensitive tests with fast response. ESR is not specific for the acute phase inflammatory response and is affected by hematocrit, red cell size and shape Recommended methods for the determination of CRP are unmarked or direct techniques, npr. immunonephelometry, immunoturbidimetry and radial immunodiffusion. In the laboratory diagnostics knowledge of the features of methods for determining the SE and CPR means a selection of a suitable sample and method, and the knowledge of the limitations of the method.

UVOD: Imunost posredovana stanicama je važna u obrani domaćina protiv virusnih infekcije i zloćudnih novotvorina. Reaktivni limfocit je ne-maligni leukocit nađen u perifernoj krvi. Pojavljuje se kao nespecifični odgovor na stres. Danas je to atipični limfocit, Downey-ova stanica, ili reaktivni limfocit te je on marker bolesti kada se pronađe u perifernoj krvi. Laboratorijska automatizacije sve se više razvija po pitanju hematoloških analizatora i algoritama kako bi na odgovarajući način otkriti abnormalne uzorke među kojima i reaktivne limfocite čiji nalaz nam jako pomaže u samoj dijagnostici stanja pacijenta. CILJ ISTRAŽIVANJA: Odrediti pouzdanost nalaza reaktivnih limfocita u 60 uzoraka pacijenata. Od ukupnog broj uzoraka, 30 uzoraka se odnosilo na zdrave uzorke u usporedbi sa 30 pozitivnih nalaza reaktivnih limfocita. MATERIJALI I METODE: Uzorci krvi (n=60) korišteni za analizu bili su podjeljeni u dvije skupine: 30 uzoraka je pripadalo zdravim ispitanicima sa negativnim nalazom reaktivnih limfocita. Preostalih 30 uzoraka je pripadalo ispitanicima s pozitivnim nalazom reaktivnih limfocita. Mjerni instrument korišten u svrhe analize je hematološki analizator ADVIA 2120. Kao kontrola korišten je mikroskopski pregled krvnog razmaza. REZULTATI: Grafičkim prikazom Passing – Bablok regresije određivali smo reaktivne limfocite metodama ADVIA- hem.analizator i mikroskopski za grupu zdravih ispitanika. Rezultati regresije za rezultate određivanja reaktivnih limfocita na analizatoru ADVIA i mikrosposki s 95% -tnom pouzdanosti nemaju značajno odstupanje među izmjerenim vrijednostima ( y=1,000+0,500 x) uz P> 0,10. Grafičkim prikazom Passing – Bablok regresije određivali smo reaktivne limfocite metodama ADVIA- hem.analizator i mikroskopski za grupu ispitanika s patofiziološkim promjenama. Rezultati regresije za rezultate određivanja reaktivnih limfocita na analizatoru ADVIA i mikrosposki s 95% -tnom pouzdanosti nemaju značajno odstupanje među izmjerenim vrijednostima ( y=1,300+0,8083 x) uz P> 0,10. ZAKLJUČAK: 1. Potvrđena je pouzdanost nalaza prisustva reaktivnih limfocita na hematološkom analizatoru ADVIA 120 provjerom reproduciblnosti- preciznost na jednom uzorku ponavljanom 10 puta uz KV =4,5%. 2. Utvrđivana je prihvatljiva razine usporedivosti udjela ( postotka ) reaktivnih limfocita određenih na hematološkom analizatoru i pregledom razmaza obojenog metodom MGG jer uz 95% pouzdanost nema bitnih odstupanja u rezultatima u grupi zdravih ispitanika. 3. Utvrđivana je prihvatljiva razine usporedivosti udjela ( postotka ) reaktivnih limfocita određenih na hematološkom analizatoru i pregledom razmaza obojenog metodom MGG jer uz 95% pouzdanost nema bitnih odstupanja u rezultatima u grupi ispitanika s patofiziološkim promjenama. 4. Utvrđena je razina osjetljivosti ( najniži broj reaktivnih limfocita ) koji analizator klasificira na vrijednost 1% .Ova vrijednost je zaokružena jer u klasifikaciji analizator izražava vrijednosti u postotku s jednom decimalom što treba uzeti u obzir pri interpretaciji rezultata. INTRODUCTION: Cell mediated immunity is important in host defense against viral infections and malignancies. Reactive lymphocyte is a non-malignant leukocytes that was found in the peripheral blood. It appears as a nonspecific response to stress. Today it is an atypical lymphocyte, Downey's cells, or reactive lymphocytes and is a marker of disease when it is found in the peripheral blood. Laboratory automation is increasingly developing in terms of hematology analyzers and algorithms in order to adequately detect abnormal patterns including reactive lymphocytes whose findings we really helpful in the diagnosis of the patient's condition. AIM OF RESEARCH: To determine the reliability of the findings of reactive lymphocytes in 60 patient samples. Of the total number of samples, 30 samples were related to healthy samples compared with 30 positive findings reactive lymphocytes. MATERIALS AND METHODS: Blood samples (n = 60) were used for the analysis were divided into two groups of 30 samples belonging to the healthy subjects with negative findings reactive lymphocytes. The remaining 30 samples belonged to subjects with positive findings reactive lymphocytes. The measuring instrument used for the purposes of analyzing is the hematology analyzer ADVIA 2120. As control was used microscopic examination of blood smears. RESULTS: Graphic display of Passing - Bablok regression was used to determine the reactive lymphocytes methods ADVIA-hem. analizator and microscopically for a group of healthy subjects. Results of regression results for the determination of reactive lymphocytes analyzer ADVIA and microscopic with 95% confidence with no significant deviation between the measured values. (Y = 1.000 +0.500 x) with P> 0.10. Graphic display of Passing - Bablok regression was used to determine the reactive lymphocytes methods ADVIA-hem. analizator and microscopically for a group of patients with pathophysiological changes. Results of regression results for the determination of reactive lymphocytes analyzer ADVIA and microscopic with 95% confidence with no significant deviation between the measured values. (Y = 1.300 +0.8083 x) with P> 0.10. CONCLUSION: 1. The reliability of the findings of the presence of reactive lymphocytes in hematology analyzer ADVIA 120 is confirmed reproducibility-checking the accuracy of a single sample repeated 10 times with CV = 4.5%. 2. Acceptable level of comparability share (percentage) of reactive lymphocytes specific to the hematology analyzer and examination of smears stained using MGG is determined because with 95% confidence there are no significant differences in the results in a group of healthy subjects. 3. Acceptable level of comparability share (percentage) of reactive lymphocytes specific to the hematology analyzer and examination of smears stained using MGG is determined because with 95% confidence there are no significant differences in the results in the group of patients with pathophysiological changes. 4. The level of sensitivity (the lowest number of reactive cells) is determined by analyzer which classifies the value of 1%. This value is rounded because the classification of the analyzer expressed as a percentage of the value to one decimal place which should be taken into account when interpreting the results.

Cilj: Cilj ovog rada je ispitati promjene biokemijskih i hematoloških parametara koje nastaju tijekom čuvanja koncentrata trombocita. Materijali i metode: Analizirano je deset koncentrata trombocita prvog dana proizvodnje, te deset koncentrata šestog dana od uzimanja krvi. U navedenim uzorcima biokemijski parametari (natrij, kalij i pH) su analizirani na analizatoru RAPIDlab 1265, Siemens. Hematološki parametri, broj leukocita, trombocita te trombocitni indeksi (MPV, PDW i PCT) su analizirani na hematološkom brojaču Advia 2120i, Siemens. Statistička obrada podataka je napravljena pomoću Microsoft Excel 2010. Za dobivene vrijednosti izračunata je aritmetička sredina, standardna devijacija, medijan te minimum i maksimum. Rezultati: Iz dobivenih rezultata utvrđene su manja promjene ispitivanih parametara u koncentratima trombocita prvog i šestog dana čuvanja. Vrijednost kalija šestog dana od dana proizvodnje porasla je za 0,15 mmol/L, natrija je pala za 0,1 mmol/L, srednji volumen trombocita je pao za 0,1 fL, a pH je pao za 0,14. Zaključak: Razlike između ispitivanih parametra (Na, K, pH, leukocita, trombocita i trombocitnih indeksa) u koncentratu trombocita prvog dana proizvodnje i šestog dana proizvodnje su minimalna. Rezultati ispitivanja pokazuju da su ispitivani parametri u prihvatljivim granicama i na kraju roka valjanosti. Ovaj rad je poticaj za istraživanje dodatnih parametara u svrhu proučavanja kvalitete i funkcije koncentrata trombocita radi produljenja roka valjanosti. Objective: The aim of this study was to examine changes in biochemical and hematological parameters that occur during storage of platelet concentrates. Materials and Methods: We analyzed ten platelet concentrates on the first day of production, as well as ten concentrate on the sixth day from the date of production. In these samples biochemical parameters (sodium, potassium and pH) were analyzed by the analyzer RAPIDlab 1265, Siemens. Hematological parameters, leukocyte count, platelet count and platelet indices (MPV, PDW and PCT) were analyzed on a hematology analyzer Advia 2120, Siemens. Statistical analysis was done using Microsoft Excel 2010th. The arithmetic mean, standard deviation, median, the maximum and minimum mean were calculated for the obtained values. Results: The results showed minor changes in the parameters examined in platelet concentrates first and sixth days of storage. Value potassium sixth days of production increased by 0.15 mmol/L, sodium level decreased by 0.1 mmol/L, mean platelet volume decreased by 0.1 fl and pH decreased by 0,14. Conclusion: The differences between the studied parameters (Na, K, pH, leukocytes, platelets and platelet index) in the platelet concentrate on the first day of production, and on the sixth day of production are minimal. Results of this study indicates that the examined parameters are still within acceptable limits at the end of the shelf life. This study have an incentive to explore some other parameters in order to study the quality and function of platelet concentrates to extend shelf life.

Uvod: Pripravci eritrocita najznačajniji su proizvod u transfuzijskoj djelatnosti. Koriste se u liječenju anemija, kirurških i životno ugroženih bolesnika. Očuvanje funkcije i kvalitete ovih priprvaka jedan je od preduvjeta uspješnog transfuzijskog liječenja. Cilj: Svrha ovog rada je utvrditi biokemijske i hematološke promjene koje nastaju tijekom čuvanja koncentrata eritrocita u optimalnim uvjetima. Materijali i metode: U deset koncentrata eritrocita proizvedenih standardiziranim postupkom ispitani su hematološki (E, Hct, Hb, MCV, MCH i MCHC) i biokemijski (AST, LDH, Na, K, Cl i pH) parametri prvog i 35. dana skladištenja. U tu svrhu korišteni su hematološki analizator Advia 2102i, biokemijski analizator Beckman Coulter AU680 te RapidLab 1265 sistem za mjerenje pH. Statistička obrada podataka izvršena je Microsoft Excel 2007 programskom podrškom. Rezultati: Tijekom skladištenja koncentrata eritrocita broj eritrocita, hemoglobin i MCH nisu se značajno promijenili. Hct i MCV su nešto porasli dok je MCHC na dan isteka roka valjanosti pripravka snižen. Posljednjeg dana čuvanja koncentrata značajno su porasli AST, LDH i K. pH je u tom razdoblju osjetno pao a zabilježen je i pad koncentracije Na i Cl. Zaključak: Najznačajnije promjene u razdoblju skladištenja eritrocitni krvni pripravci pokazuju u porastu koncentracija K, LDH i u sniženju pH. To je važna spoznaja koju treba uzeti u obzir kod liječenja pacijenata koncentratima eritrocita pred kraj roka valjanosti. Introduction: RBC concentrates are the most important product in the transfusion activities. They are used to treat anemia, surgical and critically ill patients. Preserving function and quality of these products is one of the conditions for a successful transfusion therapy. Objective: The purpose of this study was to determine the biochemical and hematological changes that occur during storage of packed red blood cells in optimal conditions Materials and Methods: In ten packed red blood cells produced by standardized procedure we examined hematological (E, Hct, Hb, MCV, MCH and MCHC) and biochemical (AST, LDH, Na, K, Cl and pH) parameters on the first and 35th day of storage. In this purpose we used Advia 2102 hematology analyzer, biochemistry analyzer Beckman Coulter AU680 and RapidLab 1265 system for measuring pH. Statistical analysis was performed by Microsoft Excel 2007 program support. Results: During the storage of packed red blood cells number of erythrocytes, hemoglobin concentration and MCH were not significantly changed. Hct and MCV were slightly increased while the MCHC on the 35th day of storage was lowered. On the last day of concentrate storage AST, LDH and K were significantly increased. It was also observed a reduction of the pH value, Na and Cl. Conclusion: The most significant changes in the storage period erythrocyte blood products show in increasing concentrations of K, LDH and decreasing of pH. This is an important finding that must be considered in the treatment of patients with erythrocyte concentrates at the end of the storage period.