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Background: The outcome of patients with advanced renal cell carcinoma (RCC) under systemic therapy shows remarkable variability, and there is a need to identify prognostic parameters that allow individual prognostic stratification and selection of optimal therapy. Artificial neural networks (ANN) are software systems that can be trained to recognize complex data patterns. In this study, we used ANNs to identify poor prognosis of patients with RCC based on common clinical parameters available at the beginning of systemic therapy., Patients and Methods: Data from patients with RCC who started systemic therapy were collected prospectively in a single center database; 175 data sets with follow-up data (median, 36 months) were available for analysis. Age, sex, body mass index, performance status, histopathologic parameters, time interval between primary tumor and detection of metastases, type of systemic therapy, number of metastases, and metastatic sites were used as input data for the ANN. The target variable was overall survival after 36 months. Logistic regression models were constructed by using the same variables., Results: Death after 36 months occurred in 26% of the patients in the tyrosine kinase inhibitors group and in 37% of the patients in the immunotherapy group (P = .22). ANN achieved 95% overall accuracy and significantly outperformed logistic regression models (78% accuracy). Pathologic T classification, invasion of vessels, and tumor grade had the highest impact on the network's decision., Conclusion: ANN is a promising approach for individual risk stratification of patients with advanced RCC under systemic therapy, based on clinical parameters, and can help to optimize the therapeutic strategy., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: The Bladder Cancer Research Consortium (BCRC) created nomograms to predict all-cause mortality (ACM), cancer-specific mortality (CSM), and recurrence after radical cystectomy (RC) for urothelial carcinoma of the bladder (UCB)., Objective: To perform a formal validation of the BCRC nomograms in a large multi-institutional patient cohort from Europe., Design, Setting, and Participants: Records of 2501 patients who underwent RC for UCB at eight European centers were reviewed. Complete information for external validation was available in 2404 patients for the ACM and CSM nomograms and in 2243 patients for the recurrence nomogram., Measurements: For the purpose of external validation, model discrimination was measured using the receiver operating characteristics derived area under the curve. Calibration plots examined the relationship between predicted and observed probabilities at 2 yr, 5 yr, and 8 yr. Decision curve analyses were applied to assess the net benefit derived from the three models., Results and Limitations: The discrimination accuracies of the BCRC nomograms for ACM and CSM at 2 yr, 5 yr, and 8 yr after RC were 71.0%, 69.1%, and 68.2%, and 74.9%, 73.1%, and 72.4%, respectively. The accuracy of discrimination for the recurrence nomogram at the same time points was 76.5%, 75.3%, and 74.9%, respectively. Calibration plots revealed slight underestimations from ideal predictions. Decision curve analyses showed an increased net benefit for the use of the BCRC nomograms in this cohort. Limitations include the retrospective study design, potential surgeon bias, and lack of a central pathologic review., Conclusions: The ACM, CSM, and recurrence nomograms showed acceptable predictive accuracies and could thus be adopted into clinical practice in UCB patients treated in Europe., (Copyright © 2011 European Association of Urology. Published by Elsevier B.V. All rights reserved.)

Objective: • To analyse the safety and efficacy of simultaneous standard anti-angiogenic therapy and stereotactic radiosurgery (SRS) in patients with spinal and cerebral metastases from renal cell carcinoma., Patients and Methods: • In all, 106 patients with spinal (n= 55) or cerebral (n= 51) metastatic lesions and an Eastern Cooperative Oncology Group status of 0 or 1 were treated with sorafenib or sunitinib and simultaneous SRS. • The primary endpoint was local control. • Secondary endpoints were toxicity and overall survival., Results: • Median follow up was 14.7 months (range 1-42 months). Forty-five patients were treated with sunitinb and 61 patients with sorafenib. Two patients had asymptomatic tumour haemorrhage after SRS. • No skin toxicity, neurotoxicity or myelopathy occurred after SRS, and SRS did not alter the adverse effects of anti-angiogenic therapy. • Local tumour control 15 months after SRS was 98% (95% confidence interval 89-99%). The median pain score before SRS was 5 (range 1-8) and was lowered to 0 (range 0-2, P < 0.01) after SRS. There were no treatment-related deaths or late complications after SRS. • Overall survival was 17.4 months in patients with spinal lesions and 11.1 month in patients with cerebral lesions (P= 0.038)., Conclusions: • Simultaneous systemic anti-angiogenic therapy and SRS for selected patients with renal cell carcinoma who have spinal and cerebral metastases is safe and effective. • Single-fraction delivery allows for efficacious integration of focal radiation treatment into oncological treatment concepts without additional toxicity. • Further studies are needed to determine the limits of SRS for renal cell carcinoma metastases outside the brain and spine., (© 2010 THE AUTHORS. BJU INTERNATIONAL © 2010 BJU INTERNATIONAL.)

Objective: To investigate the pattern of DNA CpG island hypermethylation in papillary renal cell carcinoma (pRCC)., Material and Methods: DNA from pRCC (n= 32) and adjacent normal tissue (n= 15) was isolated. A quantitative methylation-specific PCR was performed to analyse the methylation pattern at APC (actin beta), CDH1 (E-cadherin), GSTP1 (glutathione S-transferase pi 1), RASSF1A (Ras association domain family member 1A) and TIMP3 (TIMP metallopeptidase inhibitor 3); a sequence of ACTB without CpG was used to normalize for DNA input and to calculate the relative amount of methylated DNA (normalized index of methylation, NIM)., Results: RASSF1A hypermethylation was observed in most pRCC and normal samples (100 vs 94.4%), but the median NIM was significantly higher in pRCC samples (2.11 vs 0.61; P < 0.001). RASSF1A hypermethylation allowed discrimination of pRCC and normal tissue with a sensitivity of 87.5% and a specificity of 73.3% as determined via receiver operator characteristic analysis (area under curve = 0.814). Hypermethylation at APC (3.0 vs 6.7%), CDH1 (15.6 vs 0%), GSTP1 (21.9 vs 6.7%) and TIMP3 (6.3 vs 0%) was infrequent in pRCC and normal tissue. CDH1 was significantly correlated with pathological stage (P= 0.015), and patients with methylated CDH1 methylation showed a trend towards shorter recurrence-free survival (log-rank P= 0.057). The number of methylated gene sites was correlated with pathological stage (P= 0.007) and lymph node metastasis (P= 0.008)., Conclusions: DNA hypermethylation at RASSF1A is common in pRCC tissue irrespective of the histological subtype, but also frequently seen at lower levels in normal adjacent tissue. Aberrant hypermethylation could be a prognostic marker for pRCC., (© 2010 THE AUTHORS. JOURNAL COMPILATION © 2010 BJU INTERNATIONAL.)

Objective: The aim of this study was to externally validate the prognostic value of concomitant urothelial carcinoma in situ (CIS) in radical cystectomy (RC) specimens using a large international cohort of bladder cancer patients., Methods: The records of 3,973 patients treated with RC and bilateral lymphadenectomy for urothelial carcinoma of the bladder (UCB) at nine centers worldwide were reviewed. Surgical specimens were evaluated by a genitourinary pathologist at each center. Uni- and multivariable Cox regression models addressed time to recurrence and cancer-specific mortality after RC., Results: 1,741 (43.8%) patients had concomitant CIS in their RC specimens. Concomitant CIS was more common in organ-confined UCB and was associated with lymphovascular invasion (p < 0.001). Concomitant CIS was not associated with either disease recurrence or cancer-specific death regardless of pathologic stage. The presence of concomitant CIS did not improve the predictive accuracy of standard predictors for either disease recurrence or cancer-specific death in any of the subgroups., Conclusions: We could not confirm the prognostic value of concomitant CIS in RC specimens. This, together with the discrepancy between pathologists in determining the presence of concomitant CIS at the morphologic level, limits the clinical utility of concomitant CIS in RC specimens for clinical decision-making., (Copyright © 2011 S. Karger AG, Basel.)

Systemic therapy of metastatic renal cell carcinoma has completely changed in the last 5 years. Although a cure for the disease is still not achieved with systemic treatment in the majority of cases immunotherapy is no longer used. The therapeutic regiments are mainly based on angiogenic inhibitors such as sunitinib, sorafenib, pazopanib, everolimus and temsirolimus as well as the combination of bevacizumab with interferon. This article gives an overview of these treatment options and the clinical setting for their usage. To achieve a prolonged progression-free survival, a continuous therapy based on the new drugs is necessary. The major goal of the treatment remains to keep the disease stable as complete remission is only seen in 2-4% of cases. With these lengthy treatment regimes a schedule for sequenced administration of drugs is necessary for most of the patients. The optimal treatment sequence is unknown and should be chosen based on the individual course of the disease and the side effects as well as comorbidities. The role of neoadjuvant and adjuvant therapies remains unclear.

Changes in the methylation pattern in particular gene promoters as well as genetic sequence mutations play an important role in carcinogenesis. Molecular methods like pyrosequencing provide the specific analysis of these epigenetic and genetic modifications. In this review the relevance of these alterations for prostate cancer and the function of pyrosequencing will be described and explained on the basis of a search of the PubMed literature database. At present, in uro-oncology only a few studies outlining methylation in prostate cancer and pyrosequencing have been published. Nevertheless, it becomes evident that epigenetic mechanisms as well as specific gene sequence alterations have an impact on the carcinogenesis of prostate cancer and knowledge of these factors might open perspectives in diagnostic approaches of the future.

Management of kidney injuries is an uncommon challenge to urologists. Therapy has evolved in recent years from mainly surgical to predominantly conservative treatment. Immediate surgical intervention for renal trauma is now only necessary in rare instances. This overview is based on the guidelines of the European Association of Urology and the Societé International d'Urologie as well as clinical experience and is intended to provide practical advice for treatment of renal trauma.

Chronic inflammation seems of importance in multiple types of cancer. Numerous reports have revealed a potential link between chronic prostatatic inflammation and prostate cancer. The aim of this paper is to review the epidemiological, moprphological, molecular and clinical aspects of prostattic inflammation and prostate cancer.

Objective: To retrospectively determine the growth rate of renal masses with a diameter of > or =4 cm at the time of surgery, as the average growth rate of untreated small (<4 cm) renal masses is assumed to be 0.1-0.7 cm/year, but little is known about the progression of large masses., Patients and Methods: Of 256 patients who had their renal tumour surgically removed between January and December 2008, we identified nine (five men and four women; median age 65.2 years, range 29.2-74.2) with solitary large renal masses (>4 cm) who had abdominal imaging with identification of renal masses >6 months before admission. In none of the patients had the initial imaging led to admission, either by accident or because the masses were overlooked. The tumour growth rate was calculated based on images taken > or =6 months before admission and actual imaging, as well as histological results., Results: All patients had surgical resection of their renal masses in 2008. The median (range) follow-up from initial diagnosis to surgery was 14.6 (6.5-58.4) months. The median observed tumour growth rate was 6.41 (2.47-8.66) cm/year. The histological diagnosis was clear cell renal cell carcinoma (RCC) in seven patients, papillary in one and clear cell RCC with portions of sarcomatoid de-differentiation in one; two patients had lymph node metastases and one had pulmonary metastases, and five had tumours of > or =T3., Conclusions: The growth rate of large tumours is much higher than of small renal masses and most patients present with advanced disease. Thus, tumours of > or =4 cm seem not to be candidates for active surveillance but require immediate therapy. Further studies are warranted to determine the clinical relevance of progressive tumour growth and the underlying mechanisms.

Background: Pancreatic cancer (PaCa) is a fatal human cancer due to its exceptional resistance to all current anticancer therapies. The cytoprotective enzyme heme oxygenase-1 (HO-1) is significantly overexpressed in PaCa and seems to play an important role in cancer resistance to anticancer treatment. The inhibition of HO-1 sensitized PaCa cells to chemo- and radiotherapy in vitro. Therefore, we investigated the effects of HO-1 and its metabolites biliverdin, carbon monoxide and iron on PaCa cells. PaCa cell lines with divergent HO-1 expression patterns were used in a murine orthotopic cancer model. HO-1 expression and activity was regulated by zinc (inhibition) and cobalt (induction) protoporphyrin. Furthermore, the influence of cellular HO-1 levels and its metabolites on effects of standard chemotherapy with gemcitabine was tested in vivo and in vitro., Results: High HO-1 expression in PaCa cell lines was associated with increased chemoresistance in vitro. Chemoresistance to gemcitabine was increased during HO-1 induction in PaCa cells expressing low levels of HO-1. The inhibition of HO-1 activity in pancreatic tumors with high HO-1 boosted chemotherapeutic effects in vivo significantly. Furthermore, biliverdin and iron promoted PaCa resistance to chemotherapy. Consequently, specific iron chelation by desferrioxamine revealed profound anticancerous effects., Conclusion: In summary, the inhibition of HO-1 and the chelation of iron in PaCa cells were associated with increased sensitivity and susceptibility of pancreatic tumors to chemotherapy in vivo. The metabolites biliverdin and iron seem to be involved in HO-1-mediated resistance to anticancer treatment. Therefore, HO-1 inhibition or direct interference with its metabolites may evolve new PaCa treatment strategies.

Background & Aims: Extracellular nucleotides are released from injured cells and bind purinergic-type 2 receptors (P2-Rs) that modulate inflammatory responses. Ectonucleotidases, such as CD39/nucleoside triphosphate diphosphohydrolase-1, hydrolyze extracellular nucleotides to integrate purinergic signaling responses. Because the role of extracellular nucleotides and CD39 in mediating inflammation and fibrosis are understood poorly, we studied the impact of CD39 gene deletion in a model of pancreatic disease., Methods: Pancreatitis was induced by cyclosporine pretreatment, followed by cerulein injections (50 mug/kg, 6 intraperitoneal injections/day, 3 times/wk); mice were killed at day 2, week 3, and week 6. Experimental parameters were correlated with cytokine levels in blood, RNA, and protein expression of purinergic and fibrosis markers in tissues. Immunohistochemistry and pancreatic morphometry of fibrosis were performed in wild-type and CD39-null mice. Effects of CD39 deletion on proliferation of primary pancreatic stellate cells (PSCs) were investigated in vitro., Results: Wild-type mice developed morphologic features of pancreatitis with the anticipated development of parenchymal atrophy and fibrosis. CD39 and P2-R became overexpressed in vascular and adventitious wild-type tissues. In contrast, CD39-null mice had inflammatory reactions but developed only minor pancreatic atrophy and limited fibrosis. Interferon-gamma became significantly increased in tissues and plasma of CD39-null mice. Wild-type PSCs expressed high levels of CD39 and P2-R. CD39-null PSCs showed decreased rates of proliferation and the expression of procollagen-alpha1 was inhibited significantly in vitro (P < .03)., Conclusions: CD39 deletion decreases fibrogenesis in experimental pancreatitis. Our data implicate extracellular nucleotides as modulators of PSC proliferation and collagen production in pancreatitis.

Through a systematic investigation of the agglutination of long chain mannolipid and glucolipid incorporated liposomes by concanavalin A (ConA) it was found that the agglutination was dependent on different factors. The studied factors reported here are (1) spacer length and (2) ground lipid matrix. The threshold and the relative saturating ConA binding concentration (saturation point to attain the binding saturation condition) of glycosides with varying spacer length for agglutination are dependent on the spacer length of the glycolipid. These concentrations decrease with the increasing number of in-built ethyleneoxy spacer length in the glycolipid and find its minimum with 6 spacer units; it increases then more and more with increasing number of spacer units (>6 units). This is supposed to be due to the requirement of a proper distance of the hydrophilic determinant from the liposome surface for the response by ConA (response invoking distance), which may be most favorable in case of 6 spacer units. Further increase in number of spacer units (>6) results to an increasing probability of the bending of the spacer chain along with the terminal polar head group more and more towards the liposome surface; this leads to a reduction of the factual distance of the terminal hydrophilic head group from the liposome surface, weakening the response for ConA binding. The threshold concentration or saturation point decreases also with the rigidity of the ground lipid matrix. Increased rigidity of the ground matrix leads to a phase separation and localized 'Domain' formation with the glycolipid inside the ground matrix layer due to their immiscibility, invoking better response resulting to a reduction of required incorporated glycolipid concentration., (Copyright 2003 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 92:2229-2235, 2003)

Purpose: To improve target specificity and uptake of liposomes by macrophages, one can improve high-affinity receptor binding to mannose determinants with their 175-kDa mannose receptor (MR), which is mainly influenced by the length and flexibility of the spacer between the carbohydrate head group and liposome surface. Liposomes containing alkylmannosides with hydrophilic spacers 0 to 8 ethyleneoxy units (EO) long (Man0...Man8) were used to investigate systematically the effects of spacer length on liposome-cell interactions., Methods: Concanavalin A (ConA)-induced liposome aggregation was studied by turbidity measurement and cell uptake using PMA-induced HL-60 cells or native human macrophages by determining 6-CF after cell lysis or NBD-fluorescence with flow cytometry. Detection of MR in native cell populations was carried out by an antibody assay using flow cytometry; MR-representing cells were selected analytically., Results: Liposomes containing mannosides with more than one EO spacer length were specifically aggregated by ConA, indicating accessibility of the carbohydrate ligands of these derivatives. Increase in EO spacer units of incorporated mannosides (two or more EO) led to suppression of cellular uptake of mannosylated liposomes by phagocytes lacking MR (HL60, U937). The extent of suppression increased with spacer length. Liposome uptake by native macrophages expressing MR was, on the contrary, improved, particularly by Man6 and Man8., Conclusions: Uptake of liposomes modified with Man6 or Man8 by native cells was enhanced but did not reach an optimum. Thus, Man6, Man8, and mannosides with even longer spacer arms are of potential use in receptor-mediated targeting.

A way to synthesize neoglycolipids with high yields and anomeric purity is described. Starting point of the synthesis strategy is the glycosylation of allyl alcohol with definite steric orientation. Introduction of the hydrophobic moiety was achieved by photoaddition of n-hexadecanethiol and 3-mercaptopropionic acid followed by amidation with n-hexadecylamine, respectively. In order to investigate the influence of different carbohydrate headgroups in the physicochemical behavior of the general glycolipid, especially the orientation of the alkyl chain, a range of neoglycolipids was synthesized. Beside the differences in the configuration between unfunctionalized glycopyranoses like D-glucose, D-galactose and D-mannose, a number of deoxy and partially O-methylated sugar derivatives was prepared. The divergences concerning the different carbohydrate headgroups and the hydrophobic moiety, respectively, can be compared to relatively simple structured glycolipids with hexadecyl residue and without spacer function.

Numerous biological, chemical and photochemical processes consume molecular oxygen, thereby forming peroxides, hydroperoxides and even hydrogen peroxide. This paper describes a new system for monitoring quasi simultaneously oxygen uptake and peroxide formation. For this purpose capillary electrophoresis with electrochemical detection was connected to a new automated injector that periodically injects a sample of the reaction mixture into the capillary electrophoresis system without any contact with the atmosphere. The device was tested with some pharmacologically relevant reactions such as enzymatic oxidation of glucose catalyzed by glucose oxidase, enzymatic lipid oxidation by lipoxygenase and the photodynamic reaction of the anti-inflammatory drug ketoprofen.

Ketoprofen (KP) is a potent nonsteroidal anti-inflammatory drug. However, application to the skin is problematic because the photosensitizing properties of the benzophenone moiety may cause phototoxic effects when the treated skin region is exposed to UVA light. Using capillary electrophoresis with electrochemical detection we are able to differentiate the peroxides formed during illumination of KP-containing solutions of linoleic acid. Contrary to other profens a high amount of hydrogen peroxide was found among the reaction products. For investigation of the skin damaging effect human keratinocytes were used as models. Cell viability, DNA synthesis efficiency and intracellular concentration of peroxides were determined. Viability and proliferation behavior was not altered under the influence of KP. While lower concentrations of KP (10-100 nM) led to a protection against the UVA-induced (8 J/cm2) cell proliferation damage, higher concentrations (10-100 microM) led to an amplification of the proliferation decrease. With UVB irradiation at relevant doses the effects were lower than using UVA. Furthermore, intracellular peroxide content was increased after UV irradiation and KP addition. In conclusion some efforts have to be done to avoid these side effects in the use of KP for topical or transdermal application.

The synthesis of two types of enantiomerically pure sn-2-deoxy-2-amido-glycero-phospholipids differing in the connection of the alkyl chain at the sn-1-position is described. Both types of lipids were prepared from L-serine-methylester as the chiral starting material.

Long chain alkylbenzenes were acylated with maleic anhydride and substituted maleic anhydrides. The resulting benzoyl acrylic acids were tested on phospholipase A2. The isomeric ratio (Z/E) of the benzoyl acrylic acids was estimated by NMR spectroscopy. A few acrylic acids cyclized to hydroxybutenolids.

Cyclization reactions of alpha-ketoarylhydrazonoyl chlorides with various dinucleophiles lead to new 1,4-benzothiazine, quinoxaline, tetrahydropyrazine, thiazole, and thiadiazoline derivatives of methyl butanoate or methyl 5-oxopentanoate. The inhibition of 5-lipoxygenase (LO) was determined by monitoring the leukotriene B4 (LTB4) formation of human polymorphonuclear leukocytes (PMNL). The IC50 values for the most active compounds with an arylhydrazone structure were found to lie between 0.7 and 7.5 microM.

Monolayers of an enantiomeric and a racemic triple-chain phosphatidylcholine (PC) at the air/water interface are studied by film balance measurements and x-ray diffraction. Although the area per three tails exceeds that per head, we observe tail ordering dependent on headgroup chirality and chain tilt. This indicates lateral headgroup interactions. The influence of the chiral carbon is suppressed at higher lateral pressures, and a centered-rectangular unit cell with tails tilted into the nearest neighbor (NN) direction is observed for both the enantiomer and the racemate. The distortion of the lattice changes at medium pressures from NN to NNN (next-nearest neighbor direction) with decreasing temperature. The phase behavior of the racemate at 15 degrees C is compared with that of a triple-chain PC with a branched chain of reduced length. Whereas the PC with the longer branched chain exhibits only a NN tilted phase at all pressures, the PC with the shorter branched chain has a rich polymorphism (NNN-NN-upright hexagonal packing) under increased lateral pressure.

alpha-Carbonyl carboxylic acid arylhydrazonochlorides obtained by Japp-Klingemann reaction are the starting substances for the synthesis of alpha-carbonyl carboxylic acid arylhydrazonoamides, -esters and -thioesters. The inhibiting activity of these compounds against 15- and 5-lipoxygenase is described. Reactions of derivatives of amidrazones with formaldehyde give triazole, triazoline and unexpected benzotriazepine derivatives.

Tetrahydro-2H-benz[b]azepin-2-ones as starting substances are synthesized by Beckmann rearrangement of Schmidt reaction. The tetrahydrobenzazepinones are transformed into the thiones, thiolactim ethers and phenylhydrazones. The compounds are tested as inhibitors of soja lipoxygenase.

The synthesis of some amidrazones as bioisosteric diazaallyl radicals is described starting from the reaction mechanism of the lipoxygenases which postulates the existence of a radical after hydrogen abstraction from the arachidonic acid, N1,N3-diaryl-, N1-aryl-N3-alkyl-benzamidrazones and N1-arylacetamidrazones are strong inhibitors of the soja lipoxygenase.

3-(4-Alkylbenzoyl)acrylic acids (ABAAs) were synthesized by acylation of alkylbenzenes with maleic anhydride and then screened in vitro for inhibition of phospholipase A2 (PLA2) from snake venom and from porcine pancreas. The inhibitory potency of ABAAs increased with the length of the alkyl residues resulting in IC50 values of between 10(-7) and 10(-4) mol/L. The most potent inhibitors of the snake venom PLA2 were the 4-(n)-hexadecyl and octadecyl (OBAA) derivatives. Kinetic experiments referred to a time-dependent inhibitory reaction. Irreversibility was examined by dilution and dialysis. A molar ratio of inactivation of OBAA of nearly 20 was estimated. Double reciprocal replots of the apparent inactivation constants to the concentration of OBAA gave a (pseudo) first order rate constant of inactivation of 2.3 min-1. For the dissociation constant of the enzyme-inhibitor intermediate, a value of 6 x 10(-6) mol/L was obtained. On the other hand, the PLA2 from porcine pancreas seemed hardly to be inhibited by ABAAs. The present data are discussed in relation to the proposed model for PLA2 inactivation by manoalide. In human PMNs leukotriene B4 and 5-HETE production was essentially reduced. In human platelets the thrombin-induced TxA2 production was reduced. Since these effects disappeared after addition of arachidonic acid, these findings refer to a PLA2 inhibition. The immunologically induced bronchospasm in guinea pigs was significantly and dose-dependently inhibited by OBAA. This indicates that ABAAs might be useful in treating allergic diseases, such as asthma, eczema, allergic shock and others.

Vesicles prepared from sucrose-palmitate-stearate and cholesterol encapsulating the anticancer drug methylglyoxal-bis-guanylhydrazone, were investigated. Treatment with drug-loaded vesicles of this composition in a q.d. 1-4 schedule resulted for murine leukemias P388 in nearly the same increase in life span as the corresponding dose of the free drug. Even very high dosages of sucrose-palmitase-stearate (up to 480 mg.kg-1.d-1) were tolerated by mice.

Similar to the chemotherapy of bacterial infections, with the advanced development of virostatics a combination of antiviral agents is supposed to be introduced into the causal treatment of viral illness. The present studies explain a method for in vitro testing of virostatic combinations in cell cultures. The principle of the method is based on the checkerboard-technique used to test antibiotic combinations. As example the combination of trisodium phosphonoformate with bromovinyl-2'-deoxyuridine, acyclovir or 2-hexadecylglycero-3-phosphocholine was tested against herpes simplex virus, type 1. For evaluation of the test results the so-called reduction dose 50 (RD50) was introduced. The RD50 corresponds to this substance concentration, which reduces alone or in combination with a second virostatic the virus concentration used in the test system to its TCID50. With analogy to the calculation of infectious doses the computation of the RD50 was performed by using the method of Spearman and Kaerber. The calculated values allow the comparability of the antiviral activity of substances and their combinations. Corresponding to testing of antibiotics the further analysis of combinations was carried out by calculation of fractional inhibitory concentration (FIC), synergy factors (SF), and the construction of isobolograms. In this way, indifferent, synergistic and antagonistic effects of substance combination should be determined.