Luigi Colacicco

Determination of copper, zinc, and selenium in human plasma and urine samples by potentiometric stripping analysis and constant current stripping analysis

Potentiometric stripping analysis and constant current stripping analysis are proposed as routine methods for analysis of copper, zinc and selenium in plasma and urine samples. The analytical performance of these methods is comparable with that reported for atomic absorption spectrometry. However the low cost, greater simplicity of the apparatus, and the facility of execution make this methodology a valid candidate for routine application in Clinical Chemistry laboratories.

Coenzyme Q10 levels, plasma lipids and peroxidation extent in renal failure and in hemodialytic patients.

Coenzyme Q10 (CoQ10), vitamin E, triglycerides and conjugated dienes were measured in a group of 48 patients on chronic hemodialysis, in 15 uremic patients and in a control group of 10 normal subjects. CoQ10 levels were significantly lower (P < 0.001) in both hemodialytic and uremic patients compared with the normal group whereas triglycerides were significantly higher (P < 0.001) with respect to both normal subjects and uremic patients. Conjugated dienes were significantly higher (P < 0.001) in both hemodialytic and uremic patients with respect to normal subjects. The predialytic values of vitamin E were higher in hemodialytic patients with respect to both normal subjects and uremic patients whereas the postdialytic values were in the normal range. A restoration mechanism of vitamin E after hemodialytic treatment was hypothesized.

Plasma levels of coenzyme Q(10), vitamin E and lipids in uremic patients on conservative therapy and hemodialysis treatment: some possible biochemical and clinical implications.

Coenzyme Q(10) (CoQ(10)), vitamin E, total cholesterol, HDL-cholesterol (HDLC) and triglycerides were measured in the plasma of 62 patients with kidney failure, 46 under hemodialysis treatment and 16 under conservative therapy, and 95 controls. The sum of LDL-cholesterol (LDL-C) and VLDL-cholesterol (VLDL-C) was also calculated for each patient. The ratio CoQ(10)/LDL-C+VLDL-C in both conservative therapy and hemodialysis populations was significantly lower (P<0.001) compared with normal controls and remained unchanged after the dialysis treatment. On the contrary the ratio vitamin E/LDL-C+VLDL-C was normal but decreased significantly (P<0.02) after each dialysis. Since coenzyme Q is the main inhibitor of the prooxidant action of vitamin E, it was hypothesized that its decrease in both the populations examined could make the lipoproteins of these patients more vulnerable to a peroxidative attack.

The Oxidative Metabolism of Antarctic Fish: Some Peculiar Aspects of Cold Adaptation

Environmental factors are key determinants in the process of adaptation and evolution of living organisms. In this respect, analysis of biological systems from Antarctic fish may provide valuable insights on the molecular and cellular strategies which have been adopted in response to the very low environmental temperature which characterizes Antarctica. Hence, the temperature of the Antarctic Ocean varies from −1.86 °C at high latitudes to +5 °C towards the Antarctic Convergence. Moreover, seasonal variations in temperature are less than 0.2 °C close to the permanent ice shelf [1] and only 2.5 °C in the northern Antarctic [2]. Very likely these conditions have existed relatively unchanged for several million years [3]. Antarctic fish have therefore been forced to become highly specialized to cold conditions [4]. This extreme specialization is outlined by the fact that a water temperature of −1.8 °C, near the freezing point of sea water, is a condition which would be lethal for virtually all other fish. Among antarctic fishes the strong challenge of this extreme environmental condition has been overcome by a number of physiological and biochemical adaptation mechanisms, the most striking of which is the synthesis of freezing-point depressing molecules (antifreeze molecules), either peptides or glycopeptides, which protect the organism from ice crystal formation in a noncolligative way [5]. In addition, hematocrit of the blood is either substantially reduced or erythrocytes and hemoglobin are absent as in Channichthyidae [6, 7, 8].

Coenzyme Q homologs and trace elements content of Antarctic fishes Chionodraco hamatus and Pagothenia bernacchii compared with the mediterranean fish Mugil cephalus

Coenzyme Q and trace element contents of two antarctic fishes (Pagothenia bernacchii and Chionodraco hamatus) were investigated and compared with that of one temperate marine fish (Mugil cephalus). In particular, muscle and erythrocyte concentrations of coenzyme Q, vitamin E, selenium, copper and zinc were determined. Antarctic fishes showed, at the level of their muscles, higher levels of selenium, zinc and copper compared with M. cepalus. In antarctic fishes, in the place of Q10, the coenzyme Q form generally observed in temperature fishes, only the homolog coenzyme Q9 was found. This particular finding has been related to the difference in the crystallization temperature existing between CoQ10 and CoQ9. Moreover, the high levels of zinc and selenium, observed in C. hamatus, correlate very well with a recent report on the presence, in the liver of this fish, of a zinc-binding protein that, in addition, is characterized by an amino acid sequence closely similar to that of a selenium-containing protein isolated from ovine heart. All together, the results indicate, for the antarctic species, a high level of antioxidant defenses with respect to the mediterranean fish and may be considered as a stimulating basis for further studies on the oxidative metabolism of Antarctic organisms.

Reactive oxygen metabolites (ROMs) are associated with cardiovascular disease in chronic hemodialysis patients

Abstract Background: The aim of our study was to measure reactive oxygen metabolites (ROMs) in chronic hemodialysis (HD) patients and evaluate the possible association with cardiovascular disease (CVD) and mortality. Methods: We measured ROMs in 76 HD patients and correlated with CVD, cardiovascular (CV) events in the follow-up and all-cause and CVD-related mortality. Results: The levels of ROMs presented a median value of 270 (238.2–303.2) CARR U (interquartile range). We created a ROC curve (ROMs levels vs. CVD) and we identified a cut-off point of 273 CARR U. Patients with ROMs levels ≥273 CARR U were significantly older, had higher C-reactive protein levels and lower creatinine concentrations. The prevalence of CVD was higher in patients with ROMs levels ≥273 (87.1%) than in those with ROMs levels <273 CARR U (17.7%; p<0.0001). ROMs levels were significantly higher in patients with CVD (317±63.8) than in those without (242.7±49.1; p<0.0001). At multiple regression analysis, age, creatinine and C-reactive protein were independent factors associated with ROMs. At multiple logistic regression analysis the association between ROMs and CVD was independent (OR: 1.02, 95% CI: 1.00–1.05; p=0.03). Twenty six patients developed cardiovascular (CV) events during the follow-up. Of these, seven were in the group with ROMs levels <273 CARR U and 19 in the group with ROMs levels ≥273 CARR U. The logistic regression analysis showed that both age (OR: 1.06, 95% CI: 1.01–1.12; p=0.013) and ROMs levels (OR: 1.10, 95% CI: 1.00–1.02; p=0.045) were independently associated with CV events in the follow-up. Conclusions: ROMs are independently associated with CVD and predict CV events in chronic HD patients.

Autoimmunity and lymphoproliferation markers in naïve HCV-RNA positive patients without clinical evidences of autoimmune/lymphoproliferative disorders

BACKGROUND HCV can lead to both chronic liver disease and B-cell lymphoproliferative disorders. A strong association exists between HCV and mixed cryoglobulinaemia (MC). METHODS Anti-nuclear antibodies (ANA), rheumatoid factor Ig-G (RF-IgG), free light chain κ and λ (FLC-κ, FLC-λ) levels and κ/λ ratio were evaluated in 50/420 subjects unexpectedly resulted anti-HCV positive after routine screenings for non-hepathological procedures. RESULTS Three/fifty patients had HCV-RNA undetectable in the serum and were excluded from the analysis. Thirty-nine/fifty patients had laboratory evidence of circulating cryoglobulins without liver disease and MC-related symptoms. Among them, 17 resulted ANA-positive. The mean cryocrit was higher in ANA-positive patients, while no other demographic/clinical differences were observed between the groups. Significantly higher levels of RF-IgG were observed in ANA-positive vs ANA-negative patients. κ and λ FLC were higher in ANA-positive patients. A ROC analysis, based on ANA-positivity vs ANA-negativity, confirmed a high sensitivity and specificity of RF-IgG test. CONCLUSIONS Published data concerning MC come mostly from symptomatic vasculitis. We analyzed HCV-patients without MC symptoms, founding cryoglobulins in the majority of them. The increased levels of FR-IgG and FLC in CGs-ANA-positive patients, suggest these test could be used to identify a state of silent autoimmune and/or lymphoproliferative condition before the transition to a frank disease in naïve HCV-patients without symptoms of extrahepatic manifestations.

Pre-analytical phase in cryoglobulin (CRG) detection: an alternative method for sample transport.

aEleonora Torti and Maria Teresa Dell’Abate contributed equally to this work. *Corresponding author: Umberto Basile, Department of Laboratory Medicine, School of Medicine, Catholic University of the Sacred Heart, Largo A. Gemelli 8, 00168 Rome, Italy, Phone: +39 06 3015 4222, E-mail: umberto.basile@rm.unicatt.it Eleonora Torti and Maria Teresa Dell’Abate: Department of Laboratory Medicine, School of Medicine, Catholic University of the Sacred Heart, Rome, Italy Luigi Colacicco and Cecilia Zuppi: Institute of Biochemistry School of Medicine, Catholic University of the Sacred Heart, Rome, Italy Francesca Gulli and Gian Lodovico Rapaccini: Institute of Internal Medicine, School of Medicine, Catholic University of the Sacred Heart, Rome, Italy Letter to the Editor

Anti-nuclear antibody detection in cryoprecipitates: distinctive patterns in hepatitis C virus-infected patients.

BACKGROUND Anti-nuclear antibodies are immunoglobulins directed against nuclear antigens. They are associated with many autoimmune disorders, but are frequently found in patients infected with hepatitis C virus, possibly indicating an underlying common origin. Likewise, mixed cryoglobulinemia often accompanies autoimmune diseases and hepatitis C infection. AIM To compare anti-nuclear antibodies and immunoglobulin content of cryoprecipitates from hepatitis C virus-positive patients in order to assess their predictive value in the onset of hepatitis C virus-driven extrahepatic disorders. METHODS Serum from 40 hepatitis C virus-positive patients and 50 controls with rheumatoid arthritis was processed for cryoglobulin detection: all subjects presented with Type III mixed cryoglobulinemia. Immunoglobulin content and immunoglobulin subclasses of cryoprecipitates were assessed by immunofixation and tested by ELISA for rheumatoid factor. Cryoprecipitates were also analysed for anti-nuclear antibodies by indirect immuno-fluorescence to identify specific patterns typical of each condition. RESULTS Anti-nuclear antibody patterns differed significantly; 26 infected subjects (65%) were IgG3 positive: of these, 25 were also anti-nuclear antibody-positive (96.1%). CONCLUSIONS IgG3 are autoreactive clones unrelated to viral recognition and possibly involved in autoimmune disorders. Altogether, these results may represent useful diagnostic device for early detection of hepatitis C virus-induced autoimmune diseases.

Relationship between circulating syndecan-1 levels (CD138s) and serum free light chains in monoclonal gammopathies

BackgroundMonoclonal gammopathies encompass a wide range of diseases characterized by the monoclonal expansion of a B-cell clone. Despite emerging therapeutic strategies, chances of survival of patients who are affected are still scarce, which implies that new tools are necessary not only for the diagnosis but also for the follow-up of patients affected by such diseases. In this context, the use of free light chains (FLCs) has been incorporated into many guidelines.Likewise, tumor microenvironment is consistently gaining importance as role player in tumor pathogenesis. Specifically, Syndecan-1 (CD138), a heparan-sulfate proteoglycan is attracting interests as it is highly expressed and shed by myeloma plasma-cells.The aim of our study was to analyze CD138 levels in the serum of patients affected by multiple myeloma or light chain only disease, and to compare the values obtained with free light chain (FLC) kappa, lambda and FLC ratio in both groups of patients.Methods84 patients affected by Multiple Myeloma and Light Chain Myeloma were recruited for this study. Serum CD138 was assessed by ELISA (Diaclone Research, France) and FLC values were quantified by nephelometry (Freelite TM Human Kappa and Lambda Free Kits, The Binding Site, UK). Data was analyzed by GraphPad Prism software and Statgraph.ResultsWe observed higher CD138 mean values in myeloma patients compared to the light chain only myeloma group. A positive linear regression of CD138 and FLC was observed in the light chain only cohort as opposed to myeloma patients which show an inverse trend.ConclusionsThe study highlighted an existing relationship between FLCs and CD138 and wishes to seek also a correlation in order to rapidly and efficiently perform diagnosis and different diagnostic schemes.