Consensus on the use and interpretation of cystic fibrosis mutation analysis in clinical practice.

It is often challenging for the clinician interested in cystic fibrosis (CF) to interpret molecular genetic results, and to integrate them in the diagnostic process. The limitations of genotyping technology, the choice of mutations to be tested, and the clinical context in which the test is administered can all influence how genetic information is interpreted. This paper describes the conclusions of a consensus conference to address the use and interpretation of CF mutation analysis in clinical settings. Although the diagnosis of CF is usually straightforward, care needs to be exercised in the use and interpretation of genetic tests: genotype information is not the final arbiter of a clinical diagnosis of CF or CF transmembrane conductance regulator (CFTR) protein related disorders. The diagnosis of these conditions is primarily based on the clinical presentation, and is supported by evaluation of CFTR function (sweat testing, nasal potential difference) and genetic analysis. None of these features are sufficient on their own to make a diagnosis of CF or CFTR-related disorders. Broad genotype/phenotype associations are useful in epidemiological studies, but CFTR genotype does not accurately predict individual outcome. The use of CFTR genotype for prediction of prognosis in people with CF at the time of their diagnosis is not recommended. The importance of communication between clinicians and medical genetic laboratories is emphasized. The results of testing and their implications should be reported in a manner understandable to the clinicians caring for CF patients.

[Importance of chemoprevention in oncology]. / Importanza della chemioprevenzione in oncologia.

The therapeutic approach to cancer is generally limited to the advanced phases of disease. The preventive strategies aim at eliminating or reducing the exposition to known carcinogenes. We act with pharmacological and/or with an accurate dietary education to induce cellular differentiation phenomena, cytostasis and apoptosis. Chemoprevention acts both on the inductive phase (metabolic activation, DNA adducts), as well as on the promotion/proliferation of the long pre-clinical period of latency (antioxidants, anti-inflammatory, retinoids, carotenoids, vitamins and micronutrients, hormones and hormonal inhibitors, polyamine inhibitors, ditholetions, isothiocyanates, telomerase inhibitors, etc). Unanimous agreement has been reached on the preventive role of retinoids in head and neck tumors and of the cervical uterus, of hormonal inhibitors in breast and prostate cancer, and of anti-inflammatory in colorectal cancer. New and more accurate parameters for the evaluation of results and individual applications of chemopreventive strategies are linked to the biological research of high-risk subjects (genetic damage) or increased individual susceptibility. Caution, instead, should be applied in the clinical trial planning. An increased risk in developing and dying of lung tumor in smokers has been shown for the use vitamin A. Many clinical studies have been started in order to establish an efficient chemoprevention in oncology, and with the early diagnostic programs, and the evaluation of genotypic and phenotypic alterations, encouraging results will be reached for the next millennium.

[Malignant lymphoma. Epidemiologic review of 150 cases]. / Linfomi maligni. Revisione epidemiologica di 150 casi.

In the present paper the incidence of lymphomatous disease in Messina and its province, with growing urbanization and rural limiting areas, is discussed: by analysing 150 cases of malignant lymphoma observed at our Institute from 1990 to 1995. The method proposed, based on data obtained from the medical files of these patients, took into consideration various parameters such as age, sex, residence, profession, clinical and bioptic diagnosis, LDH and disease presentation. The final results showed an increase of the risk for NHL in the rural province where the main profession is agriculture or handicraft (ceramics, forged iron, glasswork, refinery), in subjects above 60 years of age; for the HL instead, over the years, a minor incidence of risk has been observed. The data obtained were partially similar to those reported in the international literature. The most present form in NHL was the lymphocytic and the centrocytic follicular form, while for HL it was the mixed cells form. The relationship between the two sexes was higher in males with HL and almost equal in NHL. The age range mostly affected by HL was between 25 and 65 years of age.

Variability between methods to determine ANA, anti-dsDNA and anti-ENA autoantibodies: a collaborative study with the biomedical industry.

This study was performed by the Italian Society of Laboratory Medicine (SIMeL) in order to establish the variability between the different analytical systems currently used in clinical laboratories for the detection of autoantibodies diagnostic of systemic autoimmune disease. Sixteen industrial, and two university laboratories participated in this study which entailed the determination of anti-nuclear (ANA), anti-dsDNA and anti-ENA antibodies in 11 sera from patients with clinically diagnosed systemic rheumatic disease, using reagents produced by these companies and different methodologies (indirect immunofluorescence, immunoenzymatic assay, counterimmunolectrophoresis, immuno and western blotting). We found 93.5% agreement between the methods used for the detection of ANA, 85.2% for anti-dsDNA antibodies, and 86.9% for anti-ENA antibodies. Among the anti-ENA antibodies, regardless of the method used, detection percentages were excellent for anti-RNP and anti-SSB/La (100%), good for anti-SSA/Ro (93%), but unacceptable for the anti-Jo-1 (67%), anti-Scl70 and anti-Sm (47%) antibodies. This further stresses the need for rigorous standardisation of commercial reagents and analytical procedures, as well as the introduction of external quality assessment (EQA) programs, and a complete definition of operative protocols adjusted to the sensitivity and specificity of the various methods.

Isolated rectal relapse in non Hodgkin lymphoma patient.

The authors report on case of isolated rectal relapse of non Hodgkin lymphoma after complete remission with chemotherapy. They analyse diagnostic and therapeutic aspects, on the grounds of literature data and personal experience.

Evaluation of a specific immunoinhibition method for the determination of pancreatic alpha-amylase.

Ten clinical centres in Italy participated in an evaluation of a pancreas-specific alpha-amylase assay using two monoclonal antibodies. Comparisons with electrophoretic methods showed good agreement in the reference range, but systematic deviations above it. The diagnostic information of the two methods appeared substantially different if the percentage values from electrophoresis were compared with activity units from the immunoinhibition method, but became similar if the two methods were compared on the basis of activity units. Reference intervals determined for serum/plasma corresponded to those previously published, but those determined for urine differed slightly from the published values. Clinical sensitivities for the assessment of acute pancreatitis (n = 134) were found as follows. Related to the upper limit of the control group (n = 141), the pancreatic alpha-amylase, total alpha-amylase and lipase showed sensitivities of 0.94, 0.87 and 0.93, respectively. When the cut-off point was set at the three-fold upper limit of the control group, sensitivities of 0.73, 0.53 and 0.70 were found, and the specificity was 1.00 for all three methods. Based on this commonly used higher cut-off point, the determination of lipase in addition to pancreatic alpha-amylase enhanced the sensitivity in the recognition of acute pancreatitis by 8%; conversely, the determination of pancreatic alpha-amylase in addition to lipase increased the number of true positive results by 13%. The high practicability and interlaboratory transferability documented in the results of the collaborative study show that the pancreatic alpha-amylase assay is very useful for recognizing pancreatic inflammations, especially in combination with lipase.

Variations in time of serum pancreatic enzyme levels in chronic pancreatitis and clinical course of the disease.

Thirty-four patients with chronic calcified pancreatitis were evaluated clinically and biochemically (at a time when painful relapses were not present) every 9 mo for 3 yr; 25 of them were also studied at 4 and 9 yr. Serum elastase-1, trypsin, lipase, and amylase in the same sera were measured at each visit; levels on entry and variations during the study were compared with the clinical and functional data of the patients. On entry, low levels of elastase-1 were found in 11.7% of the patients, high levels in 41.1%; in contrast, high levels of trypsin and lipase were found in only a small number of patients (5.8 and 11.7%, respectively), whereas low levels were present in a substantial number (47.8 and 32.3% for trypsin and lipase, respectively). Over time, we found a significant (p = 0.000002) reduction in elastase-1 levels. Such reduction was not found for trypsin, lipase, or amylase. The reduction of serum elastase-1 was significantly (p less than 0.003) more frequent in patients presenting a reduction in painful relapses than in patients with a stable or increased attach rate; this association was weaker (p less than 0.05) for lipase and trypsin, and absent for amylase. No correlation was found between circulating enzymes and either alcohol consumption or age of patients. In patients with severe exocrine impairment, low levels of elastase were found in only 20% of the cases, whereas trypsin and lipase were reduced in 73.3 and 53.3% of the cases, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Fractional urinary clearance of immunoreactive lipase in chronic pancreatic disease.

Using an immunoenzymatic method, we studied lipase in the serum and urine of 23 controls, 22 chronic pancreatitis patients in symptomatic remission, and in 9 patients with proven pancreatic cancer. Serum and urine lipase and its fractional urinary clearance were compared with those of amylase and immunoreactive trypsin. Lipase immunoreactivity was detectable in the urine of 81.5% of the studied subjects (controls: 82%, chronic pancreatitis: 86%, pancreatic cancer: 66%); its output was higher than the upper limit of controls in 31.8% of chronic pancreatitis and in only 1 of pancreatic cancer, and it was significantly correlated with the urinary output of trypsin (r = 0.487, P less than 0.001), but not with that of amylase. A significant correlation was found between urinary output and serum levels for lipase, but not for trypsin or amylase. Fractional clearance of lipase was of the same magnitude as that of trypsin but only 0.1% that of amylase. 19% of chronic pancreatitis and pancreatic cancer showed a fractional clearance of lipase above the upper limit of controls, compared with 45% for trypsin and 3.2% for amylase. No difference in urinary clearance of the three enzymes was found between chronic pancreatitis and pancreatic cancer. In conclusion, although of no diagnostic relevance in pain-free patients with chronic pancreatic disease, this measurement can provide information on the mechanisms of renal excretion of pancreatic enzymes.

A clinical evaluation of monoclonal (CA19-9, CA50, CA12-5) and polyclonal (CEA, TPA) antibody-defined antigens for the diagnosis of pancreatic cancer.

We measured in 193 patients, admitted to our wards for symptoms and signs suggestive of pancreatic or digestive malignancy, the serum levels of five tumor-associated antigens (CA 19-9, CA 50, CA 125, TPA, CEA) and we evaluated their diagnostic accuracy both when used alone and in combination. For CA 19-9 and CA 50 a sensitivity for pancreatic cancer as high as 92 and 88%, respectively, and specificity of 91.8% were found. A lower sensitivity vs. pancreatic cancer was found for the other tumor markers, and vs. the other digestive and nondigestive malignancies for all tumor markers (apart for CA 19-9 and CA 50 vs. biliary carcinomas). As for the combined assays, the best figures were found vs. pancreatic cancer for CA 19-9 plus CA 50, CA 50 plus CEA, CA 50 plus CA 125; a sensitivity by far worse vs. the other gastrointestinal cancers was found for all the possible combinations. We conclude that in selected symptomatic patients some tumor-marker determinations can be useful in identifying those with a high probability of harboring a pancreatic cancer, to be further studied or operated upon. The clinical relevance of this in patients already symptomatic is at present unclear.

Enzyme immunoassay for pancreatic lipase: comparison with turbidimetric method in pancreatic diseases.

We report the results of the analytical and clinical evaluation of a specific enzyme immunoassay for determination of human pancreatic lipase, in comparison with a turbidimetric method, in pancreatic pathology. Under standardized conditions of incubation time and temperature we found intraassay coefficient of variation (CV) of 1.3, 3.2, 2.1% at means = 18.7, 43.7, 224 micrograms/L and interassay CV of 2.8, 4.6, 3.0% at means = 19, 42.6, 230 micrograms/L, respectively. In general, a good correlation (r = 0.97) was found between lipase determined as a protein or through its catalytic activity. No significant correlation (r = 0.38) was observed with samples containing low concentration of lipase (up to 18 micrograms/L). We conclude that the turbidimetric method is reliable for routine determinations in the diagnosis of acute pancreatic pathology. However, the better sensitivity of the immunochemical assay should provide additional information for monitoring pancreatic insufficiency.

Contribution of four markers of tubular proteinuria in detecting upper urinary tract infections. A multivariate analysis.

The presence of tubular involvement, as a marker for the detection of urinary tract infection (UTI) site, was examined in 19 patients with pyelonephritis and in 15 patients with cystitis or asymptomatic bacteriuria. The urinary excretion of four markers of tubular proteinuria, beta 2-microglobulin (beta 2M), lysozyme (LZ), lactic dehydrogenase isoenzyme V (LAD-5) and N-acetyl-beta D-glucosaminidase (NAG), was investigated. LAD-5 appeared particularly valuable for the early detection of upper UTI. However, the overall diagnostic accuracy appeared to be further strengthened using, besides LAD-5, one additional variable. A set of simple and noninvasive biochemical tests on urine samples can reliably help to identify the site of UTI.

Serum immunoreactive trypsin in cystic fibrosis.

Immuno-reactive trypsin (IR-TRY) level has been measured by RIA in 185 patients with cystic fibrosis (CF) aged between 4 days and 34 years. The correlation of IR-TRY values with age and functional state of pancreas have been studied. The interpretation of the data have been put forward on the basis of possibilities of neonatal screening.

Biochemical and cytochemical effects of cytosine arabinoside (ARA C) and hydrocortisone on HeLa cells.

The biochemical and cytochemical effects of cytosine arabinoside (Ara C) and of hydrocortisone on HeLa cell in vitro have been studied. In cultures treated with Ara C, a relationship exists between the cytocidal effects of the drug and an increase in alkaline phosphatase. Although hydrocortisone had no influence on the proliferative rhythm, it induced an increase in alkaline phosphatase. Results obtained with the cytochemical technique were evaluated, particularly in relation to enzyme localization.