Florian Seseke

Signal propagation via gap junctions, a key step in the regulation of liver metabolism by the sympathetic hepatic nerves

Cell‐to‐cell communication via gap junctions has been proposed to be involved in the metabolic actions of sympathetic liver nerves in the rat. The effects of hepatic nerve stimulation and noradrenaline‐, PGF2α‐ and glucagon infusion on glucose metabolism and perfusion flow were studied in perfused rat liver in the absence and presence of the gap junctional inhibitors, heptanol, carbenoxolone and (4β)phorbol 12‐myristate 13‐acetate (4βPMA). (i) Stimulation of the hepatic nerve plexus increased glucose output, decreased flow and caused an overflow of noradrenaline into the hepatic vein. (ii) Heptanol completely inhibited not only the nerve stimulation‐dependent metabolic and hemodynamic alterations but also the noradrenaline overflow. Thus the heptanol‐dependent inhibitions were caused primarily by a strong impairment of transmitter release. (iii) Carbenoxolone inhibited the effects of neurostimulation on glucose metabolism partially by about 50%, whereas it left perfusion flow and noradrenaline overflow essentially unaltered. (iv) 4βPMA reduced the nerve stimulation‐dependent enhancement of glucose release by about 80% but the noradrenaline‐dependent increase in glucose output only by about 30%; the increase in glucose release by PGF2α and by glucagon remained essentially unaltered. 4βPMA reduced the nerve stimulation‐dependent decrease in portal flow by about 35% but did not affect the noradrenaline‐and PGF2α‐elicited alterations, nor did it alter noradrenaline overflow. The results allow the conclusion that gap junctional communication plays a major role in the regulation of hepatic carbohydrate metabolism by sympathetic liver nerves, but not by circulating noradrenaline, PGF2α or glucagon.

Treatment of Iatrogenic Postoperative Ureteral Strictures with Acucise Endoureterotomy

OBJECTIVES To determine factors influencing the outcome of Acucise endoureterotomy in patients with iatrogenic postoperative ureteral strictures after different open surgical procedures. MATERIAL AND METHODS Acucise endoureterotomy was performed in 18 patients with ureteral strictures after pyeloplasty (n = 5), renal transplantation (n = 5), ureteroenteric anastomosis (n = 3), calicoureterostomy (n = 1), ureterocystoneostomy (n = 1), hysterectomy (n = 1), ureterorenoscopy (n = 1) and transurethral resection of the ureteral orifice (n = 1). Success was determined as relief of clinical symptoms, improvement of renal function or improvement of radiographic findings. RESULTS The overall success rate was 61% (mean follow-up: 21.5 months). Six out of 18 patients showed relevant side effects. Neither the localization of the stricture nor the duration of postoperative ureteral stenting but the length of the stricture had influence on the postoperative outcome. Decreased renal function to less than 25% of the total function was always associated with failure of the treatment. The time period between the ureteral injury and the appearance of the ureteral stricture had influence on the outcome of the treatment. CONCLUSIONS Acucise endoureterotomy is effective in the treatment of postoperative ureteral strictures, but only in selected cases. The selection criteria are the time period from the primary operation to the appearance of the stricture (>6 months), the length of the stricture (<1.5 cm) and the renal function (>25% of the total function). In other cases, open surgical treatment of the ureteral stricture may provide better results.

Metastasis from renal cell carcinoma to the cervix uteri.

We report on a patient with clear cell adenocarcinoma of the left kidney and a solitary metastasis to the cervix uteri. Metastases from renal cell carcinoma to the female genitalia are uncommon and metastatic involvement of the uterus is very rare. To our knowledge, no more then five cases have been published. A review of the literature is given.

Phytoestrogens from Belamcanda chinensis regulate the expression of steroid receptors and related cofactors in LNCaP prostate cancer cells

To investigate the changes in expression underlying the marked reduction of tumour growth in vivo, by analysing the effect of Belamcanda chinensis extract (BCE) on LNCaP cells in vitro, as phytoestrogens are chemopreventive in prostate cancer, and in previous studies we examined the effects of the isoflavone tectorigenin isolated from B. chinensis on LNCaP prostate cancer cells, and a BCE consisting of 13 phytoestrogenic compounds on tumour‐bearing nude mice.

Histologic and molecular evidence of obstructive uropathy in rats with hereditary congenital hydronephrosis.

Abstract Partial obstruction of the upper urinary tract, a frequent challenge for the pediatric urologist, leads to renal damage, if deobstruction is delayed. Several but sometimes unsatisfactory animal models have been developed to study this phenomenon. Obstruction created by surgical manipulation lacks adequate correlation with a developing congenital obstruction. In some animals with congenital hydronephrosis, evidence of renal obstruction is absent. A study of the renal morphology of rats with hereditary unilateral hydronephrosis has exhibited clear evidence of renal obstruction distinguishable from renal dilatation. The renal mRNA expression of renin and transforming-growth factor- β1 (TGF-β1) was measured by a semiquantitative RT-PCR technique. In hydronephrotic kidneys, a marked loss of parenchyma, atrophy and dilation of tubuli and collecting ducts and interstitial fibrosis was observed. The mRNA expression of renin was increased significantly in comparison to controls, whereas the contralateral kidneys showed renin activity below control levels. TGF-β1 expression was markedly increased in hydronephrotic kidneys, whereas contralateral kidneys did not differ significantly from control values. These data suggest the presence of renal obstruction and not only renal dilatation in these rats with congenital hydronephrosis. This colony seems to be a representative animal model to study congenital renal obstruction even in the fetal period without the need of surgical manipulation.

IMPAIRED NEPHROGENESIS IN RATS WITH CONGENITAL OBSTRUCTIVE UROPATHY

PURPOSE Alterations caused by renal obstruction in developing kidneys are of particular interest in basic research of congenital obstructive uropathy. In rats nephrogenesis mainly occurs 7 to 10 days postnatally. Therefore, surgically induced neonatal ureteral obstruction in rats has been suggested to be analogous to congenital obstruction in the fetus. An attempt less prone to surgical artifacts and assessing even earlier developmental stages is to monitor the development of obstructed kidneys in rats with congenital obstructive uropathy. MATERIALS AND METHODS Rats from an inbred strain with congenital renal obstruction in 70% of their littermates were observed. Morphologically, significant hydronephrosis was not detected before day 5 post partum and progressed with age. Unilateral obstructed kidneys were compared with contralateral kidneys and kidneys from healthy control animals at ages of 1, 5, 10, 18 and 32 days. A total of 72 renal units were investigated. The renal messenger RNA expression of renin and transforming growth factor-beta1 (TGF-beta1) was quantified by competitive quantitative reverse transcription polymerase chain reaction using a gene specific complementary RNA standard. RESULTS In controls the gene expression of renin decreased from day 1 to day 18 and remained stable. TGF-beta1 expression increased during the first 10 days and then decreased again. Renin expression of the obstructed kidneys was reduced (p <0.05) on day 1, increased to a maximum versus controls (p <0.01) on day 10 and decreased to an unchanged elevated level (p <0.01) on days 18 and 32. Renin expression of the contralateral kidneys showed no significant alterations to control kidneys. Messenger RNA expression of TGF-beta1 of obstructed kidneys stayed decreased during the first 10 days (p <0.05), then increased excessively on day 18 (p <0.01) and slightly decreased on day 32. TGF-beta1 expression of the contralateral kidneys was parallel to controls on a slightly elevated level, increased on day 18 and returned to control level on day 32. CONCLUSIONS Within the postpartum period of nephrogenesis gene expression of renin and TGF-beta1 was decreased in obstructed kidneys compared to controls. As the renin angiotensin system and TGF-beta1 have important functions in normal kidney development, these results suggest impaired nephrogenesis of congenital obstructed kidneys even before the onset of morphological signs of hydronephrosis. These features differ from surgical induced unilateral ureteral obstruction at birth and promise new insights into the pathophysiology of congenital obstructive uropathy.

Pharmacological potential of phytoestrogens in the treatment of prostate cancer

ZusammenfassungEinleitungPhytoöstrogene sind Wirkstoffe pflanzlichen Ursprungs mit antiproliferativem Potential bei Prostatakarzinom-Zellen, dessen Grundlage im Detail ungeklärt ist. Proliferation von PCa-Zellen wird vor allem durch den Androgenrezeptor (AR) und seinen Co-Aktivator, dem Prostate-derived Ets Factor (PDEF) vermittelt, die wiederum den IGF-1 Rezeptor (IGF-1-R) steuern. Phytoöstrogene greifen in diesen Prozess ein. Die aktuelle Studie verdeutlicht dies exemplarisch anhand der Wirkung von Tectorigenin aus Belamcanda chinensis. Material und MethodeAndrogen-sensitive PCa-Zellen der LNCaP-Linie wurden über 24 Stunden mit Tectorigenin 100 µM behandelt und die mRNA-Expression von AR, PSA, PDEF, Telomerase Reverse Transcriptase (hTERT), Tissue-Inhibitor of Matrix-Metalloproteinase-3 (TIMP-3) und des IGF-1-R durch Real-Time-PCR bestimmt. Die Aktivität von PSA, Telomerase und IGF-1-R wurde auch auf Proteinebene bestimmt. Weiterhin wurde in-vivo der Einfluss einer Diät mit Rohextrakt Belamcanada chinensis im Vergleich zu sojafreier Diät auf das Wachstum von subkutan injizierten Prostatakarzinomzellen in Nacktmäusen untersucht.ErgebnisseBei den LNCaP bewirkte Tectorigenin eine Herunterregulierung der Genexpression von PDEF und PSA. Die Aktivität der hTERT war vermindert. Es zeigte sich eine verminderte Genexpression des IGF-1-R. Auf Proteinebene fand sich eine verminderte Aktivität von PSA, hTERT und IGF-R-1. Die Genexpression von TIMP-3 war unter dem Einfluss von Tectorigenin deutlich erhöht. Bei den Nacktmäusen fand sich neben einer verminderten Inzidenz ein verringertes Tumorvolumen unter Diät mit Belamcanda chinensis.SchlussfolgerungTectorigenin zeigte eine Hemmung der IGF-1-R-vermittelten Proliferation bei PCa-Zellen, durch eine vom AR unabhängige Modulation der Aktivität von PDEF. Über Aktivierung von TIMP-3 hat Tectorigenin proapoptotische Effekte und hemmt die Gewebeinvasivität des Tumors. Phytoöstrogene könnten somit eine interessante Option in der Therapie insbesondere des fortgeschrittenen Prostatakarzinoms darstellen.AbstractIntroductionPhytoestrogenes are plant-derived compounds that have been shown to exert an antiproliferative potential on prostate cancer cells, although the exact mechanisms are still unclear. In prostate cancer cells proliferation is regulated by modulation of the IGF-1 receptor (IGF-R-1) by the androgen receptor (AR) and its co-activator prostate derived Ets factor (PDEF). Phytooestrogenes interact with these mechanisms as demonstrated exemplarily in the presented study with the isoflavone tectorigenin derived from Belamcanda chinensis.Material and methodsCultured androgen-sensitive LNCaP prostate cancer cells were treated with tectorigenin of 100 µM for 24 hours. The mRNA-expression of AR, PSA, PDEF, hTERT, TIMP-3 and IGF-R-1 were quantified by real-time RT-PCR. Furthermore, the expression or activity of PSA, telomerase and IGF-R-1 was measured on the protein level. In addition, we investigated in nude mice the influence of a diet of extracts of Belamcanda chinensis on the growth of subcutaneously injected LNCaP cells versus a control group of animals fed with a soy-free diet.ResultsIn cultured LNCaP cells treatment with tectorigenin resulted in a significant down-regulation of the gene expression of AR, PDEF, PSA, IGF-R-1 and hTERT. On the protein level PSA secretion and the activity of telomerase and IGF-R-1 expression was also decreased. The gene expression of TIMP-3 was distinctly up-regulated by tectorigenin. Nude mice fed with Belamcanda chinensis extract showed a significantly decreased incidence and tumor growth compared to controls.ConclusionsTectorigenin shows an inhibition of the IGF-1-R modulated cell proliferation of PCa-Cells, due to modulation of the activity the co-activator PDEF independently from the AR. Furthermore, tectorigenin has pro-apoptotic effects and decreases tissue invasion by up-regulation of TIMP-3. Therefore, phytooestrogenes are an interesting option in the therapy of prostate especially advanced prostate cancer.INTRODUCTION Phytoestrogenes are plant-derived compounds that have been shown to exert an antiproliferative potential on prostate cancer cells, although the exact mechanisms are still unclear. In prostate cancer cells proliferation is regulated by modulation of the IGF-1 receptor (IGF-R-1) by the androgen receptor (AR) and its co-activator prostate derived Ets factor (PDEF). Phytooestrogenes interact with these mechanisms as demonstrated exemplarily in the presented study with the isoflavone tectorigenin derived from Belamcanda chinensis. MATERIAL AND METHODS Cultured androgen-sensitive LNCaP prostate cancer cells were treated with tectorigenin of 100 microM for 24 hours. The mRNA-expression of AR, PSA, PDEF, hTERT, TIMP-3 and IGF-R-1 were quantified by real-time RT-PCR. Furthermore, the expression or activity of PSA, telomerase and IGF-R-1 was measured on the protein level. In addition, we investigated in nude mice the influence of a diet of extracts of Belamcanda chinensis on the growth of subcutaneously injected LNCaP cells versus a control group of animals fed with a soy-free diet. RESULTS In cultured LNCaP cells treatment with tectorigenin resulted in a significant down-regulation of the gene expression of AR, PDEF, PSA, IGF-R-1 and hTERT. On the protein level PSA secretion and the activity of telomerase and IGF-R-1 expression was also decreased. The gene expression of TIMP-3 was distinctly up-regulated by tectorigenin. Nude mice fed with Belamcanda chinensis extract showed a significantly decreased incidence and tumor growth compared to controls. CONCLUSIONS Tectorigenin shows an inhibition of the IGF-1-R modulated cell proliferation of PCa-Cells, due to modulation of the activity the co-activator PDEF independently from the AR. Furthermore, tectorigenin has pro-apoptotic effects and decreases tissue invasion by up-regulation of TIMP-3. Therefore, phytooestrogenes are an interesting option in the therapy of prostate especially advanced prostate cancer.

Push and smash increases success rates in treatment of ureteric calculi by ESWL

During a 3-year period 105 patients underwent attempted retrograde manipulation for ureteric calculi into the renal pelvis prior to extracorporeal shock wave lithotripsy (ESWL). The success rate of this group was compared to 93 patients receiving ESWL of ureteric calculi in situ.The method of retrograde manipulation was recorded prospectively. Retrograde flushing with a 5 F Tiemann ureteral catheter was performed after coating the proximal ureter and renal pelvis with a mixture of saline and lidocaine gel.Success rate of calculi push was 91.4%. Mean number of ESWL treatments in this group of patients was 1.4, compared to 2.1 in patients with in situ treatment.Reposition of ureteric calculi prior to ESWL treatment increases successful initial treatment and can be performed easily.

Clear Cell Adenocarcinoma of the Male Urethra in Association with So-Called Nephrogenic Metaplasia

A clear cell adenocarcinoma of the urethra associated with so-called nephrogenic metaplasia occurring in a 57-year-old male patient is presented. Ten months following total urethrectomy, multiple pulmonary metastases had developed. The patient died 2.5 years after surgery. The possible histogenesis of clear cell adenocarcinoma of the lower urinary tract – reported to develop in the male urethra only in a very few cases – is reviewed briefly. We favor an origin from preexisting nephrogenic metaplasia as one of the possible histogenetic pathways.

Alterations of selected genes of the Wnt signal chain in rat kidneys with spontaneous congenital obstructive uropathy

OBJECTIVE To determine the role of the Wnt signal cascade in the pathophysiology of congenital obstructive uropathy in rats, we assessed the expression patterns of selected genes. METHODS Total cellular mRNA of complete obstructed, contralateral and healthy control kidneys of rats with hereditary, spontaneous congenital hydronephrosis at the age of 14 and 32days was extracted and pooled. mRNA expression was assessed by real-time reverse transcription-polymerase chain reaction. RESULTS Significant (P<0.05) differences in gene expression levels of obstructed versus contralateral and control kidneys were evaluated. In 14-day-old animals, Wnt-4 expression was decreased. Secreted frizzled-related protein (sFRP-1, sFRP-2), beta-catenin and Jun N-terminal kinase (JNK) expression was increased. The gene expression of Wnt-7b, Friz-1, Friz-2, calcineurin and Wnt inhibitory factor-1 (WIF-1) was unaltered. In 32-day-old animals, Wnt-4, Wnt-7b, Friz-2, sFRP-1 and WIF-1 expression was increased, whereas levels of Friz-1, sFRP-2, beta-catenin, JNK and calcineurin were unaltered. CONCLUSIONS The results suggest an age-dependent role of Wnts in the pathophysiology of congenital renal obstruction in rats. Down-regulated Wnt signalling at the end of nephrogenesis in 14-day-old rats might indicate impaired tubulogenesis. Up-regulation of beta-catenin and JNK points to stressed cell-cell adhesion due to progressive hydronephrosis. Increased Wnt signalling in older animals probably contributes to interstitial fibrosis and tubular apoptosis. This study should be considered as a starting point to correlate in detail the observed alterations in gene expression with the pathophysiology of congenital obstructive uropathy.