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I saggi immunometrici qualitativi: valutazione e controllo
Guest Editor: Renzo Malvano

Biochimica Clinica: 2005; 29(3): 297-308 [Article in italian]

ABSTRACT: Nel corso del loro sviluppo le tecnologie analitiche applicate nel laboratorio clinico hanno sempre più assunto il carattere delle così dette “analisi strumentali”. Cuore di ogni analisi strumentali è un “principo analitico”, ossia una reazione che consente di “riconoscere” la molecola misuranda, generando di conseguenza un “segnale” misurabile, di intensità proporzionale alla concentrazione della molecola medesima. Nel corso degli anni i principi analitici sono passati dal chimico (e/o chimico-fisico), all’enzimatico, all’immunochimico, alla ibridazione. I primi tre principi sono applicabili alla misurazione di una ampia varietà di molecole; l’ibridazione è riservata agli acidi nucleici. Nel medesimo ordine, passando dal primo all’ultimo principio, si è ottenuto un incremento della sensibilità e della specificità dei procedimenti di misura, con evidenti vantaggi analitici e clinici. Nella stragrande maggioranza dei procedimenti, opportuni accorgimenti associamo la reazione di riconoscimento alla generazione di un segnale ottico, ossia di assorbimento o di emissione di energia luminosa.

Specie Reattive dell’Ossigeno (ROS): biochimica, patologia, prevenzione, laboratorio
Marcello Ciaccio, Stefano Miccichè

Biochimica Clinica: 2005; 29(3): 309-330 [Article in italian]

ABSTRACT: Il Convegno si inquadra nell’ambito delle attività formative, scientifiche e di aggiornamento che la Cattedra e la Scuola di Specializzazione in Biochimica Clinica della Facoltà di Medicina e Chirurgia dell’Università degli Studi di Palermo congiuntamente alla Sezione Sicilia della Società Italiana di Biochimica Clinica e Biologia Molecolare Clinica organizza periodicamente. Il Convegno si articolerà in 2 Sessioni Scientifiche dal titolo, rispettivamente, “ROS e Patologia” e “ROS: prevenzione e diagnostica”, e vedrà la partecipazione di Docenti e Ricercatori che terranno relazioni sulle più recenti acquisizioni inerenti la biochimica e la fisiopatologia delle Specie Reattive dell’Ossigeno e della loro importanza in alcune patologie di larga incidenza come il processo arteriosclerotico e le malattie cardiovascolari, la neurodegenerazione, le neoplasie ed in alcuni processi fisiologici come la senescenza cellulare e l’invecchiamento.

Attività della Glutatione S-transferasi (gst) nel fegato di ratti diabetici sottoposti a trattamento antiossidante con n-Acetilcisteina e Taurina
Renata Cordara
ASL 3 Genovese - Unità Operativa Igiene e Sanità Pubblica - Dipartimento di Prevenzione

Biochimica Clinica: 2005; 29(3): 331-336 [Article in italian]

ABSTRACT: The role of oxidative stress in diabetes is currently under discussion, but several alterations in diabetic are oxidative in nature or may depend on increase oxidative oxidative mechanism. It also well known that glycation, which many authors consider to underlie the pathogenesis of diabetic sequelae, is strictly interconnected with diabetes and induce structural and functional modifications of many enzymes. Previous studies suggested that peroxidative aldehydes (MDA =malondiadehyde, HNE = 4-hydroxy-2-nonenal, ecc) accumulation in diabetic liver may depend on both mechanism: increased lipoperoxidation and decreased enzymatic removal. The main enzyme capable of metabolizing peroxidative aldehydes in liver and other tissues is Glutathione S-transferase (GST), wich catalyses the coniugation of these to Glutathione (GSH), the major cellular thiol partecipating in cellular redox reactions and thioether formation. In many studies is showed that GST activity is reduced in liver of diabetic rats and peroxidative aldehydes accumulation can generated vicious circles able to amplify the oxidative damage. The results of recents studies suggest that oral supplementation with Thiol- containing antioxidants such as N-Acetylcysteine (NAC) and with Taurine (TAU) to be beneficial for both mantaining natural antioxidant system and protecting against a number of degenerative disease associated with free radical damage and toxin exposure. This study investigate wether NAC and TAU supplementation, improving GSH availability and reducing lipoperoxidation, have influence upon activity of GST.

Valutazione dell'attività proliferativa del fattore timico umorale
Arpino Anna
Specialista in Biochimica clinica

Biochimica Clinica: 2005; 29(3): 337-339 [Article in italian]

ABSTRACT: The Thymic Humoral Factor (THF - ?2) is a peptide extracted from calf 's thyme. It seems to cause various lymphocyte reactions, development of the transplant 's rejection, as well as development toxic answer 's. The purpose of this study was to estimated biological activity of the THF - ?2. Blood cells extracted from umbilical cord has been utilized for this experiment.

Apolipoprotein E Gene Polymorphism as a Risk Factor for Atherosclerosis in Egyptians
Olfat G. Shaker*, Mohamed A. Ay El Deen
*Departments of Medical Biochemistry and Vascular Surgery, Cairo University

Biochimica Clinica: 2005; 29(3): 340-345 [Article in english]

ABSTRACT: Apolipoprotein E (apo E) plays a role in the regulation of lipid metabolism by mediating cellular uptake of lipoprotein particles via apo E and apo B/E-specific receptors in liver and other tissues. It is classified into 3 major isoforms in human (E2, E3 and E4) according to the differences of amino acids in position 112 and 158. In this study, 100 persons were classified into 3 groups: control group (26), atherosclerosis group (36) and high risk group (diabetic, hypertensive and obese patients) (38). Apo E genotyping was done using the PCR. The frequency of the E2 allele was relatively high in the control group (30.8%) and in the persons with normal serum cholesterol level (46.6%). Meanwhile, the frequency of the E4 allele was relatively high in the atherosclerosis group (29.2%) and also in hypercholesterolemics (61.3%). We can conclude that E2 allele is protective against the development of atherosclerosis, while E4 allele is associated with an increased risk of atherosclerosis.

Effetti della somministrazione di granulocyte colony stimulating factor sui livelli di insulin-like growth factor e sulla beta2-microglobulina nel liquido di lavaggio broncoalveolare di neonati prematuri
Ettore Capoluongo*, Giovanni Vento, Franco Ameglio, Paola Lulli, Concetta Santonocito, Stefano Angelo Santini, Sandro Rocchetti, Mirca Antenucci, Cinzia Carrozza, Costantino Romagnoli, Bruno Giardina, Cecilia Zuppi
*Biochemistry and Clinical-Biochemistry Institute, S. Giovanni Calibita" Fatebenefratelli/AFAR, Rome

Biochimica Clinica: 2005; 29(3): 346-351 [Article in italian]

ABSTRACT: We studied the effects of the therapy with recombinant Granulocyte Colony Stimulating Factor (rhG-CSF) on the IGF-I and b2-microglobulin (B2M) levels in bronchoalveolar lavage fluids (BALF) of six neutropenic preterm infants who developed chronic lung disease (CLD) and we compared the results with eleven non-neutropenic neonates with CLD, and 13 non-neutropenic whitout CLD. With the exception of the first day values, there was a clear similarity between non-neutropenic and neutropenic CLD patients. In contrast, non-neutropenic patients without CLD showed significantly lower values of free IGF-I and B2M. These findings suggest that 1) the mechanisms leading to CLD might be mediated by high levels of IGF-family molecules soon after birth 2) B2M could be a marker of BALF cellularity with potential inflammatory properties 3) G-CSF treatment induces an increased synthesis of IGF-I molecules by cells recruited in the lung, with possible enhancement of the fibrogenic mechanisms.

Nell'amiloidosi da catene leggere immunoglobuliniche monoclonali la riduzione delle catene leggere libere circolanti si associa a diminuzione della concentrazione sierica del pro-peptide natriuretico di tipo B
Francesca Lavatelli*, Giovanni Palladini, Tiziana Bosoni, Gianvico Melzi d'Eril, Remigio Moratti, Giampaolo Merlini
*Centro per lo Studio delle Amiloidosi Sistemiche, Laboratorio di Biotecnologie - Dipartimento di Biochimica, Università dell'Insubria, 21100 Varese

Biochimica Clinica: 2005; 29(3): 352-354 [Article in italian]

ABSTRACT: AL amyloidosis is caused by tissue deposition of fibrils derived from monoclonal immunoglobulin light-chains. Heart involvement is the main prognostic determinant and its severity can be evaluated by the quantitation of N-terminal pro-natriuretic peptide type B. The measurement of the circulating free light-chains makes it possible to estimate the concentration of the amyloidogenic precursor. The modifications of the concentration of these two markers measured before and after therapy were significantly correlated (p <0.02). Reducing the synthesis of the amyloidogenic light-chain promptly translates into an improvement of myocardial dysfunction and increases survival, despite the amount of amyloid deposited (i.e. echo-pattern) remains unaltered.

Interferenza da acido niflumico/morniflumato nella determinazione dei cannabinoidi su autoanalizzatore ILAB 600
Arcangelo de Gironimo, Vincenzo Musolino
Ospedale Principale Marittimo "G. Venticinque" - Laboratorio Analisi - Servizio di Tossicologia -Taranto

Biochimica Clinica: 2005; 29(3): 352-354 [Article in italian]

ABSTRACT: We describe an experimental investigation showing that some drugs give false positive results when metabolites of cannabinoids are measure in urine samples by an immunochemical method, with the ILAB 600 instrument. The observation of some samples positive to the immunochemical assay but not confirmed by chromatography led us to consider the possibility that this could be due to drug interference in the immunochemical method. By means of GC-mass spectrometry it was shown that niflumic acid and its beta-morphinoethilic ester (Morniflumate) are responsible for false-positive results in the immunochemical assay. This finding was confirmed by the assay of urine samples from subjects free from drug of abuse, but using therapeutic drugs containing niflumic acid/Morniflumate as the active principle.

Recenti sviluppi degli esami antidoping. Presupposti giuridici e ricadute pratiche
Giuseppe Lippi*, Giuseppe Banfi, Alberto Dolci, Camilla Mattiuzzi, Gian Cesare Guidi
Per il Gruppo di Studio della SIBioC sulla Medicina di Laboratorio dello Sport
*Istituto di Chimica e Microscopia Clinica, Dipartimento di Scienze Morfologico-Biomediche, Università degli Studi di Verona, Verona

Biochimica Clinica: 2005; 29(3): 360-366 [Article in italian]

ABSTRACT: Athletes have traditionally taken enormous benefits from dietary supplements, drugs and pharmacological therapies, most of which were originally proposed for improving fitness and treating pathologies. The adoption of such measures, inducing an artificial improvement of the athletic performances, might disguise severe risk for the health, corrupts the ethic of competitive sports and ultimately represents a crime, as sanctioned by the current Italian jurisprudence. Since the promulgation of the Italian law 376/2000 on doping, antidoping testing on athletes has been substantially intensified. However, the Italian legislation has as yet some gaps and ambiguities, which paradoxically narrow an effective preventive intervention. Aim of this document is to provide an overview on the current Italian legislation on this topic, analyzing results of antidoping testing on athletes since the year 2000.

Sensitive, rapid and simple method for multi-samples HPLC determination of urinary 2,5-hexanedione, suitable for automatic implementation
Lucia Scalini*, Patrizia Villa, Augusta Brega, Vincenzo Palomba
*Laboratorio Eurolab di Analisi Cliniche, Via Roma 68, Collebeato, Brescia (Italy)

Biochimica Clinica: 2005; 29(3): 367-370 [Article in italian]

ABSTRACT: In this paper we study the determination of urinary levels of 2,5-hexanedione (2,5-HD), an active metabolite of n-hexane by using high-performance liquid chromatography, DNPH derivatization and UV detection. We propose a novel method for 2,5-HD detection where the derivatization with DNPH is performed directly in the ethereal urinary extract. This allows to obtain only the mono-derivative of 2,5-HD wich, injected in a C18 column, originates in the chromatogram a peak at 7 min. without interferences.

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