RESUMEN
In Angola during 2003-2012, we detected Haemophilus influenzae in 18% of 2,634 and 26% of 2,996 bacteriologically positive pleural or cerebrospinal fluid samples, respectively, from children. After vaccination launch in 2006, H. influenzae empyema declined by 83% and meningitis by 86%. Severe H. influenzae pneumonia and meningitis are preventable by vaccination.
Asunto(s)
Empiema/epidemiología , Vacunas contra Haemophilus/inmunología , Haemophilus influenzae , Meningitis por Haemophilus/epidemiología , Adolescente , Angola/epidemiología , Niño , Preescolar , Empiema/microbiología , Vacunas contra Haemophilus/administración & dosificación , Humanos , Lactante , Meningitis por Haemophilus/microbiología , Vigilancia de la Población , Estudios Prospectivos , VacunaciónRESUMEN
BACKGROUND: In childhood acute bacterial meningitis, the level of consciousness, measured with the Glasgow coma scale (GCS) or the Blantyre coma scale (BCS), is the most important predictor of outcome. The Herson-Todd scale (HTS) was developed for Haemophilus influenzae meningitis. Our objective was to identify prognostic factors, to form a simple scale, and to compare the predictive accuracy of these scales. METHODS: Seven hundred and twenty-three children with bacterial meningitis in Luanda were scored by GCS, BCS, and HTS. The simple Luanda scale (SLS), based on our entire database, comprised domestic electricity, days of illness, convulsions, consciousness, and dyspnoea at presentation. The Bayesian Luanda scale (BLS) added blood glucose concentration. The accuracy of the 5 scales was determined for 491 children without an underlying condition, against the outcomes of death, severe neurological sequelae or death, or a poor outcome (severe neurological sequelae, death, or deafness), at hospital discharge. RESULTS: The highest accuracy was achieved with the BLS, whose area under the curve (AUC) for death was 0.83, for severe neurological sequelae or death was 0.84, and for poor outcome was 0.82. Overall, the AUCs for SLS were ≥0.79, for GCS were ≥0.76, for BCS were ≥0.74, and for HTS were ≥0.68. CONCLUSIONS: Adding laboratory parameters to a simple scoring system, such as the SLS, improves the prognostic accuracy only little in bacterial meningitis.
Asunto(s)
Meningitis Bacterianas/diagnóstico , Angola , Preescolar , Bases de Datos Factuales , Países en Desarrollo , Femenino , Humanos , Lactante , Masculino , Meningitis Bacterianas/fisiopatología , Oportunidad Relativa , Valor Predictivo de las Pruebas , Pronóstico , Curva ROC , Factores de Riesgo , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
BACKGROUND: Helicobacter pylori is mainly acquired in childhood. Although adult studies reported a high prevalence of H. pylori infection in Portugal, the actual rate in children remains unknown. This study aimed to determine the prevalence and the incidence of H. pylori infection in an asymptomatic pediatric population of the Lisbon area and to correlate prevalence with sociodemographic determinants. MATERIALS AND METHODS: Helicobacter pylori infection was determined by stool antigen test in 844 asymptomatic children (age 0-15 years; 49.4% boys). For the incidence study, H. pylori-negative children in the prevalence study were followed-up every 6 months over a 3-year period. RESULTS: The global prevalence of H. pylori infection was 31.6%, increasing with age (19.9, 37.0 and 51.5%, in age groups 0-5, 6-10, and 11-15, respectively), but was similar among genders (34.5% in boys and 28.4% in girls). Older age and attendance of nursery/kindergarten during preschool constituted independent risk factors. The overall estimated incidence was 11.6 per 100 child-years (CY). Although 47.5% of children acquired H. pylori infection before 5 years of age, the mean age of acquisition was 6.3. The incidence of infection was similar among the three age groups (11.5, 13.0, and 10.5 per 100 CY, in age groups 0-5, 6-10, and 11-15, respectively). CONCLUSIONS: The prevalence of H. pylori infection in the Portuguese pediatric population is still high. Although this study confirmed that the highest acquisition rate occurs at young age, it showed that in high-prevalence populations, older children can also acquire H. pylori infection at a rate similar to that of young children.
Asunto(s)
Infecciones por Helicobacter/epidemiología , Helicobacter pylori/aislamiento & purificación , Adolescente , Factores de Edad , Niño , Preescolar , Femenino , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Portugal/epidemiología , PrevalenciaRESUMEN
We report a morality rate of 33% among 403 children with bacterial meningitis in Angola. A fatal outcome was associated with impaired consciousness, severe dyspnea, and seizures, and severe neurological sequelae (found in 25% of our patients) was associated with delayed presentation to the hospital, impaired consciousness, and seizures. Being underweight was of secondary importance. Treatment with ceftriaxone, rather than with penicillin plus chloramphenicol, did not improve outcome.
Asunto(s)
Meningitis Bacterianas/complicaciones , Meningitis Bacterianas/mortalidad , África del Sur del Sahara/epidemiología , Angola , Antibacterianos/uso terapéutico , Niño , Preescolar , Trastornos de la Conciencia/etiología , Disnea/etiología , Humanos , Lactante , Modelos Logísticos , Meningitis Bacterianas/tratamiento farmacológico , Meningitis Bacterianas/fisiopatología , Análisis Multivariante , Factores de Riesgo , Convulsiones/etiología , Resultado del TratamientoRESUMEN
BACKGROUND: The homB gene is a Helicobacter pylori disease-marker candidate, strongly associated with peptic ulcer disease, while homA, its paralogue gene with 90% sequence identity, is correlated with non-ulcer dyspepsia. The HomB encoded outer membrane protein was shown to contribute to the proinflammatory properties of H. pylori and also to be involved in bacterial adherence.This study investigated the distribution of homB and homA genes in 455 H. pylori strains from East Asian and Western countries, and carried out sequence comparison and phylogenetic analyses. RESULTS: Both homB and homA genes were heterogeneously distributed worldwide, with a marked difference between East Asian and Western strains.Analysis of homB and homA sequences revealed diversity regarding the number of copies and their genomic localization, with East Asian and Western strains presenting different genotypes. Moreover, homB and homA sequence analysis suggests regulation by phase variation. It also indicates possible recombination events, leading to gene duplication or homB/homA conversion which may as well be implicated in the regulation of these genes. Phylogenetic reconstruction of homB and homA revealed clustering according to the geographic origin of strains. Allelic diversity in the middle region of the genes was observed for both homB and homA, although there was no correlation between any allele and disease. For each gene, a dominant worldwide allele was detected, suggesting that homB/homA allelic variants were independent of the geographical origin of the strain. Moreover, all alleles were demonstrated to be expressed in vivo. CONCLUSION: Overall, these results suggest that homB and homA genes are good candidates to be part of the pool of H. pylori OMPs implicated in host-bacteria interface and also contributing to the generation of antigenic variability, and thus involved in H. pylori persistence.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/genética , Variación Genética , Infecciones por Helicobacter/microbiología , Helicobacter pylori/clasificación , Helicobacter pylori/genética , Filogenia , Factores de Virulencia/genética , Alelos , Secuencia de Aminoácidos , Proteínas de la Membrana Bacteriana Externa/química , Helicobacter pylori/patogenicidad , Humanos , Datos de Secuencia Molecular , Alineación de SecuenciaRESUMEN
The articles published this last year in the field of Helicobacter pylori diagnosis reported the development of in vivo histology, small improvements in some invasive methods (urease test, culture, and histology) and new kits for the stool antigen tests. They also contributed to increasing our knowledge, by further exploration into specific conditions for the urea breath test and into the significance of cagA antibodies. The role of serum markers of atrophy was also confirmed. Molecular methods are still being developed for direct genotyping, detection of H. pylori and its clarithromycin resistance, either by polymerase chain reaction or fluorescent in-situ hybridization. For the first time, there was a report on a possible interest of magnetic resonance spectroscopy.
Asunto(s)
Técnicas y Procedimientos Diagnósticos , Infecciones por Helicobacter/diagnóstico , Helicobacter pylori/aislamiento & purificación , Anticuerpos Antibacterianos , Infecciones por Helicobacter/inmunología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/genética , Helicobacter pylori/inmunología , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
Non-invasive tests are needed to assess Helicobacter pylori infection, especially to screen a pediatric population. Assure H. pylori Rapid Test (Genelabs Diagnostics, Singapore) is an immunochromatographic assay device intended for the rapid detection of antibodies to H. pylori in human serum, plasma or whole blood. The aim of this study was to evaluate the performance of the rapid test, Assure H. pylori, in the diagnosis of H. pylori infection in children, using a Portuguese pediatric population. The study group included 130 children with age ranging from 1 to 14 years old (average age 9.2+/-3.1 years). According to the gold standard, 70 of the 130 patients studied were H. pylori positive and 60 were H. pylori negative. Using Assure H. pylori Rapid Test (Genelabs Diagnostics, Singapore), 53 sera had a positive result after 15 min (resulting in 17 false negatives) and 57 sera a negative result (resulting in 3 false positives). The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the test were 75.7%, 95.0%, 94.6% and 77.0% respectively. When a longer read time of 45 min is considered, the rapid test revealed a good performance (sensitivity 98.6% and specificity 95%) in the evaluation of the H. pylori infection in a pediatric population. In conclusion, the test showed a good performance, suggesting its applicability as a screening method for the H. pylori infection.
Asunto(s)
Infecciones por Helicobacter/diagnóstico , Helicobacter pylori/aislamiento & purificación , Juego de Reactivos para Diagnóstico/microbiología , Gastropatías/microbiología , Adolescente , Anticuerpos Antibacterianos/sangre , Niño , Preescolar , Reacciones Falso Negativas , Reacciones Falso Positivas , Infecciones por Helicobacter/sangre , Infecciones por Helicobacter/microbiología , Humanos , Lactante , Portugal , Juego de Reactivos para Diagnóstico/normas , Sensibilidad y Especificidad , Gastropatías/sangre , Gastropatías/diagnósticoRESUMEN
BACKGROUND: Data concerning the effectiveness of Helicobacter pylori eradication regimens based in antibiotic susceptibility testing are scanty in children. AIMS: To identify the prevalence of antibiotic resistance in H. pylori strains isolated from Portuguese children in 1999-2003; to evaluate eradication rate after antibiotic susceptibility testing-based treatment; and to identify factors associated with resistance and eradication outcome. METHODS: Included were 109 children with a gastric biopsy culture positive for H. pylori. First treatment (amoxicillin, omeprazole and clarithromycin or metronidazole) was guided by susceptibility testing (E test), and eradication was assessed by [C]urea breath test. RESULTS: Strains were susceptible to amoxicillin and tetracycline; 39.4% were resistant to clarithromycin, 16.5% to metronidazole and 4.5% to ciprofloxacin. No significant association was found between resistance and sex, age, clinical status, gastritis scores, H. pylori density scores and genotype. Clarithromycin resistance was significantly associated with European origin [odds ratio (OR), 3.9], previous H. pylori empiric therapy (OR 2.8) and amoxicillin minimal inhibitory concentration, > or =0.016 (OR 6.0). Eradication rate after susceptibility-based treatment was 74.7% (59 of 79; 95% confidence interval, 65.9-82.9), and a significant association was found between eradication failure and presence of resistance to 1 or more antibiotics (P < 0.05). CONCLUSIONS: The prevalence of H. pylori antibiotic resistance was high in the studied population. The modest therapeutic success of clarithromycin and metronidazole susceptibility-based regimens suggests that in addition to resistance, other factors may be involved. The need of susceptibility-based treatment studies in children and of antimicrobial resistance surveillance in high prevalence areas for H. pylori are emphasized.
Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/epidemiología , Helicobacter pylori/efectos de los fármacos , Adolescente , Distribución por Edad , Antibacterianos/uso terapéutico , Pruebas Respiratorias , Niño , Preescolar , Estudios de Cohortes , Femenino , Gastroscopía/métodos , Infecciones por Helicobacter/diagnóstico , Helicobacter pylori/aislamiento & purificación , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Análisis Multivariante , Oportunidad Relativa , Portugal/epidemiología , Prevalencia , Probabilidad , Pronóstico , Medición de Riesgo , Índice de Severidad de la Enfermedad , Distribución por SexoRESUMEN
OBJECTIVES: Helicobacter pylori infection has not been well studied in older people, especially in hospitalized, frail patients. The aim of our study was to evaluate the prevalence of the infection in this population using five H. pylori diagnostic tests. DESIGN: Prospective observational study. SETTING: Geriatric acute care unit of the Department of Geriatrics (Hôpital Xavier Arnozan, Pessac, France). PARTICIPANTS: One hundred seven consecutively hospitalized patients with a diagnostic indication for upper gastrointestinal endoscopy. MEASUREMENTS: Geriatric assessment, information on drug intake, indication/results of gastric endoscopy, and results of H. pylori infection diagnostic tests (culture and histological analysis on biopsy specimens, serology, 13carbon-urea breath test (13C-UBT), detection of H. pylori stool antigens (HpSA)) were assessed for each included patient. RESULTS: Fifty-one patients (47.7%) were H. pylori positive with at least one test. 13C-UBT was more frequently positive than the other four tests, with a significant difference from culture, histological analysis, and HpSA (P <.05). Positive 13C-UBT results were significantly associated with H. pylori presence using histological analysis and neutrophil activity of the antrum and corpus. Antibiotic treatments significantly decreased the positivity rate of all of the tests performed, and severe corpus atrophy decreased the positivity rate of culture, histological analysis, and HpSA. CONCLUSIONS: Almost one-third of the H. pylori-positive patients would have remained undetected without performing the 13C-UBT. The low prevalence of H. pylori detection in these hospitalized, frail patients may be explained by the high frequency of current and previous antibiotic treatments.
Asunto(s)
Anciano Frágil , Infecciones por Helicobacter/epidemiología , Helicobacter pylori/aislamiento & purificación , Úlcera Péptica/microbiología , Anciano , Anciano de 80 o más Años , Pruebas Respiratorias , Endoscopía Gastrointestinal , Femenino , Infecciones por Helicobacter/diagnóstico , Hospitalización , Humanos , Masculino , Úlcera Péptica/epidemiología , Úlcera Péptica/patología , Prevalencia , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Giardia duodenalis is a widespread parasite of mammalian species, including humans. The prevalence of this parasite in children residing in Portugal is currently unknown. This study intended to estimate G. duodenalis infection prevalence and identify possible associated risk factors in a healthy paediatric population living in the District of the Portuguese capital, Lisbon. METHODS: Between February 2002 and October 2008, 844 children were randomly selected at healthcare centres while attending the national vaccination program. A stool sample and a questionnaire with socio-demographic data were collected from each child. Giardia infection was diagnosed by direct examination of stools and antigen detection by ELISA. RESULTS: The population studied revealed a gender distribution of 52.8% male and 47.2% female. Age distribution was 47.4% between 0-5 years and 52.6% between 6-15 years.The prevalence of Giardia infection was 1.9% (16/844) when estimated by direct examination and increased to 6.8% (57/844) when ELISA results were added. The prevalence was higher among children aged 0-5 years (7.8%), than among older children (5.8%), and was similar among genders (6.9% in boys and 6.5% in girls). The following population-variables were shown to be associated risk factors for G. duodenalis infection: mother's educational level (odds ratio (OR)= 4.49; confidence interval (CI): 1.20-16.84), father's educational level (OR = 12.26; CI: 4.08-36.82), presence of Helicobacter pylori infection (OR = 1.82; CI: 1.05-3.15), living in houses with own drainage system (OR = 0.10; CI: 0.02-0.64) and reported household pet contact, especially with dogs (OR = 0.53; CI: 0.31-0.93). CONCLUSION: The prevalence of giardiasis in asymptomatic children residing in the region of Lisbon is high. Several risk factors were associated with Giardia prevalence and highlight the importance of parents' education and sanitation conditions in the children's well being. The association between G. duodenalis and H. pylori seems an important issue deserving further investigation in order to promote prevention or treatment strategies.
Asunto(s)
Giardia lamblia/inmunología , Giardiasis/epidemiología , Infecciones por Helicobacter/epidemiología , Helicobacter pylori , Adolescente , Distribución por Edad , Animales , Niño , Preescolar , Coinfección , Perros , Escolaridad , Heces/parasitología , Femenino , Giardia lamblia/aislamiento & purificación , Giardiasis/complicaciones , Giardiasis/diagnóstico , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/diagnóstico , Vivienda , Humanos , Lactante , Masculino , Padres , Mascotas/microbiología , Portugal/epidemiología , Prevalencia , Factores de Riesgo , Saneamiento , Factores SocioeconómicosRESUMEN
BACKGROUND: homB encodes a Helicobacter pylori outer membrane protein. This gene was previously associated with peptic ulcer disease (PUD) and was shown to induce activation of interleukin-8 secretion in vitro, as well as contributing to bacterial adherence. Its 90%-similar gene, homA, was previously correlated with gastritis. The present study aimed to evaluate the gastric disease association with homB and homA, as well as with the H. pylori virulence factors cagA, babA and vacA, in 415 H. pylori strains isolated from patients from East Asian and Western countries. The correlation among these genotypes was also evaluated. RESULTS: Both homB and homA genes were heterogeneously distributed worldwide, with a marked difference between East Asian and Western strains. In Western strains (n = 234, 124 PUD and 110 non-ulcer dyspepsia (NUD), homB, cagA and vacA s1 were all significantly associated with PUD (p = 0.025, p = 0.014, p = 0.039, respectively), and homA was closely correlated with NUD (p = 0.072). In East Asian strains (n = 138, 73 PUD and 65 NUD), homB was found more frequently than homA, and none of these genes was associated with the clinical outcome. Overall, homB was associated with the presence of cagA (p = 0.043) and vacA s1 (p < 0.001), whereas homA was found more frequently in cagA-negative (p = 0.062) and vacA s2 (p < 0.001) strains. Polymorphisms in homB and homA copy number were observed, with a clear geographical specificity, suggesting an involvement of these genes in host adaptation. A correlation between the homB two-copy genotype and PUD was also observed, emphasizing the role of homB in the virulence of the strain. CONCLUSION: The global results suggest that homB and homA contribute to the determination of clinical outcome.
RESUMEN
Incidence, morbidity and mortality of bacterial meningitis in developing countries are manifold greater than those in the industrialized world. We reviewed retrospectively children with meningitis treated in the paediatric hospital of Luanda in 2004. Among the 555 children, median age 11.0 months, the leading agents were Haemophilus influenzae type b (Hib), pneumococcus, and meningococcus in 60%, 24%, and 10%, respectively. The median length of illness before admission was 7 d. 65% had convulsed. Altered level of consciousness was observed in 61% and blood haemoglobin lower than 8 g/dl in 36% of cases. Case fatality was 35% and, of survivors, 24% were left with severe neurological sequelae. Blood transfusion appeared beneficial since fatality of children with and without transfusion was 23% versus 39% (p=0.003). While awaiting large-scale vaccinations, tools to improve the prognosis of meningitis in Angola comprise generating better awareness to reduce the delay, better fluid treatment and monitoring and active use of blood transfusions.
Asunto(s)
Meningitis por Haemophilus/epidemiología , Meningitis Meningocócica/epidemiología , Meningitis Neumocócica/epidemiología , Angola/epidemiología , Antibacterianos/uso terapéutico , Transfusión Sanguínea , Preescolar , Femenino , Humanos , Incidencia , Lactante , Masculino , Meningitis por Haemophilus/mortalidad , Meningitis por Haemophilus/terapia , Meningitis Meningocócica/mortalidad , Meningitis Meningocócica/terapia , Meningitis Neumocócica/mortalidad , Meningitis Neumocócica/terapia , Oxígeno/uso terapéutico , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
BACKGROUND: homB codes for a putative Helicobacter pylori outer membrane protein and has previously been associated with peptic ulcer disease (PUD) in children. METHODS: A total of 190 H. pylori strains isolated from children and adults were studied to evaluate the clinical importance of the homB gene. In vitro experiments were performed to identify HomB mechanisms of bacterial pathogenicity. RESULTS: Characterization of the isolates demonstrated that homB was significantly associated with PUD in 86 children (odds ratio [OR], 7.64 [95% confidence interval {CI}, 2.65-22.05]) and in 32 adults < or =40 years of age (OR, 11.25 [95% CI, 1.86-68.13]). homB was correlated with the presence of cagA, babA2, vacAs1, hopQI, and oipA "on" genotype (P< .001) The HomB protein was found to be expressed in the H. pylori outer membrane and was noted to be antigenic in humans. H. pylori homB knockout mutant strains presented reduced ability to induce interleukin-8 secretion from human gastric epithelial cells, as well as reduced capacity to bind to these cells. Both of these functions correlated with the number of homB copies present in a strain. CONCLUSION: homB can be considered a comarker of H. pylori strains associated with PUD. Moreover, results strongly suggest that HomB is involved in the inflammatory response and in H. pylori adherence, constituting a novel putative virulence factor.
Asunto(s)
Proteínas de la Membrana Bacteriana Externa/genética , Marcadores Genéticos , Helicobacter pylori/genética , Úlcera Péptica/microbiología , Adulto , Antígenos Bacterianos , Adhesión Bacteriana , Proteínas de la Membrana Bacteriana Externa/metabolismo , Línea Celular Tumoral , Niño , Células Epiteliales/metabolismo , Femenino , Regulación Bacteriana de la Expresión Génica , Genes Bacterianos , Variación Genética , Helicobacter pylori/patogenicidad , Humanos , Interleucina-8/metabolismo , Modelos Logísticos , Masculino , Oportunidad Relativa , VirulenciaRESUMEN
Peptic ulcer disease (PUD) occurs after a long-term Helicobacter pylori infection. However, the disease can develop earlier, and rare cases have been observed in children, suggesting that these H. pylori strains may be more virulent. We used suppressive subtractive hybridization for comparative genomics between H. pylori strains isolated from a 5-year-old child with duodenal ulcer and from a sex- and age-matched child with gastritis only. The prevalence of the 30 tester-specific subtracted sequences was determined on a collection of H. pylori strains from children (15 ulcers and 30 gastritis) and from adults (46 ulcers and 44 gastritis). Two of these sequences, jhp0562 (80.0% versus 33.3%, P = 0.008) and jhp0870 (80.0% versus 36.7%, P = 0.015), were highly associated with PUD in children and a third sequence, jhp0828, was less associated (40.0% versus 10.0%, P = 0.048). Among adult strains, none of the 30 sequences was associated with PUD. However, both jhp0562 and jhp0870 were less prevalent in adenocarcinoma strains than in PUD strains from children and adults, the difference being statistically significant for jhp0870. In conclusion, two H. pylori genes were identified as being strongly associated with PUD in children, and their putative roles as an outer membrane protein for jhp0870 and in lipopolysaccharide biosynthesis for jhp0562, suggest that they may be novel virulence factors of H. pylori.
Asunto(s)
Helicobacter pylori/aislamiento & purificación , Hibridación in Situ , Úlcera Péptica/metabolismo , Úlcera Péptica/microbiología , Adulto , Biomarcadores/metabolismo , Niño , Preescolar , Femenino , Biblioteca de Genes , Helicobacter pylori/genética , Helicobacter pylori/patogenicidad , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la PolimerasaRESUMEN
OBJECTIVES: Both bacterial and host determinants underlying differences in histopathology and clinical outcome in H. pylori pediatric infection, as compared to adults, are still poorly documented. Pediatric studies may provide important insights on H. pylori infection immunopathogenesis, particularly in high gastric cancer risk populations. The present study concerns H. pylori genotypic diversity of isolates in children from a population with high gastric cancer risk, and its association with demographic and clinical variables, including gastroduodenal endoscopic and histopathological features. METHODS: A total of 119 subjects (mean age 10.3 yr, 1.5-18.0 yr) with H. pylori infection were studied. H. pylori vacA, cagA, and iceA genotypes were determined (PCR) in antral-obtained primary cultures; histopathological evaluation was performed in corpus, antrum, and duodenum biopsy specimens. RESULTS: cagA-, vacA s2m2, and iceA2 were the most prevalent genotypes. No association was observed between H. pylori genotypes and subject demographic and clinical variables, with the exception of a significant association between vacA s2 genotype and lower corpus inflammation score (p< 0.03). CONCLUSIONS: In this pediatric cohort, H. pylori genotype profiles were distinct from those reported in adult subjects in the same area, with a lower prevalence of the putative more virulent genotypes. Moreover, they were not associated with clinical expression of gastroduodenal disease, suggesting the potential role of host and/or environmental factors for the development of clinical disease at a later age.
Asunto(s)
ADN Bacteriano/genética , Infecciones por Helicobacter/microbiología , Helicobacter pylori/genética , Antro Pilórico/microbiología , Neoplasias Gástricas/complicaciones , Adolescente , Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Biopsia , Niño , Preescolar , Duodeno/microbiología , Duodeno/patología , Endoscopía Gastrointestinal , Femenino , Estudios de Seguimiento , Genotipo , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/patología , Helicobacter pylori/aislamiento & purificación , Humanos , Lactante , Masculino , Reacción en Cadena de la Polimerasa , Prevalencia , Estudios Prospectivos , Antro Pilórico/patología , Factores de Riesgo , Índice de Severidad de la Enfermedad , Neoplasias Gástricas/epidemiología , Neoplasias Gástricas/patologíaRESUMEN
OBJECTIVE: Serum pepsinogens and gastrin have been proposed as markers of gastritis, but have seldom been studied in children. In this study the aim was to identify host- and Helicobacter pylori-related factors linked to variations in serum gastrin, PGI, PGII, and to evaluate the potential of these biomarkers for diagnosing gastritis, whether H. pylori-associated or not. MATERIAL AND METHODS: Ninety-two dyspeptic children referred for endoscopy (peptic ulcer exclusion) were included in the study. H. pylori status (urease, culture, histology) was assessed, and genotype determined (PCR) in H. pylori-positive subjects. Serum gastrin, PGI and PGII levels were measured by standard radioimmunoassay (RIA). RESULTS: PGI and PGII levels were significantly higher in H. pylori-positive subjects (p=0.007; p=0.012, respectively). Gastrin levels were significantly higher in H. pylori-negative subjects (p=0.035). PGI and PGII were associated significantly with higher antrum inflammation scores (p=0.002; p=0.016, respectively); only PGI was associated with age, after controlling for inflammation (p=0.033) and for activity (p=0.037). The contribution of virulence factors could not be assessed owing to the low number of virulent strains. After multivariate analysis, only antrum inflammation was independently associated with PGI level (p=0.012). Receiver operating characteristic (ROC) analysis showed a low PGI and PGII discriminant power for predicting antrum inflammation. CONCLUSIONS: Pepsinogen levels as measured in this study seem predominantly to reflect antral inflammation, but they are not an effective screening test for gastritis (H. pylori-positive or -negative) in dyspeptic children.
Asunto(s)
Mucosa Gástrica/patología , Gastrinas/sangre , Helicobacter pylori/patogenicidad , Pepsinógeno A/sangre , Pepsinógeno C/sangre , Adolescente , Niño , Duodenoscopía , Gastroscopía , Humanos , Inflamación , Curva ROCRESUMEN
Preclimacteric 'Rocha' pears stored under chilling conditions, had a larger increase of ACO (1-aminocyclopropane-1-carboxylate oxidase) activity and softened faster than those treated with ethylene. Non-treated fruit did not ripen or soften, acquired a rubbery texture, and showed barely detectable levels of ACO activity. The transcript accumulation of seven genes encoding cell wall modifying enzymes was followed during fruit growth, ripening, and senescence, and in fruit that failed to ripen, by quantitative real-time PCR. Transcripts from 'Rocha' pear polygalacturonase1 and 2 (PcPG1, PcPG2), beta-galactosidase (PcbetaGAL) and beta-xylosidase (PcXYL) genes accumulated up to 1000-fold at the climacteric onset, while low transcript levels were detected in growing fruit. In fruit that did not ripen, this transcript accumulation was lower compared with fruits that ripened normally. Transcripts for expansin1 and 2 (PcEXPA1, PcEXPA2) accumulated in growing fruit, but about 10-fold more in fruit after rewarming. Xyloglucan endotransglucosylase/hydrolase (PcXTH) had the highest basal expression levels in all samples, showing only a small increase during fruit growth and ripening. PcEXPA2 and PcXTH transcripts accumulated in untreated fruit, 21 d after harvest, to levels similar to those of fruit that ripened normally. Since in untreated fruit ACO activity was barely detectable, it is likely that the activation of these genes might occur at very low ethylene levels. Results suggest that PcXTH and PcEXPA2 gene induction might be associated with cell wall maintenance during 'Rocha' pear development and ripening, while PcEXPA1, PcPG1, PcPG2, PcbetaGAL, and PcXYL expression is likely to be related to cell wall disassembly and loosening.
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Aminoácido Oxidorreductasas/metabolismo , Pared Celular/enzimología , Frío , Frutas/enzimología , Regulación de la Expresión Génica de las Plantas/fisiología , Pyrus/enzimología , Etilenos/biosíntesis , Etilenos/farmacología , Frutas/efectos de los fármacos , Reguladores del Crecimiento de las Plantas/biosíntesis , Proteínas de Plantas/metabolismo , Pyrus/efectos de los fármacos , Factores de Tiempo , Activación TranscripcionalRESUMEN
Helicobacter pylori infection is one of the most common gastrointestinal infections worldwide and almost invariably causes chronic gastritis in the infected host. A predominant Th1 profile has been demonstrated in H. pylori-infected mucosa from adults, but no previous study has evaluated in situ cytokine expression in children. We therefore examined expression of proinflammatory, anti-inflammatory, and regulatory cytokines by immunohistochemistry in cryopreserved antral biopsy specimens from 10 H. pylori-infected and 10 uninfected children and correlated expression of cytokines with histology scores. Concomitant expression of interleukin-8 (IL-8), gamma interferon (IFN-gamma), IL-4, transforming growth factor beta, and tumor necrosis factor alpha was seen in 8/10 H. pylori-infected cases and in 5/10 noninfected cases; all H. pylori-infected subjects showed staining for at least two of the cytokines. The proportion of epithelial cytokine-specific staining did not differ significantly between the groups, either in surface or glandular epithelium. Furthermore, no significant differences were noticed between intraepithelial or lamina propria lymphocyte staining in the groups. There was, however, a tendency of higher numbers of IFN-gamma- and IL-8-positive cells in the H. pylori-infected group. IFN-gamma and IL-8 lamina propria lymphocyte expression correlated significantly with antrum chronic inflammation, but there was no correlation between histology scores and epithelial cytokine expression. When the same techniques were used, the cytokine response appeared to be smaller in H. pylori-infected children than in adults, and there was no clear Th1 dominance. These results therefore suggest a different mucosal immunopathology in children. It remains to be determined whether the gastric immune response is downregulated in children with H. pylori infection and whether this is relevant to the outcome of infection.
Asunto(s)
Citocinas/biosíntesis , Mucosa Gástrica/inmunología , Infecciones por Helicobacter/inmunología , Adolescente , Biopsia , Niño , Preescolar , Células Epiteliales/inmunología , Células Epiteliales/patología , Mucosa Gástrica/patología , Genotipo , Infecciones por Helicobacter/sangre , Infecciones por Helicobacter/patología , Helicobacter pylori/clasificación , Helicobacter pylori/genética , Humanos , Inmunohistoquímica , Linfocitos/inmunologíaRESUMEN
The main cause of failure of Helicobacter pylori eradication therapy is resistance to clarithromycin. The resistance is due to three point mutations in two positions on the 23S rRNA (A2142C, A2142G, and A2143G). Our aim was to develop a rapid and accurate method to detect these mutations directly on biopsy specimens. We developed a real-time PCR that included a simultaneous detection of the amplicons by hybridization of two probes labeled with LC-Red and fluorescein by using the fluorescence resonance energy transfer (FRET) technology and melting curve analysis with the LightCycler thermocycler. The assay was first applied successfully on reference strains, reference plasmids, and H. pylori-negative biopsies. Biopsies from 200 patients having failed a first eradication attempt and for whom the H. pylori strain was available were then tested with the new assay. A result was obtained in 199 cases; a single genotype was detected in 157 cases, two genotypes were detected in 41 cases, and three genotypes were detected in one case. There were, in total, seven discrepancies between the real-time PCR and the phenotypic method of determination of clarithromycin susceptibility, and in an additional four cases the two phenotypic methods were in disagreement. PCR-restriction fragment length polymorphism was applied to a sampling of biopsies, including all of the cases with multiple genotypes and all the cases with discrepant results. Finally, in four cases with discrepant results, the real-time PCR detected the resistant population at a concentration so low that it could not be detected by the phenotypic method, while in three cases other mutations could be involved. This assay had an accuracy at least as satisfactory as that of the phenotypic tests and could be performed within 2 h, allowing it to be used before the administration of therapy in the case of a first H. pylori eradication.
Asunto(s)
Antibacterianos/farmacología , Claritromicina/farmacología , Farmacorresistencia Bacteriana/genética , Helicobacter pylori/efectos de los fármacos , Reacción en Cadena de la Polimerasa/métodos , ADN Bacteriano/análisis , Helicobacter pylori/genética , Humanos , Mutación Puntual , Polimorfismo de Longitud del Fragmento de Restricción , Estómago/microbiologíaRESUMEN
BACKGROUND: Noninvasive diagnostic tests are useful as screening tools for Helicobacter pylori infection in pediatric populations. The aim of this study was to evaluate performance of the immunoblot assay, Helico Blot 2.1, for the diagnosis of H. pylori infection in symptomatic children. MATERIALS AND METHODS: Immunoblot assay was used for detection of IgG antibodies to specific H. pylori proteins and to a recombinant H. pylori antigen, CIM marker. The study was performed on sera collected from 134 symptomatic, untreated children (mean age, 9.1 +/- 3.2 years; range, 1-14 years). H. pylori infection status was determined by culture, histology and rapid urease test. RESULTS: Immunoblot assay yielded a positive result in 71 of the 72 infected patients (sensitivity 98.6%) and in eight of the 62 noninfected ones (specificity 87.1%). The predictive values for a positive and a negative result were 89.9% and 98.2%, respectively. The performance of the CIM band alone, as a marker for H. pylori infection status, was also evaluated. This band was present on the blot of 71 infected patients and on four of the 62 H. pylori-negative patients. The sensitivity, specificity, PPV and NPV of the CIM antigen were 98.6%, 93.5%, 94.7% and 98.3%, respectively. CONCLUSIONS: The immunoblot assay Helico Blot 2.1 is a suitable noninvasive test for the serodiagnosis of H. pylori infection in children. The good level of performance demonstrated by the novel recombinant antigen CIM suggests it may be a useful contribution to the qualitative and quantitative performance of the Helico Blot 2.1 in pediatric populations.