RESUMO
Oligonucleotides are important therapeutic approaches, as evidenced by recent clinical successes with antisense oligonucleotides (ASOs) and double-stranded short interfering RNAs (siRNAs). Phosphorothioate (PS) modifications are a standard feature in the current generation of oligonucleotide therapeutics, but generate isomeric mixtures, leading to 2n isomers. All currently marketed therapeutic oligonucleotides (ASOs and siRNAs) are complex isomeric mixtures. Recent chemical methodologies for stereopure PS insertions have resulted in preliminary rules for ASOs, with multiple stereopure ASOs moving into clinical development. Although siRNAs have comparatively fewer PSs, the field has yet to embrace the idea of stereopure siRNAs. Herein, it has been investigated whether the individual isomers contribute equally to the in vivo activity of a representative siRNA. The results of a systematic evaluation of stereopure PS incorporation into antithrombin-3 (AT3) siRNA are reported and demonstrate that individual PS isomers dramatically affect in vivo activity. A standard siRNA design with six PS insertions was investigated and it was found that only about 10 % of the 64 possible isomers were as efficacious as the stereorandom control. Based on this data, it can be concluded that G1R stereochemistry is critical, G2R is important, G21S is preferable, and G22 and P1/P2 tolerate both isomers. Surprisingly, the disproportionate loss of efficacy for most isomers does not translate into significant gain for the productive isomers, and thus, warrants further mechanistic studies.
Assuntos
Antitrombinas/química , Hepatócitos/efeitos dos fármacos , Oligonucleotídeos Fosforotioatos/química , RNA de Cadeia Dupla/genética , RNA Interferente Pequeno/genética , Animais , Antitrombinas/metabolismo , Células Cultivadas , Hepatócitos/metabolismo , Camundongos , RNA de Cadeia Dupla/administração & dosagem , RNA de Cadeia Dupla/química , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/químicaRESUMO
We investigated how many patients with a diagnosis of nonceliac wheat sensitivity (NCWS) still experienced wheat sensitivity after a median follow-up time of 99 months. We collected data from 200 participants from a previous study of NCWS, performed between July and December 2016 in Italy; 148 of these individuals were still on a strict wheat-free diet. In total, 175 patients (88%) improved (had fewer symptoms) after a diagnosis of NCWS; 145 of 148 patients who adhered strictly to a gluten-free diet (98%) had reduced symptoms, compared with 30 of 52 patients who did not adhere to a gluten-free diet (58%) (P < .0001). Of the 22 patients who repeated the double-blind, placebo-controlled challenge, 20 reacted to wheat. We conclude that NCWS is a persistent condition. Clinicaltrials.gov registration number: NCT02823522.
Assuntos
Cooperação do Paciente , Hipersensibilidade a Trigo/dietoterapia , Hipersensibilidade a Trigo/diagnóstico , Adulto , Doença Crônica , Dieta Livre de Glúten , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Inquéritos e Questionários , Avaliação de SintomasRESUMO
OBJECTIVE: To evaluate the long-term validity and safety of pure oats in the treatment of children with celiac disease. STUDY DESIGN: This noninferiority clinical trial used a double-blind, placebo-controlled, crossover design extended over 15 months. Three hundred six children with a biopsy-proven diagnosis of celiac disease on a gluten-free diet for ≥2 years were randomly assigned to eat specifically prepared gluten-free food containing an age-dependent amount (15-40 g) of either placebo or purified nonreactive varieties of oats for 2 consecutive 6-month periods separated by washout standard gluten-free diet for 3 months. Clinical (body mass index, Gastrointestinal Symptoms Rating Scale score), serologic (IgA antitransglutaminase antibodies, and IgA anti-avenin antibodies), and intestinal permeability data were measured at baseline, and after 6, 9, and 15 months. Direct treatment effect was evaluated by a nonparametric approach using medians (95% CI) as summary statistic. RESULTS: After the exclusion of 129 patients who dropped out, the cohort included 177 children (79 in the oats-placebo and 98 in the placebo-oats group; median, 0.004; 95% CI, -0.0002 to 0.0089). Direct treatment effect was not statistically significant for clinical, serologic, and intestinal permeability variables (body mass index: median, -0.5; 95% CI, -0.12 to 0.00; Gastrointestinal Symptoms Rating Scale score: median, 0; 95% CI, -2.5 to 0.00; IgA antitransglutaminase antibodies: median, -0.02; 95% CI, -0.25 to 0.23; IgA anti-avenin antibodies: median, -0.0002; 95% CI, -0.0007 to 0.0003; intestinal permeability test: median, 0.004; 95% CI, -0.0002 to 0.0089). CONCLUSIONS: Pure nonreactive oat products are a safe dietary choice in the treatment of children with celiac disease. TRIAL REGISTRATION: ClinicalTrials.gov: NCT00808301.
Assuntos
Avena/efeitos adversos , Doença Celíaca/dietoterapia , Doença Celíaca/imunologia , Criança , Estudos Cross-Over , Dieta Livre de Glúten , Método Duplo-Cego , Feminino , Humanos , Mucosa Intestinal/imunologia , MasculinoRESUMO
BACKGROUND & AIMS: There is much interest in wheat sensitivity among people without celiac disease (CD), but little is known about any risks associated with the condition. We evaluated the prevalence of autoimmune diseases (ADs) among patients with nonceliac wheat sensitivity (NCWS), and investigated whether they carry antinuclear antibodies (ANA). METHODS: We performed a retrospective study of 131 patients diagnosed with NCWS (121 female; mean age, 29.1 years) at 2 hospitals in Italy from January 2001 through June 2011. Data were also collected from 151 patients with CD or irritable bowel syndrome (IBS) (controls). Patient medical records were reviewed to identify those with ADs. We also performed a prospective study of 42 patients (38 female; mean age, 34 years) diagnosed with NCWS from July 2011 through March 2014 at 3 hospitals in Italy. One hundred age- and sex-matched subjects with CD or IBS served as controls. Serum samples were collected from all subjects and ANA levels were measured by immunofluorescence analysis. Participants completed a questionnaire and their medical records were reviewed to identify those with ADs. RESULTS: In the retrospective analysis, similar portions of subjects with NCWS (29%) and CD (29%) developed ADs (mainly Hashimoto's thyroiditis, 29 cases), compared with a smaller proportion of subjects with IBS (4%) (P < .001). In the prospective study, 24% of subjects with NCWS, 20% of subjects with CD, and 2% of subjects with IBS developed ADs (P < .001). In the retrospective study, serum samples tested positive for ANA in 46% of subjects with NCWS (median titer, 1:80), 24% of subjects with CD (P < .001), and 2% of subjects IBS (P < .001); in the prospective study, serum samples were positive for ANA in 28% of subjects with NCWS, 7.5% of subjects with CD (P = .02), and 6% of subjects with IBS (P = .005 vs patients with NCWS). ANA positivity was associated with the presence of the HLA DQ2/DQ8 haplotypes (P < .001). CONCLUSIONS: Higher proportions of patients with NCWS or CD develop autoimmune disorders, are ANA positive, and showed DQ2/DQ8 haplotypes compared with patients with IBS.
Assuntos
Anticorpos Antinucleares/sangue , Doenças Autoimunes/imunologia , Doença Celíaca/imunologia , Hipersensibilidade a Trigo/imunologia , Adulto , Doenças Autoimunes/sangue , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/epidemiologia , Doenças Autoimunes/genética , Biomarcadores/sangue , Doença Celíaca/sangue , Doença Celíaca/diagnóstico , Doença Celíaca/epidemiologia , Doença Celíaca/genética , Feminino , Antígenos HLA-DQ/genética , Antígenos HLA-DQ/imunologia , Haplótipos , Humanos , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Estudos Prospectivos , Estudos Retrospectivos , Fatores de Risco , Hipersensibilidade a Trigo/sangue , Hipersensibilidade a Trigo/diagnóstico , Hipersensibilidade a Trigo/epidemiologia , Hipersensibilidade a Trigo/genéticaRESUMO
BACKGROUND: Non-celiac gluten sensitivity (NCGS) or 'wheat sensitivity' (NCWS) is included in the spectrum of gluten-related disorders. No data are available on the prevalence of low bone mass density (BMD) in NCWS. Our study aims to evaluate the prevalence of low BMD in NCWS patients and search for correlations with other clinical characteristics. METHODS: This prospective observation study included 75 NCWS patients (63 women; median age 36 years) with irritable bowel syndrome (IBS)-like symptoms, 65 IBS and 50 celiac controls. Patients were recruited at two Internal Medicine Departments. Elimination diet and double-blind placebo controlled (DBPC) wheat challenge proved the NCWS diagnosis. All subjects underwent BMD assessment by Dual Energy X-Ray Absorptiometry (DXA), duodenal histology, HLA DQ typing, body mass index (BMI) evaluation and assessment for daily calcium intake. RESULTS: DBPC cow's milk proteins challenge showed that 30 of the 75 NCWS patients suffered from multiple food sensitivity. Osteopenia and osteoporosis frequency increased from IBS to NCWS and to celiac disease (CD) (P <0.0001). Thirty-five NCWS patients (46.6%) showed osteopenia or osteoporosis. Low BMD was related to low BMI and multiple food sensitivity. Values of daily dietary calcium intake in NCWS patients were significantly lower than in IBS controls. CONCLUSIONS: An elevated frequency of bone mass loss in NCWS patients was found; this was related to low BMI and was more frequent in patients with NCWS associated with other food sensitivity. A low daily intake of dietary calcium was observed in patients with NCWS.
Assuntos
Índice de Massa Corporal , Densidade Óssea , Glutens/efeitos adversos , Enteropatias/complicações , Adulto , Doenças Ósseas Metabólicas/complicações , Doenças Ósseas Metabólicas/epidemiologia , Doença Celíaca/complicações , Método Duplo-Cego , Feminino , Humanos , Síndrome do Intestino Irritável/complicações , Pessoa de Meia-Idade , Osteoporose/complicações , Osteoporose/epidemiologia , Prevalência , Estudos Prospectivos , Risco , Adulto JovemRESUMO
Peptide conjugates represent an emerging class of therapeutics. However, in contrast to that of small molecules and peptides, the discovery and optimization of peptide conjugates is low in throughput, resource intensive, time-consuming, and based on educated decisions rather than screening. A strategy for the parallel synthesis and screening of peptide conjugates is presented that (1) reduces variability in the conjugation steps; (2) provides a new method to rapidly and quantitatively measure conversion in crude conjugation mixtures; (3) introduces a purification step using an immobilized chemical scavenger that does not rely on protein-specific binding; and (4) is supported by robust analytical methods to characterize the large number of end products. Copper-free click chemistry is used as the chemoselective ligation method for conjugation and purification. The productivity in the generation and screening of peptide conjugates is significantly improved by applying this strategy as is demonstrated by the optimization of the anti-Angiopoietin-2 (Ang2) CovX-body, CVX-060, a peptide-antibody scaffold conjugate that has advanced in clinical trials for oncology indications.
Assuntos
Peptídeos/síntese química , Anticorpos/química , Química Click , Estrutura Molecular , Peptídeos/química , Peptídeos/isolamento & purificaçãoRESUMO
AIM: Functional abdominal pain (FAP) is a frequent condition affecting 10-20% of children and can be considered within the classification of functional gastrointestinal disorders (FGID). The objective of this study was to determine the effect of daily supplementation with the probiotic Lactobacillus reuteri DSM 17938 in children with FAP. METHODS: The children (aged 6-16 years) were screened for FAP as defined in the Rome III criteria and 60 patients were recruited in this double-blind, randomised, placebo-controlled trial. The children were randomly allocated to receive either L. reuteri (2×10(8) CFU/day) or identical placebo for 4 weeks followed by a 4-week follow-up period without supplementation. Frequency and intensity of pain was self-recorded by the subjects. RESULTS: The L. reuteri-supplemented children had significantly lower pain intensity compared with the placebo controls. CONCLUSIONS: Supplementation with L. reuteri reduced perceived abdominal pain intensity, which may encourage clinicians to use this probiotic in children with FAP.
Assuntos
Dor Abdominal/tratamento farmacológico , Gastroenteropatias/tratamento farmacológico , Limosilactobacillus reuteri , Probióticos/uso terapêutico , Dor Abdominal/diagnóstico , Adolescente , Distribuição de Qui-Quadrado , Criança , Suplementos Nutricionais , Feminino , Seguimentos , Gastroenteropatias/diagnóstico , Humanos , Masculino , Medição da Dor , Valores de Referência , Medição de Risco , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
OBJECTIVES: Non-celiac wheat sensitivity (NCWS) is a newly described clinical entity characterized by symptoms, which can involve the gastrointestinal tract, the nervous system, the skin, and other organs. There is little data on the pathogenesis of NCWS and it is probable that different pathogenic mechanisms are involved in the different clinical manifestations of the disease. The only common denominator of NCWS "syndrome" is wheat consumption: the symptoms disappear on exclusion of wheat from the diet, and reappear on wheat consumption. The objective of this study was to review our prior data regarding NCWS and to review relevant medical literature regarding NCWS, with particular attention to the hypothesis that NCWS patients could suffer from non-immunoglobulin E (IgE)-mediated wheat allergy. METHODS: We reviewed our data on 276 patients diagnosed with NCWS by means of double-blind placebo-controlled (DBPC) wheat challenge. The data indicating a possible wheat allergy diagnosis were examined and other data in the literature were reviewed; we review the role of serum immunoglobulin G antibodies and the basophil activation assay in food allergy, and the histology findings in the food allergy diagnosis. RESULTS: The comparison between patients suffering from NCWS and presenting with irritable bowel syndrome (IBS) and controls with IBS not due to NCWS showed that NCWS was characterized by: a personal history of food allergy in the pediatric age (0.01), coexistent atopic diseases (0.0001), positive serum anti-gliadin (0.0001) and anti-betalactoglobulin (0.001) antibodies, positive cytofluorimetric assay revealing in vitro basophil activation by food antigens (0.0001), and a presence of eosinophils in the intestinal mucosa biopsies (0.0001). CONCLUSIONS: Patients with NCWS and multiple food sensitivity show several clinical, laboratory, and histological characteristics that suggest they might be suffering from non-IgE-mediated food allergy. However, other pathogenic mechanisms need to be considered.
Assuntos
Hipersensibilidade Alimentar/imunologia , Síndrome do Intestino Irritável/imunologia , Triticum/imunologia , Doença Celíaca/imunologia , Método Duplo-Cego , Feminino , Gastroenteropatias/imunologia , Humanos , Imunoglobulina G/imunologia , MasculinoRESUMO
OBJECTIVES: Patients with chronic constipation due to food hypersensitivity (FH) had an elevated anal sphincter resting pressure. No studies have investigated a possible role of FH in anal fissures (AFs). We aimed to evaluate (1) the effectiveness of diet in curing AFs and to evaluate (2) the clinical effects of a double-blind placebo-controlled (DBPC) challenge, using cow's milk protein or wheat. METHODS: One hundred and sixty-one patients with AFs were randomized to receive a "true-elimination diet" or a "sham-elimination diet" for 8 weeks; both groups also received topical nifedipine and lidocaine. Sixty patients who were cured with the "true-elimination diet" underwent DBPC challenge in which cow's milk and wheat were used. RESULTS: At the end of the study, 69% of the "true-diet group" and 45% of the "sham-diet group" showed complete healing of AFs (P<0.0002). Thirteen of the 60 patients had AF recurrence during the 2-week cow's milk DBPC challenge and 7 patients had AF recurrence on wheat challenge. At the end of the challenge, anal sphincter resting pressure significantly increased in the patients who showed AF reappearance (P<0.0001), compared with the baseline values. The patients who reacted to the challenges had a significantly higher number of eosinophils in the lamina propria and intraepithelial lymphocytes than those who did not react to the challenges. CONCLUSIONS: An oligo-antigenic diet combined with medical treatment improved the rate of chronic AF healing. In more than 20% of the patients receiving medical and dietary treatment, AFs recurred on DBPC food challenge.
Assuntos
Constipação Intestinal/complicações , Constipação Intestinal/prevenção & controle , Eosinófilos , Comportamento Alimentar , Fissura Anal/etiologia , Hipersensibilidade Alimentar/complicações , Hipersensibilidade Alimentar/dietoterapia , Linfócitos , Proteínas do Leite/imunologia , Triticum/imunologia , Adulto , Doença Crônica , Constipação Intestinal/etiologia , Método Duplo-Cego , Feminino , Hipersensibilidade Alimentar/imunologia , Humanos , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Contagem de Leucócitos , Contagem de Linfócitos , Masculino , Manometria , Pessoa de Meia-Idade , Hipersensibilidade a Leite/complicações , Hipersensibilidade a Leite/dietoterapia , Proteínas do Leite/administração & dosagem , Proteínas do Leite/efeitos adversos , Recidiva , Resultado do Tratamento , Triticum/efeitos adversosRESUMO
BACKGROUND: The diagnosis of food hypersensitivity (FH) in adult patients with gastrointestinal symptoms, beyond the immediate IgE-mediated clinical manifestations, is very often difficult. The aims of our study were to: 1) evaluate the frequency of FH in patients with irritable bowel syndrome (IBS)-like clinical presentation; and 2) compare the diagnostic accuracy of two different methods of in vitro basophil activation tests. METHODS: Three hundred and five patients (235 females, age range 18-66 years) were included and underwent a diagnostic elimination diet and successive double-blind placebo-controlled (DBPC) challenges. Two different methods of in vitro basophil activation tests (BAT) (CD63 expression after in vitro wheat or cow's milk proteins stimulation) were evaluated: one was performed on separated leukocytes, and the other on whole blood. RESULTS: Ninety patients of the 305 studied (29.5%) were positive to the challenges and were diagnosed as suffering from FH. BAT on separate leukocytes showed a sensitivity of 86% and a specificity of 91% in FH diagnosis. BAT on whole blood showed a sensitivity of 15%-20% and a specificity of 73% in FH diagnosis (p<0.0001 compared to the other method). CONCLUSIONS: About one third of the IBS patients included in the study were suffering from FH and were cured on the elimination diet. The BAT based on CD63 detection on whole blood samples did not work in FH diagnosis and showed a significantly lower sensitivity, specificity and diagnostic accuracy than the assay based on separated leukocytes.
Assuntos
Basófilos/imunologia , Glutens/imunologia , Síndrome do Intestino Irritável/imunologia , Hipersensibilidade a Leite/diagnóstico , Proteínas do Leite/imunologia , Triticum/imunologia , Adolescente , Adulto , Idoso , Doença Celíaca/diagnóstico , Doença Celíaca/imunologia , Feminino , Humanos , Síndrome do Intestino Irritável/diagnóstico , Masculino , Pessoa de Meia-Idade , Hipersensibilidade a Leite/imunologia , Adulto JovemRESUMO
OBJECTIVES: Non-celiac wheat sensitivity (WS) is considered a new clinical entity. An increasing percentage of the general population avoids gluten ingestion. However, the real existence of this condition is debated and specific markers are lacking. Our aim was thus to demonstrate the existence of WS and define its clinical, serologic, and histological markers. METHODS: We reviewed the clinical charts of all subjects with an irritable bowel syndrome (IBS)-like presentation who had been diagnosed with WS using a double-blind placebo-controlled (DBPC) challenge in the years 2001-2011. One hundred celiac disease (CD) patients and fifty IBS patients served as controls. RESULTS: Two hundred and seventy-six patients with WS, as diagnosed by DBPC challenge, were included. Two groups showing distinct clinical characteristics were identified: WS alone (group 1) and WS associated with multiple food hypersensitivity (group 2). As a whole group, the WS patients showed a higher frequency of anemia, weight loss, self-reported wheat intolerance, coexistent atopy, and food allergy in infancy than the IBS controls. There was also a higher frequency of positive serum assays for IgG/IgA anti-gliadin and cytometric basophil activation in "in vitro" assay. The main histology characteristic of WS patients was eosinophil infiltration of the duodenal and colon mucosa. Patients with WS alone were characterized by clinical features very similar to those found in CD patients. Patients with multiple food sensitivity were characterized by clinical features similar to those found in allergic patients. CONCLUSIONS: Our data confirm the existence of non-celiac WS as a distinct clinical condition. We also suggest the existence of two distinct populations of subjects with WS: one with characteristics more similar to CD and the other with characteristics pointing to food allergy.
Assuntos
Autoanticorpos/sangue , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , Gliadina/imunologia , Triticum/imunologia , Adulto , Idoso , Anemia Hipocrômica/etiologia , Doença Celíaca/diagnóstico , Doença Celíaca/imunologia , Diagnóstico Diferencial , Método Duplo-Cego , Feminino , Hipersensibilidade Alimentar/complicações , Humanos , Hipersensibilidade Imediata/complicações , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Síndrome do Intestino Irritável/diagnóstico , Síndrome do Intestino Irritável/imunologia , Masculino , Pessoa de Meia-Idade , Projetos de Pesquisa , Fatores de Risco , Índice de Gravidade de Doença , Redução de PesoRESUMO
BACKGROUND & AIMS: Some patients with irritable bowel syndrome (IBS)-like symptoms suffer from food hypersensitivity (FH); their symptoms improve when they are placed on elimination diets. No assays identify patients with FH with satisfactory levels of sensitivity. We determined the frequency of FH among patients with symptoms of IBS and the ability of fecal assays for tryptase, eosinophil cationic protein (ECP), or calprotectin to diagnose FH. METHODS: The study included 160 patients with IBS, 40 patients with other gastrointestinal diseases, and 50 healthy individuals (controls). At the start of the study, patients completed a symptom severity questionnaire, fecal samples were assayed, and levels of specific immunoglobulin E were measured. Patients were observed for 4 weeks, placed on an elimination diet (without cow's milk and derivatives, wheat, egg, tomato, and chocolate) for 4 weeks, and kept a diet diary. Those who reported improvements after the elimination diet period were then diagnosed with FH, based on the results of a double-blind, placebo-controlled, oral food challenge (with cow's milk proteins and then with wheat proteins). RESULTS: Forty of the patients with IBS (25%) were found to have FH. Levels of fecal ECP and tryptase were significantly higher among patients with IBS and FH than those without FH. The ECP assay was the most accurate assay for diagnosis of FH, showing 65% sensitivity and 91% specificity. CONCLUSIONS: Twenty-five percent of patients with IBS have FH. These patients had increased levels of fecal ECP and tryptase, indicating that they might cause inflammation in patients with IBS. Fecal assays for ECP could be used to identify FH in patients with IBS.
Assuntos
Fezes/química , Síndrome do Intestino Irritável/complicações , Hipersensibilidade a Leite/diagnóstico , Hipersensibilidade a Trigo/diagnóstico , Adolescente , Adulto , Dieta/métodos , Método Duplo-Cego , Proteína Catiônica de Eosinófilo/análise , Feminino , Glutens/imunologia , Humanos , Complexo Antígeno L1 Leucocitário/análise , Masculino , Pessoa de Meia-Idade , Placebos/administração & dosagem , Triptases/análise , Adulto JovemRESUMO
BACKGROUND: In the diagnosis of celiac disease (CD), serum assays for anti-endomysium (EMA) and anti-transglutaminase (anti-tTG) antibodies have excellent diagnostic accuracy. However, these assays are less sensitive in young pediatric patients. Recently, a new ELISA test using deamidated gliadin peptides (DGP) as antigen has proved to be very sensitive and specific even in pediatric patients. In addition, anti-actin IgA antibodies (AAA) is another test that can be used in CD patients because antibody concentrations correlate with the degree of villous atrophy. This study evaluated the clinical accuracy of anti-tTG, EMA, AGA, anti-DGP and AAA and the effectiveness of these in different combinations for diagnosing CD in a large cohort of pediatric patients. METHODS: Sera of 150 children under 6 years of age were tested: 95 patients had a diagnosis of CD, while 55 patients who did not suffer from CD were used as controls. Anti-DGP IgA/IgG and AAA were assayed with ELISA kits, while anti-tTG IgA/IgG and AGA IgG/IgA were assayed using a quantitative fluoroimmunoassay. The EMA test was conducted by indirect immunofluorescence. RESULTS: Seventy-six of 95 (80%) CD patients were positive for DGP IgA and/or tTG IgA. Eighty of 95 (84.2%) patients were positive for DGP IgG and/or tTG IgA. None of the controls were positive for these antibodies. Eighty-four of 95 (88.4%) patients and 8/55 (14.5%) controls were positive for AAA and/or anti-tTG IgA. CONCLUSIONS: In very young children, association of anti-tTG IgA with anti-DGP IgG is the best test combination for diagnosing CD, yielding a cumulative sensitivity of 84.2% and a specificity of 100%.
Assuntos
Doença Celíaca/diagnóstico , Técnicas de Laboratório Clínico/normas , Estudos de Casos e Controles , Doença Celíaca/sangue , Pré-Escolar , Técnicas de Laboratório Clínico/economia , Ensaio de Imunoadsorção Enzimática , Feminino , Imunofluorescência , Humanos , Lactente , Masculino , Padrões de Referência , Sensibilidade e EspecificidadeRESUMO
BACKGROUND & AIMS: A percentage of patients with symptoms of irritable bowel syndrome (IBS) suffer from food hypersensitivity (FH) and improve on a food-elimination diet. No assays have satisfactory levels of sensitivity for identifying patients with FH. We evaluated the efficacy of an in vitro basophil activation assay in the diagnosis of FH in IBS-like patients. METHODS: Blood samples were collected from 120 consecutive patients diagnosed with IBS according to Rome II criteria. We analyzed in vitro activation of basophils by food allergens (based on levels of CD63 expression), as well as total and food-specific immunoglobulin (Ig)E levels in serum. Effects of elimination diets and double-blind food challenges were used as standards for FH diagnosis. RESULTS: Twenty-four of the patients (20%) had FH (cow's milk and/or wheat hypersensitivity); their symptom scores improved significantly when they were placed on an elimination diet. Patients with FH differed from other IBS patients in that they had a longer duration of clinical history, a history of FH as children, and an increased frequency of self-reported FH; they also had hypersensitivities to other antigens (eg, egg or soy). The basophil activation assay diagnosed FH with 86% sensitivity, 88% specificity, and 87% accuracy; this level of sensitivity was significantly higher than that of serum total IgE or food-specific IgE assays. CONCLUSIONS: A cytometric assay that quantifies basophils after stimulation with food antigens based on cell-surface expression of CD63 had high levels of sensitivity, specificity, and accuracy in diagnosing FH. This assay might be used to diagnose FH in patients with IBS-like symptoms.
Assuntos
Basófilos/imunologia , Técnicas Citológicas/métodos , Hipersensibilidade Alimentar/diagnóstico , Síndrome do Intestino Irritável/complicações , Adolescente , Adulto , Alérgenos/imunologia , Animais , Antígenos CD/análise , Células Cultivadas , Feminino , Humanos , Imunoglobulina E/sangue , Masculino , Pessoa de Meia-Idade , Glicoproteínas da Membrana de Plaquetas/análise , Sensibilidade e Especificidade , Tetraspanina 30 , Adulto JovemRESUMO
BACKGROUND: Celiac disease (CD) is a common disease in the Saharawi population of Arab-Berber origin. Saharawi patients with CD and their families were invited to participate in a follow-up study aimed at checking the CD serology status in patients being treated with the gluten-free diet (GFD) and investigating the prevalence of CD in first-degree relatives. PATIENTS AND METHODS: We investigated 975 subjects (62.8% females and 37.2% males, age range 0.7 to 75.4 years, median age 13.4 years) belonging to 212 families, by determining the serum immunoglobulin A anti-transglutaminase and anti-endomysial antibody levels. Thirty-two first-degree relatives were already receiving GFD when tested. RESULTS: Overall, 42.2% of the 244 treated subjects with CD showed a borderline/positive anti-transglutaminase determination, and 36.6% were also anti-endomysial positive. The serologic family screening detected 33 previously undiagnosed CD cases. The 65 affected first-degree relatives were sibling (42), mother (12), son/daughter (7), and father (4). The overall prevalence of CD among first-degree relatives was 65 of 763 (8.5%, 95% confidence interval 6.5-10.5). Based on our previous estimate of CD prevalence in the pediatric Saharawi population (5.6%), the sibling relative risk was 1.5. CONCLUSIONS: The risk of CD in the Saharawis is only modestly increased in first-degree relatives compared with the general population, probably because of the higher frequency of CD predisposing genes in the general population. Saharawi patients with CD receiving GFD showed poor adherence to the treatment, which could contribute to residual CD-related morbidity and mortality.
Assuntos
Autoanticorpos/sangue , Doença Celíaca/genética , Família , Transglutaminases/genética , Adolescente , Adulto , África do Norte/epidemiologia , Idoso , Árabes , Doença Celíaca/diagnóstico , Doença Celíaca/epidemiologia , Criança , Pré-Escolar , Dieta Livre de Glúten , Feminino , Seguimentos , Predisposição Genética para Doença , Humanos , Imunoglobulina A/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Fibras Musculares Esqueléticas/imunologia , Fatores de Risco , Testes Sorológicos , Transglutaminases/imunologia , Adulto JovemRESUMO
In this study, a biodiesel was produced from blending vegetable and animal sources with diesel and diesel-ethanol using a motor-generator set to evaluate its performance and emission characteristics. Fifteen and twenty percent of animal-vegetable biodiesel were added to each diesel-ethanol blend. A motor-generator test was conducted for each mixture; each sample was subjected to resistive loads from 2 to 5â kW with six repetitions. The physicochemical properties met the national standard guidelines, while the best specific fuel consumption (SFC) was observed for the 15% biodiesel-1% ethanol (B15E1) blend at the load of 5â kW with 327.069â gâ kW-1â h-1, followed by diesel (334.875â gâ kW-1â h-1). The exhaust gas temperature behaved differently depending on the ethanol concentration; it was lower when the concentration of added ethanol was higher. The NO emissions decreased while the SO2 emissions increased as the ethanol concentration increased.
Assuntos
Biocombustíveis , Etanol , Gasolina , Emissões de VeículosRESUMO
BACKGROUND & AIMS: Rectal bleeding and lymphonodular hyperplasia (LNH) in children can be caused by food hypersensitivity (FH). Our aim was to verify whether similar clinical and endoscopy presentations in adults can be due to FH. METHODS: Consecutive adult patients with rectal bleeding were enrolled. All underwent routine assays, colonoscopy, and histology study. RESULTS: Ten of 64 (15%) patients showed LNH as the unique sign at colonoscopy. An oligoantigenic diet resolved the rectal bleeding in 9 patients, and the reintroduction of several foods caused symptom reappearance. Double-blind placebo-controlled challenges with cow's milk and wheat protein confirmed the FH; symptoms reappeared 1-96 hours after the challenge. None of the patients were positive for IgE-mediated assays. In patients with LNH and FH, histology of the ileum and colon mucosa showed a higher number of lymphoid follicles and intraepithelial and lamina propria eosinophils compared with the other patients with rectal bleeding. CONCLUSIONS: Recurrent rectal bleeding can be caused by FH in adult patients. Endoscopic evidence of LNH characterizes these cases.
Assuntos
Hipersensibilidade Alimentar/complicações , Hemorragia/etiologia , Doenças Retais/etiologia , Adulto , Idoso , Criança , Colonoscopia , Método Duplo-Cego , Histocitoquímica , Humanos , Hiperplasia , Imunoglobulina E/análise , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Linfonodos/patologia , Pessoa de Meia-Idade , Hipersensibilidade a Leite/diagnóstico , Placebos/administração & dosagem , Recidiva , Hipersensibilidade a Trigo/diagnósticoRESUMO
BACKGROUND AND AIM: We evaluated the diagnostic performance of an ELISA test for anti-gliadin IgA and IgG antibodies, which uses synthetic deamidated gliadin peptides (anti-gliadin antibodies, AGAs) as coating; the results were compared with a test that uses extracted gliadin (AGAe). METHODS: The study was conducted on the sera of 144 patients suffering from celiac disease (CD), including 20 patients with IgA deficiency and 9 who were following a gluten-free diet (GFD), and 129 controls. RESULTS: In the 115 CD patients (without IgA deficiency), the sensitivity of AGAe IgA and IgG was 32.2 and 60.9%, whereas that of AGAs IgA and IgG was 59.1 and 72.2%. The specificity for AGAe IgA and IgG, and AGAs IgA and IgG was 93.8 and 89.9%, and 96.9% and 99.2%, respectively. Of the 20 patients with CD and IgA deficiency, 7 tested positive for AGAe IgG and 14 for AGAs IgG. The test using deamidated gliadin peptides performed better in terms of sensitivity and specificity than the AGA tests with extracted antigen. CONCLUSIONS: The very high specificity of the AGAs IgG test (99.2%) also suggests that patients who test positive with this assay require a thorough followup, even if the anti-tissue transglutaminase antibodies (anti-tTG) and anti-endomysial autoantibodies (EMA) assays are negative.
Assuntos
Anticorpos/sangue , Anticorpos/imunologia , Doença Celíaca/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Gliadina/imunologia , Adolescente , Adulto , Idoso , Autoanticorpos/sangue , Autoanticorpos/imunologia , Estudos de Casos e Controles , Doença Celíaca/sangue , Doença Celíaca/imunologia , Criança , Pré-Escolar , Reações Falso-Negativas , Reações Falso-Positivas , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina A/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade , Peptídeos/imunologia , Curva ROC , Sensibilidade e Especificidade , Testes Sorológicos , Transglutaminases/imunologia , Adulto JovemRESUMO
OBJECTIVE: Persistent villous atrophy in patients with celiac disease (CD) on a gluten-free diet (GFD) is reported with increasing frequency. The aim of this study was to evaluate a possible association between persistent damage of the villi and "atypical" gastrointestinal symptoms in CD patients on a GFD. MATERIAL AND METHODS: Sixty-nine CD patients on a GFD were divided into two groups: Group A included 42 patients (6 M, 36 F, age range 17-62 years) undergoing esophagogastroduodenoscopies (EGDs) due to the presence of symptoms; Group B included 27 control patients (6 M, 21 F, age range 24-71 years) who were asymptomatic at the time of the study. Both groups underwent EGDs and a duodenal histologic study. RESULTS: Persistent endoscopic lesions were more frequent in Group A (30/42) than in Group B (12/27; p=0.01). Villous atrophy was significantly more frequent in Group A than in Group B: 85% versus 33% (p<0.0001; odds ratio (OR)=12; 95% CI 3.7-38.9). Gastrointestinal symptoms in the Group A patients were different from those present at CD diagnosis: anemia/diarrhea/weight loss in 6 cases; gastroesophageal reflux disease (GERD)-like symptoms in 12 cases; abdominal pain/constipation in 24 cases. In Group A there was no difference in gender distribution, age and duration of GFD between subjects with normal villi and those with persistent partial villous atrophy. Patients with persistent symptoms showed a higher intraepithelial eosinophil count (p=0.005) than the asymptomatic patients (p=0.01). CONCLUSIONS: Persistent intestinal villous atrophy in CD patients on a GFD is associated with gastrointestinal symptoms considered "atypical" for CD and not present at CD diagnosis.
Assuntos
Doença Celíaca/dietoterapia , Doença Celíaca/patologia , Dieta Livre de Glúten , Mucosa Intestinal/patologia , Adolescente , Adulto , Idoso , Endoscopia Gastrointestinal , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
BACKGROUND: Treatment of celiac disease (CD) is based on the avoidance of gluten-containing food. However, it is not known whether trace amounts of gluten are harmful to treated patients. OBJECTIVE: The objective was to establish the safety threshold of prolonged exposure to trace amounts of gluten (ie, contaminating gluten). DESIGN: This was a multicenter, double-blind, placebo-controlled, randomized trial in 49 adults with biopsy-proven CD who were being treated with a gluten-free diet (GFD) for > or =2 y. The background daily gluten intake was maintained at < 5 mg. After a baseline evaluation (t0), patients were assigned to ingest daily for 90 d a capsule containing 0, 10, or 50 mg gluten. Clinical, serologic, and histologic evaluations of the small intestine were performed at t0 and after the gluten microchallenge (t1). RESULTS: At t0, the median villous height/crypt depth (Vh/Cd) in the small-intestinal mucosa was significantly lower and the intraepithelial lymphocyte (IEL) count (x 100 enterocytes) significantly higher in the CD patients (Vh/Cd: 2.20; 95% CI: 2.11, 2.89; IEL: 27; 95% CI: 23, 34) than in 20 non-CD control subjects (Vh/Cd: 2.87; 95% CI: 2.50, 3.09; IEL: 22; 95% CI: 18, 24). One patient (challenged with 10 mg gluten) developed a clinical relapse. At t(1), the percentage change in Vh/Cd was 9% (95% CI: 3%, 15%) in the placebo group (n = 13), -1% (-18%, 68%) in the 10-mg group (n = 13), and -20% (-22%, -13%) in the 50-mg group (n = 13). No significant differences in the IEL count were found between the 3 groups. CONCLUSIONS: The ingestion of contaminating gluten should be kept lower than 50 mg/d in the treatment of CD.