Perioperative immunomodulation with interleukin-2 in patients with renal cell carcinoma: results of a controlled phase II trial.

We conducted a non-randomised controlled phase II trial to investigate the role of preoperative administration of interleukin-2 (IL-2) in patients with renal cell carcinoma undergoing tumour nephrectomy. A total of 120 consecutive patients were allocated alternately to the two study groups: perioperative immunomodulation with IL-2 (IL-2 group; n=60) and perioperative immunomonitoring without immunomodulation (control group; n=60). Patients from the IL-2 group received four doses of 10 x 10(6) IU m(-2) twice daily subcutaneously a week before operation followed by a daily maintenance dose of 3 x 10(6) IU m(-2) subcutaneously until a day before the operation. Parameters of cellular and humoral immunity (leucocytes, T-cell markers CD3, CD4, and CD8, B-cell marker CD19, monocyte marker CD14, natural killer (NK) cell markers CD16, CD56, and CD57, activation markers CD6, CD25, CD28, and CD69, progenitor cell marker CD34, as well as IL-2, IL-6, IL-10, soluble IL-2 receptor, IL-1 receptor antagonist, transforming growth factor-beta1, and vascular endothelial growth factor) were measured in peripheral venous blood at various intervals. Interleukin-2-related toxicity was WHO grade 1 (24%), 2 (67%), and 3 (9%). In the postoperative period, T-cell markers, activation markers, and NK cell markers decreased, and IL-6 and IL-10 increased. However, all these alterations were significantly less accentuated in patients who had been pretreated with IL-2. Median follow-up was 40 months. Tumour-specific survival in the IL-2 group and the control group was 98 vs 81% after 1 year and 86 vs 73% after 5 years (P=0.04). A similar effect was found for progression-free survival. We conclude that IL-2 can be safely administered in the perioperative period and modulates immunological parameters. However, to validate the survival data, a larger randomised phase III trial is needed.

Adjuvant treatment with interleukin-2- and interferon-alpha2a-based chemoimmunotherapy in renal cell carcinoma post tumour nephrectomy: results of a prospectively randomised trial of the German Cooperative Renal Carcinoma Chemoimmunotherapy Group (DGCIN).

We conducted a prospectively randomised clinical trial to investigate the role of adjuvant outpatient immunochemotherapy administered postoperatively in high-risk patients with renal cell carcinoma. In total, 203 renal carcinoma patients' status post radical tumour nephrectomy were stratified into three risk groups: patients with tumour extending into renal vein/vena cava or invading beyond Gerota's fascia (pT3b/c pN0 or pT4pN0), patients with locoregional lymph node infiltration (pN+), and patients after complete resection of tumour relapse or solitary metastasis (R0). Patients were randomised to undergo either (A) 8 weeks of outpatient subcutaneous interleukin-2 (sc-rIL-2), subcutaneous interferon-alpha2a (sc-rIFN-alpha2a), and intravenous 5-fluorouracil (iv-5-FU) according to the standard Atzpodien regimen (Atzpodien et al, 2004) or (B) observation. Two-, 5-, and 8-year survival rates were 81, 58, and 58% in the treatment arm, and 91, 76, and 66% in the observation arm (log rank P=0.0278), with a median follow-up of 4.3 years. Two, 5-, and 8-year relapse-free survival rates were calculated at 54, 42, and 39% in the treatment arm, and at 62, 49, and 49% in the observation arm (log rank P=0.2398). Stage-adapted subanalyses revealed no survival advantages of treatment over observation, as well. Our results established that there was no relapse-free survival benefit and the overall survival was inferior with an adjuvant 8-week-outpatient sc-rIL-2/sc-rIFN-alpha2a/iv-5-FU-based immunochemotherapy compared to observation in high-risk renal cell carcinoma patients following radical tumour nephrectomy.

Calreticulin and calnexin in the endoplasmic reticulum are important for phagocytosis.

Calreticulin and calnexin are Ca2+-binding proteins with chaperone activity in the endoplasmic reticulum. These proteins have been eliminated by gene replacement in Dictyostelium, the only microorganism known to harbor both proteins; family members in Dictyostelium are located at the base of phylogenetic trees. A dramatic decline in the rate of phagocytosis was observed in double mutants lacking calreticulin and calnexin, whereas only mild changes occurred in single mutants. Dictyostelium cells are professional phagocytes, capable of internalizing particles by a sequence of activities: adhesion of the particle to the cell surface, actin-dependent outgrowth of a phagocytic cup, and separation of the phagosome from the plasma membrane. In the double-null mutants, particles still adhered to the cell surface, but the outgrowth of phagocytic cups was compromised. Green fluorescent protein-tagged calreticulin and calnexin, expressed in wild-type cells, revealed a direct link of the endoplasmic reticulum to the phagocytic cup enclosing a particle, such that the Ca2+ storage capacity of calreticulin and calnexin might directly modulate activities of the actin system during particle uptake.

Sonolytic, photolytic, and photocatalytic decomposition of atrazine in the presence of polyoxometalates.

Aqueous solutions of atrazine [2-chloro-4-(isopropylamino)-6-(ethylamino)-s-triazine] (CIET) decompose upon illumination with a low-pressure Hg-arc lamp (254 nm). However, no decomposition takes place with lambda > 300 nm. On the other hand, addition of polyoxometalates (POM), PW12O40(3-) or SiW12O40(4-), into a solution of atrazine photodecomposes the substrate within a few minutes (cutoff fiter 320 nm). Ultrasound (US) treatment also decomposes aqueous solutions of atrazine within a few minutes. Both methods, sonolysis and photolysis with POM, give common intermediates, namely, 2-hydroxy-4-(isopropylamino)-6-amino-s-triazine (OIET), 2-chloro-4-(isopropylamino)-6-amino-s-triazine (CIAT), 2-chloro-4-amino-6-(ethylamino)-s-triazine (CAET), 2-hydroxy-4,6-diamino-s-triazine (OAAT), and 2-hydroxy-4-hydroxy-6-amino-s-triazine (OOAT) among others. The final products for both methods, US and photolysis with POM, were cyanuric acid (OOOT), NO3-, Cl-, CO2, and H2O. OOOT showed no signs of decomposition by sonication and/or photolysis with POM. It also resisted degradation upon photolysis with plain UV light (254 nm). However, it has been reported to decompose upon photolysis with lambda > 200 nm. Combination of US and photolysis with POM produces only a cumulative effect.

Sugar- and nitrogen-dependent regulation of an Amanita muscaria phenylalanine ammonium lyase gene.

The cDNA of a key enzyme of secondary metabolism, phenylalanine ammonium lyase, was identified for an ectomycorrhizal fungus by differential screening of a mycorrhizal library. The gene was highly expressed in hyphae grown at low external monosaccharide concentrations, but its expression was 30-fold reduced at elevated concentrations. Gene repression was regulated by hexokinase.

Three-dimensional patterns and redistribution of myosin II and actin in mitotic Dictyostelium cells.

Myosin II is not essential for cytokinesis in cells of Dictyostelium discoideum that are anchored on a substrate (Neujahr, R., C. Heizer, and G. Gerisch. 1997. J. Cell Sci. 110:123-137), in contrast to its importance for cell division in suspension (DeLozanne, A., and J.A. Spudich. 1987. Science. 236:1086-1091; Knecht, D.A., and W.F. Loomis. 1987. Science. 236: 1081-1085.). These differences have prompted us to investigate the three-dimensional distribution of myosin II in cells dividing under one of three conditions: (a) in shaken suspension, (b) in a fluid layer on a solid substrate surface, and (c) under mechanical stress applied by compressing the cells. Under the first and second conditions outlined above, myosin II does not form patterns that suggest a contractile ring is established in the furrow. Most of the myosin II is concentrated in the regions that flank the furrow on both sides towards the poles of the dividing cell. It is only when cells are compressed that myosin II extensively accumulates in the cleavage furrow, as has been previously described (Fukui, Y., T.J. Lynch, H. Brzeska, and E.D. Korn. 1989. Nature. 341:328-331), i.e., this massive accumulation is a response to the mechanical stress. Evidence is provided that the stress-associated translocation of myosin II to the cell cortex is a result of the dephosphorylation of its heavy chains. F-actin is localized in the dividing cells in a distinctly different pattern from that of myosin II. The F-actin is shown to accumulate primarily in protrusions at the two poles that ultimately form the leading edges of the daughter cells. This distribution changes dynamically as visualized in living cells with a green fluorescent protein-actin fusion.

Protection against osmotic stress by cGMP-mediated myosin phosphorylation.

Conventional myosin functions universally as a generator of motive force in eukaryotic cells. Analysis of mutants of the microorganism Dictyostelium discoideum revealed that myosin also provides resistance against high external osmolarities. An osmo-induced increase of intracellular guanosine 3',5'-monophosphate was shown to mediate phosphorylation of three threonine residues on the myosin tail, which caused a relocalization of myosin required to resist osmotic stress. This redistribution of myosin allowed cells to adopt a spherical shape and may provide physical strength to withstand extensive cell shrinkage in high osmolarities.

A MAP kinase necessary for receptor-mediated activation of adenylyl cyclase in Dictyostelium.

Analysis of a developmental mutant in Dictyostelium discoideum which is unable to initiate morphogenesis has shown that a protein kinase of the MAP kinase/ERK family affects relay of the cAMP chemotactic signal and cell differentiation. Strains in which the locus encoding ERK2 is disrupted respond to a pulse of cAMP by synthesizing cGMP normally but show little synthesis of cAMP. Since mutant cells lacking ERK2 contain normal levels of both the cytosolic regulator of adenylyl cyclase (CRAC) and manganese-activatable adenylyl cyclase, it appears that this kinase is important for receptor-mediated activation of adenylyl cyclase.

Purification, functional characterization, and cDNA sequencing of mitochondrial porin from Dictyostelium discoideum.

Porin of Dictyostelium discoideum was extracted from mitochondria with Genapol X-80 and was purified by hydroxyapatite and CM-cellulose chromatography. The purified protein displayed a single band of 30 kDa in SDS-polyacrylamide gel electrophoresis. The formation of channels in artificial lipid bilayer membranes defined its function as a channel-forming component. Its average single-channel conductance was 3.9 nanosiemens in 1 M KCl, which suggested that the effective diameter of the channel is approximately 1.7 nm at small transmembrane potentials. The channel displayed a characteristic voltage dependence for potentials higher than 20 mV. It switched to substates of smaller conductance and a selectivity different to that of the open state. The closed state was stabilized at low ionic strength. The cDNA sequence of mitochondrial porin from D. discoideum was determined. It showed little sequence similarities to other known mitochondrial porins. The functional similarity, however, was striking. Localization of the porin in the mitochondrial outer membrane was confirmed by immunogold labeling of cryosections of fixed cells.

Stage-specific tyrosine phosphorylation of actin in Dictyostelium discoideum cells.

A 45 kDa protein in Dictyostelium discoideum cells that was recognized by a phosphotyrosine-specific antibody was identified by its binding activity to DNase I and its 2D-electrophoretic behavior as actin. The reactivity of actin with the antibody was transiently enhanced for about 30 minutes shortly after starving cells were reintroduced into nutrient medium. This effect indicates a modification of actin that is regulated under physiological conditions. A similar effect was obtained when growing cells were treated with phenylarsine oxide (PAO), an inhibitor of phosphotyrosine phosphatases. This effect was reversed and the cells fully recovered upon addition of the PAO antagonist 2,3-dimercaptopropanol. Starved cells did not show this enhancement of antibody labelling, which indicates that the response to PAO depends on the developmental stage. Phosphorylated amino acid residues were identified after in vivo labelling with [32P]phosphate in the presence of PAO. Part of the radioactivity in the actin band was recovered as phosphotyrosine, another part as phosphoserine. PAO caused the cells to form elongated blebs, to round up and finally to become immobilized. Fluorescence labelling with phalloidin of cells that were fixed at different times of PAO treatment revealed a progressive decrease in the staining for actin filaments and showed that these alterations in cytoskeleton organization were readily reversible, in accordance with the reversal of tyrosine phosphorylation at actin.

[The value of computerized tomography and renovasography in kidney injuries]. / Stellenwert von Computertomographie und Renovasographie bei Nierenverletzungen.

Computerized tomography (CT) and renal angiography are compared in the staging of traumatic renal lesions. After evaluation of 44 angiographic and 74 CT investigations in 111 patients it was concluded, that CT is superior to angiography in the diagnosis of blunt renal injuries. However, if vascular injuries are suspected an angiographic investigation should be done.

[Partial kidney resection in kidney tumors--indications and results]. / Nierenteilresektion bei Nierentumoren--Indikation und Ergebnisse.

With the use of ultrasonography synchronous and asynchronous bilateral renal tumors and tumors in single kidneys are diagnosed more frequently also in the stage I. Organ-preserving operations as partial kidney resection or tumor enucleation combined with contralateral radical nephrectomy are used stage-related successfully. Hypothermia in vivo or ex vivo seems to be advantageous in some cases. The tumor stage (according to Robson) and an optimal operative technique are of significant prognostic importance. In the own material of 15 bilateral renal tumors and 5 tumors in single kidneys only 5 patients do survive currently without tumor signs. The other 15 patients were at the time of operation in the tumor stages III and IV. Therefore, the diagnosis should made earlier, especially by more frequently and qualified ultrasonographic examinations.

Inverted Y-ureteral duplication with an uterine ectopy as cause of ureteric enuresis.

Presentation of the casuistry of a girl at the age of 3 years with an inverted Y-ureteral duplication in connection with a uterine ectopy and 'urinary incontinence'. Reference to cases of this extremely seldom occurring malformation published so far and embryonic background are given.

[Reactions of tumorous urinary bladders following local and systemic therapy with cytostatic drugs]. / Reaktionen an tumortragenden Harnblasen nach lokaler bzw. systemischer Zytostatikatherapie.

In 22 patients with bladder cancer after local chemotherapy and 13 patients after systemic chemotherapy, respectively, multiple biopsy specimens were examined histologically. Manifold hyperplastic, inflammatory, dysplastic, metaplastic and toxic alterations on the urothel, tumor cells and the lamina propria were found. This wide spectrum is the consequence of various etiological factors. On such treated cancer bladders the therapy-, age- and TuR-conditioned pathomorphological findings are running up with the known panurothelial reactions of a primary tumor in the urinary tract. We have classified the occurrence of multiple single cell necroses within the tumors as obligatory therapeutic effects. The granulomatous and degenerative stromal findings be conditioned by the therapy, if they are localized safely outside the area of previous TuR. The other reactive hyper- and dysplastic reactions be conditioned facultatively by the therapy.

[Generalized angiomyolipomatosis within the framework of Bournville-Pringle disease--urological aspects]. / Die generalisierte Angiomyolipomatose im Rahmen des Morbus Bourneville-Pringle--Urologische Aspekte.

2 cases of angiomyolipomatosis of the kidneys are taken as the basis for presenting the symptoms, diagnosis and therapy of this rare disease. In one case there was also an angiomyolipomatous change in the retroperitoneal lymph nodes. In differential diagnosis this gave rise to a suspicion of metastasis or a malignant system disease. In another case there was a coincidence with cystic degeneration of the kidneys.

[Correction of hypospadias using the Cecil-Michalowski method--report of clinical experiences]. / Die Korrektur der Hypospadie nach Cecil/Michalowskiein klinischer Erfahrungsbericht.

A report is given on our own experience with the operative correction of hypospadia. The method of Cecil/Michalowski and slight variations are explained and presented. The rate of complication was extremely low in a total of 45 patients.

[Development of sigmoid carcinomas after ureterosigmoidostomy]. / Zur Entwicklung von Sigmakarzinomen nach Ureterosigmoidostomie.

It is reported on a case of sigma carcinoma 15 years after ureterosigmoidostomy. A survey table contains the cases of sigma tumours after passing of the urine into the sigma hitherto reported in the literature of the world. Short discussion of frequency, latency period and diagnostic and therapeutic measures after ureterosigmoidostomy.