The antioxidative protecting role of the Mediterranean diet [Antioksidativno protektivno djelovanje mediteranske dijete]

Recent meta-analysis shows that adherence to a Mediterranean diet (MD) can significantly decrease the risk of overall mortality, mortality from cardiovascular diseases, as well as incidence of mortality from cancer, and incidence of Parkinson's and Alzheimer's disease. All of these diseases could be linked to oxidative stress (OS) as antioxidative effect of MD is getting more attention nowadays. Although a lot of research has been done in this area and it suggests antioxidative protective role of MD, the presented evidence is still inconclusive. The aim of this paper is to review studies investigating the effect of MD on OS, as well as to identify the areas for further research

The Role of Endothelin-1 and Nitric Oxide in the Pathogenesis of Hypertension in Diabetic Patients

The pathogenesis of renal hypertension has not yet been fully clarified. As the potential role of endothelin-1 (ET-1) and nitric oxide (NO) has been postulated, their concentrations were determined in plasma and urine of diabetic patients. The study included 30 diabetic patients (both IDDM and NIDDM) with initial or advanced diabetic nephropathy (decreased endogenous creatinine clearance, proteinuria) and 20 healthy control subjects. The correlation with blood pressure and other renal function parameters was monitored and compared with the control group. Also, the effect of ACE inhibitors (ACEI) on ET-1 and NO patterns was monitored in correlation with arterial hypertension. In diabetic patients that did not receive ACEI therapy, the increase in plasma ET-1 was associated with both systolic and diastolic blood pressure elevation, whereas in those administered ACEI the increase in plasma ET-1 was associated with a systolic blood pressure decline. In addition, the increase in plasma NO was accompanied by a statistically significant decline of both systolic and diastolic blood pressure in diabetic patients receiving ACEI

Red blood cell distribution width as a prognostic marker of mortality in patients on chronic dialysis: a single center, prospective longitudinal study

Aim To determine if red cell distribution width (RDW) is associated with all-cause mortality in patients on chronic dialysis and to evaluate its prognostic value among validated prognostic biomarkers. Methods This is a single center, prospective longitudinal study. At the time of inclusion in January 2011, all patients were physically examined and a routine blood analysis was performed. A sera sample was preserved for determination of NT-pro-brain natriuretic peptide (NT-pro-BNP) and eosinophil cationic protein. Carotid intima media thickness (IMT) was also measured. Following one year, all-cause mortality was evaluated. Results Of 100 patients, 25 patients died during the follow- up period of one-year. Patients who died had significantly higher median [range] RDW levels (16.7% [14.3-19.5] vs 15.5% [13.2-19.7], P < 0.001. They had significantly higher Eastern Cooperative Oncology Group (ECOG) performance status (4 [2-4] vs 2 [1-4], P < 0.001), increased intima-media thickness (IMT) (0.71 [0.47-1.25] vs 0.63 [0.31-1.55], P = 0.011), increased NT-pro-BNP levels (8300 [1108-35000] vs 4837 [413-35000], P = 0.043), and increased C-reactive protein (CRP) levels (11.6 [1.3-154.2] vs 4.9 [0.4-92.9], P < 0.001). For each 1% point increase in RDW level as a continuous variable, one-year all cause mortality risk was increased by 54% in univariate Cox proportional hazard analysis. In the final model, when RDW was entered as a categorical variable, mortality risk was significantly increased (hazard ratio, 5.15, 95% confidence interval, 2.33 to 11.36) and patients with RDW levels above 15.75% had significantly shorter survival time (Log rank P < 0.001) than others. Conclusions RDW could be an additive predictor for allcause mortality in patients on chronic dialysis. Furthermore, RDW combined with sound clinical judgment improves identification of patients who are at increased risk compared to RDW alone

Comparison of sensitivity of nested PCR and quantitative PCR in Bcr-Abl p210 transcript detection in chronic myelogenous leukemia

Uvod: Za otkrivanje minimalne ostatne bolesti u bolesnika s kroničnom mijeloičnom leukemijom (KML) koji su postigli potpunu kliničku remisiju i potpun citogenetski odgovor može se primijeniti ugnježđena PCR (engl. nestedPCR, nPCR) i kvantitativna PCR u stvarnom vremenu (engl. quantitative real-time PCR, qPCR). Cilj liječenja je postizanje molekularne remisije, pa postizanje visoke osjetljivosti molekularne pretrage ima presudnu ulogu i kliničku primjenu. Usporedili smo razinu osjetljivosti nPCR i qPCR u otkrivanju BCR-ABL p210 prijepisa u modelu razrjeđenja Bcr/Abl-pozitivnih stanica. Materijal i metode: Za određivanje razine osjetljivosti načinjena su serijska razrjeđenja stanične linije K562 (Bcr-Abl pozitivne) sa staničnom linijom NB4 (Bcr-Abl negativnom) (raspon razrjeđenja pozitivnih stanica: 10-3-10-7). Izolirani uzorci RNA prepisani su u cDNA i testirani na p210 prijepis pomoću nPCR i qPCR. Objema metodama također su ispitani uzorci koštane srži i periferne krvi dvoje bolesnika s KML na terapiji imatinib-mesilatom. Rezultati: U testu staničnog razrjeđenja nPCR je pokazala osjetljivost za otkrivanje Bcr-Abl pozitivnih stanica od 10-5, dok je qPCR pokazala osjetljivost od 10-6. Obje su metode otkrile p210 prijepise u uzorcima koštane srži bolesnika s KML. Međutim, qPCR je uz to otkrila prijepise i u uzorcima periferne krvi, no uz nižu razinu prijepisa u usporedbi s uzorcima koštane srži. Zaključak: Iako se često navodi kako je nPCR za otprilike 1 log osjetljivija od qPCR, u našem pokusu s razrjeđenjem na biljeg pozitivne stanične linije K562 dokumentirali smo višu osjetljivost za standardiziranu metodu qPCR, koja je razvijena za potrebe Europskog programa za borbu protiv karcinoma.Background: For minimal residual disease detection in chronic myelogenous leukemia (CML) patients who have achieved complete clinical remission and complete cytogenetic response, nested PCR (nPCR) and quantitative realtime PCR (qPCR) can be used. Achieving of molecular remission is the goal of therapy, so it is of critical importance and clinical utility to obtain high sensitivity of molecular testing. We compared the level of sensitivity of nPCR and qPCR in the detection of BCR-ABL p210 transcripts in a Bcr/Abl-positive cell dilution model. Materials and Methods: For determination of sensitivity level, serial dilutions of K562 cell line (Bcr-Abl-positive) in NB4cell line (Bcr-Abl-negative) were made (range of dilution of positive cells: 10-3-10-7). Isolated RNA samples were transcribed into cDNA and tested for p210 transcript by nPCR and qPCR. Bone marrow and peripheral blood samples of two CML patients on imatinib mesylate therapy were also tested by both methods. Results: In the cell dilution test, nPCR showed sensitivity for detecting Bcr-Abl positive cell of 10-5, and qPCR showed a sensitivity of 10-6. Both methods detected p210 transcripts in bone marrow samples of CML patients. However, qPCR also detected transcripts in peripheral blood samples, with a lower transcript level in comparison to bone marrow samples. Conclusion: Although frequently quoted as nPCR being by approximately 1 log more sensitive than qPCR, in our marker-positive cell line K562 dilution experiment we documented higher sensitivity of the standardized Europe Against Cancer Program developed qPCR method

Comparability of Pathromtin SL, Dade Actin FS i STA Cephascreen reagens for activated partial thromboplastin time measurement

Cilj: Ispitati korelaciju vrijednosti APTV dobivenih korištenjem triju reagensa: Pathromtin SL i Dade Actin FS na analizatoru Berichrom Coagulation System (BCS), odnosno STA Cephascreen na analizatoru STA Compact. Materijali i metode: APTV je određen u 114 uzoraka svježe plazme ispitanika, od kojih je 46 liječeno niskomolekularnim heparinom, a ostalih 68 nije bilo na antikoagulantnoj terapiji. Vrijednosti APTV određene su koagu-lometrijskom metodom usporedno pomoću sva tri reagensa neposredno nakon donošenja uzoraka u koagulacijski laboratorij. Vrijednosti APTV određene su i u komercijalnim kontrolnim pripravcima Lypocheck Coagulation Control proizvođača Bio-Rad (Bio-Rad Laboratories, SAD), koji obuhvaćaju tri različita raspona vrijednosti. Rezultati: Vrijednosti komercijalnih kontrolnih uzoraka Bio-Rad kretale su se unutar raspona što ga navodi proizvođač za sva tri ispitivana reagensa. Korelacijom reagensa Pathromtin SL i Dade Actin FS, Pathromtin SL i STA Cephascreen, te Dade Actin FS i STA Cephascreen dobivene su izvrsne povezanosti (R = 0,9485,0,9353 i 0,9072). Ispitana je razlika između dobivenih koeficijenata korelacije i nađena je statistički značajna razlika između koeficijenata korelacije rezultata dobivenih reagensima Pathromtin SL i Dade Actin FS. Passing-Bab-lok regresijom vrijednosti nagiba i odsječka na osi y obuhvaćale su 1 odnosno 0 samo u kombinaciji reagensa Dade Actin FS i STA Cephascreen. Zaključak: Ispitivanje je pokazalo da su najbolje usporedive vrijednosti dobivene reagensima Dade Actin FS i STA Cephascreen. Unatoč izvrsnoj povezanosti Passing-Bablok regresija pokazala je da su reagensi Pathromtin SL i Dade Actin FS te Pathromtin SL i STA Cephascreen slabije usporedivi. Kako dobiveni rezultati potvrđuju opažanja iz svakodnevnog rada, bitno je da se bolesnici izloženi heparinskom liječenju ne prate istodobno različitim reagensima zbog različite osjetljivosti reagensa prema terapiji niskomolekularnim heparinom.Aim: Aim of the study was to investigate correlation of activated partial thromboplastin time (APTT) measured with three different reagents: Pathromtin SLand Dade Actin FS reagents on Berichrom Coagulation System analyzer, and by STA Cephascreen reagent on STA Compact analyzer. Material and methods: APTT was determined in parallel by Pathromtin SL and Dade Actin FS reagents, and by STA Cephascreen reagent in 114 fresh plasma samples from subjects administered low-molecular-weight heparin and in subjects known to receive no anticoagulant therapy. APTT values were determined by coagulometry method immediately upon the material receipt at coagulation laboratory. APTT values were also determined in Bio-Rad Lypocheck Coagulation Control samples (Bio-Rad Laboratories, USA) covering three different value ranges. Results: The values recorded in Bio-Rad controls were within the range declared by the manufacturer for all three reagents used. Correlation of APTT values obtained by use of Pathromtin SL vs. Dade Actin FS, Pathromtin SL vs. Cephascreen and Dade Actin FS vs. STA Cephascreen yielded strong correlations (R = 0.9485,0.9353 and 0.9072, respectively). A statistically significant difference was recorded between the results obtained by Pathromtin SL and Dade Actin FS reagents. Passing-Bablok regression slope and y-axis intercept included 1 and 0 only for Dade Actin FS vs. STA Cephascreen. Conclusion: Study results showed the best compliance of values obtained by Dade Actin FS and STA Cephascreen reagents. In spite of strong correlations, Passing-Bablok regression indicated lower comparability between Pathromtin SL and Dade Actin as well as between Pathromtin SL and STA Cephascreen reagents. As the study results confirmed the observations from daily routine, it is of utmost importance that individual patients receiving heparin therapy be not monitored by use of different reagents due to the variable reagent sensitivity to low molecular weight heparin

Oxidative Stress Markers in Patients with Post-Traumatic Stress Disorder

Recent study data support the role of oxidative stress in diverse psychiatric disorders. Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants and/or a depletion of antioxidants. There are numerous studies that indicate that free radicals (FRs) damage neurons, and then play an important role in the pathophysiology of schizophrenia and depression. Active oxygen can cause considerable damage and disrupt the important physiological functions of proteins, lipids, enzymes and DNA. The aim of our study was to investigate the possible differences in the concentration of tromboxane B2, 8-OHdG and protein carbonyls, as significant markers of oxidative damage, and urate, albumin and total protein concentrations as antioxidative molecules in PTSD patients in comparison to the healthy control group. The study included 74 male participants who were active soldiers in the Croatian armed forces from 1991 to 1995. 46 subjects with chronic and current PTSD were recruited from the Department of Psychiatry of Dubrava University Hospital during 2010, 28 healthy subjects were recruited in the same period during the regular medical examination at the Dubrava University Hospital. Study results have shown that there is no statistically significant difference in urinary concentrations of 8-OHdG, serum thromboxane B2, and serum urates between two studied groups. Statistically significant difference of the protein carbonyl concentrations was examined. Concentrations were significantly lower in the PTSD group than in the control group. The clinical significance of these results was examined using ROC analysis. The obtained ROC curves did not separate the groups in a satisfactory manner

Correlation of Endothelin-1 Concentration and Angiotensin-Converting Enzyme Activity with the Staging of Liver Fibrosis

Increased serum angiotensin-converting enzyme (SACE) activity and serum concentration of endothelin-1 (ET-1) were found in liver cirrhosis. We investigated a correlation between the different stages of liver fibrosis and SACE activity and serum ET-1 concentration. Seventy patients with pathohistologically established chronic liver disease were divided in three groups according to Ishak criteria for liver fibrosis: minimal fibrosis (Ishak score 0–1, n=20), medium fibrosis (Ishak score 2–5, n=20) and cirrhosis (Ishak score 6, n=30). SACE activity and ET-1 concentration were determined using commercial ELISA kits. SACE activity and ET-1 concentrations were proportional to the severity of disease, the highest being in patients with liver cirrhosis. Maximal increase in SACE activity was found between minimal and medium fibrosis while maximal increase in ET-1 concentration was revealed between medium fibrosis and cirrhosis. The analysis of the Receiver Operating Characteristic (ROC) curve for SACE activity suggested a cut-off value to separate minimal from medium fibrosis at 59.00 U/L (sensitivity 100%, specificity 64.7%). The cut-off value for serum ET-1 concentration to separate medium fibrosis from cirrhosis was 12.4 pg/mL (sensitivity 96.8%, specificity 94.4%). A positive correlation between SACE activity and ET-1 concentration was registered (Spearman’s ñ=0.438, p=0.004). Both SACE activity and ET-1 concentration were increased in all stages of liver fibrosis. Cut-off points for SACE activity and ET-1 concentration could be a biochemical marker for the progression of fibrosis. Positive correlation between SACE activity and ET-1 concentration might indicate their interaction in the development of liver cirrhosis

Comparison of two immunoassays for CA19-9, CEA and AFP tumor markers

Uvod: Monoklonska antitijela koriste se za otkrivanje antigena u serumu koji su povezani sa specifičnim zloćudnim oboljenjima. Ti su tumorski biljezi najkorisniji za praćenje odgovora na terapiju i otkrivanje ranog recidiva, međutim, rezultati dobiveni različitima analizama razlikuju se te promjena metode tijekom praćenja može biti uzrokom problema. Cilj ove studije bio je provesti analitičku evaluaciju usporedivosti analiza za tumorske biljega CA19-9, CEA i AFP na dva različita automatizirana kemijska analizatora. Materijali i metode: Koncentracije CA19-9, CEA i AFP su određene na ana-lizatorima Vitros ECi (Ortho Clinical Diagnostics, Johnson & Johnson, Buckinghamshire, V. Britanija) i Cobase 411 (Hitachi High Technologies Corporation, Tokyo, Japan). Korelacija među metodama ispitana je na 38 uzoraka seruma za CA19-9 i AFP te 39 uzoraka seruma za CEA. Rezultati: Vrijednosti komercijalnih kontrolnih uzoraka bile su unutar raspo-na navedenih od proizvođača za sve tumorske biljege uključene u istraživanje na oba analizatora. Najveće odstupanje od deklariranih kontrolnih vrijednosti nađene su za CA19-9 na analizatoru Vitros ECi te za AFP na analizatoru Cobas e411. Visoka je korelacija utvrđena među metodama za sva tri ispitana tumorska biljega (r = 0,978 za CA19-9; r = 0,995 za CEA, te r = 0,999 za AFP). Nagib i odsječ ak na osi Y iznosili su 1, odnosno 0, samo za usporedbenu analizu AFP. Zaključak: Rezultati istraživanja pokazali su najbolje podudaranje vrijednosti za AFP dobivene na dva ispitana analizatora za imunoanalize. Bez obzira na snažne korelacije, regresija po Passing-Babloku ukazala je na nižu usporedivost dviju imunoanaliza za CEA i CA19-9. S obzirom da su rezultati studije potvrdili zapažanja iz svakodnevnog rutinskog rada, za svakoga je pacijenta od najveće važnosti da ga se prati primjenom iste imunoanalize i reagensa na istom analizatoru.Introduction: Monoclonal antibodies are used to detect serum antigens associated with specific malignancies. These tumor markers are most useful for monitoring response to therapy and detecting early relapse, but results obtained by different assays vary, and a change of method during follow-up may cause problems. The aim of our study was to perform the analytical evaluation of the inter-assay comparability for tumor markers CA19-9, CEA and AFP on two different automated chemistry analyzers. Materials and methods: CA19-9, CEA and AFP concentrations, using Vitros ECi (Ortho Clinical Diagnostics, Johnson and Johnson, Buckinghamshire, UK), and Cobas e 411 (Hitachi High Technologies Corporation, Tokyo, Japan) immu-noassay analyzers, were determined. Between-method correlation was studied in 38 serum samples for CA19-9 and AFP and 39 serum samples for CEA. Results: The values of commercial controls were within the range declared by the manufacturer for all tumor markers included in the study on both analyzers. The highest deviation from the declared control values was found for CA19-9 on Vitros ECi and for AFP on Cobas e411 analyzer. High correlation was found between methods for all of three tumor markers studied (R = 0.978 for CA19-9; R = 0.995 for CEA and R = 0.999 for AFP). Passing-Bablok regression slope and y-axis intercept included 1 and 0 only for AFP comparative assays. Conclusion: Study results showed the best compliance of values for AFP obtained on two studied immunoassay analyzers. Regardless of high correlations, Passing-Bablok regression indicated lower comparability between two immunoassays for CEA and CA19-9. As the study results confirmed the observations from daily routine, it is of utmost importance for individual patients to be monitored using the same immunoassay and reagents on the same analyzer