A rapid test for the visual detection of anti-hepatitis C virus antibodies in whole blood.

A rapid test for the visual detection of anti-HCV antibodies in whole blood was evaluated in its accuracy when compared with EIA method. The rapid test was performed blind on 50 HCV EIA-positive (adsorbance greater than 3.0) and on 50 HCV EIA-negative samples. Each whole blood sample was 1:20, 1:50 and 1:100 diluted with saline solution for a total of 400 samples. Results showed a sensitivity of 100% when whole blood was tested, 96% when 1:20 diluted blood was tested, 30% when 1:50 diluted blood was tested and 4% when 1:100 diluted blood was tested. The specificity gave also better results and only one false-positive was found in all samples tested. The test took less than 3 min and only a mechanical pipette was required. In conclusion, the HCV Ab rapid test showed a very high accuracy and could be very useful for the detection of HCV-positive subjects in situations where rapid results are required or technical expertise is limited.

Plasma catecholamines in pre- and in postmenopausal women with mild to moderate essential hypertension.

The sympathetic nervous system (SNS) is thought to play an important role in the pathogenesis of essential hypertension and many studies have established a relationship between plasma levels of norepinephrine (NE) and epinephrine (E) and sympathetic nervous activity (SNA). Furthermore, it has been suggested that climacteric women are more exposed to psychosocial stress which can produce a transient rise in blood pressure (BP) and, with time, determine a hypertensive state. Plasma NE and E levels were measured at rest and after physiological stimulation (head-up tilt test) in 20 hypertensive (BP: 146 +/- 13/101 +/- 4 mm Hg) and in 20 normotensive women (BP: 132 +/- 7/85 +/- 4 mm Hg). Women in each of these two groups were further subdivided according to their climacteric status (10 premenopausal and 10 postmenopausal women). No difference in NE values at rest was found between groups and subgroups. During head-up tilt test, Ln NE plasma values increased in normotensive and hypertensive groups; the rise was significantly higher in hypertensive than in normotensive women (P < 0.01). In climacteric subgroups, Ln NE appeared markedly increased above resting levels in pre- and postmenopausal hypertensive women when their position was changed from supine to upright (P < 0.01). Since high plasma NE levels after stimulation (head-up tilt) are associated with sympathetic overactivity, we conclude that SNA is involved in the pathogenesis of essential hypertension in climacteric women.

De novo hepatitis B and C viral infection after liver transplantation.

Hepatitis B (HBV) and hepatitis C (HCV) viral infections often recur after orthotopic liver transplantation (OLT), but viral infections acquired with OLT have not been widely investigated. The aim of the study was to evaluate the incidence, evolution, and diagnostic problems of de novo HBV and HCV infections in liver transplant recipients with long-term follow-up. Altogether 121 transplant recipients entered the study. HBV, HDV, and HCV infections were diagnosed by means of serology and the polymerase chain reaction (PCR). Three patients became hepatitis B surface antigen (HBsAg)-positive after OLT, all of whom showed signs of persistent viral replication. Twelve patients became anti-HCV-positive after OLT: After clearance of passive antibodies, active anti-HCV seroconversion was usually delayed. The viral genome was detected in 9 of 12 patients, with fluctuations of viremia during their follow-up. The other three patients, who were HBsAg-positive before and after OLT, were repeatedly HCV-RNA-negative despite persistent anti-HCV reactivity. Four pre-OLT HBsAg-positive patients had evidence of HBV-related liver transplant disease. The remaining 8 of 12 patients experienced repeated alanine aminotransferase increases more than two times normal after transplant. De novo infections due to primary hepatotropic viruses were frequent after OLT in our experience. Early diagnosis of infection, especially when the HCV is involved, may be problematic and should be taken into account in patients showing persistent aminotransferase abnormalities. Monitoring viral markers and accurate evaluation of biopsy specimens are mandatory. The interference between HBV and HCV might play a role in the replicative cycle of one or both viruses in co-infected patients.

Serum interleukin-6, interleukin-8, and beta 2-microglobulin in early assessment of severity of acute pancreatitis. Comparison with serum C-reactive protein.

The aim of this study was to compare the sensitivity, specificity, and diagnostic accuracy of serum interleukin-6, interleukin-8, beta 2-microglobulin, and C-reactive protein in the assessment of the severity of acute pancreatitis using commercial kits for their respective assays. Thirty-eight patients with acute pancreatitis (25 men, 13 women, mean age 59 years, range 16-97) were studied; the diagnosis was based on prolonged upper abdominal pain associated with a twofold increase of serum lipase, and it was confirmed by imaging techniques. According to the Atlanta criteria, 15 patients had severe illness and 23 had mild disease. The four serum markers were determined in all patients on admission, as well as daily for the following five days. On the first day of the disease, the sensitivity (calculated on patients with severe pancreatitis), specificity (calculated on patients with mild pancreatitis), and the diagnostic accuracy of these serum markers for establishing the severity of acute pancreatitis were 100%, 86%, and 91% for interleukin-6 (cutoff level 2.7 pg/ml); 100%, 81% and 88% for interleukin-8 (cutoff level 30 pg/ml); 58%, 81%, and 73% for beta 2-microglobulin (cutoff level 2.1 mg/liter); and 8%, 95%, and 64% for C-reactive protein (cutoff level 11 mg/dl). The results of our study indicate that, when assayed during the first 24 hr of disease onset, interleukin-6 and interleukin-8 are better markers than beta 2-microglobulin or C-reactive protein for evaluating the severity of acute pancreatitis.

Serum CA 242 in pancreatic cancer. Comparison with CA 19-9 and CEA.

Serum CA 242, CA 19-9 and CEA concentrations were determined in 94 subjects divided into 5 groups: Group 1 consisted of 22 healthy subjects; Group 2 consisted of 40 patients with pancreatic adenocarcinoma; according to Cubilla and Fitzgerald's classification, 11 tumours were Stage I, 4 were Stage II, and 25 were Stage III. Group 3 consisted of 10 chronic pancreatitis patients, group 4 of 10 acute pancreatitis patients, group 5 of 12 patients with nonpancreatic digestive carcinomas. Ten of these 12 patients had distant metastases. The sensitivity of CA 19-9 in the diagnosis of pancreatic cancer was higher than that of CEA and CA 242 (p < 0.05 and p < 0.005, respectively). In Stage I cancer patients the sensitivity of the markers studied was less than 50% (45% for CA 19-9, 18% for CEA, and 9% for CA 242) whereas most of the 25 patients with metastatic tumours of the pancreas had elevated serum levels of all 3 markers. The various combinations of the three markers did not significantly improve the sensitivity in diagnosing pancreatic cancer. No relationship was found between the localization of the tumour and the serum levels of the 3 markers studied. Similarly, no differences were found between patients with cholestasis and those without. The specificity of the 3 markers, evaluated in patients with benign pancreatic diseases, was 100% for CA 242, 90% for CA 199 and 70% for CEA.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum beta 2-microglobulin in chronic diseases of the pancreas.

Elevated serum concentrations of beta 2-Microglobulin (beta 2-MG) has been reported in a variety of chronic diseases and solid tumors. We determined serum beta 2-MG concentrations in 140 subjects divided into five groups: group 1, 34 patients with proven chronic pancreatitis, 8 of whom were studied during a painful relapse; group 2, 40 patients with pancreatic cancer staged according to the Cubilla-Fitzgerald classification; group 3, 40 healthy subjects; group 4, 10 patients with digestive nonpancreatic carcinomas; group 5, 16 patients with benign digestive nonpancreatic diseases. Serum soluble interleukin-2 receptor (sIL-2R) was also determined in all patients with pancreatic diseases as an index of immune system activation. In addition, serum CA 19-9 was assayed in patients of groups 2 and 4, and C-reactive protein (CRP) of groups 1 and 5. Renal function, evaluated by serum creatinine determination, was normal in all subjects studied. Patients with pancreatic cancer and those with chronic pancreatitis had serum concentrations of beta 2-MG significantly higher than those of healthy subjects (p < 0.001 and p < 0.005, respectively). Patients with stage I and stage III pancreatic cancer had similar serum levels of beta 2-MG, and these concentrations were significantly lower than those of patients with stage II tumors (p < 0.002 and p < 0.05, respectively). In chronic pancreatitis patients, those studied during painful relapse of the disease had serum concentrations of beta 2-MG similar to those studied during clinical remission.(ABSTRACT TRUNCATED AT 250 WORDS)

Indomethacin enhances serum 2'5'-oligoadenylate synthetase in patients with hepatitis B and C virus chronic active hepatitis.

Activation of the arachidonic acid metabolism seems to be one of the post-receptor mechanisms by which interferon-alpha induces antiviral protein synthesis. In this study, we evaluated the changes on serum 2'5'-oligoadenylate synthetase levels induced by acute administration of interferon-alpha, indomethacin and interferon-alpha plus indomethacin in 21 patients with hepatitis B or C virus chronic active hepatitis. Serum samples for 2'5'-oligoadenylate synthetase determination were collected in basal conditions and 8 h after the administration of interferon-alpha, indomethacin and interferon-alpha plus indomethacin. Compared to control value (mean +/- SE) (40.2 +/- 7.9 pg/dl) serum 2'5'-oligoadenylate synthetase concentration was significantly increased 1.5-fold after interferon-alpha (63.4 +/- 11, p < 0.001), 2.8-fold after indomethacin (115.5 +/- 21, p < 0.001) and 3.7-fold after interferon-alpha plus indomethacin (148.9 +/- 25.1, p < 0.001). When patients with different viral infections were considered separately, basal 2'5'-oligoadenylate synthetase concentrations were similar in both HBeAg- and HBeAb-positive patients, but about 2-fold higher in hepatitis C virus. Compared to the control value, interferon-alpha caused a 1.5-fold increase in HBeAg- and hepatitis C virus-positive and a 2-fold increase in HBeAb-positive patients. Indomethacin led to a 1.8-fold increase in HBeAg, a 2-fold increase in hepatitis C virus-positive and surprisingly a 6.8-fold increase in HBeAb-positive patients. Simultaneous administration of the two drugs had an additive effect on the 2'5'-oligoadenylate synthetase increase in HBeAg-positive (2.4-fold increase) and a synergistic effect in hepatitis C virus- and HBeAb-positive patients (2.7- and 10.2-fold increase, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

[Models of the organization of neonatal screening]. / Modelli di organizzazione degli screening neonatali.

The authors evaluate the different organizational strategies of a congenital hypothyroidism screening program. Positive and negative aspects of laboratory screening tests (TSH only, T4-supplemental TSH, TSH and T4), organization strategies (centralization or decentralization), recall and first follow-up criteria are examined. The authors consider that the necessity for an early diagnostic confirmation can be associated with a precise etiologic diagnosis and an evaluation of the prenatal severity of congenital hypothyroidism factors. Some European and North-American experiences are compared with the activity of a regional Italian screening center.

Morphologic and functional evaluation of the exocrine pancreas in beta-thalassemia major.

Thirty-nine consecutive patients with beta-thalassemia major and iron overload were studied by ultrasonography and serum pancreatic enzyme determination to assess the frequency and characteristics of pancreatic damage in this condition. The results were compared with those of a matched group of healthy controls. Most of the thalassemic patients had a markedly increased echogenicity of the pancreas and decreased size of the gland (p < 0.001 for both features as compared with controls). Both echogenicity and decreased size of the gland were significantly correlated (p < 0.001) with patient age and duration of transfusional therapy. Serum concentrations of trypsin and lipase were significantly lower (p < 0.02) in patients than in controls. The lowest enzyme values were found in older patients with longer duration of transfusional therapy, who also had the most marked sonographic changes. The results suggest that the exocrine pancreas is structurally and functionally damaged in the majority of thalassemic patients with iron overload.

Anti-hepatitis C virus antibodies amongst Italian homo-bisexual males.

The authors report on an anti-hepatitis C virus antibody (HCV Ab) prevalence (6.9%) in 622 homo-bisexual males from Northern Italy, voluntarily attending an HIV and STDs screening program in the period 1984-89. The anti-HCV antibody prevalence shows a significant correlation with: i) presence of serological markers for HBV (O.R. = 3.12, 95% C.I. = 1.53-6.52) and HIV (O.R. = 12.09; C.I. = 6.52-22.52) infection; ii) a stable relationship with an anti-HCV antibody positive partner (O.R. = 7.79; 95% C.I. = 2.50-23.90); iii) more than twenty different male partners per year (O.R. = 2.55; 95% C.I. = 1.17-5.66). These data demonstrate the existence of a sexual transmission of HCV among homosexuals. This route might contribute in maintaining endemic levels of HCV infection in the homo-bisexual population and it might represent an important way of spreading the virus in the general population too.

Serum pancreatic enzymes in HIV-seropositive patients.

Serum concentrations of trypsin and elastase I were determined in 109 HIV Ab-positive patients (52 asymptomatic HIV-infected patients, 25 with lymphadenopathy syndrome, and 32 with acquired immunodeficiency syndrome) to assess the prevalence of possible pancreatic damage in these patients. Serum trypsin was abnormally elevated in 46 of the 109 patients (42.2%): 19 of the 52 asymptomatic HIV-infected patients (36.6%), 9 of the 25 with lymphadenopathy syndrome (36%), and 18 of the 32 with acquired immunodeficiency syndrome (56.3%). Serum elastase 1 was elevated in 14 of the 109 HIV Ab-positive patients (12.8%): 3 of the 52 asymptomatic HIV-infected patients (5.8%), 3 of the 25 with lymphadenopathy syndrome (12%), and 8 of the 32 with acquired immunodeficiency syndrome (25%). None of the patients with abnormally high serum pancreatic enzyme concentrations had clinically evident pancreatic disease. There was no statistically significant difference in serum levels of trypsin and elastase I between drug addicts and nonaddicts, between alcoholics and nonalcoholics, or between those with cytomegalovirus infection and those without. A significant inverse relationship was found between serum enzyme concentrations and the number of CD4+ lymphocytes. The results of this study show that high levels of serum trypsin and elastase are present in an elevated percentage of patients with acquired immunodeficiency syndrome, suggesting that the pancreas is frequently damaged in this disease. The finding of abnormally high serum enzyme concentrations not only in patients with AIDS, but also in asymptomatic carriers and in patients with lymphadenopathy syndrome suggests an association between HIV infection and the development of pancreatic lesions.

Epidemiology of hepatitis C in a population of hemodialysis patients.

A search for antibodies against hepatitis C virus (HCV) was performed in 185 patients on chronic hemodialysis by means of 1st and 2nd generation ELISA tests. Immunoblot assays were performed on positive sera. This study shows a 38% prevalence of HCV-positive patients in our dialysis population according to the 2nd generation ELISA test which shows a higher specificity and sensitivity when compared to the 1st generation one (38 vs. 20%). A correlation was found between the prevalence of HCV-positive patients and how long they had been on dialysis and how many blood transfusions they had received.

Serum lipase assay. A test of choice in acute pancreatitis.

We evaluated the clinical accuracy of an automated turbidimetric assay for serum lipase determination in order to screen for acute pancreatic damage. Seventy patients with pancreatic and thirty with nonpancreatic digestive diseases were studied. Fifty-two healthy subjects were also studied as controls. Serum lipase concentrations were abnormally high in all patients with acute pancreatitis and in 3 (10%) in the group of 30 patients with nonpancreatic acute abdomen. In the 35 patients with chronic pancreatitis studied during clinical remission, serum lipase levels were abnormally high in 8 (23%), and abnormally low in 3 (9%). In the 9 patients with pancreatic cancer, 4 (44%) had abnormally elevated serum lipase values and 1 (11%) abnormally low. The results indicate that serum lipase determination is useful in the emergency diagnosis of acute pancreatic damage because of its high sensitivity and specificity. In patients with chronic pancreatitis and in patients with pancreatic carcinoma serum lipase determination is of limited value.

Improvements in automated analysis of catecholamine and related metabolites in biological samples by column-switching high-performance liquid chromatography.

Previously two fully automated methods based on column switching and high-performance liquid chromatography have been described, one for plasma and urinary catecholamines and the other for catecholamine urinary metabolites. Improvements in these methods, after 3 years of routine application, are now reported. The sample processing scheme was changed in order to eliminate memory effects and, in the procedure for plasma catecholamines, a pre-analytical deproteinization step was added which enhances the analytical column lifetime. The applied voltages for the electrochemical detector have been optimized, resulting in an automated method, suitable for the simultaneous determination of vanillylmandelic acid, 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid. The sensitivity of the methods allows the detection of 2-3 ng/l of plasma catecholamines and 0.01-0.06 mg/l of urinary metabolites. Also, it is possible to switch from one method to the other in only 30 min. The normal values obtained from 200 healthy people are reported, together with a list of 57 potential interfering substances tested.

[Immunoenzymatic method for the quantitative determination of specific IgE antibodies]. / Metodica immunoenzimatica per la determinazione qualitativa di anticorpi IgE specifici.

We describe a qualitative immunoenzymatic method for the determination of specific IgE antibodies in serum or plasma, that can be used as screening in case of suspected IgE-mediated allergy. The comparative analysis of results obtained with this new method and with standard methods for in vitro determination of specific IgE shows a perfect correspondence of the results for both sera with levels of specific IgE or non detectable levels of specific IgE.

Full automation of catecholamine metabolite determination by column switching and high-performance liquid chromatography.

The urinary catecholamine metabolites, vanimandelic acid, homovanillic acid, 3,4-dihydroxyphenylacetic acid and 5-hydroxyindoleacetic acid, were extracted on a silica-bonded strong-anion-exchanger cartridge (SAX) and then injected into an high-performance liquid chromatographic (HPLC) system by column switching. Chromatography was performed on a reversed-phase analytical column with electrochemical detection. Full automation was obtained by coupling two devices: a solid-phase automatic sampler and intelligent autosampler. For each substance the recovery was greater than 95% and the coefficient of variation was ca. 3%; the analysis takes 11 min. Substance instability problems are overcome, because the samples are extracted and injected in rapid succession. The normal values and correlation with manual HPLC were established for a large number of samples.

Role of serum pancreatic enzyme assays in diagnosis of pancreatic disease.

The serum behavior of amylase, pancreatic isoamylase, lipase, trypsinogen, and elastase 1 was studied in 145 patients with pancreatic disease and in 66 patients with abdominal pain of nonpancreatic origin, for the purpose of evaluating the relative diagnostic utility of their assays. In 34 patients with acute pancreatitis, serum lipase, trypsinogen, and elastase 1 were elevated in all 34, pancreatic isoamylase in 33 (97%) and amylase in 30 (88%). Ten of these acute pancreatitis patients were followed sequentially for seven days: the variations in their serum enzyme levels were parallel, although the lipase, trypsinogen, and particularly the elastase 1 elevations persisted longer than did those of amylase and pancreatic isoamylase. Among the patients with chronic pancreatitis, either in painful relapse (N = 19) or with pancreatic cysts (N = 15), the respective percentages of enzymes elevations were: 79 and 80% for elastase 1, 68 and 67% for trypsinogen, 63 and 73% for pancreatic isoamylase, 58 and 60% for lipase, 53 and 60% for amylase. In the 52 chronic pancreatitis patients studied during clinical remission, serum enzyme behavior varied greatly, and a majority of the assays (60%) were normal; even in the case of severe pancreatic exocrine insufficiency, normal as well as abnormally high and low enzyme values were seen. Highly variable enzyme behavior was also seen in the 40 patients with pancreatic cancer, and elastase I was the most frequently (35%) elevated enzyme in this group as well. Among the patients with abdominal pain of nonpancreatic origin, abnormally high enzyme levels were present in percentages ranging from 6% for lipase to 21% for trypsinogen.(ABSTRACT TRUNCATED AT 250 WORDS)