Laboratory of Clinical Biochemistry

Città della Pieve, Italy

Laboratory of Clinical Biochemistry

Città della Pieve, Italy
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Zalesak M.,Slovak Academy of Sciences | Blazicek P.,Laboratory of Clinical Biochemistry | Pancza D.,Slovak Academy of Sciences | Gablovsky I.,Slovak Academy of Sciences | And 2 more authors.
Physiological Research | Year: 2016

Several studies have shown that diabetes mellitus modulates heart resistance to ischemia and abrogates effectivity of cardioprotective interventions, such as ischemic preconditioning (IP). The aim of this study was to evaluate whether the effect of hyperglycemic conditions on the severity of ischemia-reperfusion (I/R) injury in preconditioned and non-preconditioned hearts (controls, C) is related to changes in osmotic activity of glucose. Experiments were performed in isolated rat hearts perfused according to Langendorff exposed to 30-min coronary occlusion/ 120-min reperfusion. IP was induced by two cycles of 5-min coronary occlusion/5-min reperfusion, prior to the long-term I/R. Hyperosmotic (HO) state induced by an addition of mannitol (11 mmol/l) to a standard Krebs-Henseleit perfusion medium significantly decreased the size of infarction and also suppressed a release of heart fatty acid binding protein (h-FABP - biomarker of cell injury) from the non-IP hearts nearly to 50 %, in comparison with normoosmotic (NO) mannitol-free perfusion. However, IP in HO conditions significantly increased the size of infarction and tended to elevate the release of h-FABP to the effluent from the heart. The results indicate that HO environment plays a cardioprotective role in the ischemic myocardium. On the other hand, increased osmolarity, similar to that in the hyperglycemic conditions, may play a pivotal role in a failure of IP to induce cardioprotection in the diabetic myocardium. © 2016 Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic.


Rom O.,Technion - Israel Institute of Technology | Grajeda-Iglesias C.,Technion - Israel Institute of Technology | Najjar M.,Technion - Israel Institute of Technology | Najjar M.,Rambam Health Care Campus | And 6 more authors.
Journal of Nutritional Biochemistry | Year: 2017

Atherosclerosis-related research has focused mainly on the effects of lipids on macrophage foam cell formation and atherogenesis, whereas the role of amino acids (AAs) was understudied. The current study aimed to identify anti- or pro-atherogenic AA in the macrophage model system and to elucidate the underlying metabolic and molecular mechanisms. J774A.1 cultured macrophages were treated with increasing concentrations of each 1 of the 20 AAs. Macrophage atherogenicity was assessed in terms of cellular toxicity, generation of reactive oxygen species (ROS) and cellular cholesterol or triglyceride content. At nontoxic concentrations (up to 1 mM), modest effects on ROS generation or cholesterol content were noted, but six specific AAs significantly affected macrophage triglyceride content. Glycine, cysteine, alanine and leucine significantly decreased macrophage triglyceride content (by 24%–38%), through attenuated uptake of triglyceride-rich very low-density lipoprotein (VLDL) by macrophages. In contrast, glutamate and glutamine caused a marked triglyceride accumulation in macrophages (by 107% and 129%, respectively), via a diacylglycerol acyltransferase-1 (DGAT1)-dependent increase in triglyceride biosynthesis rate with a concurrent maturation of the sterol regulatory element-binding protein-1 (SREBP1). Supplementation of apolipoprotein E-deficient (apoE−/−) mice with glycine for 40 days significantly decreased the triglyceride levels in serum and in peritoneal macrophages (MPMs) isolated from the mice (by 19%). In contrast, glutamine supplementation significantly increased MPM ROS generation and the accumulation of cholesterol and that of triglycerides (by 48%), via enhanced uptake of LDL and VLDL. Altogether, the present findings reveal some novel roles for specific AA in macrophage atherogenicity, mainly through modulation of cellular triglyceride metabolism. © 2017 Elsevier Inc.


Prometti P.,Salvatore Maugeri Foundation | Olivares A.,Laboratory of Cardiovascular Pathophysiology | Gaia G.,Laboratory of Clinical Biochemistry | Bonometti G.,Salvatore Maugeri Foundation | And 2 more authors.
Medicine (United States) | Year: 2016

The aim of this study was to evaluate if the Biodex Fall Risk Assessment could provide an age-adjusted index useful for classifying patients at "risk of fall." This was a cohort study conducted on 61 chronic patients, in stable conditions, having a history of ataxia, difficulty in walking or loss of balance, and aged >64 years. These patients were coming from home to our Institute undergoing a period of in-hospital standard rehabilitation. Assessment of clinical parameters was performed at entry. Functional scales (Functional Independence Measure [FIM] for motor and cognitive function, Barthel G, Tinetti POMA), and the Biodex Fall Risk Index (FRI) were performed at entry and discharge. The Normalized FRI, obtained adjusting FRI to the reported maximum predictive FRI for the relevant age, identified 2 types of patients: those with a greater risk of fall than expected for that age, labeled Case 1 (Normalized FRI>1); and those with an equal or even lesser risk of fall than expected for that age, labeled Case 0 (Normalized FRI-1). FRI, Normalized FRI as well as independent variables as age, sex, pathology group, FIM, BarthelG, were considered in a multiple regression analysis to predict the functional improvement (i.e., delta Tinetti Total score) after rehabilitation. Normalized FRI is useful in assessing patients at risk of falls both before and after rehabilitation. At admission, the Normalized FRI evidenced high fall risk in 46% of patients (Case 1) which decreased to 12% after rehabilitation, being greater than age-predicted in 7 patients (Case 1-1) despite the functional improvement observed after the rehabilitation treatment. Normalized FRI evidenced Case 1-1 patients as neurological, "very old" (86% in age-group 75-84 years), and with serious events at 18 to 24 months' follow-up. Normalized FRI, but not FRI, at admission was a predictor of improvement in Tinetti Total scores. The normalized FRI effectively indicated patients at higher risk of fall, in whom health deterioration, falls, or cognitive decline was later documented at follow-up. The normalized FRI could be a standardized measure for identifying frailer patients becoming a further criterium of discharge home and marker of fall risk at home. Copyright © 2016 Wolters Kluwer Health, Inc. All rights reserved.


Carobbio S.,University of Cambridge | Hagen R.M.,University of Cambridge | Lelliott C.J.,Astrazeneca | Slawik M.,University of Cambridge | And 28 more authors.
Diabetes | Year: 2013

The epidemic of obesity imposes unprecedented challenges on human adipose tissue (WAT) storage capacity that may benefit from adaptive mechanisms to maintain adipocyte functionality. Here, we demonstrate that changes in the regulatory feedback set point control of Insig1/SREBP1 represent an adaptive response that preserves WAT lipid homeostasis in obese and insulinresistant states. In our experiments, we show that Insig1 mRNA expression decreases in WAT from mice with obesity-associated insulin resistance and from morbidly obese humans and in in vitro models of adipocyte insulin resistance. Insig1 downregulation is part of an adaptive response that promotes the maintenance of SREBP1 maturation and facilitates lipogenesis and availability of appropriate levels of fatty acid unsaturation, partially compensating the antilipogenic effect associated with insulin resistance. We describe for the first time the existence of this adaptive mechanism in WAT, which involves Insig1/SREBP1 and preserves the degree of lipid unsaturation under conditions of obesity-induced insulin resistance. These adaptive mechanisms contribute to maintain lipid desaturation through preferential SCD1 regulation and facilitate fat storage in WAT, despite ongoing metabolic stress. © 2013 by the American Diabetes Association.


Calcagno A.,University of Turin | Cusato J.,University of Turin | Marinaro L.,University of Turin | Simiele M.,University of Turin | And 8 more authors.
AIDS | Year: 2016

Objective: To assess if tenofovir (TFV) clearance is associated with urinary retinal-binding protein (RBP) in HIV-positive patients with normal estimated filtration rate. Design: A cross-sectional diagnostic study. Methods: HIV-positive patients with estimated creatinine clearance above 60ml/min, on tenofovir disoproxil fumarate (TDF)-containing combination since at least 6 months, taking TDF at night, and without significant comorbidities (diabetes, untreated hypertension, known renal malformations, recurrent nephrolithiasis) and nephrotoxic drugs were included. TFV plasma and urinary concentrations were measured 12h after drug intake (C 12). RBP was measured through enzyme immunoassay kit on spot urines and corrected per urinary creatinine (uRBP/uCr); normality ranges were below 130μg/g (in patients aged <50 years) and below 172μg/g (in patients aged ≥50 years). Results: Two hundred and eighty-nine patients were included (median age of 45.8 years, 71.6% male and 85.4% whites); patients were concomitantly treated with nonnucleoside reverse transcriptase inhibitors (155, 53.6%), protease inhibitors (118, 40.8%), or integrase inhibitors (16, 5.5%)-containing regimens. Estimated creatinine clearance was 89.4ml/min (78.6-105.9). Urinary RBP (uRBP) and uRBP/uCr were 204.6ng/ml (92-380) and 169.7μg/g (85.8-318.3), respectively; abnormally high uRBP/uCr was observed in 157 patients (54.3%). A multivariate binary logistic regression confirmed that both ethnicity (P=0.004, β 2.93, 95% confidence interval 1.41-6.10) and TFV urinary C 12 less than 21mg/ml (P=0.006, β 2.04, 95% confidence interval 1.12-3.41) were significantly associated with abnormal uRBP/uCr. Conclusion: HIV-positive TDF-treated patients showed a high prevalence of proximal tubular impairment: ethnicity (whites) and low urinary TFV concentrations were significantly associated with elevated uRBP. © 2016 Wolters Kluwer Health, Inc.


Kaplan M.,Laboratory of Clinical Biochemistry | Hamoud S.,Rambam Health Care Campus | Tendler Y.,Laboratory of Clinical Biochemistry | Meilin E.,Lipid Research Laboratory | And 4 more authors.
Journal of Inflammation (United Kingdom) | Year: 2014

Background: Atherosclerosis is a complex disease involving different cell types, including macrophages that play a major role in the inflammatory events occurring in atherogenesis. C-Reactive Protein (CRP) is a sensitive systemic marker of inflammation and was identified as a biomarker of cardiovascular diseases. Histological studies demonstrate CRP presence in human atherosclerotic lesions, and we have previously shown that macrophages express CRP mRNA. CRP could be locally secreted in the atherosclerotic lesion by arterial macrophages and local regulation of CRP could affect its pro-atherogenic effects. Moreover, human blood derived macrophages (HMDM) expression of CRP could reflect atherosclerotic lesion secretion of CRP. Methods. Ten type 2 diabetic patients and ten non-diabetic patients scheduled to undergo carotid endarterectomy were enrolled in this study, and their blood samples were used for serum CRP, lipid determination, and for preparation of HMDM further analyzed for their CRP mRNA expression and CRP content. Carotid lesions obtained from the patients were analyzed for their CRP and interleukin 6 (IL-6) content by immunohistochemistry. Results: Lesions from diabetic patients showed substantially higher CRP levels by 62% (p = 0.05) than lesions from non diabetic patients, and CRP staining that co-localized with arterial macrophages. CRP carotid lesion levels positively correlated with CRP mRNA expression (r§ssup§2§esup§ = 0.661) and with CRP content (r§ssup§2§esup§ = 0.611) in the patient's HMDM. Conclusions: Diabetes up-regulated carotid plaques CRP levels and CRP measurements in HMDM could reflect atherosclerotic lesion macrophages secretion of CRP. Understanding the regulation of locally produced macrophage CRP in the arterial wall during atherogenesis could be of major importance in identifying the underlying mechanisms of inflammatory response pathways during atherogenesis. © 2014 Kaplan et al.; licensee BioMed Central Ltd.


Ulvik A.,Laboratoriebygget | Midttun O.,Laboratoriebygget | Pedersen E.R.,University of Bergen | Eussen S.J.P.M.,University of Bergen | And 4 more authors.
American Journal of Clinical Nutrition | Year: 2014

Background: Plasma concentrations of PL 5'-phosphate (PLP), which is the active coenzyme form of vitamin B-6, are reduced during inflammation. The underlying mechanisms may include altered tissue distribution or increased catabolism via pyridoxal (PL) to pyridoxic acid (PA). Recently, we showed that catabolic enzyme activity could be assessed by substrate product ratios measured in plasma. Objective: We evaluated the ratios PA:PL, PA:PLP, and PA:(PL + PLP) as possible markers of vitamin B-6 catabolism. Design: Cross-sectional and longitudinal data were derived from the Western Norway B-Vitamin Intervention Trial. We analyzed associations of ratios with inflammatory markers and other clinical variables by using multiple linear regression and partial correlation. In addition, intraclass correlation coefficients (ICCs) were used to assess the ability of plasma indexes to differentiate between subjects. Results: PA:(PL + PLP) had the highest ICC of all vitamin B-6 metabolites and ratios tested. In regression models, the inflammatory markers C-reactive protein, white blood cell count, neopterin, and kynurenine:tryptophan collectively accounted for 28% of the total and > 90% of the explained variation in PA:(PL + PLP). For individual B-6 metabolites, corresponding numbers were 19-25% and 20-44%, respectively, with vitamin supplement intake, smoking, and kidney function (estimated glomerular filtration rate) as additional predictors. In an analysis of receiver operating characteristics, PA:(PL + PLP) discriminated high inflammatory concentrations with an area under the curve (95% CI) of 0.85 (0.81, 0.89). Conclusions: Broad-specificity enzymes upregulated to reduce oxidative and aldehyde stress could explain increased catabolism of vitamin B-6 during inflammation. The ratio PA:(PL + PLP) may provide novel insights into pathologic processes and potentially predict risk of future disease. © 2014 American Society for Nutrition.


Naffaa M.,Rambam Health Care Campus | Naffaa M.,Technion - Israel Institute of Technology | Makhoul B.F.,Rambam Health Care Campus | Makhoul B.F.,Technion - Israel Institute of Technology | And 9 more authors.
American Journal of Emergency Medicine | Year: 2014

Background: Procalcitonin and interleukin 6 (IL-6) are well-known predictors of blood culture positivity in patients with sepsis. However, the association of procalcitonin and IL-6 with blood culture positivity was assessed separately in previous studies. This study aims to examine and compare the performance of procalcitonin and IL-6, measured concomitantly, in predicting blood culture positivity in patients with sepsis. Methods: Forty adult patients with sepsis were enrolled in the study. Blood cultures were drawn before the institution of antibiotic therapy. The area under the curve (AUC) of the receiver operating characteristic curve was estimated to assess the performance of procalcitonin and IL-6 in predicting blood culture positivity. Results: Positive blood cultures were detected in 10 patients (25%). The AUC of procalcitonin and IL-6 was 0.85 and 0.61, respectively. The combined performance of procalcitonin and IL-6 was similar to that of procalcitonin alone, AUC of 0.85. On univariate analysis, only procalcitonin and IL-6 were associated with blood culture positivity. Multivariate logistic regression analysis showed that only procalcitonin was associated with blood culture positivity (odds ratio, 12.15 [1.29-114.0] for levels above the median compared with levels below the median). Using procalcitonin cut points of 1.35 and 2.14 (nanogram per milliliter) enabled 100% and 90% identification of positive blood cultures and reduced the need of blood cultures by 47.5% and 57.5%, respectively. Conclusions: Compared with IL-6, procalcitonin better predicts blood culture positivity in patients with sepsis. Using a predefined procalcitonin cut points will predict most positive blood cultures and reduce the need of blood cultures in almost half of patients with sepsis. © 2014 Elsevier Inc. All rights reserved.


Makhoul B.F.,Rambam Health Care Campus | Makhoul B.F.,Technion - Israel Institute of Technology | Khourieh A.,Technion - Israel Institute of Technology | Kaplan M.,Laboratory of Clinical Biochemistry | And 4 more authors.
International Journal of Cardiology | Year: 2013

Background: Increased red blood cell distribution (RDW) has been associated with adverse outcomes in patients with heart failure. We studied the association between baseline RDW and changes in RDW during hospital course with clinical outcomes in acute decompensated heart failure (ADHF) patients. Methods and results: We prospectively studied 614 patients with ADHF. Baseline RDW and RDW change during hospital course were determined. The relationship between RDW and clinical outcomes after hospital discharge was tested using Cox regression models, adjusting for clinical characteristics, echocardiographic findings and brain natriuretic peptide levels. During follow up (1 year), 286 patients (46.6%) died and 84 were readmitted for ADHF (13.7%). Median RDW was significantly higher among patients who died compared to patients who survived (15.6% interquartile range [14.5 to 17.1] vs 14.9% mg/L interquartile range [14.1 to 16.1], P < 0.0001). Compared with patients in the 1st RDW quartile, the adjusted hazard ratio [HR] for death or rehospitalization was 1.9 [95% CI 1.3-2.6] in patients in the 4th quartile. Changes in RDW during hospitalization were strongly associated with changes in mortality risk. Compared with patients with persistent normal RDW (< 14.5%), the adjusted HR for mortality was 1.9 [95% CI 1.1-3.1] for patients in whom RDW increased above 14.5% during hospital course, similar to patients with persistent elevation of RDW (HR was 1.7, 95% CI 1.2-2.3). Conclusion: In patients hospitalized with ADHF, RDW is a strong independent predictor of greater morbidity and mortality. An increase in RDW during hospitalization also portends adverse clinical outcome. © 2012 Elsevier Ireland Ltd.


Kushlinskii N.E.,Laboratory of Clinical Biochemistry | Nemtsova M.V.,Barrikadnaya Street
Vestnik Rossiiskoi Akademii Meditsinskikh Nauk | Year: 2014

The review presents the main and additional features that distinguish tumor cells from normal tissue cells. They include sustained proliferative signaling, evasion from growth suppressors, resisting cell death, enabling replicalive immortality, angiogenesis induction, and invasion and metastasis activation. Basis for the formation of these features is provided by tumor genome instability. Tumors are complex tissues that consist of different cell types interacting with each other as well as with normal cells. An important characteristic of tumor cells is the ability to interact with the tumor microenvironment and the formation of tumor stroma.

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