Characterization of human, mouse, and rat cultures of enteric glial cells and their effect on intestinal epithelial cells.

BACKGROUND: Enteric glial cells (EGC) are major regulators of neuronal and intestinal epithelial cell (IEC) functions. Simple isolation methods of EGC, especially human tissues, remain scarce and limit their study. We present herein a method to isolate EGC and we characterize EGC phenotype and their functional impact on IEC. METHODS: Longitudinal muscle and myenteric plexus preparations of rat, mouse, or human intestine were obtained by microdissection. After mechanical and enzymatic dissociation, individual ganglionic or interganglionic structures were seeded into plates, maintained in culture several weeks and passaged up to 4 times. Purity of cultures was assessed by immunocytochemistry using antibodies against glial fibrillary acidic protein (GFAP), S100ß and Sox10 or smooth muscle actin. Effects of adenosine triphosphate (ATP) on intracellular Ca²âº signaling in EGC were studied. Co-cultures of EGC with IEC line, Caco-2, were performed for 2-6 days to analyze their impact on monolayer resistance, cell proliferation, and cell spreading. KEY RESULTS: More than 80% of DAPI-positive cells were GFAP, S100ß, and Sox10-immunoreactive. EGC expressed these glial markers over 4 consecutive passages, and the majority of them responded to ATP by an increase in intracellular Ca²âº concentration. In addition, rat, mouse, and human EGC increased intestinal barrier resistance, IEC size, and reduced IEC number. CONCLUSIONS & INFERENCES: We have developed a simple method to isolate and culture human, rat, or mouse EGC. EGC exhibit similar functional properties on the intestinal barrier independently of the species. This study sets the basis for exploring glial biology and functions in human health and diseases.

Postnatal development of myenteric neurochemical phenotype and impact on neuromuscular transmission in the rat colon.

Profound changes in intestinal motility occur during the postnatal period, but the involvement of the enteric nervous system (ENS), a key regulator of gastrointestinal (GI) motility, in these modifications remains largely unknown. We therefore investigated the postnatal development of the ENS phenotype and determined its functional repercussion on the neuromuscular transmission in the rat colon. Sprague-Dawley rats were euthanized at postnatal day (P) 1, P3, P5, P7, P14, P21, and P36. Whole mounts of colonic myenteric plexus were stained with antibodies against choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), and HuC/D. Colonic contractile response induced by electrical field stimulation (EFS) was investigated in organ chambers in absence or presence of N-nitro-l-arginine methyl ester (l-NAME) and/or atropine. In vivo motility was assessed by measurement of the colonic bead latency time. Randomly occurring ex vivo contractions appeared starting at P5. Starting at P14, rhythmic phasic contractions occurred whose frequency and amplitude increased over time. In vivo, bead latency was significantly reduced between P14 and P21. Ex vivo, EFS-induced contractile responses increased significantly over time and were significantly reduced by atropine starting at P14 but were sensitive to l-NAME only after P21. The proportion of ChAT-immunoreactive (IR) neurons increased time dependently starting at P14. The proportion of nNOS-IR neurons increased as early as P5 compared with P1 but did not change afterward. Our data support a key role for cholinergic myenteric pathways in the development of postnatal motility and further identify them as putative therapeutic target for the treatment of GI motility disorders in the newborn.

High-flow priapism in children: immediate treatment by selective embolization.

We present a case of a high-flow priapism related to a posttraumatic aneurysm in a ten-year-old boy treated by selective embolization. Patient history and thorough physical examination often lead to the diagnosis of high-flow priapism. Colour-coded duplex ultrasonography confirms the diagnosis, and treatment consists of angiographic embolization.

[Comparison of 3 self lubrificated urethral catheters: prospective study on 27 patients]. / Comparaison de trois sondes urétrales droites autolubrifiées: étude prospective chez 27 patients.

Clean intermittent catheterization has become a usual procedure in urologic practice. Self lubrificated catheters are nowadays accepted as the most efficient. This randomised and prospective study compared tolerance and efficiency in a group of 27 patients of three well known catheters (12 charriere, straight): Flocath (Ruschcare), Lofric (Astratech) and Easycath (Coloplast). The systematic comparison of this three catheters did not allow to put in a prominent position one of these. The three catheters are extremely close in terms of tolerance and performance.

[Recurrent peri-stomal varix hemorrhage after diversion using the Bricker's technique]. / Hémorragies récurrentes par des varices péri-stomiales après dérivation selon Bricker.

Peristomal varices can occur in patients with gastrointestinal or urinary diversions associated with portal hypertension. It is now no longer rare to propose radical surgery for invasive bladder tumours in patients also suffering from hepatic cirrhosis, responsible for specific subsequent complications. Less than ten cases of varicose haemorrhages have been described on ileal bladders. The authors report the case of a patient with cirrhosis (Child B) treated surgically (radical cystoprostatectomy and Bricker transileal cutaneous diversion) for invasive bladder tumour. Episodes of bleeding varices occurred two months after surgery. Repeated and abundant haemorrhage led to the placement of an intrahepatic shunt (TIPS) allowing reduction of the portal hypertension and the severity of the bleeding. When local control of the bleeding is no longer possible, reduction of the portosystemic pressure gradient is required. TIPS is an effective alternative to surgical shunts, responsible for high morbidity and mortality in these debilitated patients.

[Long-term results of interferential current stimulation in the treatment of bladder instability]. / Résultats à long-terme de la stimulation par courants interférentiels dans le traitement de l'instabilité vésicale.

OBJECTIVE: To study the late results of interferential current stimulation in the treatment of detrusor instability. MATERIAL AND METHODS: 62 patients with detrusor instability refractory to medical treatment by anticholinergic drugs were treated by interferential current stimulation. This technique combines the advantages of retraining stimulation with external application. This retrospective study was based on 62 patients (43 children, 11 men and 8 women) presenting with detrusor instability between January 1990 and December 1997. All patients were assessed clinically and by a radiological, bacteriological and urodynamic work-up prior to treatment. The mean follow-up was 5 years (range: 18 months to 10 years). RESULTS: The results of this technique were excellent, with 80.9% of cures at one year, but they tended to fade over time to 40% of cures at 5 years. However, results which deteriorate after one year can generally be maintained by performing 5 maintenance sessions every 12 or 18 months. CONCLUSION: Treatment of detrusor instability by interferential current is a reliable technique which constitutes an alternative to the other methods of retraining stimulation and can be performed in cases of instability refractory to anticholinergic drugs, before considering neuromodulation or surgery. Five to 10 maintenance sessions every 12 or 18 months ensure stable long-term results in the majority of cases.