Udo Rolle

Interstitial cells of Cajal in the normal gut and in intestinal motility disorders of childhood

Interstitial cells of Cajal (ICCs) are pacemaker cells which are densely distributed throughout the whole gastrointestinal tract. ICCs have important functions in neurotransmission, generation of slow waves and regulation of mechanical activities in the gastrointestinal tract, especially for the coordinated gastrointestinal peristalsis. Therefore, a loss of ICCs could result in gastrointestinal motor dysfunction. In recent years c-kit labeling has been widely used to study pathological changes of ICCs in gastrointestinal motility disorders. Paediatric gastrointestinal motility disorders such as hypertrophic pyloric stenosis, Hirschsprung’s disease, total colonic aganglionosis, hypoganglionosis, intestinal neuronal dysplasia, internal anal sphincter achalasia, megacystis microcolon intestinal hypoperistalsis syndrome have been reported to be associated with loss or deficiency of ICCs networks. This review describes the distribution of ICCs in the normal gastrointestinal tract and its altered distribution in intestinal motility disorders of childhood.

C-Kit receptor (CD117) in the porcine urinary tract.

C-Kit positive interstitial cells of Cajal (ICC) play an important role in the regulation of the smooth muscle motility, acting as internal pacemakers to provide the slow wave activity within various luminal organs. Recently c-Kit-(CD117)-positive interstitial cells (IC) have been shown in the genitourinary tract, but systematic studies on the distribution and density of IC within the urinary tract are still lacking. Therefore the aim of the present study was to analyze systematically the localization and distribution of the c-Kit receptor in the urinary tract of the pig using immunohistochemical and molecular methods. Tissue samples were harvested from the porcine urinary tract including renal calices and pelvis, ureteropelvic junction, proximal, middle and distal ureter, ureteral orifice, fundus, and corpus of the bladder and the internal urethral orifice. Small and large intestine specimen served as controls. Immunohistochemistry (APAAP, IF) was applied on serial frozen sections using four monoclonal and polyclonal antibodies recognizing CD117. Whole mounts of the porcine upper urinary tract were prepared and investigated using conventional and confocal fluorescence microscopy followed by three-dimensional reconstruction. UV-laser microdissection and RT-PCR were applied to confirm the immunohistochemical results. CD117-immunoreactivity labeled bipolar IC and round-shaped mast cells (MC) throughout the adventitia, tunica muscularis and submucosa within the whole porcine urinary tract. While MC were found continuously in all investigated segments, a gradient of bipolar IC was evident. The whole mount preparations gave a detailed cytomorphology of IC within the various layers of the porcine urinary tract. Whole mount preparations revealed closed apposition of bi- and tripolar c-Kit positive IC parallel to the smooth muscle bundles and to veins of the tunica muscularis and adventitia. In the urothelium single CD117-positive interepithelial cells were found. The highest density of CD117-positive cells was found at the ureteropelvic junction, however the differences in between the segments were minimal. Microdissection and RT-PCR confirmed the results uncovered by immunohistochemistry. The ubiquitous distribution of IC and their close relationship to smooth muscle provides strong evidence that IC could contribute to the intrinsic pacemaker activity within the porcine (upper and lower) urinary tract. The role of the interepithelial CD117-positive cells as mechanosensors or as a precursor cell in the regeneration of the urothelium, is conceivable.

Experimental study evaluating the effect of a barrier method on postoperative intraabdominal adhesions.

The purpose of this animal study was to determine if tissue glue-coated collagen sponge is an effective barrier method to prevent localized adhesions in a modified rabbit sidewall model. Rabbits were divided into two groups and underwent laparotomy with subsequent creation of a cecal wound according to the rabbit sidewall model. Rabbits of group I (treatment group; n = 10) were treated with a TachoComb H patch placed on the defect, whereas group II animals (control group; n = 6) did not receive further treatment. All animals were sacrificed 2 weeks postoperatively and adhesions were evaluated using special adhesion score. A further six rabbits underwent TachoComb H application at the cecum to investigate the histological changes during a course of 12 weeks. The average adhesion scores were significantly (P < 0.05) reduced in the treatment group compared to the controls. Histologically the TachoComb H patch was surrounded by granulation tissue without signs of infection. Tissue glue-coated collagen sponge (TachoComb H) is effective to prevent localized intraabdominal adhesions in the modified rabbit sidewall model.

Structural basis of voiding dysfunction in megacystis microcolon intestinal hypoperistalsis syndrome

PURPOSE Megacystis microcolon intestinal hypoperistalsis syndrome (MMIHS) is a rare, congenital and usually fatal condition of unknown etiology. It is characterized by abdominal distension caused by a distended, non-obstructed urinary bladder and intestinal hypoperistalsis with functional intestinal obstruction. Previous studies reported vacuolar degenerative changes in the smooth muscle cells of bowel and bladder suggesting that MMIHS may be due to a visceral myopathy. The aim of this study was to examine the expression of contractile, cytoskeletal and extracellular matrix proteins in the detrusor muscle of MMIHS patients. MATERIAL AND METHODS Bladder specimens were obtained from six MMIHS patients. Normal bladder specimens were obtained during partial cystectomy and served as controls. Single fluorescence immunohistochemistry for alpha-smooth muscle actin (SMA), desmin, dystrophin, vinculin and collagen types I and III was carried out. Specific connective tissue stains (trichrome Masson, van Gieson) and electron microscopical investigations were also performed. RESULTS Trichrome Masson and van Gieson staining demonstrated markedly increased dense connective tissue between the layers of the detrusor muscle in MMIHS compared to controls. Collagen type I immunoreactivity was markedly increased and SMA, desmin and dystrophin immunoreactivity was markedly reduced in the bladder muscle of MMIHS compared to controls. Electron microscopy revealed vacuolar degenerative changes in smooth muscle cells and an abundance of connective tissue between these cells. CONCLUSION These data suggest that the detrusor muscle in MMIHS is strikingly abnormal and is the likely cause of voiding dysfunction.

C-kit receptor in the human vas deferens: distinction of mast cells, interstitial cells and interepithelial cells

The molecular mechanisms underlying the regulation of vas deferens (VD) motility and semen emission are still poorly understood. Interstitial cells of Cajal (ICC), which harbour the c-kit receptor (CD117), provide the basis of coordinated gut motility. We investigated whether c-kit receptor-positive cells also exist in the normal human VD. Enzyme and fluorescence immunohistochemical techniques were applied on serial sections of human proximal, middle, and distal VD segments (n=49) employing 13 different monoclonal and polyclonal antibodies recognizing the c-kit receptor. The c-kit receptor was detected in either round- or spindle-shaped cells. On account of their antigenic profile, the round- and oval-shaped c-kit receptor-positive cells were identified as mast cells (MC) occurring in all layers of the VD except the epithelium. In contrast, two distinct populations of exclusively c-kit receptor-positive spindle-shaped cells were found within the lamina propria and, rarely, in the inner and outer smooth muscle layers, as well as within the epithelium. Different shaped c-kit receptor-positive MC and IC were present in all layers of the human VD. Our findings demonstrate the presence of different c-kit receptor-positive cells also in the human VD. Their rather ubiquitous distribution within the lamina propria and muscle layers suggests that IC and MC may modulate the neuromuscular transmission and the propagation of electrical signals in multiple systems involved in the draining of fluids. The importance of the c-kit receptor-positive interepithelial cells remains unclear.

Universal expression of cell adhesion molecule NCAM in neuroblastoma in contrast to L1: implications for different roles in tumor biology of neuroblastoma?

PurposeNeuroblastoma is a biological, genetic and morphological heterogeneous tumor with a variable clinical course. NCAM is a cell adhesion molecule belonging to the immunoglobulin superfamily with structural similarities to cell adhesion molecule L1. The aim of this study was to determine the expression of NCAM in neuroblastoma and to compare the results to the findings of a previous study which examined L1 expression in the same group of patients.Materials and methodsNCAM expression was investigated on a tissue array with 66 surgically resected neuroblastoma samples by immunohistochemistry with a monoclonal antibody clone 1B6 and peroxidase method.ResultsStrong expression of NCAM was detected in all of the 66 (100%) neuroblastoma tumors in contrast to L1 which was not expressed in all tumors.ConclusionIn contrast to L1, which was found to predict favorable outcome, NCAM is universally expressed in neuroblastoma. Therefore NCAM represents a marker for neuroblastomas irrespectively of their stages whereas L1 as an indicator for developing neuronal cells seems to identify more mature stages of this tumor. The high grade of NCAM expression might present a prerequisite for establishment of antibody-based therapies.

Pulmonary artery sling and congenital tracheal stenosis in another patient with Mowat–Wilson syndrome

Sibylle Strenge,* Wolfram Heinritz, Christiane Zweier, Anita Rauch, Udo Rolle, Andreas Merkenschlager, and Ursula G. Froster Medical Faculty, Institute of Human Genetics, University of Leipzig, Leipzig, Germany Institute of Human Genetics, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nuremberg, Erlangen, Germany Department of Pediatric Surgery, University of Leipzig, Leipzig, Germany Hospital for Children and Adolescents, University of Leipzig, Leipzig, Germany

Combination of Spit Fistula Advancement and External Traction for Primary Repair of Long-Gap Esophageal Atresia

Primary repair of long-gap esophageal atresia with almost complete absence of thoracic esophagus was usually believed to be impossible. Thus, esophageal replacement with colon or gastric interposition seemed inevitable. Esophageal lengthening techniques could be an alternative approach. Herewith we describe for the first time the successful combination of the stepwise subcutaneous advancement of the upper esophageal segment (Kimuras technique) with transthoracic traction on the lower esophageal segment (Fokers technique). This combined lengthening technique leads to the primary repair of a long-gap esophageal atresia.

Postnatal development of the mucosal plexus in the porcine small and large intestine

Knowledge regarding the foetal and postnatal development of the enteric nervous system is crucial for the understanding of congenital disorders. While lot of information exists regarding the myenteric and submucosal plexuses, the development of the mucosal plexus has not been previously studied. The mucosal innervation seems to play an important role in the local reflex activity of the gut. In this study, we examined the development of enteric mucosal innervation in the pig at various ages of life. Small and large bowel paraffin-embedded specimens were stained with PGP 9.5 and neurofilament protein in three piglets from six age groups (60 and 90 days gestation, newborn, 4 and 12 weeks old, and adult pigs). Small and large bowel demonstrated identical innervation patterns. Myenteric and submucosal plexuses were stained with PGP 9.5 at 60 days gestation. However, the mucosal staining was first noted clearly at the newborn period. By 4 weeks, PGP 9.5 staining was noted in small amounts within the mucosa. Inner proprial and villous fibres were seen ahead in time to the subepithelial fibres. Both inner proprial and villous staining became quiet prominent by 12 weeks of age and remained unchanged into adulthood. However, the subepithelial fibres appear to increase in adulthood. This study demonstrates for the first time that enteric mucosal innervation first appears only at birth. The immaturity of the mucosa generated reflex activity, and secretory functions may have implication in the management of functional intestinal obstruction in the premature infant.

Bladder outlet obstruction causes fetal enterolithiasis in anorectal malformation with rectourinary fistula

Extraluminal calcified meconium is found frequently by prenatal ultrasound in cases with bowel perforation and meconium peritonitis. Intraluminal intestinal meconium calcifications are rarely seen in prenatal sonography. Meconium calcifications result from a mixture of meconium and urine that indicates a connection between intestinal and urinary tract. We report a case of a male newborn prenatally diagnosed with intraluminal echogenic calcifications at 23 weeks of gestation, suggesting an anorectal malformation (ARM) with rectourinary fistula. At birth, the child presented with a complex ARM including high anal atresia with both perineal and rectourethral fistula. Furthermore, a bladder outlet obstruction due to a urethral stenosis was diagnosed. Vesicostomy was performed as an emergency procedure followed by colostomy during neonatal period. Posterior sagittal anorectoplasty was performed at the age of 4 months. Prenatal echogenic calcifications within bowel should raise the suspicion of ARM with rectourinary fistula and bladder outlet obstruction.