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Metabolismo glucidico alterato e fenomeni di proteolisi in mioblasti condizionati da cellule di cancro del pancreas: analisi di espressone genica mediante tecnica microarray su una piattaforma di 5000 geni di muscolo scheletrico*

Eliana Greco*, Daniela Basso, Caterina Millino, Chiara Romualdi, Paola Fogar, Anna Valerio, Milena Bellin, Carlo-Federico Zambon, Filippo Navaglia, Noemi Dussini, Angelo Avogaro, Sergio Pedrazzoli, Gerolamo Lanfranchi, Mario Plebani
*Dipartimento di medicina di Laboratorio, Università di Padova

Biochimica Clinica: 2004; 28(4): 443-453 [Article in italian]

ABSTRACT. Background & Aims: We verified whether pancreatic cancer cell lines (MIAPaCa2, CAPAN-1, PANC-1, BxPC3) conditioned media (CM) alter glucose metabolism and gene expression profile (microarray experiment with a platform of 5000 skeletal muscle cDNA) in mice myoblasts. Methods: Myoblasts were incubated with control or pancreatic cancer CM for 24 and 48 hrs. Results: Lactate significantly increased in CM as compared to non-conditioned myoblasts. No variations in the expression levels of the main genes involved in glycolysis were found in CM myoblasts. Propionil Coenzyme A carboxylase and isocitrate dehydrogenase 3 beta (IDH3B) genes, which encode enzymes of the tricarboxylic acid cycle were overexpressed, while IGFIIR and VAMP5 genes were underexpressed in CM myoblasts. PAFAH1B1 and BCL-2 genes (intracellular signal transduction) and the serine protease cathepsin G (proteolysis), were overexpressed in CM myoblasts. Tyrosine accumulation in CM myoblasts suggested that proteolysis overcomes protein synthesis. Sorcin, actin alpha, troponin T1 and filamin A were underexpressed in CM myoblasts. Conclusions: Our findings demonstrate that pancreatic cancer cell conditioned media enhanced lactate production and induced proteolysis, possibly by altering the expression levels of a large number of genes, not only those involved in protein biosynthesis and degradation or glucose metabolism, but also those involved in the trycarboxylic acid cycle and in vesicle traffic.

Profil protéique immunitaire au cours du portage de parasites intestinaux chez l'enfant ivoirien scolarisé

Houphouet F. Yapi*, Hugues Ahiboh, Kouame Ago, Dagui Monnet
*UFR de sciences pharmaceutiques et biologiques de Cocody, Laboratoire de Biochimie et biologie moléculaire

Biochimica Clinica: 2004; 28(4): 454-456 [Article in franch]

ABSTRACT. Blood immunoglobulins A,G and M determination has been done by the method of radial immuno - diffusion of Mancini at the children schooled in tropical environment. A transverse investigation and a drawing to 2 levels have been achieved on 262 samples. Blood immunoglobulins have been determined among 194 intestinal interference carriers and among 68 witnesses.It has been shown that the blood immunoglobulins A were lowered respectively to the course carriage of the Ascaris lumbricoides and the Hookworms. Blood immunoglobulins M were increased during the carriage of Giardia intestinalis and Trichomonas intestinalis. The hypergammaglobulinaemia G has been observed as well among the children carriers or no of intestinal interferences because of the infectious tropical environment.

Fisiopatologia della coagulazione: nuove acquisizioni

Franco Manzato*, Giuseppe Lippi, Massimo Franchini, Gian Cesare Guidi
*Laboratorio di Patologia Clinica, Azienda Ospedaliera Carlo Poma, Mantova

Biochimica Clinica: 2004; 28(4): 457-469 [Article in italian]

ABSTRACT. Haemostasis is a field of clinical biochemistry in constant evolution; technical progresses joined to recent scientific discoveries, promote the comprehension and characterization of the mechanisms that regulate the delicate balance of the haemostatic system. After endothelial damage, following activation of primary haemostasis, secondary haemostasis, known also as blood coagulation, develops. Blood coagulation involves a series of coordinated and calcium-dependent conversions of proenzymes to the respective serine proteases, culminating in the conversion (activation) of prothrombin to thrombin. Activation, amplification of the coagulation cascade and stabilization of the blood clot are rigorously modulated by a balanced activity of specific proteases and respective allosteric or enzymatic inhibitors, which proper function is essential to ensure the correct development of the system.

Valutazione multicentrica del metodo Tosoh AIA-Pack‰ di 2^ generazione per la determinazione della troponina I cardiaca

Franca Pagani*, Annalisa Iervasi, Francesca Stefini, Gianmatteo Micca, Paolo Hoffer, Marco Caputo, Gian Carlo Zucchelli, Mauro Panteghini
*Laboratorio Analisi Chimico Cliniche 1, Azienda Ospedaliera "Spedali Civili", Brescia

Biochimica Clinica: 2004; 28(4): 470-474 [Article in italian]

ABSTRACT. Cardiac troponin I (cTnI) is a powerful tool for diagnosis of acute myocardial infarction. This necessitates the availability of highly specific, sensitive, and precise assays, thoroughly validated in well-designed studies. Here we describe the performance characteristics of the Tosoh AIA-Pack 2nd-generation cTnI immunoassay evaluated in a multicenter study. The protocol consisted of following sections evaluating analytical and preanalytical aspects: calibration stability, detection limit and imprecision, linearity, method comparison, sample stability, and anticoagulant interference. Reference values and the 10% CV cutoff for detecting myocardial necrosis were also defined. The minimum detectable cTnI concentration was 0.038 mg/L. Linearity of response was demonstrated along the entire dynamic range of the assay. The cTnI concentration corresponding to a total CV of 10% was 0.13 mg/L. cTnI values in heparin and EDTA plasma were on average ~15% and ~13% higher than in matched serum, respectively. cTnI, measured in 120 healthy individuals, was always undetectable. In conclusion, the AIA cTnI assay revealed acceptable analytical performance making it suitable for clinical applications.

The future of Laboratory Medicine: understanding the new pressures (*)

Mauro Panteghini
Laboratorio Analisi Chimico Cliniche 1, Azienda Ospedaliera "Spedali Civili", Brescia

Biochimica Clinica: 2004; 28(4): 475-481 [Article in english]

ABSTRACT. Since the future role of Laboratory Medicine is strongly and equally challenged by economical and new technological pressures, it is essential to take a broad view of the discipline and present to the administrators and other decision makers the full spectrum of activities and benefits Laboratory Medicine can provide. In particular, the importance and the true impact of Laboratory Medicine can only be achieved by assuring an appreciable added value to laboratory tests, represented by their effectiveness in influencing the management of patients and related clinical outcomes.

(*) This paper is based on the David Curnow Plenary Lecture held at the 10th Asian Pacific Congress of Clinical Biochemistry, 18-23 September 2004, Perth, Australia

Valutazione del test immunocromatografico rapido ToxSTATus¿ per lo screening urinario di oppiacei, metadone, cocaina, cannabinoidi e amfetamine

Lucio Marchioro*, Silvia Ponchia, Anna Tommasi, Mario Plebani
Dipartimento di Medicina di Laboratorio, Azienda Ospedaliera - Università di Padova

Biochimica Clinica: 2004; 28(4): 482-487 [Article in italian]

ABSTRACT. We evaluated the ToxSTATus¿ Multiple Drug Dipcard for the qualitative determination of several drugs of abuse in urine samples. We compared the immunochromatographic test with the results obtained by the KIMS technique (Kinetic Interaction of Microparticles in Solution). ToxSTATus¿ involves the use of specific antibodies for several drugs as Cocaine, Opiates, Cannabinoids, Amphetamines and Methadone. We analyzed nÉ45 samples and we verified sensitivity, specificity and efficiency (according to Insook) of all parameters. In the complex we obtained good results for samples and controls: each result must be then confirmed with screening assays.

HER-2 m-RNA ed HER-2 ECD (dominio extracellulare) circolanti nel sangue di pazienti con tumore precoce della mammella(*)

Benedetta Salvadori*, Claudio Orlando, Pamela Pinzani, Vito Distante, Donato Casella, Simonetta Bianchi, Milena Paglierani, Vania Vezzosi, Rainer Neumann, Luigi Cataliotti, Mario Pazzagli
*Unità di Biochimica Clinica, Dipartimento di Fisiopatologia Clinica, Università di Firenze

Biochimica Clinica: 2004; 28(4): 488-491 [Article in italian]

ABSTRACT. The HER-2 gene encodes a 185kDa transmembrane glycoprotein member of the type I family of growth factor receptors. HER-2 gene activation has been found in 20% to 40% of breast cancers, analysed for gene amplification or overexpression. This has been associated with poor prognosis, high risk of relapse and reduced patient survival. HER-2 status assessment is usually studied in advanced breast cancer. We investigated a group of patients affected by early breast cancer and the role of circulating HER-2 extracellular domain (ECD) and HER-2 mRNA, potentially suitable for monitoring HER-2 status in the tumor. Circulating ECD/HER-2 was determined, before and after surgery, in 50 patients with early stage infiltrating breast cancer. In the same blood samples we measured HER-2 mRNA with a quantitative real-time RT-PCR assay. These results were compared with HER-2 status in the tumor, evaluated by immunohistochemistry and real-time RT-PCR. Although the blood levels of ECD/HER-2 before surgery were in the normal range (below 15 ng/ml), we observed a significant decrease (p<0.001) of levels of this circulating protein in post-surgery samples. Blood HER-2 mRNA was significantly higher (p=0.001) in pre-surgery cancer patients than in healthy subjects. After surgery, HER-2 mRNA decreased significantly (p=0.04) reaching the level of the control group. In tissue samples, HER-2 mRNA expression was significantly higher (p<0.001) in tumor specimens than in the corresponding normal tissue of the same patient. Interestingly, only patients that showed overexpression in the tumor, measured both by RT-PCR and immunohistochemistry, had a significant decrease in the circulating levels of both ECD/HER-2 and HER-2 mRNA after surgery (p=0.001 and p=0.04, respectively). We conclude that pre- and post-surgery measurements of circulating ECD/HER-2 and HER-2 mRNA are useful in defining HER-2 status and may identify a subgroup of early breast cancer patients which could benefit of a selected therapy.

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