1,721,005 research outputs found
Organoidi gastrointestinali pediatrici e fetali: modello di cultura tridimensionale in vitro
Full text linkObiettivi
Il COVID-19 è una malattia principalmente respiratoria nell’età adulta; nei bambini, al contrario, i sintomi gastrointestinali sono più frequenti. Inoltre, non si conoscono i meccanismi di infezione in epoca fetale, epoca in cui i pazienti sono raramente colpiti da COVID-19. Gli organoidi sono uno strumento relativamente nuovo per stabilire in vitro colture di lunga durata che assomigliano tridimensionalmente al tessuto di origine e possono sia mantenere la staminalità che differenziarsi completamente in tutti i tipi di cellule.
Abbiamo quindi mirato a sviluppare un sistema di coltura per organoidi gastrointestinali (GIO) per studiare l'infezione da SARS-CoV-2 nell'epitelio gastrico lungo tutta la durata della vita.
Metodi
I GIO sono stati derivati da feti di 8-21 settimane e da tessuti pediatrici e adulti. Sono stati coltivati utilizzando un terreno chimicamente definito, per testare la loro capacità di mantenere la forma e di differenziarsi completamente. GIO sono stati analizzati in correlazione all'ECM circostante.
Gli organoidi sono stati quindi indotti a differenziarsi nella forma a polarità cellulare inversa (RP-GO) e quindi incubati con SARS-CoV-2. Tutti gli esperimenti sono stati analizzati mediante qPCR, immunofluorescenza e analisi qualitativa a seconda dei casi.
Risultati
Gli organoidi gastrointestinali possono essere isolati da tutte le età gestazionali, dimostrando la normale morfologia dell'epitelio gastrico ed esprimendo tipi di cellule mature, comprese le cellule della nicchia, del villo/ghiandola e di tipo enteroendocrino. Queste colture possono essere mantenute indefinitamente in vitro e coltivate in condizioni conformi alle indicazioni GMP. Gli RP-GO mostrano polarità apicale, esponendo ACE2 sulla superficie esterna, ottimizzando le condizioni per l'infezione virale. La nucleoproteina virale è stata dimostrata nelle cellule in fase di apoptosi, con gli RP-GO pediatrici più suscettibili ed infettati in modo efficiente rispetto agli organoidi fetali e adulti.
Conclusioni
Abbiamo stabilito con successo un efficiente sistema di coltura di organoidi gastrointestinali per tutte le età, dalla vita fetale all'età adulta. La tecnologia basata sugli organoidi può essere utilizzata per modelli di malattie in vitro, test sui farmaci o terapia cellulare. L'applicazione di regole conformi alle GMP rende la traduzione clinica più vicina.Aims
Adult COVID-19 is mainly respiratory illness, but in children GI symptoms are more frequent. Furthermore, fetuses are rarely affected by COVID-19. Organoids are a relatively new tool to in vitro establish long-living culture that three-dimensionally resemble the tissue of origin and may both maintain the stemness and fully differentiate in all cell types.
As a proof of concept, we aimed to develop a culture system for gastrointestinal organoids (GIOs) to investigate SARS-CoV-2 infection in gastric epithelium across the lifespan.
Methods
GIO were derived from 8-21 week fetuses and from pediatric and adult tissues. They were cultured using chemically-defined medium, to test their ability to maintain stemness and to fully differentiate. GIO were analyzed in correlation to the surrounding ECM.
Reverse cellular polarity Organoids (RP-GOs) were induced and incubated with SARS-CoV-2. All experiments were analyzed by qPCR, immunofluorescence and qualitative analysis as appropriate.
Results
Gastrointestinal organoids can be isolated from all gestational ages, demonstrating normal gastric epithelial morphology and expressing mature gastric cell types including, the niche, secretive, and enteroendocrine cells. These cultures may be maintained indefinitely in vitro and cultured in GMP-compliant conditions. RP-GOs exhibit apical-out polarity, exposing ACE2 on the external surface, optimizing conditions for viral infection. Viral nucleoprotein was demonstrated in cells undergoing apoptosis, with pediatric RP-GOs most susceptible and efficiently infected compared to fetal and adult organoids.
Conclusions
We have successfully established an efficient gastrointestinal organoid culture systems for all ages, from fetal life to adulthood. Organoid-based technology can be used for in vitro disease modelling, drug testing or cell therapy. The application of GMP compliant rules makes the clinical translation closer
Newborn with incarcerated inguinal hernia and complete androgen insensitivity syndrome
Full text linkDisorders of sex development represent a large and heterogeneous group of diseases that have an important impact on physical and mental well-being of patients and their families. An early diagnosis and a multidisciplinary approach are fundamental to guarantee the highest standard of care.
We describe a case of complete androgen insensitivity syndrome (CAIS) in a six-days-old female newborn brought to our Emergency Department for intestinal occlusion due to an incarcerated inguinal hernia. During the operation, male-appearing gonads were identified in both inguinal canals and the histology confirmed the testicular features. Blood tests validated the diagnosis.
The diagnosis of CAIS is usually made during elective inguinal hernia repair or for delayed puberty. To our knowledge, this is the first case diagnosed in a neonatal urgent setting. Being able to immediately recognize this clinical picture, it allowed us to offer the best standard of care for the patient and the family
Approccio al bambino neuropatico con disfagia: l’alleanza necessaria tra anestesista e chirurgo.
No full textThe fate of implant after endoscopic injection of dextranomer/hyaluronic acid in vesicoureteral reflux: time to partial reabsorption and stabilization
No full textVesicoureteral reflux (VUR) potentially leads to renal damage, scarring, and eventually end-stage renal disease. Endoscopic treatment is well tolerated in children, it has reduced costs, and it effectively prevents urinary tract infections (UTIs), while avoiding long term antibiotics use
Multiple Congenital Colonic Stenosis: Case Report and Review of the Literature
No full textCongenital malformations affecting the colon are rare pediatric conditions often presenting as obstruction.
Colonic atresia accounts for 5-‐10% of atresia in newborns, stenosis is even more rare.Since 1968, only
16 cases of CCS have been reported in literature (Table 1). Because of the rarity of the disease, little is known about this uncommon condition and management is still controversial. We present a case of multiple congenital colonic stenosis and review the literature with a special focus on management of CCS. To the authors' knowledge, multiple congenital colonic stenosis has not been reported previously
in the literature
Eosinophilic Esophagitis in Esophageal Atresia: Is It Really a New Disease?
Full text linkEosinophilic esophagitis (EoE) is a chronic, immune-mediated esophageal disease. Symptoms are related to mucosal eosinophilic-predominant inflammation that leads to esophageal dysfunction. Recent data suggest that esophageal atresia (EA) patients may have an increased incidence of EoE compared to the general population. As EoE symptoms may be confused with EA-related symptoms, they may significantly worsen morbidity in this specific group of patients. We investigated specific characteristics of patients with AE and EoE compared to those with EoE only. We conducted an observational retrospective monocentric study including all patients diagnosed with EoE from 1 January 2010 to 31 December 2021. For each patient, demographic, clinical and histopathological data were collected and then compared between the two cohorts (EA-EoE vs. EoE only). During the study period, 62 patients were included: 17 children were in the follow-up because of EA (18.1% of 94 EA patients screened in that period), while the other 45 presented EoE only. The demographic and clinical features of EA-EoE patients demonstrate a lower prevalence of allergic subjects (23.5% vs. 80%, p < 0.05), a lower age of presentation (3.1 vs. 12.2 years, p < 0.05), non-specific symptoms and a higher resolution rate with PPI therapy (64.7% vs. 17.8%, p < 0.05) compared to EoE-only patients. Our data confirm that EA patients are at high risk for developing EoE. As symptoms may overlap with the EA spectrum, early recognition of EoE may prevent patients from receiving unnecessary invasive therapeutic interventions and from developing complications from untreated EoE
Comparison of outcomes following radiological reduction of intussusception with or without sedation
No full textBackground: Non-operative radiological reduction (NORR) is usually the first line treatment in pediatric ileo-colic intussusception. The aim of our study was to compare outcomes of NORR with or without sedation. Methods: All patients undergoing to contrast enema NORR for intussusception between 01.01.2015 to 31.12.2020 in two hospitals were included: in one centre patients were sedated (A) while in the other patients were awake (B). Primary outcome was the rate of radiological reduction. Secondary outcomes were length-of-stay, complications and recurrence rate. Results: Seventy-seven and 49 patients were included in group A and B respectively. Successful reduction rate was 72.7% in group A and 61.2% in group B (P>0.05). There were no complications related to the procedure among the 2 groups. Adverse events to sedation were observed in 3 patients. Conclusions: NORR has similar success rate when performed under sedation or awake, despite the former being graved by additional anesthesiologic risks and thus warrant careful indications
Intraoperative ultrasound-assisted approach for endoscopic treatment of vesicoureteral reflux in children
Full text linkPurpose Despite minimal invasiveness and high success rate, guidelines still prescribe voiding Cystourethrogram (VCUG) after endoscopic treatment for vesicoureteral reflux (VUR) in children. The aim of this paper was to analyze whether intraoperative ultrasound (IO-US) could improve surgical accuracy and perioperative counseling, thus potentially decreasing the need for postoperative VCUG. Methods We selected children treated for moderate to high grade VUR, renal scarring or repeated infections under antibiotic prophylaxis from January to December 2015. Endoscopic injection was combined with IO-US to detect optimal needle placement and to guide mound formation. IO-US findings were compared to surgeon opinion and postoperative VCUG, performed 3 months after surgery. All patients were followed-up for 1 year. Results A significant relationship was found between IO-US mound height (p = 0.003) or localization (p < 0.0005) and VCUG. Success of endoscopic treatment vs persistence of reflux groups had a mean mound height of 10.62 ± 1.36 mm and 8.39 ± 1.08 mm respectively (p < 0.0005). Height maintained a significant correlation with success in simple and multivariable regression analysis. ROC curve determined ⥠9.8 mm as predictor of reflux resolution (95% CI 0.825 to 0.998; p < 0.0001). Conclusions IO-US facilitates pediatric urologists to find an optimal location, to reach a volcano mound morphology and height, thus increasing intraoperative accuracy. IO-US also helps evaluating high-risk recurrence and guiding prognostic counseling. Type of study Treatment study. Level of evidence II
Long-Term Analysis of Respiratory-Related Complications Following Gastrostomy Placement with or without Fundoplication in Neurologically Impaired Children: A Retrospective Cohort Study
Full text linkGastrostomy placement is crucial in neurologically impaired (NI) children to ensure an adequate food intake and a safe route for drugs administration and to reduce the risk of primary aspiration. NI patents are more prone to gastroesophageal reflux. The association with fundoplication is deemed to reduce reflux-related respiratory complications. However, long-term benefits of this approach are not clear. We therefore aimed to compare long-term reflux-related respiratory complications of gastrostomy only (GO) to gastrostomy with fundoplication (GF). We retrospectively reviewed 145 consecutive NI children managed from 2008 to 2018. As long-term outcomes, we analyzed number and length of hospital admissions (Reflux-Related-Hospitalization, RRH) and emergency department accesses (Reflux-Related-Accesses, RRA) due to respiratory problems. Results were analyzed with appropriate statistical method. Median age at referral and at gastrostomy placement were 2.2 and 3.4 years (SD 5.6), respectively. Median follow-up was four years (range 1–12). Anti-reflux procedures were performed in 26/145 patients (18%); tracheotomy in 23/145 (16%). RRH following surgery showed lower number of admissions/year (0.32 vs. 1 for GO vs. GF, p < 0.005) and days hospitalization/year (3 vs. 13, p = 0.08) in GO compared to GF; RRA was similar (0.60 vs. 0.65, p = 0.43). Gastrostomy placement alone appeared not to be inferior to gastrostomy plus fundoplication with respect to long-term respiratory-related outcomes for NI children in our center
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