Hook-effect in MAGLUMI immunoassay for serum anti-GAD antibodies in neurological disorders: When "wrong" matrix is the right choice.

Antibodies against glutamic acid decarboxylase (anti-GAD) are a valuable diagnostic tool to detect severe autoimmune conditions as type 1 diabetes mellitus (T1DM) and anti-GAD related neurological disorders, having the latter more often anti-GAD concentrations in serum multiple times higher than in the former. Automated immunoassays, either with ELISA or chemiluminescent technology, are validated for diagnostic use in serum with analytical ranges suitable for T1DM diagnosis. In a patient presenting with a suspected autoimmune ataxia, anti-GAD testing on an automated chemiluminescent immunoassay (CLIA) resulted in slightly abnormal concentrations in serum (39.2 KIU/L) and very high concentrations in CSF (>280 KIU/L), thus prompting to proceed to serum dilutions to exclude a false negative result and a misdiagnosis. Different dilutions of serum resulted in nonlinear concentrations with endpoint result of 276,500 KIU/L at dilution 1:1000. CSF dilution was instead linear with endpoint result of 4050 KIU/L. In this case report we found that anti-GAD testing in CSF was essential to establish the clinical diagnosis and to suspect hook-effect in serum due to the excess of autoantibodies in this severe autoimmune condition.

A non-classical presentation of APECED in a family with heterozygous R203X AIRE gene mutation.

PURPOSE: Biallelic loss-of-function mutations of AIRE cause the autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) syndrome. However, single nucleotide mutations may cause a milder phenotype. In this paper, we describe an unusual and mild phenotype in a mother and her two children (son and daughter) who carry a rare heterozygous mutation of AIRE. METHODS AND RESULTS: The son presented with alopecia and subclinical hypothyroidism due to Hashimoto's Thyroiditis (HT); the daughter had alopecia, vaginal mycosis, stomach pains and subclinical hypothyroidism due to HT; and the mother had alopecia, vaginal mycosis and stomach pains. Organ- and non-organ-specific autoantibodies were evaluated as well as antibodies against interleukin-17A, -17F, -22 (IL-Abs) and interferon -α and -ω (IFN-Abs). The organ- and non-organ-specific autoantibodies screening was negative in the son, while the daughter was positive for liver-kidney microsomal antibodies (LKMAbs) and the mother was positive for glutamic acid decarboxylase antibodies (GADAbs). Daughter and mother were also positive for IFN-Abs. Analysis of the AIRE gene identified a rare heterozygous R203X mutation in all three family members. CONCLUSIONS: We describe for a first time a family with heterozygous R203X AIRE mutation causing an APECED-like condition, as confirmed by presence of IFN-Abs. The unusual mild phenotype should be reassuring for the patients and assist in their clinical management.

Autoimmune polyendocrine syndrome type 1 (APECED) in the Indian population: case report and review of a series of 45 patients.

BACKGROUND: Autoimmune polyendocrinopathy-candidiasis-ectodermal-dystrophy (APECED) or autoimmune polyglandular syndrome type 1 (APS-1) is a rare autosomal recessive genetic disease due to mutations in the AIRE (AutoImmune REgulator) gene. The clinical diagnosis is classically based on the presence of at least two of the three main components: chronic mucocutaneous candidiasis, hypoparathyroidism and primary adrenal insufficiency. Patients often suffer from other endocrine or non-endocrine autoimmune conditions throughout life. APECED etiopathogenesis is mediated by T lymphocytes. Autoantibodies against proteins of the affected organs are found in the serum of APECED patients as well as neutralizing antibodies against cytokines. We report here the clinical and genetic characteristics of 45 Indian APECED patients in comparison to Finnish, Sardinian, Turkish and North/South American cohorts from their published results. We also report a new case of APECED of Indian origin, a 2-year old child suffering from chronic mucocutaneous candidiasis since the age of 8 months, with confirmatory AIRE homozygous mutation c.274C > T (p.R92W). CONCLUSION: With the inherent limitations of a retrospective study, analysis of Indian APECED patients suggested that compared to classic criteria, application of Ferre/Lionakis criteria validated in North/South American patients could help in earlier diagnosis in 3 of 8 (37.5%) patients for whom adequate information for evaluation was available.

Portadores faríngeos de Streptoccocus pneumoniae en dos grupos poblacionales. Ciudad Bolivar, Estado Bolívar, Venezuela

Resumen Introducción Streptococcus pneumoniae es un patógeno para el ser humano que precisa de una previa colonización faríngea para causar enfermedad, cuya morbimortalidad se acentúa en los menores de 6 años y los mayores de 65 años de edad. Objetivo Determinar la prevalencia de portadores faríngeos de Streptococcus pneumoniae en dos grupos poblacionales en Ciudad Bolívar, Estado Bolívar. Metodología Durante los años 2009 a 2010, se evaluaron dos grupos: uno que incluyó a 66 individuos de la tercera edad institucionalizados en el Asilo "San Vicente de Paúl" y Geriátrico "Carlos Fragachan", con una edad promedio de 75 años ± 6 años y otro que abarcó a preescolares y escolares de 2 a 12 años hospitalizados en los Servicios de Pediatría y del área de emergencia del Complejo Hospitalario Universitario "Ruíz y Páez". A cada individuo se le tomó una muestra faríngea, la cual fue procesada según los lineamientos establecidos por la Sociedad Americana de Microbiología. Se estudiaron 80 exudados de origen faríngeo. Resultados En el grupo de los individuos de la tercera edad se identificaron 10 casos (15,15%) de portadores faríngeos de S. pneumoniae. El 70% (n=7) correspondía al género masculino. En todas las edades se diagnosticaron casos, frecuentemente observados en el grupo entre 71 a 80 años de edad (n=4; 40%). El 46,2% de los portadores faríngeos de la tercera edad refirieron antecedentes tabáquicos. Se observaron diferencias estadísticas significativas entre el estado de portador de neumococo y la diabetes mellitus tipo II y estado de vacunación contra la bacteria, En el grupo de los pacientes preescolares y escolares hospitalizados, se aislaron 3 cepas de S. pneumoniae que representó el 3,75% del total. El mayor porcentaje de muestras positivas se obtuvo en el grupo de 2 a 4 años, con predominio del género masculino (2,5%). En el grupo de la población infantil hospitalizada, se observaron diferencias estadísticamente significativas entre el estado de portador y antecedentes personales como asma, resfriado común a repetición, infección urinaria e infección de piel y tejido blando. No estaban vacunados contra el neumococo o solo cumplieron una sola dosis. En ambas investigaciones se determinaron 13 cepas de Streptococcus pneumoniae, las cuales mostraron un perfil de resistencia a la Penicilina, por método del disco de Oxacilina, del 50%; mientras que la totalidad de las cepas aisladas resultaron con alta resistencia a Macrólidos, Clindamicina y Sulfamidas, y sensibles a Vancomicina. Conclusión Se identificó una baja prevalencia de individuos colonizados por Streptococcus pneumoniae, sin embargo, se debe considerar la investigación de colonización faríngea por neumococo como un buen método por ser simple y fácil de obtener muestras bacterianas y poder reflejar la progresión de la resistencia bacteriana en grupos de riesgo.

Abstract Introduction Streptococcus pneumonia is a pathogen for humans that requires a prior pharyngeal colonization to cause disease, whose mortality is accentuated in children 6 years and older than 65 years of age. Objective To assess the prevalence of pharyngeal carriage of S pneumonia in two population groups in Ciudad Bolívar, Bolívar State. Methodology . During the years 2009 and 2010, two groups were evaluated: one which included 66 senior individuals institutionalized in the asylum "San Vicente de Paúl" and the geriatric "Carlos Fragachan", with an average of 75 years ± 6 years age and another that it comprised preschool and school aged 2 to 12 hospitalized in Pediatrics and the area of the University Hospital emergency services "Ruiz and Páez". Each individual took a pharyngeal sample, which was processed according to the guidelines established by the American society for of microbiology. 80 exudates from pharyngeal origin were studied. Results Between elderly individuals 10 cases were identified (15.15%) of S. pneumonia pharyngeal carriers. 70% (n = 7) corresponded to the male gender. In all the ages were diagnosed cases, often observed in the group between 71 to 80 years of age (n = 4; 40%). 46.2% of pharyngeal carriers concerned smoking history. Were observed significant statistical differences between the pneumococcus`s state carrier and type II diabetes mellitus and vaccination´s state against the bacterium, In the group of preschool and school inpatients, 3 strains of S. pneumonia that accounted for 3.75% of the total were isolated. The highest percentage of positive specimens was obtained in the group of 2 to 4 years, with a predominance of the male gender (2.5%). Statistically significant differences were observed in the hospitalized children, between the carrier´s state and personal history as asthma, cold common, urinary tract infection and infection of skin and tissue soft. They were not vaccinated against the pneumococcus or met only a single dose. Both studies identified 13 strains of Streptococcus pneumoniae, which showed a profile of resistance to penicillin, by method of disk Oxacillin, of 50%; While all of the isolates were with high resistance to macrolides and clindamycin, sulfonamides, and sensitive to Vancomycin. Conclusion We identified a low prevalence of individuals colonized by Streptococcus pneumoniae, however should be consider the research of colonization of the pharynx by Pneumococcus as a good method to be simple and easy to obtain bacterial samples and to reflect the progression of bacterial resistance in risk groups.

Fish Ecology and Evolution in the World's Oxygen Minimum Zones and Implications of Ocean Deoxygenation.

Oxygen minimum zones (OMZs) and oxygen limited zones (OLZs) are important oceanographic features in the Pacific, Atlantic, and Indian Ocean, and are characterized by hypoxic conditions that are physiologically challenging for demersal fish. Thickness, depth of the upper boundary, minimum oxygen levels, local temperatures, and diurnal, seasonal, and interannual oxycline variability differ regionally, with the thickest and shallowest OMZs occurring in the subtropics and tropics. Although most fish are not hypoxia-tolerant, at least 77 demersal fish species from 16 orders have evolved physiological, behavioural, and morphological adaptations that allow them to live under the severely hypoxic, hypercapnic, and at times sulphidic conditions found in OMZs. Tolerance to OMZ conditions has evolved multiple times in multiple groups with no single fish family or genus exploiting all OMZs globally. Severely hypoxic conditions in OMZs lead to decreased demersal fish diversity, but fish density trends are variable and dependent on region-specific thresholds. Some OMZ-adapted fish species are more hypoxia-tolerant than most megafaunal invertebrates and are present even when most invertebrates are excluded. Expansions and contractions of OMZs in the past have affected fish evolution and diversity. Current patterns of ocean warming are leading to ocean deoxygenation, causing the expansion and shoaling of OMZs, which is expected to decrease demersal fish diversity and alter trophic pathways on affected margins. Habitat compression is expected for hypoxia-intolerant species, causing increased susceptibility to overfishing for fisheries species. Demersal fisheries are likely to be negatively impacted overall by the expansion of OMZs in a warming world.

Prevalencia de patología cardíaca en la enfermedad de Chagas: años 2004-2012. Programa y Red de Chagas de CABA / Prevalence of cardiac pathology in Chagas disease: 2004 -2012. Program and Chagas CABA network

El Instituto Universitario de Ciencias de las Salud ha mostrado un particular compromiso con la formación de sus estudiantes en la estrategia de Atención Primaria de la Salud, con las prácticas asistenciales dedicadas al 1er nivel de atención ambulatoria y a las patologías prevalentes en ese ámbito. Del mismo modo se han desenvuelto las actividades de formación en investigación. Como exponente de esa orientación, la revista Ciencias de la Salud publicó en el Vol. 2, N°1, 2011:4-9, el artículo “Prevalencia de la Enfermedad de Chagas” de Érica G. Morais, que había obtenido el premio “Futuros Líderes”, otorgado por el Curso Anual Internacional de Investigación en Ciencias de la Salud (IUCS-AMA, Prof. Carlos Álvarez Bermúdez). Aquella investigación formaba parte de un proyecto más amplio realizado en el Hospital Teodoro Álvarez entre 2004 y 2012, en el que participaron un conjunto de investigadores, que compartieron la autoría de la actual publicación. El Dr. Jorge Mitelman, Prosecretario de Ciencia y Técnica del IUCS e integrante de ese equipo, preparó además una reseña sobre la jornada del INCOSUR, realizada en abril del presente año, describiendo asimismo el proceso de desarrollo de la Ciudad de Buenos Aires, como área no endémica, para encarar las consecuencias de la enfermedad de Chagas

[Risk management: the opinions of nursing staff and coordinators in a local health district in the Piemonte region (Italy)]. / Il rischio clinico: opinioni di infermieri e coordinatori dell'ASL 17 della Regione Piemonte.

Clinical risk management includes a set of clinical and administrative activities performed to identify, evaluate and reduce risks for patients, staff and visitors as well as the organization itself. The first fundamental step in risk management is to evaluate risk factors; it is impossible to implement corrective actions and modify and eliminate risk factors if these are not known. The aim of this study was to evaluate the degree to which selected sentinel events were perceived by nursing staff as being severe and whether the degree of perceived severity of an event was associated with specific variables such as nursing work area (medical, surgical, intensive care), years of experience, degree and position. The study also aimed to evaluate the level of knowledge of clinical risk management, identify the main categories of errors within the organization, and evaluate nursing staff opinions on the use of an anonymous system to report errors. A sample of 98 nurses (91 female, 7 male), working in three hospitals in a local health district in the Piemonte region (Italy) participated in the study. The mean age of participants was 37 years (range 22-61). Participants were interviewed between 30 October and 6 November 2006, by using a structured questionnaire. Most participants (93%) were aware of the definition of risk management but over 60% did not use any tool for identifying clinical errors. Nurses perceived infections to be the most serious error, followed by medication errors and surgery and post-operative complications. Almost all participants (99%) considered it right to report errors and 59% admitted to having made errors, most frequently medication errors. Over 90% of participants agreed that an anonymous report form should be used for reporting. Healthcare professionals' willingness to cooperate and their ability to not play down the importance of sentinel events but rather voluntarily bring these to light are essential to the success of risk management in an organization. Clearly, patient safety does not depend on the single individual but rather on an interdisciplinary approach to problem notification and solving and collaboration among interdisciplinary team members.

DNA microarray analysis for the detection of mutations in hemophilia A.

BACKGROUND: Congenital deficiency of factor (F) VIII results in the inherited X-linked bleeding disorder hemophilia A. More than 900 different mutations are reported in the hemophilia A mutation database with the largest number of mutations being single nucleotide substitutions distributed throughout the gene. Complicating the molecular characterization of this disease is the complexity of the F8 gene, the mutational heterogeneity, and technical limitations of the current mutation detection techniques. OBJECTIVE: Development of a DNA oligonucleotide microarray-based technique for F8 gene analysis to detect hemophilia A mutations. METHODS: To construct the oligonucleotide DNA microarray system: a total of 720, one base pair overlapping, 25-mer perfect match probes were designed from six exons of the F8 gene. Twenty-two different F8 gene mutations previously identified by CSGE and DNA sequence analysis were tested by using a loss-of-signal analysis approach. Differentially labeled wild type and hemophilic samples were co-hybridized to the array. Sequence alterations were detected by quantifying relative losses of test sample hybridization signals to the perfectly matched probes. RESULTS: A total of 22 different F8 mutations were tested. To test the sensitivity of the system, a blinded study was performed on 16 of the samples. F8 gene mutations can be detected with 96% efficiency with this microarray system. CONCLUSION: This proof-of-principle study has demonstrated that a F8 DNA microarray platform is an alternative gene mutation analysis approach that has a high sensitivity, and reproducibility. The methodology is, however, expensive and time consuming, and with the reduction in sequencing costs, direct sequencing is now the most cost and time efficient strategy for hemophilia A mutation analysis.

Suicide gene therapy with HSV-TK in pancreatic cancer has no effect in vivo in a mouse model.

AIM: To study in vivo whether pancreatic cancer tumour growth and metastasis can be modified by a gene construct with HSV-TK suicide gene and IL2 co-expression. METHODS: Seventy-eight female SCID mice were i.p. inoculated with retrovirally transduced or control MIA PaCa 2, CAPAN-1 and PANC-1 cell lines. The animals were then randomly selected for saline or ganciclovir (GCV) treatment from the second week, for a total of two weeks. RESULTS: Most inoculated mice developed tumour nodules and spleen metastases. The liver was colonized by control CAPAN-1 and MIA PaCa 2, but not by PANC-1. Tumours in transduced MIA PaCa 2 cell injected mice were smaller, and in transduced CAPAN-1 injected mice larger, than in control-inoculated mice. There were increased pancreatic and decreased spleen metastases from transduced CAPAN-1, and diminished liver involvement from transduced MIA PaCa 2. No differences were found between mice inoculated with transduced and control PANC-1 cell lines. GCV treatment had no effect on tumour's size or metastases. CONCLUSIONS: The HSV-TK suicide gene does not confer GCV sensitivity to pancreatic cancer in this in vivo model. Different pancreatic cancer cell lines cause different growth and metastasis patterns after inoculation in SCID mice, possibly because of variations in their inherent characteristics. The different effects of our vector on cell growth and metastasis may be attributable to the effects of the immunostimulatory cytokine IL2.

Helicobacter pylori virulence genes and host IL-1RN and IL-1beta genes interplay in favouring the development of peptic ulcer and intestinal metaplasia.

Helicobacter pylori infection outcome might depend on genotypic polymorphisms of both the bacterium and the host. We ascertained: (1) the functionality of H. pylori oipA gene; (2) the polymorphism of the hostinterleukin (IL-1beta) gene (-31 C/T) and of the IL-1RN gene (intron 2 VNTR); (3) the association between the above genes and the histological and pathological outcome of H. pylori infection. One hundred and sixty-five H. pylori positive and 137 H. pylori negative subjects (23 gastric adenocarcinoma, 58 peptic ulcer, 221 gastritis) were studied. oipA was sequenced, IL-1beta was RFLP analysed. Antral and body mucosal biopsies were histologically evaluated. Functional oipA genes were correlated with cagA gene; both genes were significantly associated with gastritis activity, peptic ulcer and gastric adenocarcinoma. In these patients heterozygousIL-1RN 1/2 and IL-1beta C/T genotypes were more frequent than in gastritis patients. Intestinal metaplasia was associated with cagA, functional oipA and IL-1RN 2 allele. In conclusion, peptic ulcer and the preneoplastic intestinal metaplasia are associated with H. pylori virulence genes and with IL-1RN 2 host allele. An interplay between bacterial virulence factors and cytokines genotypes, is probably the main route causing H. pylori infection to lead to benign mild disease, benign severe disease or preneoplastic lesions.

[Diagnosis of Helicobacter pylori infection: comparison of techniques]. / Diagnostica dell'infezione da Helicobacter pylori: tecniche a confronto.

UNLABELLED: Several diagnostic assays are available for evaluating Helicobacter pylori infection: histological examination, culture of gastric biopsies, urea breath test and serology. Recently a new enzyme immunoassay has been introduced for the detection of H. pylori antigens in stool samples (HpSA). The aim of our study was to evaluate and compare diagnostic efficacy of HpSA with histological examination, culture, urea breath test and serology in a group of 95 patients. Patients were classified H. pylori positive (43) or negative (52) on the basis of histology, culture and urea breath test. HpSA optical densities were significantly higher in infected patients compared to those obtained in H. pylori-negative patients (t = 5.47, p < 0.001). Overall, with a fixed cut-off of 0.1 unit of optical density, the sensitivity was 79% and the specificity 100%. In the H. pylori positive patients, HpSA optical density correlated with bacterial load histologically evaluated in the gastric antrum (r = 0.405, p < 0.05) and was inverse correlated with levels of serum IgG elicited against H. pylori (r = -0.315, p < 0.05). Considering patients with a positive HpSA finding and/or levels of anti-H. pylori antibodies upper than 30 U/mL, sensitivity in detecting infected patients was 98%. IN CONCLUSION: (1) immunodetection of H. pylori antigens in stools is a good alternative of breath test; (2) a reduction in H. pylori density grade might be accompanied by low HpSA optical density, leading to a false negative result and (3) combining the HpSA determination with the serum detection of anti-H. pylori antibodies a better clinical sensitivity is obtained.

Role of anti-transglutaminase (anti-tTG), anti-gliadin, and anti-endomysium serum antibodies in diagnosing celiac disease: a comparison of four different commercial kits for anti-tTG determination.

The aims of this study were: (1) to compare the diagnostic efficacy for celiac disease (CD) diagnosis of serum determination of anti-gliadin (AG) (IgA and IgG) and anti-endomysium (AE) with that of anti-transglutaminase (AtTG); and (2) to compare the accuracy of four different assays to measure AtTG. We studied 72 children: the histological diagnosis of CD was made in 38 cases and excluded in the remaining 34 children. In fasting sera we measured AE, AG-IgA and IgG, and AtTG, the latter with four different commercial kits (Eurospital, Medipan, Inova, Arnika). Moreover AtTG was measured in a group of 58 CD children after a gluten-free diet. AE was positive in all but 1 case of CD patients (sensitivity = 97%); false positive results were found in 1/34 controls (specificity = 97%). When a specificity of 95% was fixed, the sensitivities were 97% for AE, 83% for AG-IgA, and 63% for AG-IgG; the sensitivities of anti-tTG were 90, 84, 84, and 75% when measured with Eurospital, Medipan, Inova, and Arnika kits respectively. The new AtTG seems to be accurate enough to be proposed as a noninvasive diagnostic tool for CD diagnosis; the 4 kits analyzed showed similar diagnostic efficacy.

Different effects of H. pylori water extracts on cytokines, pepsinogen C and gastrin mucosal release in patients with or without duodenal ulcer.

In the present study we ascertained whether cagA positive and negative H. pylori strains release water soluble products that can influence the production of gastric mucosal cytokines and endocrine (gastrin) or exocrine (pepsinogen C) secretion in 23 H. pylori positive and 19 H. pylori negative patients. Antral biopsies were obtained to classify inflammation, activity, atrophy, intestinal metaplasia and H. pylori density grade. The cagA gene was identified by means of the polymerase chain reaction (PCR) in H. pylori positive colonies after culture of mucosal samples. Three antral biopsies from each patient were incubated with (1.) Water extracts from cagA positive, (2.) Water extracts from cagA negative strains or (3.) H2O (control) at 37 degrees C in a CO2 incubator for 24 hrs. Gastrin, pepsinogen C, IL-1 beta, IL-8, GMCSF, and TNF alpha were measured in the supernatants and mucosal homogenates. H. pylori infection was significantly associated with an increased antral inflammation and activity (chi 2 = 21.7, p < 0.001 and chi 2 = 42.0, p < 0.001), and increased mucosal levels of IL-1 beta, IL-8 and TNF alpha. Water extracts from cagA positive strains enhanced the release of PGC in mucosal biopsy supernatants (p < 0.05) when patients were considered overall and the release of TNF alpha (p < 0.05) when only patients with duodenal ulcer were considered. Water extracts from cagA negative strains stimulated gastrin secretion (p < 0.05). None of the remaining cytokines were influenced by H. pylori water extracts. In conclusion, pepsinogen C and TNF alpha can be induced by cagA positive water extracts and may contribute to damage the gastric and duodenal mucosa. Our findings indicate that in patients with H. pylori infection the increase of the mucosal levels of IL-1 beta and IL-8 does not depend on H. pylori water soluble products, but probably depends on the entire bacterium.

ME-PCR for the identification of mutated K-ras in serum and bile of pancreatic cancer patients: an unsatisfactory technique for clinical applications.

Our aim was to assess the clinical reliability of mutated K-ras detection in serum or bile for the diagnosis of pancreatic cancer using ME-PCR. DNA was extracted from 1 ml serum obtained from 29 patients with pancreatic cancer and 12 control subjects. ME-PCR was optimized using a mixture of normal DNA added with different amounts of mutated DNA. The analysis of sera obtained from the 29 patients and of bile obtained from 11 pancreatic cancer patients demonstrated the presence of mutated K-ras in two (6.9%) and four cases (36%). By contrast K-ras was not amplifiable in any of the 12 serum samples obtained from healthy controls. In conclusion the DNA obtained from pancreatic cancer patients' sera is suitable for K-ras amplification and for the identification of codon 12 point mutations. However ME-PCR alone has an unsatisfactory sensitivity for the detection of pancreatic cancer using serum DNA as starting template.

CEA mRNA identification in peripheral blood is feasible for colorectal, but not for gastric or pancreatic cancer staging.

OBJECTIVE: It has been suggested that the molecular identification of cancer cells in the circulation may be useful in predicting the presence of micrometastasis in several cancer types. The aim of the present study was therefore to assess the feasibility of CEA mRNA identification in blood for diagnosing and staging colorectal, gastric and pancreatic cancer. METHODS: We studied 16 control subjects, 69 patients with colorectal (CRC), 30 with gastric (GC), 27 with pancreatic cancer (PC) and 8 with benign diseases of the pancreatobiliary tree. At diagnosis CEA mRNA was identified in peripheral blood by means of a RT-PCR procedure. RESULTS: The specificity of this test in control subjects was 94%, and its sensitivity in identifying CRC, GC and PC were 34, 37 and 41%, respectively. False-positive findings were recorded in 25% patients with benign diseases. No association was found between CEA mRNA and stage in patients with GC or PC. In CRC patients, positive CEA mRNA findings were correlated with local spread (chi(2) = 14.6, p<0.01), lymph node (chi(2) = 18.95, p<0.001) and distant metastasis (chi(2) = 11.3, p<0.001). In these cases, CEA mRNA, but not CEA, was entered in stepwise discriminant analysis to classify the presence of lymph node metastasis. CONCLUSIONS: The molecular detection of micrometastasis in the blood by means of CEA mRNA identification is feasible for colorectal, but not for gastric or pancreatic cancer staging. Further studies are needed in order to define the clinical utility of this marker also in follow-up protocols.

Antigastric autoantibodies in Helicobacter pylori infection: role in gastric mucosal inflammation.

UNLABELLED: The aim of the study was to ascertain whether there is an association between the presence of serum parietal cell autoantibodies (PCA) and: (1) Helicobacter pylori infection; (2) the presence and degree of gastritis and intestinal metaplasia; and (3) the H. pylori infecting strain. Gastric mucosal biopsies were obtained from 49 consecutive patients in order to assess and grade gastritis, make a histological diagnosis, and culture and genotype H. pylori. H. pylori infection was present in 26 patients (group 1), had been present in 17 patients (group 2), and the remaining 6 (group 3) had never had the infection. The infecting strain was cagA positive in 21 of 26 group 1 patients. Positive PCA results were found in 84%, 76%, and 14% of patients in groups 1, 2, and 3, respectively. PCA results were correlated with anti-H. pylori antibody titers (P<0.05). In group 2 patients, PCA were associated with the degree of antral gastritis (Fisher's exact test P<0.05). cagA status was not associated with the presence of PCA (chi2=0.68, NS). The frequency of positive findings for PCA in group 2 was higher in patients with (90%) than in those without (50%) intestinal metaplasia. IN CONCLUSION: (1) H. pylori infection is associated with the production of PCA, which, after eradication of the infection, persist and might contribute to the persistent antral chronic gastritis and intestinal metaplasia; (2) the gastric lesions associated with infections sustained by the more-virulent H. pylori strains do not appear to be due to the induction of antigastric autoantibodies.

Effect of cagA status on the sensitivity of enzyme immunoassay in diagnosing Helicobacter pylori-infected children.

BACKGROUND: The aims of our study were twofold. First, we sought to evaluate in symptomatic children the influence of the Helicobacter pylori genotype on gastritis, abdominal pain, and circulating anti-H. pylori IgG antibodies (anti-H. pylori IgG) or pepsinogen A (PGA) and C (PGC). Additionally, we sought to assess anti-H. pylori IgG, PGA, and PGC patterns in a large cohort (N = 921) of asymptomatic children. MATERIALS AND METHODS: In 183 symptomatic children, H. pylori infection and the presence of gastritis were evaluated by histology. In a subgroup of 20 H. pylori-positive children, the H. pylori genotype was evaluated also by polymerase chain reaction. Nine hundred and twenty-one asymptomatic children, aged 11 to 14 years, were studied by anti-H. pylori IgG, PGA, and PGC serum determination. RESULTS: The infection was found in 33 of 183 symptomatic children; among the 20 H. pylori-positive children for which the H. pylori genotype was available, cagA was present or absent in equal percentages. H. pylori infection was associated with more severe gastritis and higher serum levels of anti-H. pylori IgG and PGC but not with abdominal pain. In infected children, higher levels of anti-H. pylori IgG and the presence of abdominal pain were associated with infections caused by cagA-positive strains. In the cohort of 921 asymptomatic children, raised levels of anti-H. pylori IgG, PGA, and PGC were found in approximately 5% of the cases. CONCLUSIONS: Infection with cagA-positive H. pylori strains can be associated with increased frequency of reported abdominal pain and higher circulating levels of anti-H. pylori IgG. The serological assessment of H. pylori IgG using H. pylori antigens containing significant amounts of cagA protein may, therefore, underestimate the true prevalence of infection.

[Biochemical markers of gastric functioning]. / Marcatori biochimici di funzionalità gastrica.

The serum determination of pepsinogen A (PGA) and pepsinogen C (PGC) might indicate gastric mucosal inflammation and atrophy. Body gastric mucosa produces both PGA and PGC, while antral mucosa produces only PGC. Therefore, diseases involving mainly the antrum, such as H. pylori infection, are mainly indicated by the variations in serum PGC than in serum PGA. In agreement, when the antral mucosa is infected by the more virulent cagA positive H. pylori strains, which cause severe inflammation, serum PGC significantly increases. Another indirect indicator of gastric inflammation is polymorphonuclear (PMN) oxidative burst after the stimulation with water extracts from H. pylori culture: this parameter is significantly increased in infected if compared to non-infected subjects. The higher oxidative burst response of peripheral PMN in infected patients, possibly consequent to the release of specific cytokines able to prime PMN towards H. pylori products, is unable to eliminate the infection, but it might concur in damaging the gastric mucosa.