Ehrlich II –2nd World Conference on Magic Bullets

Conclusion: Substantial CYP3A4 enzyme activity was indicated in jejunum and ileum while such activity was insignificant in the colon.

Neurotoxicity Related to Lithium Combination Treatment in a Patient with Schizoaffective Disorder SEIDL A, SCHOPPER C, BRANDENBERGER M, BOEKER H University Hospital Zürich, Switzerland Background: The therapeutic efficacy of lithium in the treatment of affective disorders is invaluable. Nevertheless neurotoxicity should be considered as a significant side effect. Neurotoxic encephalopathy has been described in the literature both in lithium mono-therapy with normal serum levels or with toxic levels and in combined treatment with other therapeutics, but particularly with neuroleptics. In the following the case of a patient with schizoaffective disorder who developed a neurotoxic encephalopathy related to lithium – risperidone combination treatment is explored. Methods: A clinical case of encephalopathy is described. Neurological examination, psychopathological state and the results of EEG investigations are presented. The relevant theoretical considerations of the aetiopathogenetic mechanisms are discussed, differential diagnostic steps and therapeutic implications are described. Results: A 60 year-old patient with schizoaffective disorder showed an acute neuropsychiatric state with severe cognitive deficits and an akinetic extrapyramidal syndrome under combined lithium risperidone treatment. An EEG investigation showed a marked change in basic activity. The clinical state slowly began to improve after withdrawl from the psychopharmaceutics, and the EEG also showed a clear improvement. The development of a neurotoxic encephalopathy under treatment with lithium and risperidone is the result of different aetiopathogenetic mechanisms. The most important hypothesis put forward is that lithium-neuroleptic treatment causes neurotoxicity by increasing dopamine receptor blockade which results in profound dopaminergic hypofunctionality reflected e.g. by extrapyramidal symptoms. A neuroleptic malignant syndrome can be said to be the most important differential diagnosis. Conclusions: 1) Neurotoxic symptoms under lithium combination treatment may be interpreted as the interaction of different aetiopathogenetic mechanisms. Interactions in the dopaminergic system are very likely to play an important role. 2) The EEG is the most important diagnostic parameter for both the acute phase and for the follow up. 3) In patients developing signs of intoxication under lithium therapy, discontinuation of lithium medication should be considered.	Silkworm infection models to evaluate the therapeutic effects of antibiotics SEKIMIZU K, HAMAMOTO H, ASAMI Y, HORIE R, MATSUMOTOY, NAGATA M, KAITO C Lab. of Microbiology, Grad. Sch. Pharm. Sci., Univ. of Tokyo Most drug candidates obtained by in vitro screening are inappropriate as medicines due to their toxicity and their pharmacodynamics in the human body. Preclinical tests using animal models are essential, however, for evaluating the therapeutic effects of drug candidates for further development. Although mammalian models have been used to examine the pharmacodynamics of drug candidates, both the high cost and the ethical issues of sacrificing mammals for drug analysis can delay the development of potentially therapeutic drugs. The use of invertebrate animals for the evaluation of drug candidates can overcome these problems. We propose the use of silkworms, Bombyx mori, as model animals to evaluate the properties of drug candidates. The lower cost and smaller space required for the maintenance of silkworms compared to mice allows for a larger number of animals to be handled in limited facilities. Because of the long history of the silk industry, the methods for taking care of silkworms are well established. Silkworms are ideal for use in a large-scale drug screening system, as they are large enough to be used in injection experiments, for making hemolymph preparations, and for isolating organs such as the midgut, which are essential processes for studying the pharmacodynamics of drugs in individual animals. The silkworm fat body functions in drug metabolism, similar to the mammalian liver, and contains a number of cytochrome P450s and sulfur or glucose conjugation enzymes, which are involved in drug detoxification. In this symposium, we report that pathogenic microorganisms, such as Staphylococcus aureus and Candida albicans, were lethal to silkworms, and clinically-used antibiotics had therapeutic effects in silkworms. Moreover, the effective doses of antibiotics in this silkworm infection model were similar to those in mammalian models. Further, the availability of antibiotics by oral administration, and the drug distribution and metabolism were similar between silkworms and mammals.
Inhibition of angiogenesis and melanoma metastasis by DisBa-01, an alphavbeta3-blocking RGD-disintegrin from Bothrops alternatus snake venom SELISTRE-DE-ARAUJO, HS^1, RIBEIRO, JU^1, COMINETTI, MR^1, RAMOS, OHP^1, PONTES, CLS^1, SOUZA, DHF^2, CREPIN, M^3, FAUVEL-LAFEVE, F^3 AND BONNEFOY, A^3,4 ^1Dep. Ciências Fisiológicas, ^2Dep. Chemistry, Universidade Federal de São Carlos, São Carlos, SP, Brazil, ^3INSERM Unit 553, Paris, France; ^4INSERM Unit 743, Montreal, QC, Canada Background: The integrin v3 is involved in multiple aspects of tumor angiogenesis and metastasis, which makes this receptor a key target for the development of anti-cancer therapies. Snake venoms are natural sources of small integrin ligands named disintegrins, which act as selective integrin inhibitors. We have previously demonstrated that DisBa-01, a recombinant RGD-disintegrin from Bothrops alternatus venom glands, is a strong inhibitor of the v3 integrin, therefore inhibiting tumor cell adhesion to vitronectin (IC50 = 225nM for B16F10 melanoma cells). Methods: Inhibition of angiogenesis: The ability of DisBa-01 to inhibit bFGF-induced angiogenesis was tested in a matrigel plug assay in athimic nude mice. Inhibition of lung metastasis: The anti-metastatic activity of DisBa-01 was evaluated by injecting the luciferase-expressing B16F10-2B8 cells mixed with DisBa-01 in the tail vein of C57BL6/j mice. The progression of pulmonary metastases was measured at day 1, 4, 7, 11 and 14 following cell inoculation by an imaging system. Flow assay: MDA-MB-231 cells labeled with cell tracker red were previously incubated with DisBa-01, mixed with whole blood labeled with calcein green and perfused at a shear rate of 1500 sec^-1 in a flow chamber on a collagen type I-coated coverslip. Adhered platelets and cells were differentially counted using the software Image J. Results: DisBa-01 dose-dependently decreased bFGF-induced angiogenesis in a matrigel plug assay (IC50= 83 nM). When injected intravenously to C57BL/6 mice together with B16F10 melanoma cells, DisBa-01 time- and dose-dependently inhibited lung metastasis. Under flow conditions, DisBa-01 (100nM) almost completely inhibited the adhesion of MDA-MB-231 cells to collagen I and to the extracellular matrix produced by endothelial cells as well. Deletion of the N-terminal up to 26 residues did not affect the inhibitory activity of DisBa-01 to the v3 integrin. Conclusions: DisBa-01 is a potent new inhibitor of v3 integrin-dependent adhesion processes involved in tumor angiogenesis and metastasis. Support: FAPESP, CNPq (Brazil) and INSERM (France)	Correlation Between Clinicopathological Features and Allelic Loss at Tp53 In Metastatic Endometrial Cancer SEMCZUK A^1, OLCHA P^1, SEMCZUK M^2, CYBULSKI M^3, PRZĄDKA-RABANIUK D^1, FILIP A^4, SKOMRA D^5, SZEWCZUK W^1 ^1II^ND Department of Gynecology, ^2Department of Obstetrics, ^3Department of Biochemistry and Molecular Biology, ^4Deparment of Human Genetics, and ^5Department of Pathology, Lublin Medical University, Lublin, Poland Adenocarcinoma of the endometrium (EC) is one of the most common malignancy of the female genital tract, with 4196 new cases diagnosed in 2005 in Poland. Although our knowledge of genetic features in primary endometrial carcinoma has been expanding, there is limited number of studies evaluated the role of molecular alterations during EC spread. Currently, we examined the allelic loss at the TP53 gene in 38 metastatic ECs, and investigated the relationship between LOH (Loss of Heterozygosity), p53 protein overexpression and clinicopathological features of cancer. Three intragenic TP53 polymorphic markers, located at intron 1, intron 4 and exon 4, were analyzed. Overexpression of p53 was evaluated immunohistochemically applying monoclonal mouse anti-human p53 antibody (clone DO-7, diluted 1:100) and the Vector Laboratories visualization systems. There was no significant association between LOH at intron 1 and clinical and pathological variables of cancer. A significant correlation existed between allelic loss at intron 4 and the presence of the neoplasm in the uterine cervix (R=0.319, p=0.049; Spearman rank correlation test). There was also a tendency for an inverse correlation between allelic loss at exon 4 and vascular space invasion, but this difference did not reach a significant value (R=-0.321, p=0.068; Spearman rank correlation test). p53 protein was overexpressed in 34% (13 out of 38) ECs, either in primary or in metastatic endometrial lesions, and was significantly related to patients’ age (p=0.043) and to the presence of the neoplasm in the fallopian tube (p=0.046). Overexpression of the protein was significantly correlated with LOH at intron 1 of the gene (R=0,599, p=0.0001; Spearman rank correlation test), and a tendency existed for the correlation between p53 overexpression and allelic loss at intron 4/exon 4. Altogether, allelic loss at the TP53 is present in a subset of advanced-stage EC patients, and is correlated with p53 protein overexpression, particularly at intron 1of the gene.
In Vivo Molecular Imaging of Capromab Pendetide in Humans and Small Animals Using Combined Dual-Modality SPECT/CT and microSPECT/CT Systems SEO Y^1 ^1University of California, San Francisco, USA Background: Capromab pendetide is a monoclonal antibody targeting an intracellular epitope of prostate specific membrane antigen (PSMA). This antibody radiolabeled with ^111In for radioimmunoscintigraphy is primarily used for detection of lymph node metastasis or recurrence of prostate cancer. The same antibody also has a potential as a radioimmunotherapeutic agent with a radiolabel that emits beta particles such as ^177Lu or ^90Y. Single photon emission computed tomography (SPECT) has been the choice of imaging radiolabeled capromab pendetide. In comparison to standalone SPECT, a combined dual-modality SPECT/CT technology can offer a) better image quality and quantitative accuracy of SPECT and b) means to localize uptake of radiolabeled capromab pendetide with anatomical details from computed tomography (CT). Over the past decade, we have improved methods to image both humans and animal models using the SPECT/CT technology and radiolabeled capromab pendetide. Methods: Patient capromab pendetide imaging studies first started in 1999 using a prototypc SPECT/CT system at UCSF. Since 2003 the studies were performed using a commercial SPECT/CT system that has a lower-resolution CT capability. Most recently in 2008, we started patient imaging studies with a high-end SPECT/CT system with a diagnostic-quality 16-slice CT capability. We started imaging LNCaP xenograft mice using ^111In- and ^177Lu- labeled capromab pendetide using a dedicated small animal pinhole SPECT combined with CT system in 2005. We quantitatively analyzed interpreter’s confidence level for patient data when SPECT/CT technology was used, and pharmacokinetics and biodistributions of ^111In- and ^177Lu- capromab pendetide in the xenograft models. Results: We found an increased confidence level in interpretation of patient imaging data of ^111In-capromab pendetide using SPECT/CT over SPECT. ^177Lu-capromab pendetide showed similar or slightly better imaging characteristics over ^111In-capromab pendetide in the LNCaP models. Conclusions: SPECT/CT imaging of capromab pendetide in humans improved the visual quality as well as our confidence in interpretation. The small animal imaging studies with both ^111In and ^177Lu as radiolabels showed a dual-role potential of ^177Lu-capromab pendetide as a radioimmunotherapy agent and as an imaging agent during radioimmunotherapy.	Illicit Drugs and Cardiac Arrhythmias in Athletes FURLANELLO F(*/**), SERDOZ LV(*), CIOFFI G(**), DE AMBROGGI L (*), CAPPATO R(*) * Arrhythmias and Electrophysiology Center, Department of Cardiology, IRCCS Policlinico San Donato, University of Milan, Italy; ** Clinica Villa Bianca – Trento, Italy The current management of athletes with arrhythmias is complicated by the large use of “illicit drugs”, taken, at any age, both by professional and non professional athletes. The World Anti-Doping Agency (WADA) yearly updates a list of prohibited substances and methods banned by the International Olympic Committee. The list includes different classes of substances namely Anabolic Androgenic Steroids, Hormones and Related Substances (Erythropoietin; Growth Hormone, Insulin like Growth Factor, Mechano Growth Factors); Gonadotrophins; Insulin; Corticotrophins, Beta-2 agonists, Diuretics and other Masking Agents, Stimulants, Narcotics, Cannabinoids, Glucocorticosteroids Alcohol, Beta-Blockers and others. The term “illicit drugs” comprises all categories of drugs banned by WADA, regardless of whether they are taken in order to improve physical performance (true “doping agents” or “performance enhancing drugs”), to mask the presence of specific doping agents during control tests (“masking agents”), or to counteract the hormonal side effects of doping agents. Several illicit drugs may cause cardiac collateral effects, through a direct or indirect cardiac action, and may provoke especially arrhythmogenic effects, during short, medium or long term. The cardiovascular effects comprehend a wide spectrum of diseases: hypertrophic, dilated, ischemic cardiomyopathies, myocarditis, thrombo-embolic diseases and also a wide range of supraventricular and/or ventricular cardiac arrhythmias, focal or reentry type, that are often symptomatic and potentially lethal even in healthy subjects. The risk of lethal arrhythmias and sudden death (SD) is very high in subjects with preexisting cardiac diseases, particularly latent arrhythmogenic substrate or primary arrhythmic disorders including some inherited cardiomyopathies at risk of SD or with “ex novo” structural disease due to assumption of the illicit drugs. Together with a continuous effort in improving the analysis of prohibited drugs it is crucial that the doping control strategies include the investigation of mechanisms of cardiac action and toxic effects of every single drug in the current “2008” WADA list. Particular attention has to pay to “recreational drugs” (or drugs of abuse) including ecstasy (MDMA) and other amphetamines and numerous very new synthetically derived formulations, several classified ad “designer drugs”.
Detection Of Lamivudine And Adefovir Resistant Hepatitis B Virus Polymerase Gene Variants During Two Years Antiviral Theraphy Of Chronic Hepatitis B Patients In The South Of Turkey SERIN MS^1, BEKIROGLU E^1, TEZCAN S^2, ASLAN G^2, SEZGIN O^3, ALTINTAS E^3, EMEKDAŞ G^2, POLAT S^1 ^1 Mersin University, Phaculty of Pharmacy, Dept. Pharmaceutical Microbiology, Mersin, TURKIYE ^2 Mersin University, Phaculty of Medicine, Dept. Medical Microbiology, Mersin, TURKIYE ^3 Mersin University, Phaculty of Medicine, Dept. Gastroenterology, Mersin, TURKIYE Background: Hepatitis B virus (HBV) belongs to the family of hepadnaviruses. The virus associated with acute hepatitis, chronic hepatitis, and development of hepatocellular carcinoma. Lamivudine (LAM) and Adefovir (ADV), are oral nucleoside/tide analogues, inhibit HBV replication and can markedly reduce serum HBV DNA levels and normalise alanine aminotransferase (ALT) levels associated with improvement in liver necroinflammatory activity, but the greatest drawback with LAM and ADV is the emergence of drug-resistant. Methods: In this study, we have examined 80 sera samples obtained from patients with chronic hepatitis B ongoing LAM and/or ADV treatment in gastroenterology department for about 24 months. A 422bp part of HBV polymerase gene was amplified by a nested PCR protocol which including B, C and D domains of viral polymerase gene. This fragment was also sequenced by a DNA cycle sequencing protocol. The sequencing gel was visualized using silver-staining method. Results: We have detected mutations related LAM resistance in HBV polymerase B and C domains in 15 sera samples (18.75%). Ten of 80 were M204I (12.5%), 3 of 80 M204V (3.75%) and 2 of 80 L179M (2.5%). We have also detected mutations related ADV resistance in B and D domains of HBV polymerase gene in 5 sera samples (6.25%). These mutations were 4 of 80 N236T (5%) and 1 of 80 P237H (1.25%). Conclusions: The study reveals that antiviral therapy with viral polymerase inhibitors is still controversial and new antiviral strategies are necessary. This study is undergoing and planned to analyze all chronic hepatitis B patients ongoing lamivudine treatment in Mersin University research hospital.	The new biocidal agents with the high sporicidal efficiency SEVERA J^1, KLABAN Vl^1, SEDLACKOVA M^1 Decomkov Prague Ltd., Laboratories Hradec Králové, Czech Republic Background: Permanent tendency of scientific workers in the area of development of the new disinfectants is to introduce the disinfection agents with the broad spectrum efficiency which are slightly or non toxic, have a good stability, they are biodegradable, easy applicable as the aerosols or as the foams. Using of foams as the carriers of chemical compounds with the disinfection efficiency has indisputable advantageous. The goal of this paper is to describe the results of the development of the two new biocidal agents with the high sporicidal efficiency. Methods: At first the mixtures from different compounds were prepared. The first agents under the name HVĚZDA (STAR) contains as a efficacious component hydrogen peroxide and the other parts are alkalised mixture of tensides (quaternary ammonium salt and anionic tenside). The other of them whith working name PTSPCH is built-up from the hypochlorite sodium, other parts are partly derivate of pyrrolidone and partly organic phosphate. Both have alkaline character.Verified stability of HVĚZDA is one year and stability of the other agent was investigated for 200 days and continued till this time. Stability was verified partly using quantitative analysis concentration of biocids and partly microbiological inquire to sporicidal efficiency in agreement with European norm. It is assumed that both agents would be used in practice as bicomponente agent. Bactericidal, fungicidal and sporicidal efficiency were carried out in according to standard operational method. Bacterial strains from the Czech collection of microorganism ( Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus subtilis) were used. Results: The development of two new biocidal agents was successfully finished. Both have high disinfection effect particularly sporicidal efficiency. Conclusions: We plan to verify these new agents in experiment with deactivation Bacillus anthracis spores in the special workplace in Czech republic.
Targeted Delivery of Cytotoxic Drugs by Means of Protein Vectors SEVERIN SE Moscow Research Institute of Medical Ecology, Moscow, Russia Background: Epidermal growth factor (EGF) and its receptor-binding fragment (EGFfr) were shown to be promising vectors for targeted delivery of cytotoxic agents to tumor cells. The use of short peptides as a vector components of the targeted preparations is more preferable owing to their high stability, easy availability and relatively low cost. The aim of the present study was design of vector peptides targeted to EGF receptor and investigating of cytotoxic activities of their conjugates with antitumor antibiotic doxorubicin (DOX). Methods: Solid phase peptide synthesis (Fmoc strategy); study of growth-stimulating activities of peptides against fibroblast cell cultures; synthesis of DOX conjugates with peptides using crosslinking reagent; study of cytotoxic activity (CTA) of conjugates in vitro against human tumor cells. Results: Two modified forms of EGFfr (EGFfr1 and EGFfr2) were synthesized. EGFfr1 differed from the original fragment by the presence of Ser instead of Lys in position 28. EGFfr2 contained Lys instead of Met in position 20 and Ser instead of Lys in position 28. Ser is one of the few amino acids forming the binding site in murine EGF. The undesirable conjugation at the -amino groups of Lys in the active center of the receptor-binding fragment can be avoided through substitution of Ser for Lys which, in its turn, prevents the inhibition of binding of the polypeptide to the EGF receptor. The substitution of Lys for Met20 in EGFfr2 was performed to improve the conjugation of the peptide to DOX. Both EGF fragments manifested biological activities in vitro which exceeded activity of native EGFfr. The conjugates manifested CTAs towards cultured human carcinoma cells HeLa which exceeded that of the free antibiotic 2-3-fold. The CTAs of the conjugates were close to that of free DOX against sensitive to this drug tumor cells MCF-7^Wt and exceeded 1.5-2-fold the CTA of DOX against resistant cells MCF-7^AdrR that characterized by hyperexpression of P170 protein. Conclusions: 1) The amino acid substitutions in the EGFfr fragment are responsible for the increase of its receptor-binding ability. 2) Both peptides can be used for targeted delivery of DOX and, perhaps, some other antitumor drugs, to tumor cells.	Spectroscopic study on the interaction of the antitumoral drug emodin with bovine serum albumin: fluorescence, circular dichroism, SERS, SEF and stopped-flow techniques SEVILLA P^1,2, GARCIA-BLANCO F^1, SANCHEZ-CORTES S^2, GARCIA-RAMOS J.V^1,2 ^1Univ. Complutense de Madrid, Madrid, ES; ^2CSIC, Madrid, ES Background: Emodin shown an anti cancer effect in neuroectodermal tumor, breast cancer cells or prostate cancer. It binds Bovine Serum Albumin (BSA) forming a complex that is very important to ensure the drug delivery. Using Surface Enhanced Raman Spectroscopy (SERS), Surface Enhanced Fluorescence (SEF), Fluorescence, Circular Dichroism (CD) and Stopped-flow we have unequivocally characterized the binding sites of emodin/BSA complex.