Microenvironmental interactions between endothelial and lymphoma cells: a role for the canonical WNT pathway in Hodgkin lymphoma.

The interaction between vascular endothelial cells (ECs) and cancer cells is of vital importance to understand tumor dissemination. A paradigmatic cancer to study cell-cell interactions is classical Hodgkin Lymphoma (cHL) owing to its complex microenvironment. The role of the interplay between cHL and ECs remains poorly understood. Here we identify canonical WNT pathway activity as important for the mutual interactions between cHL cells and ECs. We demonstrate that local canonical WNT signaling activates cHL cell chemotaxis toward ECs, adhesion to EC layers and cell invasion using not only the Wnt-inhibitor Dickkopf, tankyrases and casein kinase 1 inhibitors but also knockdown of the lymphocyte enhancer binding-factor 1 (LEF-1) and ß-catenin in cHL cells. Furthermore, LEF-1- and ß-catenin-regulated cHL secretome promoted EC migration, sprouting and vascular tube formation involving vascular endothelial growth factor A (VEGF-A). Importantly, high VEGFA expression is associated with a worse overall survival of cHL patients. These findings strongly support the concept that WNTs might function as a regulator of lymphoma dissemination by affecting cHL cell chemotaxis and promoting EC behavior and thus angiogenesis through paracrine interactions.

WNT5A: a motility-promoting factor in Hodgkin lymphoma.

Classical Hodgkin lymphoma (cHL) has a typical clinical manifestation, with dissemination involving functionally neighboring lymph nodes. The factors involved in the spread of lymphoma cells are poorly understood. Here we show that cHL cell lines migrate with higher rates compared with non-Hodgkin lymphoma cell lines. cHL cell migration, invasion and adhesion depend on autocrine WNT signaling as revealed by the inhibition of WNT secretion with the porcupine inhibitors Wnt-C59/IWP-2, but did not affect cell proliferation. While application of recombinant WNT5A or WNT5A overexpression stimulates HL cell migration, neither WNT10A, WNT10B nor WNT16 did so. Time-lapse studies revealed an amoeboid type of cell migration modulated by WNT5A. Reduced migration distances and velocity of cHL cells, as well as altered movement patterns, were observed using porcupine inhibitor or WNT5A antagonist. Knockdown of Frizzled5 and Dishevelled3 disrupted the WNT5A-mediated RHOA activation and cell migration. Overexpression of DVL3-K435M or inhibition of ROCK (Rho-associated protein kinase) by Y-27632/H1152P disrupted cHL cell migration. In addition to these mechanistic insights into the role of WNT5A in vitro, global gene expression data revealed an increased WNT5A expression in primary HL cells in comparison with normal B-cell subsets and other lymphomas. Furthermore, the activity of both porcupine and WNT5A in cHL cells had an impact on lymphoma development in the chick chorionallantoic membrane assay. Massive bleeding of these lymphomas was significantly reduced after inhibition of WNT secretion by Wnt-C59. Therefore, a model is proposed where WNT signaling has an important role in regulating tumor-promoting processes.

Right esophageal lung in a preterm child with VACTERL association and Mayer-Rokitansky-Kuster-Hauser syndrome.

We report on a preterm girl (birth weight 1,200 g) with a right esophageal lung in esophageal atresia type VIag (according to the extended classification of Kluth). Additionally, the child suffered from an atrioseptal defect, a dextrocardia with a left descending aorta, a duodenal atresia, a high type of anal atresia (VACTERL association), agenesis of the left kidney, and agenesis of the vagina, uterus, and ovarian tubes (Mayer-Rokitansky-Kuster-Hauser syndrome or incomplete MURCS association). The child was treated with an emergency gastrostomy because of increasing abdominal dilatation. Thereafter, the parents refused further surgical treatment, and the child was maintained on basic therapy. After an uneventful period of 4 weeks, the child died of an acute massive aspiration. This case shows that sufficient spontaneous ventilation is possible in esophageal lung as long as a gastrostomy is kept on suction to prevent overinflation of the affected lung and the stomach. Ethical aspects have to be considered when treatment is planned in cases of prematurity and associated malformations when a chance of good survival is rather limited. The stepwise approach as proposed in the present case appears to be the only possible therapeutic regimen that can be offered in this complicated condition.

Sonographic prognostic factors in prenatal diagnosis of SCT.

OBJECTIVE: A subset of fetuses with sacrococcygeal teratomas (SCT) develops hydrops caused by high-output heart failure. Identification of fetuses at risk for hydrops is important because surgical intervention may reverse the pathophysiology of the disease. The aim of this study was to evaluate sonographic prognostic factors regarding tumor morphology and vascularity associated with the development of hydrops in utero. METHODS: Over a 10-year period, we identified 7 fetuses with SCT diagnosed antenatally and managed at the University of Mainz. We retrospectively reviewed the charts of mothers and infants and recorded data on prenatal diagnosis, tumor size and localization, perinatal management, neonatal care, and fetal outcome. RESULTS: The diagnosis of SCT was made in all cases by ultrasound. The median gestational age at the time of initial diagnosis was 23 weeks. In 3 cases, signs of fetal heart failure were detected by ultrasound. Pathological blood flow in the venous system was further noted in 2 cases. One fetus developed hydrops. The mean gestational age at delivery was 35 weeks, depending on the presence or absence of maternal or fetal complications. Six infants were delivered by cesarean section, and 1 by vaginal delivery. After fetal stabilization, surgery was performed in 5 of 7 cases. Inadequate ventilation secondary to prematurity was a contributing factor to death in 1 fetus. One fetal intrauterine death occurred at 27 weeks of gestation. CONCLUSION: Pregnancies with antenatally diagnosed fetal SCT necessitate frequent monitoring to ensure the detection of fetal/maternal complications by ultrasound and Doppler ultrasound. The most important prognostic criteria were cardiomegaly, fetal hydrops, and increased preload indexes of the fetal venous system as sign of fetal heart failure. Many studies show that the occurrence of pulsations in the umbilical vein of a hydropic fetus correlates with a poor fetal outcome. The decision on the optimal time of delivery should therefore be made by a multidisciplinary team of specialists.

Ellipsometric microscopy: developments towards biophysics.

Ellipsometric microscopy is a novel technique that combines the merits of ellipsometry and light microscopy, i.e. it allows noninvasive, label-free measurements of thin film thickness and refractive index at high lateral resolution. Ellipsometric microscopy has been successfully applied to silicon-air interfaces. However, typical biological systems require immersion in an aqueous buffer. Thus the authors have adapted the instrument for the observation of the interface between glass and water. In particular, the comparatively small differences in refractive indices between substrate and ambient media proved to be a challenge for instrument design. The first experiments with this new instrument are presented.

The clinical relevance of non-classified dysganglionoses and implications for a new grading system.

In addition to the classified types of dysganglionosis, certain non-classified dysganglionoses (NCD) (types 1-4) were introduced by Meier-Ruge in 1992. Clinical data on these conditions are limited. Among 134 children with intestinal dysganglionoses (ID) treated from 1979 to 1999, 12 were identified to have a NCD. Their clinical course is presented. The existence of mild ID (type 1) is difficult to demonstrate. Current definitions and data on clinical relevance are not convincing. An indication for surgical treatment is not present. Isolated hypogenesis of the submucous plexus (SMP) (type 2, n = 8) is clinically a more severe kind of intestinal neuronal dysplasia type B and often requires early surgical intervention, but not resection. When associated with aganglionosis, its recognition is important for surgical strategy, to avoid complicated clinical courses, which are frequent if total or nearly-total resection is not performed. Hypogenesis of the myenteric plexus (MP) (type 5, n = 1) has received little attention so far. The sporadic appearance of heterotopic nerve cells of the SMP in the mucosa (type 3, n = 1) is physiologic; clusters of such cells, however, are probably of pathologic value, especially in combination with other types of ID in the same patient. Heterotopic nerve cells of the MP (type 4, n = 3) in the circular and longitudinal muscle layers are highly pathologic. This clearly-defined type is of major clinical relevance and requires complete resection. A severe disturbance of the migration process is the underlying cause. To simplify the terminology of IDs, a grading system based on the anatomic structures and clinical findings is proposed: innervation disturbances of the mucosa (grade I) are of limited clinical significance. Isolated malformations of the SMP (grade II) may require an enterostomy, but do not require resection except in certain cases associated with distal aganglionosis. Dysganglionosis of the MP (grade III) usually exhibits more severe symptoms and resection is indicated, especially with associated hypo- or aganglionosis. In aganglionic bowel (grade IV) resection is mandatory.

Complex long-segment intestinal dysganglionosis.

A case is reported with aganglionosis of the rectum, sigma, and descending colon; dysganglionosis with heterotopic ganglionic cells in the muscularis propria of the hypoganglionic transverse colon; and extreme hypoganglionosis (without detection of ganglionic cells) of the ascending colon and distal ileum. The ileum showed a transition zone with hypoganglionosis and intestinal neuronal dysplasia (IND) type B. As to the etiology of such complex intestinal innervation defects, pre- and perinatal perfusion deficits must be considered because their localization seems to be linked to the vascular anatomy of the colon. Early diagnosis may be difficult, causing a delay in operative treatment and multiple operations. Different manifestations of dysganglionosis may be found in the same patient. The classical continuum of distal aganglionosis followed by proximal hypo- or dysganglionosis and then normally innervated bowel may not always be present. Therefore, in children with recurrent (sub-)ileus after resection of an aganglionic bowel segment, additional dysganglionosis such as IND or hypoganglionosis or even complex intestinal dysganglionosis should be excluded by full-thickness colon and small bowel biopsies.

[Erectile dysfunction secondary to hyperprolactinemia. Apropos of 13 cases]. / Dysérection secondaire à une hyperprolactinémie. A propos de 13 cas.

Hyperprolactinemia is the cause of erectile dysfunction in less than 1% of cases. From 1989 to 1996, 13 patients consulted for erectile disorders associated with hyperprolactinemia. The mean age was 47.5 years. 10 patients complained of decreased libido. 3 patients had gynecomastia. Plasma prolactin levels ranged from 31.3 ng/ml to 1,300 ng/ml. 7 patients had a plasma testosterone less than 4 ml/ng. 7 patients had a micro- or macroadenoma of the sella turcica visualized by MRI. After drug treatment, plasma prolactin levels returned to normal in all patients in whom assays were performed. 6 patients considered that their erectile function was restored. 5 of the 6 patients with no improvement of their sexual function had a concomitant disease able to explain the impotence. Hyperprolactinemia is a rare cause of erectile dysfunction, but it must be considered in any patient presenting with idiopathic erectile dysfunction associated with decreased libido, gynecomastia, and decreased plasma testosterone. Drug treatment is effective and MRI of the sella turcica should be performed looking for a pituitary adenoma.

Postoperative intussusception in childhood.

Over a period of 10 years, five children developed postoperative intussusception after intra-abdominal procedures at the Department of Pediatric Surgery of the Johannes Gutenberg University Mainz. Two appendectomies, one ileal resection for a Meckel's diverticulum, one operative procedure for Hirschsprung's disease plus intestinal neuronal dysplasia type B, and one hiatoplasty with jejunostomy preceded the intussusception. Three of the five children were older than 2 years. The clinical symptoms consisted primarily of abdominal distension, diffuse abdominal pain, bilious vomiting, and rectal bleeding in one case. Preoperative diagnosis was achieved in four cases by abdominal ultrasound. Plain abdominal radiographs demonstrated dilated loops of small intestine with air-fluid levels in four of the five cases. In the case without radiographic findings, the jejunojejunal intussusception was missed even by a bowel follow-through. The intussusceptions were ileocolic (3), ileoileal (1), and jejunojejunal (1). A hydrostatic procedure to reduce an ileocolic intussusception was not successful. Operative treatment of the intussusception was performed in three cases within 5 days, once at 32 days, and once 3 months after the primary operation, in all cases by laparatomy and simple manual reduction without intestinal resection. In contrast to idiopathic intussusception, noninvasive hydrostatic procedures are not indicated in postoperative intussusception, since protection of intestinal anastomoses from hydrostatic pressure and exclusion of other causes of postoperative ileus are mandatory.

[Diagnosis and surgical therapy of thyroid gland diseases in childhood]. / Diagnostik und operative Therapie von Schilddrüsenerkrankungen im Kindesalter.

Among children, thyroid disease rarely points to a particular surgical therapy and therefore requires a very specific surgical approach as well as close interdisciplinary cooperation between pediatricians, pediatric surgeons, and nuclear medicine practitioners, in particular regarding the follow-up. As a consequence, excellent functional and long-terms results can be achieved. The extent of the radical surgery pertaining to the small, encapsulated papillary carcinoma is still being discussed and investigated.

[Diagnostic possibilities and treatment approaches for fracture of the of the intercondylar eminence in the growth period]. / Diagnostische Möglichkeiten und Behandlungsstrategien für die Fraktur der Eminentia intercondylica im Wachstumsalter.

Between 1970 and 1993 29 fractures of the intercondylar eminence were seen in children with almost the same number of cases showing no or minor displacement as those requiring open reduction for major dislocation. Combined knee injury must be suspected the higher the patient's age and the more pronounced the displacement of the fragment. Magnetic resonance imaging (MRI) has gained diagnostic predominance. Arthroscopy remains important for surgical management, especially of associated meniscus and cartilage damage. Early puncture of hemarthrosis is essential. Closed management is the therapy for nondisplaced fragments or those with bone contact or only tuberculum fractures--while displaced fragments require operative reduction. Arthroscopy-aided wire fixation is the preferred technique, followed by open reduction using absorbable suture material (wire in special cases). Traction screws should be used only in older children with large fragments. Our series show good late results because occasional function impairment with knee instability tends to normalize with growth. Corrective orthopedic procedures--if at all--are required only for juveniles or adults.

[New aspects in the treatment of complete and isolated diaphyseal fracture of the forearm in childhood]. / Neue Gesichtspunkte zur Behandlung der kompletten und isolierten diaphysären Unterarmfrakturen im Kindesalter.

From 1984 to 1992 153 children were treated at the pediatric surgical department of the University of Mainz, of whom 19 had fractures of the radius, 9 of the ulna and 125 had a combination of both in the diaphysis of the forearm. Factors like patient's age, type of fracture and therapeutic methods were analyzed. One hundred and forty-one children were treated conservatively, 12 by operation. One hundred and one of these patients were healthy, when discharged (about 8 weeks after accident). Follow-up was possible in 69 cases. We found that occasional a remaining dislocation angle does not impair function: in 65 cases the result was "good" and "very good", but only "moderate" in 4 cases. These 4 children had been treated conservatively by reposition and plaster cast; 2 of these 4 children showed bone reconstruction without dislocation. Twenty-two of the 56 X-ray follow-up's showed persisting dislocation. To prevent functional problems (2 of our cases) it is essential to obtain a very exact reposition of diaphyseal forearm fractures. Therefore we would recommend a more generous indication for operation, preferably using elastic-stable Nancy pins.

Immunochemical studies on the nature of the serum component (SAA) related to secondary amyloidosis.

The relative occurrence of the amyloid-related serum protein SAA in various disease states and in healthy subjects has been compared by both solid phase radioimmunoassay ((RIA) and immunodiffusion techniques which employ antibodies to purified amyloid fibril proten AA of hemogeneous size and charge. SAA levels were elevated above normal in certain categories of neoplastic, inflammatory, and infectious diseases as well as in secondary amyloidosis. The lowest median value, 8 ng./ml., was observed for approximately 150 normal sera, with no age-related increase in subjects ranging in age from 16 to 70 years. The results are consistent with several recent observations that SAA is normal acute phase reactant, and hence the RIA for SAA has no prognostic or diagnostic significance for secondary amyloidosis. The sensitivity of RIA for the detection of SAA is lower than would be expected when AA cross-reactivity values for sera are correlated with their reaction with anti-AA antibodies in immunodiffusion. This observation, along with others reported elsewhere suggests that those determinants which cross-react with anti-AA antibodies are relatively hidden in native SAA. Myeloma sera were less reactive than other groups of pathologic sera in immunodiffusion, although they were similar to other patients' sera when analyzed by RIA. Antibodies to highly purified AA were also used to investigate the structue of SAA by a double-antibody immunoprecipitation method. Precipitated SAA was partially dissociated during sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis to a 12,500 molecular weight moiety designated, SAAL. Multiple radiolabeled species of molecular weight intermidiate to SAA and SAAL were also detected and appeared to represent imcompletely dissociated SAA. The results suggest the SAA is an aggregate of several SAAL chains.