RESUMEN
OBJECTIVE: To describe the clinical, endoscopic, histologic, and treatment outcomes of Helicobacter heilmannii (H. heilmannii) associated gastritis in children in the New England region of the United States. METHODS: Retrospective study of children (1-18 years) with H. heilmannii identified on gastric mucosal biopsies from two pediatric centers over a 21-year period, January 2000-December 2021. Cases were identified by querying pathology databases at each institution. Demographic and clinical data were obtained from the medical record. Endoscopic and histologic findings were extracted from endoscopy and pathology reports, respectively. RESULTS: Thirty-eight children were diagnosed with H. heilmannii-associated gastritis during the study period. The mean age at diagnosis was 10.1 ± 5.3 years, and 25/38 (66%) cases were male. Abdominal pain (32%) and nausea with or without vomiting (26%) were the most common symptoms. Thirty-two children (84%) had endoscopic findings including gastric nodularity (55%) and erythema (26%). All children had histologic signs of chronic gastritis, including those with normal endoscopic exams. Antibiotic regimens used for treating Helicobacter pylori were frequently prescribed. Of the 17 children who underwent a follow-up endoscopy (range 2-68 months), 15 (88%) did not have H. heilmannii identified on gastric biopsies. CONCLUSION: H. heilmannii was an infrequent but potential cause of epigastric abdominal pain and nausea in our cohort of New England children. While morphologically distinct from H. pylori, the bacteria can result in similar endoscopic and histologic findings of nodularity and chronic gastritis, respectively. The rate of eradication, as assessed by histology following treatment with H. pylori therapies, was below the 90% recommended goal for antimicrobial therapies.
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Gastritis , Infecciones por Helicobacter , Helicobacter heilmannii , Helicobacter pylori , Niño , Humanos , Masculino , Femenino , Estudios Retrospectivos , Gastritis/diagnóstico , Gastritis/tratamiento farmacológico , Gastritis/microbiología , Infecciones por Helicobacter/diagnóstico , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/microbiología , New England , Náusea , Dolor AbdominalRESUMEN
Cyclic dinucleotides (CDNs) are secondary messengers used by prokaryotic and eukaryotic cells. In mammalian cells, cytosolic CDNs bind STING (stimulator of IFN gene), resulting in the production of type I IFN. Extracellular CDNs can enter the cytosol through several pathways but how CDNs work from outside eukaryotic cells remains poorly understood. Here, we elucidate a mechanism of action on intestinal epithelial cells for extracellular CDNs. We found that CDNs containing adenosine induced a robust CFTR-mediated chloride secretory response together with cAMP-mediated inhibition of Poly I:C-stimulated IFNß expression. Signal transduction was strictly polarized to the serosal side of the epithelium, dependent on the extracellular and sequential hydrolysis of CDNs to adenosine by the ectonucleosidases ENPP1 and CD73, and occurred via activation of A2B adenosine receptors. These studies highlight a pathway by which microbial and host produced extracellular CDNs can regulate the innate immune response of barrier epithelial cells lining mucosal surfaces.
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Adenosina/metabolismo , Células Epiteliales/metabolismo , Inmunidad Innata , Inmunidad Mucosa , Nucleótidos Cíclicos/metabolismo , 5'-Nucleotidasa/metabolismo , Línea Celular Tumoral , Cloruros/metabolismo , Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Células Epiteliales/inmunología , Proteínas Ligadas a GPI/metabolismo , Humanos , Interferón beta/metabolismo , Mucosa Intestinal/citología , Hidrolasas Diéster Fosfóricas/metabolismo , Poli I-C/inmunología , Pirofosfatasas/metabolismo , Receptor de Adenosina A2B/metabolismo , Transducción de Señal/inmunologíaRESUMEN
OBJECTIVES: To assess if the distribution of villous intraepithelial lymphocytes (IELs) in a pediatric cohort with Marsh I histopathology is specific to celiac disease (CeD). METHODS: Multicenter, retrospective case-control study between January 2001 and December 2019 in children (<18 years) with and without CeD with intraepithelial lymphocytosis and normal villous architecture. Pathology specimens were reviewed by 2 study pathologists who were blinded to the final diagnosis. Morphologic features (villous height to crypt depth ratio [Vh:Cd]) and IELs in the villous tip, top, or bottom half of the villus were quantified. RESULTS: Of the 97 children with Marsh I histopathology identified during the study period, 63 were excluded due to an insufficient number of well-oriented villous-crypt complexes or a Vh:Cd less than 2. Villous IELs were measured in 34 cases (14 CeD, 20 non-CeD controls). There was no difference between the non-CeD and CeD groups in the mean IELs at the villous tip (14.0 ± 7.1 vs 11.7 ± 6.0, P = .31), top (46.4 ± 18.4 vs 38.3 ± 10.8, P = .11), or bottom (29.8 ± 16.8 vs 28.5 ± 12.8, P = .80) half of each villus, respectively. CONCLUSIONS: The distribution of IELs in Marsh I lesions is not specific for CeD.
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Enfermedad Celíaca , Linfocitos Intraepiteliales , Linfocitosis , Humanos , Niño , Enfermedad Celíaca/diagnóstico , Enfermedad Celíaca/patología , Estudios Retrospectivos , Estudios de Casos y Controles , Linfocitos Intraepiteliales/patología , Cadmio , Humedales , Linfocitosis/diagnóstico , Linfocitos/patología , Duodeno/patología , Mucosa Intestinal/patología , BiopsiaRESUMEN
BACKGROUND: Individuals with esophageal atresia (EA) have lifelong increased risk for mucosal and structural pathology of the esophagus. The use of surveillance endoscopy to detect clinically meaningful pathology has been underexplored in pediatric EA. We hypothesized that surveillance endoscopy in pediatric EA has high clinical yield, even in the absence of symptoms. STUDY DESIGN: The medical records of all patients with EA who underwent at least 1 surveillance endoscopy between March 2004 and March 2023 at an international EA referral center were retrospectively reviewed. The primary outcomes were endoscopic identification of pathology leading to an escalation in medical, endoscopic, or surgical management. Logistic regression analysis examined predictors of actionable findings. Nelson-Aalen analysis estimated optimal endoscopic surveillance intervals. RESULTS: Five hundred forty-six children with EA underwent 1,473 surveillance endoscopies spanning 3,687 person-years of follow-up time. A total of 770 endoscopies (52.2%) in 394 unique patients (72.2%) had actionable pathology. Esophagitis leading to escalation of therapy was the most frequently encountered finding (484 endoscopies, 32.9%), with most esophagitis attributed to acid reflux. Barrett's esophagus (intestinal metaplasia) was identified in 7 unique patients (1.3%) at a median age of 11.3 years. No dysplastic lesions were identified. Actionable findings leading to surgical intervention were found in 55 children (30 refractory reflux and 25 tracheoesophageal fistulas). Significant predictors of actionable pathology included increasing age, long gap atresia, and hiatal hernia. Symptoms were not predictive of actionable findings, except dysphagia, which was associated with stricture. Nelson-Aalen analysis predicted occurrence of an actionable finding every 5 years. CONCLUSIONS: Surveillance endoscopy uncovers high rates of actionable pathology even in asymptomatic children with EA. Based on the findings of the current study, a pediatric EA surveillance endoscopy algorithm is proposed.
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Atresia Esofágica , Esofagitis , Reflujo Gastroesofágico , Humanos , Niño , Atresia Esofágica/diagnóstico , Atresia Esofágica/cirugía , Estudios Retrospectivos , Esofagitis/complicaciones , Esofagitis/diagnóstico , Reflujo Gastroesofágico/etiología , Reflujo Gastroesofágico/complicaciones , EndoscopíaRESUMEN
Classically considered a disease of early childhood characterised by malabsorption and failure to thrive, coeliac disease is now recognised to arise in genetically susceptible individuals at any age. Although permissive HLA genotypes are the strongest predictor of coeliac disease, they are not sufficient. Several prospective cohort studies enrolling genetically at-risk infants have investigated the role of potential triggers of coeliac disease autoimmunity, such as timing of gluten introduction, viral infections and dietary patterns. Much less is known about triggers of coeliac disease in adulthood. Better understanding of factors leading to coeliac disease may be helpful in the management of those with potential coeliac disease (elevated serum celiac antibodies without villous atrophy in the small intestine), many of whom initiate a gluten-free diet without demonstration of villous atrophy. There are a range of clinical presentations of celiac disease in childhood and patterns of coeliac serology, including fluctuation and spontaneous reversion on a gluten-containing diet, vary. There is a current debate over best strategies to manage adults and children with potential coeliac disease to avoid over-treatment and under-treatment. Childhood and adolescence carry unique issues pertaining to the diagnosis and management of coeliac disease, and include nutrition and growth, rescreening, repeat biopsy, dietary adherence concerns and transition to adult care. In conclusion, while coeliac disease has similar pathogenesis and general clinical manifestations in paediatric and adult populations, diagnostic and management approaches need to adapt to the developmental stages.
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Enfermedad Celíaca , Adolescente , Adulto , Atrofia/patología , Enfermedad Celíaca/diagnóstico , Enfermedad Celíaca/terapia , Niño , Preescolar , Dieta Sin Gluten , Glútenes , Humanos , Lactante , Mucosa Intestinal/patología , Estudios ProspectivosRESUMEN
Polarized epithelial cells form an essential barrier against infection at mucosal surfaces. Many pathogens breach this barrier to cause disease, often by co-opting cellular endocytosis mechanisms to enter the cell through the lumenal (apical) cell surface. We recently discovered that the loss of the cell polarity gene PARD6B selectively diminishes apical endosome function. Here, we find that in response to the entry of certain viruses and bacterial toxins into the epithelial cells via the apical membrane, PARD6B and aPKC, two components of the PARD6B-aPKC-Cdc42 apical polarity complex, undergo rapid proteasome-dependent degradation. The perturbation of apical membrane glycosphingolipids by toxin- or virus-binding initiates degradation of PARD6B. The loss of PARD6B causes the depletion of apical endosome function and renders the cell resistant to further infection from the lumenal cell surface, thus enabling a form of cell-autonomous host defense.
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Toxinas Bacterianas , Virus , Toxinas Bacterianas/metabolismo , Polaridad Celular/fisiología , Endosomas/metabolismo , Células Epiteliales , Proteína Quinasa C/metabolismo , Virus/metabolismoRESUMEN
Epithelial cells lining mucosal surfaces distinctively express the inflammatory bowel disease risk gene INAVA. We previously found that INAVA has dual and competing functions: one at lateral membranes where it affects mucosal barrier function and the other in the cytosol where INAVA enhances IL-1ß signal transduction and protein ubiquitination and forms puncta. We now find that IL-1ß-induced INAVA puncta are biomolecular condensates that rapidly assemble and physiologically resolve. The condensates contain ubiquitin and the E3 ligase ßTrCP2, and their formation correlates with amplified ubiquitination, suggesting function in regulation of cellular proteostasis. Accordingly, a small-molecule screen identified ROS inducers, proteasome inhibitors, and inhibitors of the protein folding chaperone HSP90 as potent agonists for INAVA condensate formation. Notably, inhibitors of the p38α and mTOR pathways enhanced resolution of the condensates, and inhibitors of the Rho-ROCK pathway induced INAVA's competing function by recruiting INAVA to newly assembled intercellular junctions in cells where none existed before.
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Proteínas Portadoras/genética , Proteínas Activadoras de GTPasa/genética , Regulación de la Expresión Génica/efectos de los fármacos , Uniones Intercelulares/efectos de los fármacos , Bibliotecas de Moléculas Pequeñas/farmacología , Proteínas con Repetición de beta-Transducina/genética , Células CACO-2 , Proteínas Portadoras/metabolismo , Línea Celular Tumoral , Proteínas Activadoras de GTPasa/metabolismo , Células HEK293 , Proteínas HSP90 de Choque Térmico/genética , Proteínas HSP90 de Choque Térmico/metabolismo , Células HeLa , Humanos , Uniones Intercelulares/metabolismo , Uniones Intercelulares/ultraestructura , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteína Quinasa 14 Activada por Mitógenos/genética , Proteína Quinasa 14 Activada por Mitógenos/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , Proteína Adaptadora de Señalización NOD2/genética , Proteína Adaptadora de Señalización NOD2/metabolismo , Proteostasis/efectos de los fármacos , Proteostasis/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Transducción de Señal , Bibliotecas de Moléculas Pequeñas/química , Bibliotecas de Moléculas Pequeñas/clasificación , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/metabolismo , Proteínas con Repetición de beta-Transducina/metabolismoRESUMEN
Retrograde membrane trafficking from plasma membrane to Golgi and endoplasmic reticulum typifies one of the key sorting steps emerging from the early endosome that affects cell surface and intracellular protein dynamics underlying cell function. While some cell surface proteins and lipids are known to sort retrograde, there are few effective methods to quantitatively measure the extent or kinetics of these events. Here we took advantage of the well-known retrograde trafficking of cholera toxin and newly defined split fluorescent protein technology to develop a quantitative, sensitive, and effectively real-time single-cell flow cytometry assay for retrograde membrane transport. The approach can be applied in high throughput to elucidate the underlying biology of membrane traffic and how endosomes adapt to the physiologic needs of different cell types and cell states.
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Bioensayo/métodos , Membrana Celular/metabolismo , Análisis de la Célula Individual/métodos , Transporte Biológico , Toxina del Cólera/metabolismo , Enfermedad , Retículo Endoplásmico/metabolismo , Fluorescencia , Células HEK293 , Humanos , Células K562RESUMEN
The D1 dopamine receptor subtype is expressed in the brain, kidney and lymphocytes. D1 receptor function has been extensively studied and the receptor has been shown to modulate a wide range of physiological functions and behaviors. The expression of D1 receptor is known to change during development, disease states and chronic treatment; however, the molecular mechanisms that mediate the changes in D1 receptor expression under these circumstances are not well understood. While previous studies have identified extracellular factors and signaling mechanisms regulating the transcription of D1 receptor gene, very little is known about other regulatory mechanisms that modulate the expression of the D1 receptor gene. Here we report that the D1 receptor is post-transcriptionally regulated during postnatal mouse brain development and in the mouse CAD catecholaminergic neuronal cell line. We demonstrate that this post-transcriptional regulation is mediated by a molecular mechanism involving noncoding RNA. We show that the 1277 bp 3'untranslated region of D1 receptor mRNA is necessary and sufficient for mediating the post-transcriptional regulation. Using deletion and site-directed mutagenesis approaches, we show that the D1 receptor post-transcriptional regulation is specifically mediated by microRNA miR-142-3p interacting with a single consensus binding site in the 1277 bp 3'untranslated region of D1 receptor mRNA. Inhibiting endogenous miR-142-3p in CAD cells increased endogenous D1 receptor protein expression levels. The increase in D1 receptor protein levels was biologically significant as it resulted in enhanced D1 receptor-mediated signaling, determined by measuring the activation of both, adenylate cyclase and, the dopamine- and cAMP-regulated phosphoprotein, DARPP-32. We also show that there is an inverse correlation between miR-142-3p levels and D1 receptor protein expression in the mouse brain during postnatal development. This is the first study to demonstrate that the post-transcriptional regulation of D1 receptor expression is mediated by microRNA-induced translational suppression.
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Regulación de la Expresión Génica , MicroARNs/metabolismo , Receptores de Dopamina D1/genética , Transcripción Genética , Regiones no Traducidas 3'/genética , Animales , Animales Recién Nacidos , Emparejamiento Base/genética , Secuencia de Bases , Encéfalo/efectos de los fármacos , Encéfalo/crecimiento & desarrollo , Encéfalo/metabolismo , Catecolaminas/metabolismo , Línea Celular , Regulación de la Expresión Génica/efectos de los fármacos , Genes Reporteros , Masculino , Ratones , MicroARNs/antagonistas & inhibidores , MicroARNs/genética , Datos de Secuencia Molecular , Oligonucleótidos Antisentido/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Dopamina D1/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transcripción Genética/efectos de los fármacosAsunto(s)
Favismo/diagnóstico , Deficiencia de Glucosafosfato Deshidrogenasa/diagnóstico , Vicia faba/efectos adversos , Preescolar , Barreras de Comunicación , Fatiga/etiología , Favismo/etiología , Alfabetización en Salud , Humanos , Lenguaje , Masculino , Metahemoglobinemia/etiología , Oxígeno/sangre , Palidez/etiologíaRESUMEN
BACKGROUND: Management of periprosthetic femoral fractures is often complex, and few studies have documented its associated mortality. METHODS: We retrospectively identified from our trauma and surgical registries 106 patients who underwent surgery for a periprosthetic femoral fracture. We then identified a contemporaneous age and sex-matched control cohort of 309 patients who had a hip fracture (femoral neck or intertrochanteric) and 311 patients who underwent primary hip or knee replacement. Mortality at one year was identified with use of the Social Security database. RESULTS: Twelve (11%) of 106 patients died within one year following surgical treatment of a periprosthetic fracture. During the same follow-up period, fifty-one (16.5%) of 309 patients died following surgery for a hip fracture and nine (2.9%) of 311 patients died following primary joint replacement. The mortality rate after a periprosthetic femoral fracture was significantly higher (p < 0.0001) compared with that for matched patients who had undergone primary joint replacement, and it was similar to the mortality rate after a hip fracture. For periprosthetic fractures, a delay of greater than two days from admission to the time of surgery was associated with an increased mortality rate at one year (p < 0.0007). Forty-nine patients underwent revision arthroplasty for the treatment of a Vancouver type-B periprosthetic fracture, and six (12%) died. In contrast, twenty-four patients with a Vancouver type-B periprosthetic fracture were treated with open reduction and internal fixation and eight (33%) died. The difference was significant (p < 0.03). CONCLUSIONS: The mortality rate within one year following surgical treatment of periprosthetic femoral fractures is high and is similar to that after treatment for hip fractures. Because revision arthroplasty for the treatment of type-B periprosthetic fractures was associated with a one-year mortality rate that was significantly less than that after surgical treatment with open reduction and internal fixation, in instances when either treatment option is feasible, revision arthroplasty may be the preferred option.