Use of a comprehensive diagnostic algorithm for Anisakis allergy in a high seroprevalence Mediterranean setting.

Summary: Background.Diagnosis of anisakis allergy (AA) is based on the skin prick test (SPT) and specific IgE (sIgE) determination. Anyway, false positivity cases are due to cross reactivity with numerous allergens. The aim of the study was to evaluate the reliability of a comprehensive diagnostic algorithm for the AA. Methods.An observational study was conducted on a sample of consecutive subjects accessing the allergology outpatient ambulatories of two hospitals located in Western Sicily. All the recruited outpatients were tested by Skin Prick Test performed using anisakis extracts by ALK-Abellò (Madrid, Spain). Specific IgE dosage for anisakis extracts was then performed by using ImmunoCAP250 (Immunodiagnostics Uppsala, Sweden). Consequently, outpatients who tested positive to first line tests underwent sIgE testing for ascaris and tropomyosin. Lastly, outpatients positive to the first line were invited to be further tested by basophil activation test (BAT) by using Flow CAST kit and anisakis commercial extract (Bühlmann Laboratories AG, Schönenbuch, Switzerland), as confirmatory analysis. Results.One hundred and eleven outpatients with an anamnesis suggestive of sensitization to anisakis (AS) and 466 subjects with chronic urticaria (CU) were recruited in the study. Of these, 22 with AS and 41 with CU showed a sensitization to anisakis allergens. The diagnostic algorithm revealed that 8.8% of outpatients who tested positive to sIgE determination were affected by CU, while 82.5% of all the sIgE positivity was related to cross-reactivity. Overall, a genuine anisakis seroprevalence of 2.3% was documented. Within a sub-sample of 15 subjects with clinical symptoms related to AA, n. 8 showed a real positivity after BAT. A greater response to A. pegreffii allergens as compared to A. simplex was reported. Conclusions.Our preliminary findings support the high clinical specificity of BAT for AA diagnosis, suggesting implementing this method in a comprehensive diagnostic algorithm.

IgA anti-actin antibodies ELISA in coeliac disease: a multicentre study.

BACKGROUND: Previous studies have demonstrated that serum anti-actin antibodies are a reliable marker of intestinal damage severity in coeliac disease. AIMS: To validate in a multicentre study the clinical usefulness of serum IgA anti-actin antibody ELISA and its possible use in monitoring intestinal mucosa lesions during gluten-free diet. PATIENTS AND METHODS: Four centres recruited 205 newly diagnosed coeliac disease patients with villous atrophy, 80 healthy controls and 81 "disease" controls. Twelve coeliac disease patients on gluten-free diet but with persistent symptoms underwent serum IgA anti-actin antibody assay and intestinal histology evaluation. IgA anti-actin antibody ELISA was performed with a commercial kit. All coeliac disease patients underwent intestinal histology study. RESULTS: IgA anti-actin antibodies showed a sensitivity of 80% and a specificity of 85% in the diagnosis of coeliac disease patients with villous atrophy. The area under the receiving operator curve for anti-actin antibodies was 0.873 [95% C.I. 0.805-0.899]. Serum anti-actin antibodies values were significantly higher in coeliac disease patients than in healthy or "disease" controls (P<0.0001). Serum anti-actin antibodies were positive in 41 of the 60 coeliac disease patients with mild intestinal histology lesions (69%) and in 123 of the 145 with severe lesions (85.3%) (P<0.05). There was a significant inverse correlation between anti-actin antibody values and the villi/crypts ratio (r=-0.423; P<0.0001). In the 12 coeliac disease patients on gluten-free diet who underwent re-evaluation as they were persistently symptomatic, intestinal histology showed three cases with persistent villous atrophy: all of these were positive for serum anti-actin antibodies ELISA, whereas both serum anti-tTG and EmAs were negative. The other nine patients showed normal intestinal villi and were negative for serum anti-actin antibodies. CONCLUSIONS: Anti-actin antibodies are a reliable marker of severe intestinal mucosa damage in coeliac disease patients and a simple ELISA technique offers an accurate method for their determination. These antibodies seem to be a very reliable marker of persistent intestinal damage in coeliac disease patients.

Usefulness of N-terminal pro-B-type natriuretic peptide levels in predicting residual myocardial ischemia in patients with ST elevation acute myocardial infarction.

AIM: N-terminal pro-b-type natriuretic peptide (NT pro-BNP) is a neurohormone synthesized predominantly in ventricular myocardium. In patients with symptoms of heart failure, elevation in NT pro-BNP accurately identifies ventricular dysfunction. However, NT pro-BNP levels are not specific for ventricular dysfunction in patients who do not have overt symptoms of heart failure, suggesting that other cardiac processes such as myocardial ischemia may also cause elevation in NT pro-BNP. The study was aimed to determine whether NT pro-BNP elevations are associated with myocardial ischemia. METHODS: One hundred and thirty patients (104 males, 26 females, mean age 61+12 years), with ST elevation acute myocardial infarction (STEMI) and preserved left ventricular ejection fraction (>45%) at echocardiography performed at entry, from February 2003 and February 2004 were enrolled. In all patients NT pro-BNP plasma levels were checked at entry and 4-5 days after symptoms onset. In addition, maximal or symptom-limited exercise treadmill test (Bruce protocol), and myocardial perfusion scintigraphy using [(99m)Tc]Tetrofosmin single photon emission computed tomography (SPECT) imaging were performed within 30 days of STEMI. Ischemia was defined as reversible perfusion abnormalities. RESULTS: Of the 130 participants, 66 (51%) had inducible ischemia. Compared with patients in the lowest tertile, those in the highest tertile of NT pro-BNP had a greater significant risk of residual ischemia (odds ratio: 8.66; 95% CI, 3.90 to 19.24). Nevertheless patients in the highest tertile were older (64.19+/-10.80 years versus 55.90+/-9.67 years, P = 0.0001), had a lower left ventricular ejection fraction (49.70+13.46% versus 59.49+/-6.58%, P = 0.0001) and had a great rate of acute myocardial infarction (anterior acute myocardial infarction = 40.63% versus 25%). CONCLUSIONS: Elevated levels of NT pro-BNP are associated with residual myocardial ischemia among patients with STEMI and preserved left ventricular ejection fraction, as demonstrated by perfusion defect on SPECT imaging, suggesting that these patients may need further evaluation for stratification of the future risk of fatal events. The observed association between NT pro-BNP levels and ischemia may explain because tests for NT pro-BNP are not specific for ventricular dysfunction among patients with coronary artery disease.

Toxoplasma gondii infections in sheep in Sicily, southern Italy.

The aim of the study was to determine the burden of Toxoplasma gondii-infections in sheep in Sicily, southern Italy and the risk factors for infection. Sera from 1961 sheep were collected just before slaughtering from 62 farms located in 8 out of 9 Sicilian administrative districts. The sera were analysed for Toxoplasma-specific IgG antibodies using commercially available enzyme-linked immunosorbent assay. Sheep less than 4 weeks old were further analysed by ELISA for Toxoplasma-specific IgM-antibodies. Data on farm size and location were obtained from slaughterhouse sanitary reports and through structured telephone interviews of the veterinary officers from public health districts. The overall seroprevalence of Toxoplasma-specific IgG-antibodies were 49.9% (937/1876) by ELISA. Eighty-seven (54/62) percent of the farms had at least one Toxoplasma-positive animal. All the farms fed the animals outdoor on pasture and only one was claiming organic farming. Having cats on the farm, age of the animals, farm size and the use of surface water sources for drinking were all significantly associated with T. gondii-infected animals on the farm. T. gondii infection in mutton used for human consumption is very prevalent, and eating unprocessed sheep and lamb meat has a high risk of transmitting infections to humans. The presence of cats on the farm, farm size and using surface water as drinking water for the animals were risk factors for infection in sheep, with age as a significant confounder.

Diagnosis of congenital toxoplasmosis: pre- and post-natal evaluation in Sicilian (Italy) epidemiological area. Preliminary data.

To evaluate the usefulness of conventional serological methods with western blot assay (WB) in congenital toxoplasmosis diagnosis, we prospectively enrolled in a clinical and serological follow-up all pregnant women with Toxoplasma gondii infection and their offspring, referred to us from October 2004. Western blot and standard serological test were performed on sera collected from mother during pregnancy and from mother and child at birth, at postpartum month 1-3-6-9 and 12. At this point in time, 22 pregnant women and 14 infants have completed the follow-up. 4 newborns were infected and 2 had specific toxoplasmosis anomalies at the birth. In mothers without seroconversion, the WB performed during pregnancy demonstrates the highest accordance with postnatal follow-up whereas in 1 case the negative result of PCR analysis was not confirmed by postnatal observation. The detection of anti-T gondii IgG against 8 kDa accessory antigenic band and against the accessory band included between 35 and 40 kDa band in immunoblot assay was useful for diagnosis of acute phase but did not improve the evaluation of comparative postnatal profile. Althougth few infants have concluded the postnatal follow-up, the preliminary results showed a greater value of using a IgM and IgA WB test than other standard method for the early diagnosis of toxoplasmosis at birth also in child born to treated mothers. The comparative anti-T gondii IgG immunoblot profile of mother and child permitted us to reduce the time of ruling out infection in newborns born to mothers with probable or possible infection and/or when prenatal diagnosis is negative or not performed.

RPBS: a web resource for structural bioinformatics.

RPBS (Ressource Parisienne en Bioinformatique Structurale) is a resource dedicated primarily to structural bioinformatics. It is the result of a joint effort by several teams to set up an interface that offers original and powerful methods in the field. As an illustration, we focus here on three such methods uniquely available at RPBS: AUTOMAT for sequence databank scanning, YAKUSA for structure databank scanning and WLOOP for homology loop modelling. The RPBS server can be accessed at http://bioserv.rpbs.jussieu.fr/ and the specific services at http://bioserv.rpbs.jussieu.fr/SpecificServices.html.

Evaluation of antibodies to thymine ROS-modified DNA poly(dT), in patients with immunologic disorders: relationships to anti n-DNA and anti ENA autoantibodies.

BACKGROUND: Hydroxyl radical, one of the most potent of all reactive oxygen species, is known to modify adenine and thymine in cellular DNA, producing some modified DNA fragments (ROS-DNA) with different antigenic properties. Despite several in vitro studies about ROS-DNA, data regarding their clinical significance are scanty. The aim of our study was to seek out the presence and clinical significance of the anti poly(dT) auto-antibodies in a group of patients suspected of autoimmune disease. METHODS: We initially evaluated more than 900 consecutive sera of hospitalized patients (range age from 6 to 70 yrs) referred to our laboratory during 18 months. Anti n-DNA, anti-ENA and poly(dT) autoantibodies were performed on 158 ANA positive sera and 28 ANA negative sera from patients strongly suspected of rheumatic diseases or affected by HCV infection. RESULTS: Anti poly (dT) were found in 22 out of 186 sera. As regards the clinical evaluation, 8 patients were affected by SLE, 5 by Scleroderma, 3 by HCV-related chronic hepatitis, 4 by recurrent abortions (without presence of the anti-cardiolipin antibodies and other clinical symptoms). In two patients the ACR criteria and the clinical aspects did not allow a definite diagnosis. Anti-Histones were detected in 18 out of 22 poly (dT) positive patients. CONCLUSIONS: Our data suggest that anti poly(dT) autoantibodies are sensitive markers of various autoimmune diseases, but with a minor specificity as compared to anti n-DNA for the diagnosis of SLE.

[Evaluation of the liberation of urinary necrosis enzymes (NAG-AAP) after administration of plasma expanders: study of dextran 40, hydroxyethyl starch and polymerized gelatin]. / Valutazione della liberazione degli enzimi urinari di necrosi (NAG-AAP) dopo somministrazione di plasmaexpanders: studio su destrano 40, amido-idrossietilico e gelatina polimerizzata.

We studied the 24-hour urinary elimination of enzymatic markers of renal tubular necrosis (NAG-AAP) in 21 patients (mean age 34.6 years old) who were treated with plasma expanders before peridural anesthesia. The patients were divided into three groups of seven subjects each: - 14 ml/Kg -1 of dextran 40 was administered to group 1 - 14 ml/Kg -1 of gelatin was administered to group 2 - 14 ml/Kg -1 of hydroxyethyl starch was administered to group 3. Urinary elimination of N-acetylglucosaminidase and of alanine aminopeptidase was determined in the 24-hour urine the day before surgery (controls), the day of surgery (G1) and the day after surgery (G2). The values of the samples, taken after plasma expander administration, did differ significantly from the control values (G1, G2). Therefore the administration of 14 ml/Kg -1 of dextran, gelatin or hydroxyethyl starch does not affect the renal tubular epithelium.