Ehrlich II –2nd World Conference on Magic Bullets

The Pharmacology, Pharmacokinetics, Clinical Efficacy, Adverse Effects And Toxicities, Drug Interactions, Dosage And Administration, And Safety Issues Related To The Use Of Prasterone Are Discussed
KOCIS P
Anticoagulation Clinic, Penn State Milton S. Hershey Medical Center, Hershey, PA USA
Prasterone is a proprietary synthetic dehydroepiandrosterone product under investigation for use in women with systemic lupus erythematosus (SLE) who are taking glucocorticoids. Initial trials investigated prasterone as a treatment to improve disease activity and symptoms in women with mild to moderate SLE. The Food and Drug Administration (FDA) did not approve prasterone's labeling for these indications. Subsequent trials have focused on prasterone as a treatment to limit bone loss in women who have SLE. A study was conducted to assess bone mineral density in patients who had been taking glucocorticoids for six months or longer. The patients in the prasterone group showed an increase in bone mineral density, while the placebo group demonstrated a loss. The most common adverse effects of prasterone therapy were acne and hirsutism. Hematuria, hypertension, and serum creatinine concentration increases have also occurred. Interactions

of prasterone potentially exist with 5-alpha reductase inhibitors and additive or antagonistic effects could possibly occur with androgens, estrogens, oral contraceptives, and progestins. In clinical trials, oral prasterone dosages of 100-200mg /day were administered. These dosages have resulted in supraphysiological hormone levels.

Conclusion:

FDA has granted orphan drug status for the prevention of loss of bone

mineral density in SLE patients taking glucocorticoids. FDA is requesting

additional Phase III trial data for the treatment of SLE and the prevention

of loss of bone mineral density.

Source:

American Journal of Health-System Pharmacy. 63(22): 2201-10, 2006 Nov 15

Targeting Cell Cycle Progression by Troglitazone KOGA H, SATA M Division of Gastroenterology, Department of Internal Medicine, Kurume University, Japan Background and Aim: Increasing evidence has confirmed that ligands for peroxisome proliferator-activated receptor γ (PPARγ) exhibit antitumoral effects through inhibition of cell proliferation and induction of cell differentiation in several malignant neoplasms. Recently, we have documented the accumulation of a cyclin-dependent kinase inhibitor, p27Kip1, as well as an unexpected accumulation in cyclin E in G1-arrested human hepatoma cells treated with the PPARγ ligand troglitazone. Simultaneous accumulations in both p27Kip1 and cyclin E are known to be characteristic phenotypes in cells derived from mice lacking Skp2, an F-box protein component of the SCF ubiquitin-ligase complex. Thus, the aim of the present study was to assess whether Skp2 might be involved in the up-regulation of p27Kip1 in troglitazone-treated human hepatoma cells. Methods: Human hepatoma cell lines were used in this study. Cell cycle was analyzed by flow cytometry and immunoblotting. The mRNA levels of p21, p27, and cyclin E in the hepatoma cells were examined by quantitative real-time RT-PCR. Skp2-overexpressing hepatoma cells were generated by the cDNA transfection. The expression levels of Skp2 and p27Kip1 in human hepatocellular carcinoma (HCC) tissues and the adjacent noncancerous liver tissues were assessed by immunoblotting. Results: A striking decrease in Skp2 expression and a reciprocal increase in p27Kip1 expression were found in troglitazone-treated hepatoma cells but not in those cells treated with other PPARγ ligands such as pioglitazone and ciglitazone. Quantitative real-time RT-PCR analysis showed that troglitazone downregulated Skp2 at the mRNA levels. Consistently, ectopic overexpression in Skp2 brought resistance to troglitazone, resulting in a decreased population of arrested cells at the G1 phase compared with that in the mock-transfected cells. In surgically resected hepatocellular carcinoma (HCC) tissue, an increased expression in Skp2 was found in both the moderately differentiated HCCs and the poorly differentiated HCCs. Conclusions: Troglitazone attenuated Skp2 expression, thereby promoting p27Kip1 accumulation in human hepatoma cells. This therapeutic potential of the ligand may lead to new cell-cycle-based antitumor strategies for advanced HCCs.	Enhanced Potency Of Antibodies Using Biologically Active Peptides KOHLER H, MULLER S, BRYAN, AJ Univ. of Kentucky, Lexingtony, USA; Background: Antibodies have evolved as effective alternative to chemical drugs to treat cancers, yet clinical response is limited by expression of tumor targets reducing the potency of antibodies. We have invented methods to enhance potency by (1) generating polyvalent (homophilic) antibodies and by (2) making membrane penetraing antbodies, The objectives of these methods is to create antibodies with enhanced anti-tumor effects and for intra-cellular targetsto control cell proliferation. Methods: To demonstrate enhanced potency of homophilic antibodies the Her2/neu specific Herceptin was crosslinked with a homophilic 26mer peptide derived from a naturally ocuuring homophlic antibody, T15, previously discovered in our laboratory. The peptide was photo-actively affinity crosslinked to Herceptin. Homophilic Herceptin was compared to naked Herceptin in FACS, apoptosis induction and in xenograft animal model using the human lung tumor H1650. Transmembrane antibodies were generated by photo-affinity crosslinking a 14mer peptide derived from human sarkosi sarcoma virus. Intracelluar targeting of life cells is demonstrated by confocal microsopy and inhibition of induced apoptosis. Results: Homophilic Herceptin poduced > 1 log stronger fluorescent intensity over naked Herceptin and increased the number of apoptotic cells to 82% compared to 12% with naked Herceptin. Homophilic Herceptin inhibited tumor growth by 60% in H1659 xenograft experiments compared to mice treated with naked Herceptin or in no-treatment controls. Trans-membrane penetrating (TMP) antibodies stained in live cells specifically actin and paxillin while naked antibodies did not. TMP antibodies also did not affect cell growth in culture. Furthermore, TMP modified anti-caspase3 antibodies blocked induction of apoptosis induced by actinomycin. Conclusions: 1) Photo-affinity crosslinking of biologically active peptides endows antibodies with properties that enhance targting and opens up novel target choices. Homophilic peptide modified Herceptin is more efficient in tumor killing in vitro and in vivo than naked Herceptin. 2) Membrane penetraing modified antibodies target intra-cellular antigens in living cells. 3) Collectively, these data provide methods to create a novel class of antibodies that are superior as diagnostic tools and therapeutic drugs.
Antioxidant and Antiglycation Potential of Some Sudanese Medicinal Plants and their Isolated Compounds KOKO WS^1, OSMAN EA^2, GALA M^1 ^1 Medicinal and Aromatic Plants Research Institute, National Center for Research, P. O. Box 2404, Khartoum, Sudan; ^2Faculty of Pure and Applied Sciences, International University of Africa, Khartoum, Sudan. Background: Free radicals or reactive oxygen species (ROS) appears to be associated with a number of human neurodegenerative disorders, inflammation, diabetes, viral infections, autoimmune pathologies and digestive system disorders. In the present some of Sudanese medicinal plants commonly used in the folk medicines against infectious diseases were investigated for their potential antioxidant, antiglycation and cytotoxicity. Methods: In the present work twenty three parts from 20 medicinal plants were extracted by 80% ethanol. These extracts were were investigated for their potential scavenging of superoxide free radicals by using PMS–NADH (phenazine methosulphate -nicotinamide adenine dinucleotide hydrogen reduced form) systems by oxidation of NADH and assayed by the reduction of NBT (nitroblue tetrazolium). The inhibition of glycated proteins was carried for the above extracts after long term icubation of glucose with BSA (bovine serum albumin). MTT {3 - (4, 5-dimethylthiazole – 2 – yl) – 2, 5 – diphenyl tetrazolium bromide} cytotoxicity assay was conducted for the above extracts against 3T3 cell line. Some of the acive extracts were subjected to phytochemical analysis for the isolation of active princibles by using column chromatographic techniques. Results: 14 extracts were revealed significant activity P < 0.5 for the scavenging of superoxide free radicals. A. nilotica barks, B. aegyptiaca barks, K. senegalensis leaves were the most potent with 74.7, 72.1, 70.5 % inhibition respectively. However, the rest revealed moderate inhibition activity. Only A. nilotica barks (78%) , K. senegalensis barks (74%), A. nilotica fruits (66%) and T. bakis. (61%) were reveled over 50% inhibition of glycation production assay, while the rest were less effective. No any significant reduction observed for tetrazolium of MTT cytotoxic test from such extracts. Sixteen compounds of different groups were isolated from acive plants and screened for the above bioassays. Conclusions: 1) A. nilotica barks extract was found to be the most potent antioxidant and antiglycation among all examined extracts 2) Bioassay guided phytochemical investigation proved catechin was the most potent isolated compound for both scavenging of superoxide free radicals and inhibition of glycation production assay3) MTT cytotoxicity against 3T3 cell line indicates the safety of all plant ethanolic extracts as well as isolated compounds.	IgY (Immunglobuline from egg-Yolk) - a “magic bullet” to fight antibiotic resistance KOLLBERG H (1), NILSSON, E(2), WEJÅKER P-E (2), LARSSON A (2). 1). CF Centre, University Children’s Hospital, Uppsala, Sweden, 2). Dep. Med sciences, University Hospital, Uppsala, Sweden. Paul Ehrlich, the father of sera therapies, searched for substances that could seek up specific disease-causing agents and link with toxins like a key in the lock. He found antibodies circulating in the body like ”magic bullets” The European Antimicrobial Resistance Surveillance system states, that the largest threat to health in Europe today is antibiotic-resistant bacteria. Each year 3 million Europeans are affected by infections in medical care, and approximately 50,000 of them die. The cause of this terrible development is excessive use of antibiotics and a continuous use of antibiotics will accelerate the rate of resistance. It is therefore extremely important to encourage a more restricted use of antibiotics (www.ecdc.europa.eu/documents/pdf/01_2006_AR_web.pdf). The increasing number of antibiotic-resistant bacteria emphasises the need to find alternative methods. Immune therapy can be used either alone or as a complementary treatment to antibiotics. Specific IgY has attracted considerable attention as an alternative to treat infectious diseases. Its biochemical properties make it attractive for passive immunotherapy. Hens vaccinated with bacteria or other antigens produce specific IgY against them. A single hen produces ~ 40 g IgY per year in their egg yolk. IgY preparations are stable for years. IgY antibodies decrease bacterial adhesion to epithelial cells and neutralise toxins Efficacy: Specific IgYs have proven efficacy against several enteric bacteria, viruses and fungi in animal and human studies. Specific IgY is effective also against antibiotic resistant microbes. In our own studies specific IgYs have been effective against Pseudomonas aeruginosa, Candida albicans and Enterobacter faecalis. Safety: Orally given immunoglobulines are not absorbed from the gastrointestinal tract. Orally administered IgY neither activates the human complement system nor reacts with any cell activators or mediators of inflammation. IgY is generally recognised as safe for oral or local use. There have not been any adverse events in our anti-pseudomonas study over 12 years to patients with Cystic fibrosis (CF). Individuals, who are allergic to eggs, must be excluded. Diminishes the use of antibiotics: In our hands IgY reduces the use of antibiotics and thereby also the severe drawbacks of antibiotics: Development of resistant strains, disturbance of the microbiologic flora, toxicity and allergenicity. There is no risk that pathogens develop resistance against IgY. State of the art: The Swedish Medical Agency has granted licence for anti-pseudomonas IgY to a group of CF patients. EMEA has approved orphan drug designation for Anti-pseudomonas IgY to treat patients with CF. Conclusion: IgY offers great opportunities to treat specific infectious diseases and thereby diminish the need of treatment with antibiotics. IgY is, indeed, one of Paul Ehrlichs “magic bullets”.
Nitric Oxide and Zidovudine Potentiate Oxidative Response of Stimulated Macrophages KOMAROV AM, WEGLICKI WB The George Washington University Medical Center, Washington, DC, USA Background: Macrophage is one of the key cell types involved in many disreases. Therefore modulation of oxidative response (respiratory burst) of stimulated macrophages by various pharmacological agents is very important. The aim of this study was to test (separately) modulation of respiratory burst by zidovudine and nitric oxide in vitro. Methods: Macrophages were isolated from the peritoneal cavity of 300-400 g male Sprague Dawley rats 3 days following injection of casein. The cells were washed and incubated with 18 μM of nitric oxide donor S-nitroso N-acetylpenicillamine (SNAP) for 3 hours, or azidothymidine (AZT) and azidothymidine monophosphate (AZT-MP) from 10 μM to 1 mM for 18 hours. The oxidation of 2’,7’-dichlorodihydrofluorescein (DCDHF; 5 μM) to 2’,7’-dichlorofluorescein by activated cells was measured with a fluorescent plate reader using a 24-well tissue culture plate. Cells were activated by adding 1 or 10 μl of a 10% suspension of polystyrene latex beads per million cells. Determination of hydrogen peroxide produced by non-activated and activated macrophages was based on horseradish peroxidase-mediated oxidation of phenol red. Results: Latex-activated cells oxidize DCDHF due to release of hydrogen peroxide and low-molecular iron complexes, which was verified in experiments using inhibitors catalase (1000 U/ml), desferal (100 μM) and peroxidase inhibitor sodium azide (2 mM). Preincubation of macrophages with SNAP (18 μM, 3 hours) increased DCDHF oxidation by latex-activated cells compared to control untreated cells. This effect was absent in macrophages incubated with SNAP decomposition products for 3 hours. SNAP also increased hydrogen peroxide release by activated cells. We have observed significant and dose-dependent increases in DCDHF oxidation in cells incubated with AZT, or AZT-MP. Catalase (100 U/ml) addition during incubation of cells with latex beads removed the effect of AZT and AZT-MP on DCDHF oxidation compared to control cells. Furthermore, AZT and AZT-MP increased hydrogen peroxide release from activated cells. Conclusions: Separate application of nitric oxide donor and zidovudine enhances oxidative potential of activated macrophages. Authors’ disclosure statement: Supported by NIH R01 HL-65178	Serine Protease Inhibitors: Cross Inhibitors of Prokaryotic and Eukaryotic Systems   KONAKLIEVA, M.I.^1 AND PLOTKIN, B.J.^2 ^1American University, Washington DC, U.S.A; ^2Midwestern University, Downers Grove, IL, U.S.A Background:-Lactams have historically been viewed as a class of antimicrobials. However, this paradigm is shifting towards a focus on their ability to function as inhibitors of bacterial enzymes, particularly those involved in broad-spectrum -lactam resistance, i.e., extended spectrum -lactamases (ESBL). This shift in focus is the result of recognition that -lactam’s acylate a broad range of enzymes that extends beyond taxonomic kingdom boundaries. These compounds have demonstrated activity against viral and mammalian serine enzymes in addition to their tradition target of bacterial enzymes. The common theme for these -lactam targets is that the majority of the enzymes have serine as nucleophile in the active site. The focus of this presentation is on the evaluation of the potential of -lactam antibiotics as inhibitors of the serine enzymes of both prokaryotic and eukaryotic origin, with specific focus on the structure-function relationship of -lactams as antimicrobial and antineoplastic agents. Methods: Molecular modeling studies and stereoselective synthesis have aided the design and synthesis of monocyclic -lactam libraries as inhibitors of prokaryotic and eukaryotic enzymes, which allowed for determination of the structure activity relationships. Results: The ability of the -lactams to act as excellent acylating agents is centered on the four-membered -lactam ring, while the substituents at the C3 and C4 positions provide certain specificity for recognition by the corresponding enzymes. Conclusions: A rigid confirmation is a common thread for -lactams functioning as enzyme inhibitors. Conformational requirements for recognition by proteases suggest a fundamental platform for the preparation of inhibitors that is dependent on developing conformationally restricted inhibitors which adopt receptor-binding conformation, and are therefore entropically advantaged for binding to a protease. However given this basic requirement for activity, selectivity has been difficult to achieve due to the similarity in the active sites. This double-edged sword will make the future development of inhibitors at once both easier and more difficult because of the potential for inflicting the very damage use of these inhibitors are hoping to block.
The Pharmaceutical Policy In Context Of Health Care System In Albania KONDAJ R Albanian Centre For Equity In Health The purpose of this study is to provide an in depth analysis on the current situation of the Pharmaceutical Sector , to identify the strengths and weaknesses in this sector and also to formulate a proposal document with the main strategic guidelines of Pharmaceutical Policy for present and future reference. My research is focused on some main problems of the pharmaceutical policy. In particular it covers pharmaceutical market, general prescribing, public private roles at the pharmaceutical sector , the approval procedure of drugs and legislation, pharmaceutical expenditure and reimbursement of drugs, price and distribution, management drug in hospital, health insurance and access to drugs , national pharmaceutical policy etc. The changes in Albanian Politics during 1990-1991, created a new atmosphere in all economic and social sectors, because Albania had inherited a list of unresolved problems and emerging complexities. Such problems are related to: Outcome ( equity, efficiency, satisfaction problems) Output ( quality and amount of services) Input structure( resource problems) On 1995, The Health Insurance Institute was established, and functioning as a “single payer” scheme. For the first time the general practitioners and essential pharmaceuticals were insured under the Health Insurance Institute. During the transition period in our country a lot of problems were faced in the pharmaceutical market, but we would like to highlight most important such as the strengths and weakness as follow: Strengths: The privatization of the pharmaceutical sector – flexibility, diversity, improvements in services and meeting patients needs. Structure of the pharmaceutical sector in a country level; Pharmaceutical distribution covers the whole country; Privatization of the pharmaceutical industry. Compilation of the pharmaceutical legislation; Formulation of the Pharmaceutical Order; All the medicine for sale have the price tag attached to the label; Weaknesses: A document of pharmaceutical politics with all its necessary components does not yet exists. Pharmaceutical sector functions like a regular market (demand- supply of pills) and not like a medical service with the health as the main concentration. The market is dominated by the symptomatic medicines instead of the etiologic ones relevant to the diseases. Almost 60% of the total Health Insurance Fund goes to the pharmaceutical sector to cover the reimbursed drugs. The production capacity of the drugs in our country is very low, and the production quality does not meet the criteria of GMP; There are no any written and approved standards by the Ministry of Health for the Pharmaceutical sector. The environment where the medicines are stored, like drug stores, warehouses and other pharmaceutical agencies do not always meet the health regulations. Lack of communication and misunderstanding between doctor-pharmacist-consumer. Uneven distribution of the drug stores in the country, most of them located in Tirana; There is missmanagement of the medicines in the hospitals; An education and training program of the staff does not exist in the institutional level; There is a lack of management and financing of the Research & Development filed. More changes are necessary in the pharmaceutical sector to insure the collaboration among all institutions, as well as the government support in improving all the necessary legislation related to the pharmaceutical sector. The pharmaceutical Policy should play an important role in Health Care System in Albania.	Experimental Approach for Growth Inhibition of Human Malignancies by the Highly Efficient Anti-tumor Peptides Delivery System KONDO E^1 ^1Dept. of Pathology, OKAYAMA Univ., Okayama,Japan Background: Molecular targeting agents have become formidable anticancerweapons, which show much promise against the refractory tumors.Functional peptides are among the more desirable of these nanobio-tools.Intracellular delivery of various functional peptides formsa basis for potent, non-invasive mode of delivery, providingdistinctive therapeutic advantages. Methods: We examined growthsuppression efficiency of aggressive human leukemia/lymphomas, glioblastomas and other human malignancies in vitro and in vivo tumor models by introducing anti-tumor peptides as a complex with the „Wr-T“ peptide transporter which serves to augment delivery of a cargo peptide. We did single, or dual peptide introduction of two tumor suppressorpeptides (p14^ARF, p16^INK4a, p21^CIP1 functionalpeptide or their combinations) using Wr-T-mediatedintracellular peptide delivery. Results: Wr-T–mediated transport of p16^INK4a functional peptidedramatically inhibits growth of highly aggressive p16-negative leukemia/lymphoma cell lines by up to 80% through restoration of p16 function. Based on this result, we further did Wr-T-mediated simultaneous introduction of two tumor suppressorpeptides, p14^ARF andp16^INK4a functional peptides, into human glioblastoma cell lines, which reversed specificloss of p14 and p16 function, thereby drastically inhibitingtumor growth by >95% within the first 72 h, whereas the growthinhibition was 40% by p14 or p16 single-peptide introduction.Additionally, the combination of p16 and p21^CIP1 peptides dramatically suppressed the growth of glioblastomaline which carries a missense mutation in p53, by>97% after 120 h. Significantly, our murine brain tumor modelfor dual-peptide delivery showed a substantial average survivalenhancement (P < 0.0001) for peptide-treated mice. The similar inhibitory effect by dual peptide introduction was also observed in p16- and p14-double negative cancer cells. Conclusions: Thus, it was demonstrated that the peptide transporter-mediated delivery of single or dual anti-tumor peptides seems to be highlyeffective against aggressive human malignancies in non-invasive manner, by singly or jointly restoringmultiple tumor suppressor functions.
pH Dependent 5-Fluorouracil Release System using Polymeric Micelles KONDO S^1, KUZUYA M^2 ^1Gifu Pharmaceutical Univ., Gifu, Japan; ^2 Matsuyama Univ., Matsuyama, Japan Background: The newly formed tumor vessels are usually abnormal in form and architecture, and tumor tissues usually lack effective lymphatic drainage. Therefore, these factors will lead to abnormal molecular and fluid transport dynamics especially for macromolecular drugs and nano carrier. On the other hand, it has been known that amphiphilic block copolymers consisting of a hydrophilic and a hydrophobic block can form core-shell micelles, referred to as “Polymeric Micelles”, in a selective solvent due to the association of the insoluble block. Polymeric micelles have attracted much attention in drug delivery systems. However, only a few works on controlled drug release from polymeric micelles has been studied. Aims: 1) To fabricate the polymeric micelle decomposed in an acidic condition. 2) To investigate the 5-fluorouracil (5FU) release profiles from the polymeric micelles with the change of pH. Methods: The amphiphilic block copolymer was synthesized by the mechano-chemical solid-state polymerization of poly 4-vinylpyridine and methacryloyl galactopyranose. Polymeric micelles were prepared by the dialysis method. Dynamic light scattering measurements were performed to observe the size change of polymeric micelles with pH. The 5FU released from polymeric micelle was monitored with UV spectrometer. Results: The number average particle diameter of polymeric micelles prepared was about 200 nm. The particle diameter of polymeric micelles steeply decreased at pH 5.6 (less than 10 nm), although its diameter unchanged from pH 7 to 5.7. When the pH was changed from pH 1 to 7, the particle diameter steeply increased at pH 5.6. It was also shown that 5FU was immediately released from the polymeric micelle at pH 5.6, although 5FU was not detected from pH 7 to 5.7 within a detectable extent. Conclusions: 1) The polymeric micelle prepared from this amphiphilic block copolymer was immeadiately deforemed or formed at pH 5.6. 2) 5-Fluorouracil incorporated in polymeric micelles was steeply released at pH 5.6. Therefore, this polymeric micelle seemes to be applicable for the pH dependent drug release system. 3) It is well-known that lysosome is acidic environment. Thus, this polymeric micelle is expected as lysosome targetting system.	Comparative Study of Dihydroartemisinin and Artesunate Safety in Healthy Thai Volunteers KONGPATANAKUL S^1, CHATSIRICHAROENKUL S^1, KHUHAPINANT A^1, ATIPAS S^1, KAEWKUNGWAL J^2 ^1Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; ^2Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand. Background: Artemisinin derivatives including artesunate and dihydroartemisinin (DHA) have been used for treatment of malaria. As part of new drug development initiatives in Thailand, a new tablet formulation of DHA has been developed. Our initial phase I bioequivalence study indicated that the new and reference DHA formulations were well tolerated; however, a significance decrease in hemoglobin (0.7-0.9 g/dl) was detected after a single 200 mg oral dose. To explore further, a clinical study with an emphasis on hematological parameters was conducted. Methods: A single-center, randomized, single-blind, cross-over clinical study was conducted in 24 healthy Thai volunteers with a dosage of 300 mg daily for 2 days. Artesunate was used as a comparator. Clinical adverse events were monitored. Laboratory assessment (CBC, RBC morphology, reticulocyte count, Coombs’test, total and direct bilirubin, LDH, AST, ALT, and hemoglobinuria) was performed on study days 0 prior to drug administration and days 2, 5, and 7 post drug administration. Results: Eighteen volunteers (10 males and 8 females) completed both rounds of the study. All adverse events were mild. Nausea and vomiting were common and occurred only in female volunteers. A statistically significant decrease in hemoglobin was detected (0.5 g/dl at study day 7, P < 0.05). Decreases in laboratory values below the normal limits were observed in some volunteers for reticulocyte and WBC counts. Other changes in laboratory values were minor. Transient, mild bone marrow suppression was evidenced by: 1) reduction of reticulocytes, 2) leukopenia, and 3) a minor drop of platelet counts. CONCLUSIONS: The present study confirmed our previous finding on a significant decrease in hemoglobin. Bone marrow suppression might be one of the causes. Considering the absence of clinically significant anemia (though decrease in hemoglobin) and its similarity in drug response profiles to artesunate, the development of the DHA should pursue.
Energetics of Cytochrome P450 Hydroxylations: Making Sense of In Silico: KOPPENOL WH, BOUNDS PL Institute of Inorganic Chemistry, Department of Chemistry and Applied Biosciences, ETH Zürich, Zürich, Switzerland Cytochrome P450 catalyzes, among others, the reaction RH + O2 +H^+ + NADPH  ROH + H2O + NADP^+, in which RH may be a drug. As most drugs interact with cytochrome P450, there are three possibilities: (a) inactivation of cytochrome P450, (b) inactivation of the drug by cytochrome P450, and (c) activation of the drug by cytochrome P450. An ideal drug, or magic bullet, is to be activated, because an effective drug that is inactivated by cytochrome P450 requires an additional drug, namely an inhibitor of cytochrome P450. Use of a pro-drug may indeed be a wise strategy as cytochrome P450 is overexpressed in some cancer cells. The thermodynamic properties of the intermediates in the cytochrome P450 cycle can be estimated with a few simple assumptions, and the result for the electrode potential of the couple Compound I/Compound II is 1.4 V (W. H. Koppenol, J. Am. Chem. Soc. 129, 9686-9690; 2007), a value in agreement with an estimated bond enthalpy D(FeOH)^3+ of ca. 410 kJ/mol (M. T. Green, J. H. Dawson, and H. B. Gray, Science 304 (5677), 1653-1656; 2004). Compound I is almost isoenergetic with the haemiron(III) – hydrogen peroxide complex that precedes it. The electrode potential of 1.4 V is quite different from that implied by the results of ab initio calculations. A higher electrode potential would imply a small association constant between the haem iron(III) and hydrogen peroxide, and a lower value would not allow hydroxylation. Compound I can thus abstract a hydrogen of a primary carbon atom; abstraction from a secondary or tertiary carbon atom is thermodynamically more facile. Thus, hydroxylation to activate a drug is thermodynamically always feasible, and is only limited by the regiospecificity of the hydroxylation. If hydroxylation is undesirable, because it inactivates the drug, see (b) above, then, rather than adding a compound that inactivates cytochrome P450, one might attempt to replace the hydrogen that is abstracted with a fluor atom.	Ab Initio Calculation Of Molecular States Of Compounds Of The Lanthanum And Yttrium Molecules KOREK M^1, FARHAT A^1, MOUSSA A^1, HAMMOUR H^1, ALLOUCHE AR^2 1-Faculty of Science, Beirut Arab University, P.O.Box 11-5020 Riad El Solh, Beirut 1107 2809, Lebanon, 2-Laboratoire de Spectrométrie Ionique et Moléculaire, CNRS et Université Claude Bernard Lyon I, Villeurbanne Cedex, France The potential energy curves of the grounds and lowest electronic states of the compounds of the Lanthanum and Yttrium Molecules have been investigated via CASSCF method. Multireference CI calculations (single and double excitation with Davidson corrections) were performed. Lanthanum and Yttrium atoms are treated in all electron scheme. In the range of internuclear distance R around the equilibrium distance of their ground states, the molecules are assumed to be mainly ionic. The potential energy curves for the considered states in the representation have been calculated in the range 2.0Å  r  3.5Å. The spectroscopic constants such as the vibrational harmonic constant e, the internuclear distance at equilibrium re, the rotation constant Be, and the electronic transition energy with respect to the ground state Te have been calculated by fitting the energy values around the equilibrium position to a polynomial in terms of the internuclear distance, the degrees of these polynomials are determined from the evaluation of the statistical error for the coefficients. By using the canonical functions approach and the cubic spline interpolation between each two consecutive points of the potential energy curves obtained from the ab initio calculation, the eigenvalue Ev, the rotational constant Bv, the centrifugal distortion constants Dv, and the abscissas of the turning point (Rmin, Rmax) have been calculated for various vibrational levels. The comparison of these values to the theoretical and experimental results available in the literature shows a good agreement. Many electronic states for the considered molecules have been studied theoretically for the first time.
Electronic Homeopathic Preparations (EHPs) as Potential “Magic Bullet”: Pilot Study on Biologic Model KORENBAUM VI^1, CHERNYSHEVA TN^2, SHIN SN^1, DEMENOK VN^1 ^1Pacific Oceanological Institute FEB RAS, Vladivostok, Russia; ^2Clinics of functional medicine “Manus”, Vladivostok, Russia Background: Medicinal and diagnostic preparations based on electronic-homoeopathic copying from parent substances have received a certain distribution in complementary medicine in spite of significant deficit of objective scientific data on their biomedical effect. EHPs specific action without side effects is declared. The aim is to test the effect of EHP with biological model of tomato seedlings. Methods: For double blinded randomized trials were handled to estimate influence of water-based EHP of organic fertilizer Biohumus and Placebo (P) on growth and weight of tomato seedlings. The electronic-homeopathic copying was carried out with "Simulator" (Metabolics Ltd, GB) apparatus by the same operator. The tomato seeds were soaked in preparations then planted out in plastic boxes by 40-49 pieces. Equal exposition of plants to external conditions was achieved. The groups of plants were fed by preparations (200-300 ml) once a week by the same researcher. On the 38-th day of each trial the plants were cut up and the length of green part of each plant was measured with a ruler. The mass of each plant was determined by electronic weighing. Nonparametric Mann-Whitney U-test and Wald-Wolfovitz runs test (*) were used to analyze differences between EHP and P. Results: Statistical significance of differences in trials is shown in the table. Direction of EHP action re P differs from trial to trial. So, in trial No.2 and No.3 the EHP action intensifies the development of plants re P, and conversely in the trial No.4 it weakens the development of plants re P. It is supposed that electronic-homoeopathic copying phenomenon may be related with operator’s body being a source of wide-band electromagnetic disturbances associated with the vital activity of cells and organs which are modulated by parent substance during copying. Conclusions: 1) Revealed in 3 independent double blind randomized trials from 4 significant differences in the results produced by EHP and P points to the reality of electronic-homoeopathic copying phenomenon. 2) The multiplicity of effects produced by EHP re P has been in different trials.	The distribution of antimicrobial resistance patterns of nasopharyngeal Haemophilus influenzae isolated from healthy preschool children KOSIKOWSKA U, MALM A Department of Pharmaceutical Microbiology, Medical University of Lublin, 1 Chodźki Str., 20-093 Lublin, Poland The ecosystem of nasopharyngeal microflora is the reservoir for bacterial pathogens, e.g. Haemophilus influenzae, involved in community-acquired respiratory tract infections, especially in young children. These infections are most frequently endogenous in origin. No literature data are available for resistance rates among nasopharyngeal H. influenzae isolated from healthy young children. In addition, day care centre attendance has been reported as a major risk factor for increased rates of carriage of some bacterial pathogens, including drug resistant strains and for increased prevalence of respiratory tract infections. The aim of the present work was to study an antimicrobial resistance of H. influenzae, isolated from throat or nose of 343 healthy preschool children (3-5 years old): 266 children from four day care centers (D group, including subgroups: D1 - 85, D2 - 62, D3 - 44, D4 - 75) and 77 children staying at home (control group, C). For identification of the bacterial isolates routine methods were used. Susceptibility of the tested bacteria to antimicrobial agents was determined by the disc diffusion procedure. Production of beta-lactamases was detected with nitrocefin test. The prevalence of H. influenzae positive children was 18.18% in C group and 21.05% in D group. 71 isolates of H. influenzae were isolated: 14 from C group and 57 from D group (D1 – 9, D2 - 15, D3 - 7, D4 – 26). 31 (43.66%) of the isolates were resistant to one or more antimicrobials: 7 (50%) from C group and 24 (42.11%) from D group: D1 - 6 (66.67%), D2 - 5 (33.33%), D3 - 2 (28.57%) or D4 - 11 (42.31%). The high rate of resistance to trimethoprim/sulfamethoxazole was found - 21.4% in C group and 29.8% in D group. 8 (11.27%) of the isolates were ampicillin-resistant (C group – 3, D group – 5), including 7 - beta-lactamase positive (C group - 3, D group – 4), and 1 from D group – beta-lactamase negative, resistant to ampicillin and amoxicillin/clavulanic acid. Some of the isolates demonstrated resistance to the other beta-lactams (e.g. II or III generation of cephalosporines or monobactams). The drug resistance pattern of nasopharyngeal H. influenzae isolated from healthy young children may be useful in prediction of drug resistance pattern of H. influenzae of clinical specimens and in consequence, for selection of proper antimicrobial agents used for empiric treatment of respiratory infections caused by haemophili rods. Besides, these data confirm that drug-resistant strains, being a part of normal microflora, can be considered as a reservoir of resistance genes.
Enveloped Virus Neutralizing Compounds (EVNCs), The Magic Bullets against a Broad Spectrum of Deadly Viruses Causing a Billion Infections Annually Around the globe KOTWAL GJ Division of Infectious Diseases, University of Massachusetts Medical School, Worcester, MA, USA, Kotwal Bioconslting, LLC, Louisville, USA and KBiotech pvt.ltd., Cape Town, South Africa. Background: Enveloped viruses like the human Immunodeficiency virus (HIV), influenza viruses including H5N1, hepatis viruses, HBV and HCV, and Herpes viruses 1 and 2,l together cause at least over a billion infections annually in humans across the globe. Most current antivirals and vaccines are specific for a certain type of virus and are susceptible to resistance due to hypermutation. In addition, a number of antiviral agents have serious and debilitating side effects that need to be carefully managed under costly state-of-the-art medical facilities and supervision that are normally not widely available in the developing world. With this in mind, my collaborators from around the world and myself, set out to screen possible natural antiviral compounds that will be a) broad spectrum, b) not susceptible to resistance, c) have minimal or no short term and long side effects and d) not require costly management. We screened several compounds including lectins and mucins and have focused our attention in characterizing the structure, efficacy, safety, stability and mechanism of action of EVNCs from fulvic acid (FA) and pomegranate juice (PJ). Methods: Our general method of testing is to treat one million virus particles with 0.5%-2% of the FA or PJ for 1-5 min at room temperature or 37 degree centigrade and to estimate viral infectivity with and without treatment. In order to determine whether the EVNCs can effectively neutralize a broad spectrum of enveloped viruses, we have tested antiviral activity of EVNCs against an attenuated vaccinia virus vGK5, cowpox virus, influenza including H5N1, SARS virus. HIV, herpes viruses 1 and 2 and are currently testing against hepatitis C virus. In order to test the short term safety we have used cell toxicity assays as well as in vivo studies and to test longterm safety we have used the Ames test for mutagenesis. In order to test heat stability we have tested the efficacy before and after autoclaving the FA and PJ solutions and to study the mechanism of action we have done kinetic studies. In order to isolate the bioactive compounds we have significantly enriched the compounds by passing through HLB copolymer columns with hydrophilic-lipohilic balance (Waters Oasis) and have analyzed the fractions for antiviral activity and structural analysis by NMR. Results: The EVNCs show antiviral activity against vGK5, SARS virus, HIV, influenza and herpes viruses at the concentration tested 0.5% to 2%. The antiviral activity is stable at the temperature of autoclaving which is 121 degree centigrade. The EVNCs can neutralize genetically diverse strains of influenza pr8, X31, H5N1 at concentrations of 0.625%, suggesting that it will be resistant to hypermutation which are charactertic of influenza. The short term toxicity studies showed that at concentrations at which the EVNCs are active there is no cell toxicity and survival and health profile of mice was not affected by a 100 fold therapeutic dose. Long-term toxicity studies showed that while the unfiltered FA had mutagenic activity, the ultrafiltered FA with 3 kDa cutoff Amicon filters did not have any long-term toxicity. Conclusions: The EVNCs have a broad spectrum antiviral activity against enveloped viruses and genetically diverse influenza viruses tested. The bioactive agent is possibly acidic (in the case of PJ, probably ellagic acid) has been enriched and its structure is being elucidated. Short erm and long term studies suggest that the EVNCs are safe and non toxic. The EVNCs has potential in the development of preventive and prophylactic vaccines against certain enveloped viruses as well as as microbicides against sexually transmitted diseases and as orally administered therapeutics. Authors’ disclosure statement: The author has a potential interest in the global production and distribution of the EVNCs and has applied for protecting the intellectual property arising from this work. The author is the President of the companies (InflaMed Inc., and kbiotech) that would be doing Research and development of the EVNCs for therapy, vaccines and microbicides, based on this work. The EVNCs and their source, FA and PJ are experimental agents at this time and therefore no claims are or can be made about actual human use and therefore not for human use.	Bacteriologic and therapeutic aspects of paediatrics osteo-articular infections. KOUAME B, OUATTARA O, ODEHOURI T, GOULI JC, DIETH A, ANOMA DS, KREH Y, TEMBELY S, DICK R Department of general paediatric surgery at the university hospital of Yopougon: Abidjan COTE D’IVOIRE Background: osteo-articular infections provided several injuries and interest all the joints bone and the paediatric skeletal. Successful osteo-articular infection treatments need effective antibiotherapy which depends of the causal bacteria and their poor treatment can lead to important functional sequela. Objective: Report the causal bacteria of the ostea-articular infections treated in our department Method: Retrospective study of 1486 osteo-articular infections treated during 18 years has been performed at the general paediatric surgery of Yopougon, Abidjan Cote d’Ivoire. The age average was 7 years ± 4, sex-ratio was 1,2. The osteo-arthritis was observed in 77% and osteomyelitis in 23%. The sickle cell disease was detected in 9% and HIV infection in 0, 01%. The first line antibiotic for medical treatment associated oxacillin to thirst generation of cephalosporin. Surgery treatment was indicated when pus collections were documented radiologically. Biologic examinations consist of the blood culture, pus culture, C reactive protein, VS and blood count cell. We studied the osteo-articular infection response to the first line antibiotic according bacteriologic characteristic of the causal bacteria (type of bacteria, the bacterial resistance and the antibiogramm). Bacteria identification was performed according classic procedure and antibiogramm was performed with Orisis Biorad with CASFM references (committee of antibiogramm of French microbiology society).