Expression of cell growth regulated genes in the fetal kidney: relevance to in utero obstruction.

Previous studies of fetal urinary tract obstruction (bladder outlet obstruction and ureteral obstruction) in lambs have shown that obstructions created relatively early in gestation (4/10 to 6/10 term) can significantly affect growth of the developing kidney. This suggests that urinary tract obstruction in utero can alter normal mechanisms of kidney growth. However, a mechanism for these effects has not yet been proposed. In this study we have used mRNA expression analysis to characterize the temporal sequence of expression of several growth-regulated genes during normal ovine kidney development. The purpose of this study was to test the hypothesis that early obstructions, such as those believed to arise in congenital obstructive uropathy in humans, might have a disproportionate effect on hyperplastic growth if the cellular growth fraction (percent of cells in the organ undergoing DNA synthesis) was greater in the second trimester than in the last. Northern blot analysis of the cell cycle-dependent genes histone H3, c-myc and ornithine decarboxylase (ODC) indicated a progressive, gradual decline in cellular proliferation in the kidney from approximately 60 to 135 days (4/10 term to term) gestation, as evidenced by decreases in the respective mRNA levels. The greatest levels of cell proliferation occurred near the midpoint of gestation. This indirect measurement of decline in cellular growth fraction was reflected in direct measurements of change in relative kidney weight. To test whether this decline in mRNA levels occurs widely among genes expressed in the fetal kidney during this period, relative expression levels of more than 300 anonymous mRNA transcripts were evaluated by differential display analysis. This method showed that genes whose expression patterns resembled the growth-regulated genes constituted less than 5% of the expressed mRNAs identified. These data indicate that intrauterine urinary tract obstructions that arise at or near the midpoint of gestation coincide with the highest rates of cell proliferation occurring in the second and third trimesters and, therefore, might adversely affect mechanisms of cell proliferation.

Pediatric laparoscopic dismembered pyeloplasty.

We performed laparoscopic dismembered pyeloplasty in a boy with right ureteropelvic junction obstruction using 4 cannula sites, and a dismembering and reanastomosis technique identical to that used in open pyeloplasty. Interrupted sutures were placed and tied intracorporeally. A nephrostomy tube was placed under direct vision for drainage but no ureteral stent was used. Total operating time was 5 hours. The patient was discharged home 36 hours after the procedure. The nephrostomy tube was removed 10 days postoperatively after radiographic demonstration of patency and 24 hours of clamping without pain. Followup excretory urography at 6 weeks showed much less hydronephrosis and a widely patent anastomosis. Our case illustrates the technical features and feasibility of laparoscopic pyeloplasty in children, and should encourage further development of pediatric urological reconstructive laparoscopic techniques.

Laparoscopic retroperitoneal lymph node dissection in stage A nonseminomatous testis cancer.

Open retroperitoneal lymph node dissection in clinical Stage A testis cancer offers the advantages of definitive staging and high cure rates. Observation is elected in various settings to avoid the morbidity and possible sequelae of this procedure. We present a case report of a new procedure, laparoscopic retroperitoneal lymph node dissection, that decreases hospital stay, recovery time, and morbidity while maintaining the advantages of open retroperitoneal lymph node dissection.

Altered extracellular matrices influence cellular processes and nuclear matrix organizations of overlying human bladder urothelial cells.

A central issue in tumor biology is the understanding of the interactions between tumor cells and their environment. Using normal and ras oncogene transfected rat fibroblast cells, we now demonstrate that the transfected cells make altered extracellular matrices (ECM) and that their resulting ECM influence the proliferation and genetic regulation of human bladder cancer EJ cells. Using Western blot analyses, we observed that the ras transfected fibroblast cells lacked the ability to produce extracellular matrix component laminin whereas the normal parental fibroblast cells were able to produce intact laminin. Both transfected and nontransfected fibroblast cells were able to synthesize other extracellular matrix molecules such as type IV collagen and fibronectin. Human bladder tumor EJ cells were grown on ECM derived from normal and transfected rat fibroblast cells, and the proliferation rate and type IV collagen mRNA expression of EJ cells were determined. We observed that EJ cells, when grown on ECM derived from the ras transfected fibroblast cells, had a higher growth rate than when grown on ECM derived from the normal fibroblast cells (P < 0.037). Furthermore, EJ cells grown on ECM derived from transfected fibroblast cells showed up-regulation of type IV collagen mRNA expression when compared with EJ cells grown on ECM derived from nontransfected fibroblast cells. Finally EJ cells grown on purified laminin but not on collagen IV coated flasks showed the same level of type IV collagen mRNA expression as when grown on ECM derived from nontransfected parental fibroblast cells. Haptotactic/motility assays with EJ cells and ECM derived from ras transfected and nontransfected fibroblast cells demonstrated that ECM of ras transfected fibroblast cells, but not the parental fibroblast cells, provided a permissive or fertile soil for EJ tumor cell invasion. Finally, two-dimensional gel electrophoresis of 35S-labeled nuclear matrix proteins of EJ cells cultured on ECM derived from ras transfected fibroblast cells revealed expression of proteins in the molecular weight range of M(r) 35,000-45,000 and isoelectric focusing pH range of 5.5 to 6.0. These proteins were not present in EJ cells cultured on ECM derived from parental nontransfected fibroblast cells. We conclude that extracellular matrices derived from transformed stroma producing cells may influence the proliferation, genetic regulation, and maintenance of the overlying urothelial tumor cells. The mechanism by which the ECM may influence cellular behavior and phenotype may be in their ability to modulate the nuclear matrix proteins of the overlying cell.

Primary renal carcinoid tumor.

We report a case of primary renal carcinoid tumor. Only 13 prior cases are documented in the literature. The tumor fulfilled both histologic and immunochemical criteria for carcinoid. In addition, we employed new diagnostic modalities (i.e., magnetic resonance imaging and chromogranin-A levels) not used in prior published reports. A review of the literature is presented.

Type IV procollagen mRNA regulation: evidence for extracellular matrix/cytoskeleton/nuclear matrix interactions in human urothelium.

The absence of basement membrane components correlates with tumor stage and progression in human bladder cancers. We have previously shown that invasive tumors possess the ability to degrade basement membrane. However, the presence of basement membrane may be affected not only by its degradation, but by its synthesis and deposition as well. Our results in the present study suggest that while the invasive human transitional carcinoma cell line EJ has an increased amount of type IV procollagen mRNA when compared to the non-invasive RT4 cell line, type IV collagen staining is absent in the invasive EJ cells and intensely present in the non-invasive RT4 cells. Moreover, when EJ cells were grown on an artificial basement membrane (Matrigel), type IV procollagen mRNA expression was down-regulated to the levels seen with the non-invasive RT4 cells. We also discovered that the invasive cells, when grown on Matrigel, appeared morphologically different from the same cells grown on plastic tissue cultures. We conclude that a deficient basement membrane in invasive cancer cells may be due not only to active proteolytic activity but also to an abnormal production and deposition of extracellular matrix components. In addition, we also demonstrated that basement membrane components may have a significant effect on epithelial cell morphology and gene regulation, and that any alterations of the extracellular matrix-cytoskeleton-nuclear matrix interactions can lead to altered gene regulations and cell function.