Ehrlich II –2nd World Conference on Magic Bullets

KASTELEIJN-NOLST TRENITÉ DGA¹, REED RC², O’BRIEN TJ³, VAN RAAY L³, HOGAN RE⁴, MORRIS MJ⁵, DEDEURWAERDERE S⁶, PICCIOLI M⁷, PARISI P¹, TISEI P¹, PIA VILLA M¹, BUTTINELLI C¹.

1. Universita di Roma, Sapienza II, Rome, Italy, 2. Abbott, Global Pharmaceutical Research & Development, Neuroscience, Abbott Park, IL, USA, 3. University of Melbourne, Australia, 4. Washington University, St Louis, USA, 5. University of New South Wales, Kensington, Australia, 6. Australian Nuclear Science & Technology Organisation, Lucas Heights, Australia, 7. San Filippo Neri hospital, Rome, Italy

In Grenoble, in 1963, H. Meunier and his staff were investigating molecules with a ring structure containing a nitrogen atom to suppress epileptic seizure activity in rabbits and mice. They noticed, however, that it was actually the solvent of the various nitrogen-containing compounds, a simple dipropyl-acid, that had protective power against cardiazol-evoked epileptic seizures. Subsequently, trials were performed with this solvent, dipropylacetic acid (valproic acid [VPA] or sodium valproate), in adult patients with generalized seizures at epilepsy and psychiatry centers. VPA’s great efficacy and safety in epilepsy patients with untreatable seizures prompted European regulatory agency approval as an anti-epileptic drug (AED) in 1967. The discovery of VPA led to a change in concept of working mechanisms for AEDs in general (an increase of the inhibitory neurotransmitter GABA). This conceptualization of the mechanism of action led to the development of new AEDs, e.g., vigabatrine, tiagabine and gabapentin.

Other indications were sought for VPA as early as 1964 by Lebreton.

VPA failed in an anxiety/stress animal model, however, it was noted that 5-10 minutes after intravenous administration of 200 mg/kg VPA in mice, activity and explorative behavior were diminished, indicating a potential role in the treatment of psychiatric disorders. It took until the 1990’s before VPA was officially recognized to be effective not only in all types of epilepsy, but also in acute mania (US FDA-approved in 1995) and migraine (US FDA-approved in 1996). At present, VPA is a candidate drug leading to interesting changes in the concepts about working mechanisms of drugs and underlying pathology of the various brain diseases and its interconnections. Recent research demonstrates this unification hypothesis: i. peak plasma VPA concentrations are not more efficacious than continuous intravenous VPA infusion; ii.VPA has proven sustained efficacy (median = 4 years, up to 12 years) in photosensitive epilepsy patients; iii. a close link exists between migraine, occipito-temporal lobe epilepsy and photosensitivity. The success of VPA therapy clinically has inspired others to search for other neuro-active drugs that can act as a “magic bullet” working across a spectrum of neuro-psychiatric diseases.

DNA Conformation in Complexes with Coordination Compounds  KASYANENKO N Faculty of Physics, St.-Petersburg State University, Russia Background: DNA is the main target for platinum-based chemotherapy drugs used to treat various types of cancers. Cisplatin was the first member of this class, which now also includes carboplatin and oxaliplatin. Platinum compounds interact with DNA in cells, prevent its replication and disrupt repair.The overcoming of the problem of high toxicity of antineoplastic agents is a main challenge in the field of cancer research. The development of drug resistance is the other obstruction that limits the efficacy of the majority of chemotherapeutic agents. The extensive research has been directed toward the seeking for new coordination compounds with the remarkably low toxic side effects compared to the utilized platinum-based drugs. Wide range of new perspective compounds demands an effective and simple method for the preliminary selection of drugs. DNA solution can be corresponded as a model system for the tentative testing of new chemicals. Material and Methods: Calf thymus DNA (Sigma) MM and plasmid DNA pFL 44 / EcoRI in linear and circular form were used. The set of experimental methods (circular dichroism, UV spectrophotometry, dynamic light scattering, electrophoresis, viscosity, flow birefringence, atomic force microscopy, NanoScope 4a, Veeco) ensures the information about DNA secondary and tertiary structure during interaction with coordination compounds. Results: DNA conformation in complexes with coordination compounds of Pd(II), Co(III), Ru(III), binuclear compounds of Pt(II) and Pt (IV) with different ligands was regarded. DNA persistence length, double-stranded structure, electrophoretic mobility, volume and shape of molecular coil were examined. It is known that complex ions can interact with DNA via strong covalent binding, van der Waals forces, hydrogen bonding or electrostatic attraction. The similarity and difference between the interactions of coordination compouns and metal ions with DNA are analyzed. The influence of pH and solution ionic strength on DNA interaction with complex ions was investigated. The consistent influence of platinum drugs and gamma irradiation on DNA structure is explored. The possibility of formation of gene vectors with coordination compounds is examined. Conclusions: The integrated approach to the analysis of DNA conformation in complexes with coordination compounds in a solution can provide the effective tentative testing of new drugs.The investigation of DNA interaction with new compounds in a solution and the comparison of experimental results with data obtained for cis-DDP, non active trans-DDP and other biological active compounds can provide the information about the molecular mechanism of anticancer activity.
Factors That Influence The Prevalence Of Drug-Drug Interactions Between Antiretroviral Drugs Prescribed To Patients Of Different Age Groups In A Section Of Private Healthcare Sector In South Africa KATENDE-KYENDA NL^1, LUBBE MS^2, SERFONTEIN JHP^3, Truter I^4 ^1Walter Sisulu University, Mthatha; ^2-3Northwest University, Potchefstroom; ^4Nelson Mandela Metropolitan University, Port Elizabeth, South Africa Background: Drug-drug interactions (DDIs) are often a serious complication due to taking multiple medications and account for 3% to 5% of all in-hospital medication errors (Leape et al., 1995). DDIs are of particular concern in HIV/AIDs patients receiving highly active antiretroviral therapy (HAART), particularly certain protease inhibitors (PIs) and nonnucleoside reverse transcriptase inhibitors (NNRTIs), for they interact with other antiretrovirals (ARVs). This is due their metabolism through the cytochrome P450 (CYP450) system. AIMS: 1) To evaluate factors that could influence the prevalence of DDIs between ARVs prescribed in different age groups. 2) To determine whether ARVs were prescribed according to the recommended prescribing guidelines in South Africa. Methods: This was a quantitative, retrospective drug utilisation study performed on 49 995 (N = 19 860 679) and 81 096 (N = 21 473 074) ARV prescriptions that were prescribed to 43 547 (N = 5 433 440) and 70 719 (N = 5 728 371) HIV patients for 2005 and 2006 and claimed through a medicines claims database. Possible DDIs between ARVs were identified according to Tatro (2005). Results: Of the 5 433 440 and 5 728 371 patients reviewed for both years, 0.80%; 1.23% was HIV positive patients, of whom 43.26%; 57.06% were males and 56.74%; 42.94% were females for 2005 and 2006 respectively. A total of 49 995; 81 096 ARV prescriptions claimed, of which 4.49; 4.07% had one item, 43.75; 43.52% two, 43.86; 49.56% three, 0.07%; 2.78% four and 0.07%; 0.06% had more than four items on the prescription. Of 811 DDIs identified for 2005, 33.54% were for two drug items, 61.90% three, 2.10% four and 2.46% had more than four items. For 2006, 1115 DDIs were identified, of which 59.64% were for 2 drug items, 27.71% three, 11.12% four and 1.52% had more than 4 items. DDIs identified in different age-groups for 2005 and 2006 were: 5.65% (2006) for patients ≤ 12 years, 74.60%; 58.75% for patients > 19 years and ≤ 45 years; 23.55%; 27.35% for patients > 45 years and ≤ 59 years; and 1.85%; 8.25% for patients ≥ 59 years. The most important interactions were identified between combinations of: Kaletra^® (Lopinavir 133.3mg/Ritonavir 33.3mg) and Stocrin^® (Efavirenz 600mg) at daily doses of 799.8mg/198mg and 600mg respectively, followed by Crixivan^® (Indinavir 400mg) and Norvir^® (Ritonavir 100mg) at daily doses of 1600mg and 200mg; and Kaletra^® (Lopinavir 133mg/Ritonavir 33.3mg) and Nevirapine (Viramune^® 200mg) at 1066.4mg/264.7mg and 400mg daily doses. All the interactions were of clinical significance level 2 (moderate effects), causing deterioration of a patient’s clinical status. Conclusions: These results demonstrate that the non-adherence of the recommended prescribing ARV drug combinations, and daily doses prescribed in different age groups could influence the prevalence of DDIs, therefore a need for more interprofessional education on the prescribing protocols for ARVs. References Tatro DS. Drug Interaction Facts. St. Louis, Facts and Comparisons, 1-1699 (2005). Leape LL, Bates DW, Cullen DJ et al. Systems analysis of adverse drug events. ADE Prevention Study Group, JAMA, 1995; 274:35-43.	EMY162 Protein As A Vaccine Candidate To Reduce Level Of Alveolar Hydatid Disease KATOH Y1), KOUGUCHI H 1), MATSUMOTO J2), SUZUKI T 1), YAGI K1) 1) Hokkaido Institute of Public Health, Sapporo, Hokkaido, Japan, 2) College of Bioresource Sciences, Nihon University, Fujisawa, Kanagawa, Japan Background: Echinococcus multilocularis is a cestode parasite. The life cycle of E. multilocularis generally occurs in foxes and rodents as intermediate hosts. Humans can be infected by accidental ingestion of the parasite eggs from an infected fox, or occasionally from infected dogs or cats. Infection in humans causes alveolar hydatid disease. The disease is a significant public health problem. We attempted to clone cDNA of secretory proteins involved in immune defense systems in order to use them in the control of alveolar hydatid disease. Method: E. multilocularis (Nemuro strain) were obtained from a dog-cotton rat life cycle maintained at the Hokkaido Institute of Public Health. Immature adult worms were collected on day 20 post-infection from dog an experimentally infected with E. multilocularis protoscoleces. Total RNAs from the adult worms were isolated. A cDNA library based on mRNA from adult worms of E. multilocularis was constructed. One of the cDNA clone, emY162, was isolated from this cDNA library. Recombinant emY162 was subcloned, and then recombinant antigen EMY162 was administrated to mice with Freud’s complete adjuvant. Antibody production was assayed by immunoblot analysis. Immunobiological reactivity was analyzed by Western blot. After the final immunization by the recombinant antigens, parasite eggs were administrated orally for vaccine trial of the recombinant EMY162. The number of alveolar cyst in each mouse was counted. Results: The putative protein from emY162 cDNA consists of 153 amino acids and has a predicted molecular weight of 17.0 kDa. The amino acid sequences of EMY162 are predicted to have single fibronectin type III-like domain. The emY162 is expressed in all four stages (protoscoleces, cultured metacestodes, immature adult worms and mature adult worms). When immunity to recombinant EMY162 was examined, strong IgG immune responses were detected in Western blots. The recombinant EMY162 antigen-specific antibody response showed a polarization toward IgG2 subclass. In addition, the recombinant EMY162 induced a significant level of host-protection (74.3%) in experimental infection with E. multilocularis eggs in mice, and showed significant reactivity to the sera from alveolar echinococcosis patients. Conclusion: 1) The EMY162 protein could target both mucosal and systemic immunity in dogs and humans. 2) The EMY162 protein will help the development of both protection against and diagnosis of alveolar hydatid disease.
Ca^2+-Signal Transducing System From The Endoplasmic Reticulum To Mitochondria Involved In The Caffeine-Inducible ATP Transport KATSURAGI T Medical Research Center, Fukuoka University, Fukuoka, Japan Adenocine triphosphate (ATP) is released as an autocrine/paracrine signal from a variety of cells. The present study was designed to clarfy the Ca^2+-signal pathway involved in the caffeine-inducible transport of ATP from cultured vas deferens smooth muscle cells. The transport of ATP induced by caffeine (3 mM) was almost completely inhibited by ryanodine and tetracaine, but not by 2-APB, thus being mediated by ryanodine receptor (RyR). The expression of messenger RNA from only RyR-2 was detected in the cells. Furthermore, the induced transport was attenuated by mitochondrial inhibitors, rotenone and oligomycin and by Cl^--channel blockers, niflumic acid and NPPB. Increase in Ca^2+-signals with fluo -4 and rhod-2 caused by caffeine were reduced by tetracaine and oligomycin plus CCCP, respectively. A close spatial relation between the endoplasmic reticulum (ER) and mitochondria was electromicroscopically observed in cells, supporting the existence of a Ca^2+-signaling bridge on both the organelli. These results suggest that caffeine stimulates ryanodine receptor (RyR-2) and facilitates a Ca^2+-signal transducing system from ER to mitochondria, and then, the signal appears to accelerate the ATP synthesis in mitochondria. In addition, the mitochondrial event may lead further cell signaling to the cell membrane and activates Cl^^-channels, resulting in the extracellular transport of cytosolic ATP. In the study with MDCK cells, we also provided evidence that such a Ca^2+-signaling pathway from ER to mitochondria mediates the transport of ATP induced by adenosine.	Structure Based Development Of Selective Inhibitors For Individual Cathepsins And Their Medical Applications For Therapeutic Purposes KATUNUMA N^1,TURK V^2 ^1Institute for Health Sciences, Tokushima Bunri University, Tokushima, Japan. ^2Jozef Stefan Institute, Ljubljana, Slovenia. Background: Cathepsins (Cath.s) are cysteine protease in lysosomes. Eleven kinds of cath.s are registered in human genome. Cath.s play an essential role in protein catabolism. Since each cath. has a different cleavage-bond, different cath.s produce different product from the same protein. Abnormal expressions of cath.s induce special diseases, therefore the specific cath. Inhibitors are useful for pathogenesis and also for the therapy. Methods: The study of cath. Inhibitors has been started from the discovery of natural inhibitors from bacteria. One is aliphatic aldehyde derivatives to bind with SH-of cath; leupeptin and antipin by Umezawa. The other is derivatives of epoxysuccinate (ES), E-64 by Katunuma and Hanada. However, they inhibited all cath. family. Using these frame inhibitors, we developed the specific inhibitors for individual cath.s, based on their different tertiary structures of their substrate binding pockets using their X-ray crystallography. Results: 1) Specific cath. Inhibitor design. (a) As the ES derivatives. Cath.B specific inhibitors having-IIe-Pro at the C-terminus; CA-074, and also cath. L specific inhibitors; CLIK-148. (b) As the aliphatic aldehyde derivatives, cath.S specificity inhibitor, CLIK-60. (c) As the pyridoxal derivatives for cath.K, CLIK-164. These inhibitors showed specific inhibition for special cath.s at the 10^-6 -10^-7 M level, in vitro and in vivo. 2) Medical applications. (a) Osteoporosis and bone metastasis of cancer were protected by cath.L or K inhibitor, CLIK-148 or CLIK-164. (b) Antigens are processed by various cath.s. T1-type and T2-type expressions were switched by antigen processing by different cath.s. (c) MHC-Class II was activated by invariant chain degradation by cath.S. (d) In auto-immune Sjögren's disease, the auto-antigen “Hodrin” was processed by cath.S, CLIK-60 suppressed the Sjögren's syndromes in the model mice. Conclusions: 1) Specific cath. inhibitors were designed and developed. 2) Osteoporosis and bone metastasis of cancer were suppressed by cath.L inhibitor. 3) Antigen processing and presentation were regulated by these inhibitors. 4) Suppression of autoantigen processing in Sjögren D. by CLIK-60 and type-1 Diabetes by CLIK-148.
Melatonin-A Possible Magic Bullet In Reducing Hypoxic Brain Injury KAUR C^1, SINGH J^2, LING EA^1 1. Department of Anatomy, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, 2.Civil Aviation Medical Board, Civil Aviation Authority of Singapore, Singapore Changi Airport, Singapore Melatonin, a neurohormone synthesized and secreted by the pineal gland is reported to have antioxidant, immunoregulatory and neuroprotective actions. Production of melatonin is regulated by light and darkness, light decreasing and darkness increasing it. Its production is also known to decline in old age and under hypoxic-ischemic conditions. Melatonin is considered the body’s chronological pacemaker and has a wide array of useful applications. It has been used in the treatment of sleep disorders, especially those associated with circadian dysrhythmicity, and is also reported to have neuroprotective effects in many central nervous system (CNS) conditions such as amyotrophic lateral sclerosis, Parkinson’s disease, Alzheimer’s disease, ischemic injury, neuropsychiatric disorders and head injury. Results from our laboratory have shown that it affords protection to the blood- brain and blood-retinal barriers in hypoxic conditions by suppressing the production of vascular endothelial growth factor and nitric oxide which are known to increase vascular permeability. Protective effects of melatonin against hypoxic damage have also been demonstrated in newborn experimental animals where it suppressed damage in many parts of the brain such as the hippocampus and choroid plexus in lateral ventricles. Along with this, exogenous administration of melatonin in newborn animals has been shown to be effective in enhancing the surface receptors and antigens on the macrophages/microglia in the CNS supporting its immunoregulatory actions. Keeping these beneficial effects in view, melatonin merits consideration as a potential ”magic bullet” for mitigating brain damage in hypoxic-ischemic injuries. This study was supported by a research grant (R-181-000-098-112) from the National University of Singapore.	Cardiac Side Effects Of Psychotrop (Antidepressant, Antipsychotic) Drugs KECSKEMETI V Semmelweis University, Budapest, Hungary Background: The most frequent cardiac side effects of psychotrop drugs (antidepressants, antipsychotics) are brady- or, tachycardia, ECG alterations (prolongation of QRS, QT interval), AV-block, ventricular arrhythmias (tachycardia, torsades de pointes, TdP) and sudden death. Aims and methods: To attempt to find relations between clinical data and the electrophysiological effects of antidepressants (fluoxetine, citalopram), antipsychotics (risperidone) obtained in isolated guinea-pig ventricular muscles and canine ventricular myocytes using the conventional microelectrode and whole cell clamp technique. Results: Fluoxetine (F) (0.5-50 mM) and citalopram (C) (10-100 μM) exhibited depressant effects on contraction and both on Ca^2+ and Na^+- dependent electrophysiological parameters of cardiac preparations and on cardiac Ca^2+ current, without modifying the K^+ currents. Risperidone (R) (0.1-10 µM) caused a concentration-dependent lengthening of action potential duration (APD) in both preparations and it blocked concentration-dependently the rapid component of the delayed rectifier K^+ current (IKr) The other K^+ currents (IK1 and Ito) and Na^+ current were not significantly modified. Conclusion: the inhibition of cardiac Ca^2+ and Na^+currents by F and C , moreover the depression of IKr currentby R may explain the cardiac side-effects observed occasionally with these drugs. Our results suggest that the new generation of antidepressants (fluoxetine, citalopram) and antipsychotics (risperidone) may have also antiarrhythmic, as well as proarrhythmic properties. Therefore, clinicians should be more vigilant about these potential adverse reactions and ECG control may be suggested during therapy, especially in patients with cardiovasular disorders.
Molecular Epidemiology of Quinolone Resistant Salmonella Typhi: South Africa 2003-2007 KEDDY KH ^1^,^2, GOVENDER N^1^,^2, SOOKA A^1, SMITH AM^1^,^2 FOR GERMS-SA* ^1Enteric Diseases Reference Unit, National Institute for Communicable Diseases, Johannesburg, South Africa and ^2University of the Witwatersrand, Johannesburg, South Africa Background: Fluoroquinolones have become the mainstay of treatment for typhoid fever in many countries, including South Africa. Resistance to the quinolones antibiotics results in treatment failures and quinolone resistance in South African strains of Salmonella enterica serotype Typhi is well documented. Methods: The molecular mechanism for quinolone resistance in Salmonella Typhi from South African clinical isolates received by the Enteric Diseases Reference Unit of the National Institute for Communicable Diseases for the period 2003-2005 was determined using pulsed-field gel electrophoresis (PFGE), multiple-locus variable-number tandem-repeats analysis (MLVA), PCR and sequencing of the quinolone resistant determining region (QRDR) genes gyrA, gyrB, parC and parE, as well as plasmid-mediated quinolone resistance determinants (PMQD) QnrA, QnrB, and QnrS. Results: PFGE showed 2 major clusters (90%) among the 20 quinolone resistance isolates. MLVA was more sensitive and grouped these same 20 isolates into 11 MLVA types, with the majority (8/20) grouped as MVLA type-16 with the other types differing by 1 allele, either TR1 or TR2. Among the 8 related and non-related PFGE isolates screened for mutations in the QRDR region 2/8 isolates had mutations in gyrA, parC and parE; 2/8 isolates had mutations in parE; 1/8 isolate had a mutation in gyrA and parE; 1/8 isolate had mutations in gyrA, gyrB, and parE, and 2/8 isolates exhibited no mutations in their QRDR regions. PCR screening for as PMQD were all negative. Conclusions: Although these results seem counter-intuitive to previously published work, the molecular mechanism of quinolone resistance for these isolates may not be attributed to a single mechanism but may be the result of a combination of mechanisms.	The impact of highly active antiretroviral therapy on cytomegalovirus retinitis: triumphs and future challenges KEDHAR SR^123 ^1The New York Eye & Ear Infirmary, New York, United States; ^2New York Medical College, Valhalla, United States; ^3The Johns Hopkins University School of Medicine, Baltimore, United States.   Background: Cytomegalovirus (CMV) is a common opportunistic infection in individuals with AIDS, with CMV retinitis representing a significant portion of end-organ disease in these patients. Before the development of highly active antiretroviral therapy (HAART), nearly one-third of people with AIDS developed CMV retinitis during their lifetime. Although effective therapies for CMV infection had been developed, treatment was often life-long due to persistent immune deficiency. Even with chronic suppressive maintenance therapy, disease relapse was nearly universal, and development of drug resistance was not uncommon. Aims: 1) To evaluate the impact of HAART on the course and complications of CMV retinitis 2) To identify continued challenges in thetreatment of CMV retinitis and propose further avenues for investigation. Methods: This study reviewed available evidence in the medical literature concerning the treatments and outcomes of CMV retinitis with particular attention to the impact of HAART and to data derived from the Longitudinal Study of the Ocular Complications of AIDS. Results/Conclusions: The widespread use of HAART has reduced the incidence and complications of CMV retinitis in patients with HIV infection. With sustained immune recovery, discontinuation of anti-CMV therapy has been possible in many patients. Still, immune recovery does not guarantee protection from recurrent disease. CMV retinitis and uveitis associated with immune recovery remain causes of vision loss in this population. Areas such as genetic susceptibility to CMV retinitis and the development of long-term drugs and drug delivery vehicles apprpriate for developing countries offer further avenues of investigation.
Political Economy and Societal Consequences of Methamphetamine Epidemic in the United States KEDIA S University of Memphis The world, beleaguered by a multitude of illnesses and syndromes, anxiously looks to pharmaceutical companies for the creation and distribution of substances that will ease pain, discomfort, and even reverse degeneration. However, a glance through pharmaceutical history tells us that a drug can be both a welcomed panacea and a despised scourge, a literal _magic bullet._ Methamphetamine (meth) is one such substance. A derivative of amphetamine, which was first synthesized in 1887, methamphetamine has roots in 1940s Japan. It mirrors the body_s natural adrenaline surge, and has been a persistent and pervasive worldwide problem, insidiously affecting national fabrics and political economies, and creating widespread societal consequences such as deterioration of public health, destruction of families, domestic violence, robbery, and murder. This presentation, _Political Economy and Societal Consequences of Methamphetamine Epidemic in the United States,_ explores history, economy, and human costs associated with this drug. The presentation will then discuss the ways in which the drug has brought about manifold changes in areas that fall under the rubric of _political economy_ including legislation, medical care, child welfare, and substance abuse treatment, changes that have come about as a reaction to and result of methamphetamine_s significant impacts on society.	Designing Drugs for Neurological Disorders: TRH-based Neurotherapeutics GREGG DJ^1, SCALABRINO GA^1, BOYLE NT^1, SLATOR GR^1, WILLIAMS CH^1, TIPTON KF^1, HOGAN N^2, O’BOYLE KM^2, BAUER K^3, KELLY JA^1 ^1School of Biochemistry and Immunology and Trinity College Institute of Neuroscience, Trinity College Dublin, Ireland; ^2UCD School of Biomolecular and Biomedical Science, Conway Institute, University College Dublin, Ireland; ^3Leibniz Institute for Age Research, Jena, Germany Background: The trend in neurotherapeutic drug design is moving from a ‘magic bullet’ to a ‘magic shotgun’ approach, yet the naturally occurring neuroactive peptide thyrotropin-releasing hormone (TRH) has potential to act as a ‘magic bullet’ in the treatment of a wide variety of CNS disorders by virtue of its multifaceted homeostatic neurobiological actions. Clinical use of TRH is restricted, however, because of its short half-life due to degradation by TRH-degrading ectoenzyme (TRH-DE). We report the development of novel compounds that offer an attractive means to overcome this constraint. Methods: The ability of novel synthetic peptides to act as TRH-DE substrates or inhibitors and/or central TRH receptor ligands was determined by kinetic analysis and radioligand binding assays. The in vivo effects of systemically injected peptides were examined in male Wistar rats (200–250 g). Results: QSAR revealed replacement of His in TRH (Glp-His-ProNH2) by Asn confers resistance to TRH-DE proteolysis and efficient TRH-DE inhibition. Addition of hydrophobic l-amino acids to the C-terminus of Glp-Asn-ProNH2 led to TRH-DE inhibitors with Ki values in the nanomolar range. Replacement of these hydrophobic residues with their d-isomers yielded a set of first-in-class compounds that potently inhibited TRH-DE and also bound to central native TRH receptors with a high affinity. Systemic injection of the lead compound of this set (Glp-Asn-Pro-d-Tyr-d-TrpNH2, 1mg/kg) antagonised barbiturate-induced narcosis; increased rat activity scores (P<0.05, vs vehicle controls, ANOVA with Dunnett’s post test) and enhanced behavioural responses compared to TRH alone (P<0.01, Bonferroni’s comparison post ANOVA) (n = 5–23). Conclusion: The development of a dual action bullet that targets two key members of the TRH signalling system and mimics and enhances TRH actions opens up a possibility for realizing the neuropharmacological potential of TRH actions and contributes useful insights to rational drug design. Funded by the Wellcome Trust, Enterprise Ireland and Health Research Board, Ireland.
Drug Absorption From The Small Intestine In Immediate Postoperative Patients KENNEDY JM^(1,2, 3), VAN RIJ AM^(2) ^(1)School of Pharmacy, and^(2)Department of Surgery, University of Otago, Dunedin, New Zealand,^(3)School of Pharmacy UCC, Cork, Ireland Background:  The effects of surgery on gastric emptying have been documented for a considerable time, but less is known about the effects in the small intestine.  It is thought that there is minimal diminution in the absorptive capacity of the small intestine postoperatively, although there is no literature on drug absorption in the early period after surgery. This study investigated drug absorption from the small bowel in patients undergoing abdominal surgery. Methods: A prospective study of patients undergoing major abdominal surgery in which patients acted as their own pre-operative controls was carried out.  Patients were administered the test drugs, paracetamol and^99mTcDTPA, pre-surgery and two days postoperatively.  Small intestine transit times, plasma concentrations and other pharmacokinetic variables were compared using Student’s paired t test.  Two complementary studies were carried out to establish pharmacokinetic parameters. Results:  There were no significant differences in the pre- and postoperative values of tmax, AUC, and AUMC pre- and postoperatively, (p > 0.05).  There were significant differences between the pre- and postoperative values of Cmax (Cmax (preop)> Cmax (postop); p < 0.05) and the pre- and postoperative values of MRT (MRT (preop) < MRT (postop) ; p < 0.01).  Conclusions: Drug absorption from the small bowel in the postoperative patient does not differ significantly from its preoperative absorptive capacity.	The ACE-Inhibitor: True Magic Bullet From Myocardium To Endocardium KER J University of Pretoria, Pretoria, South Africa The following outcome data with the use of angiotensin converting enzyme inhibitors will be presented: Atherosclerosis: Primary and secondary prevention of myocardial infarction. Peripheral vascular disease: Primary and secondary prevention data Retinopathy: Data on the prevention of diabetic retinopathy with the use of ACE-inhibitors Heart failure: Outcome data in ischaemic and non-ischaemic causes of heart failure Rhythm disturbances: Data on the prevention and treatment of atrial and ventricular rhythm disturbances Renal disease: Data on the prevention and treatment of renal disease Prevention of type 2 diabetes mellitus
ODAM As A Diagnostic And Therapeutic Target For Human Breast Cancer Kestler DP, Foster JS, Patton AL, Siddiqui S, Bell JL, Panella TJ, Murphy CL, Macy SD, Wall J, Solomon A University of Tennessee Graduate School of Medicine, Knoxville, USA Background: We have previously reported that the structurally novel Odontogenic Ameloblast Associated Protein (ODAM), expressed in ameloblasts during late tooth developments and in odonotogenic tumors is also found in human breast cancer (Kestler et. al. 2008 Mol Med 14:318-326). Methods: To investigate the possible role of ODAM as a biomarker of breast cancer, as well as a potential diagnostic and therapeutic target, we have tested the capabilities of our anti-ODAM mAbs to immunostain a) mouse mammary tissue from different stages of development; b) human breast cancer arrays, and c) biopsy specimens obtained from patients with stages I-IV breast cancer. Additionally, we have used dual micro-SPECT/CT to image mouse mammary tumor xenografts with an ^125I-labelled anti-ODAM mAb. Results: Tissues from all stages of mouse mammary development except lactation expressed ODAM; further, a significant number of specimens contained in human breast carcinoma arrays also were stained by these reagents. Among 60 patient samples analyzed in a retrospective study, ODAM expression was significantly greater in advanced (stage IV) than in early (stages I-III) disease. The ^125I-anti-ODAM mAb was also capable of imaging a murine mammary xenograft. Furthermore, we found (using an ELISA-based procedure) that the sera of patients with metastatic breast cancer contained elevated titers of anti-ODAM antibodies. Conclusions: Our finding that ODAM expression in human breast cancer correlates with disease stage has prognostic import. The presence of anti-ODAM antibodies in the sera of patients with metastatic disease also is of note, given that autoantibodies to growth regulatory factors have been detected in individuals with other types of malignancies in which the titers of these components have correlated with survival and other clinicopathological parameters. Additionally, radiolabeled anti-ODAM mAbs may prove useful to document the presence of tumor metastasis or relapse. Based on our data, we posit that ODAM has a functional role in breast development and in the pathogenesis of breast cancer where it could serve as a novel diagnostic and therapeutic target.	Treatment Prospects for Breast Cancer: Lessons Learnt from a Decade of Research on Maspin KHALKHALI-ELLIS Z, HENDRIX MJC Children’s Memorial Research Center, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, USA Background: Maspin is a multifaceted protein, interacting with a diverse group of intra- and extra-cellular proteins, regulating cell adhesion, motility, apoptosis, angiogenesis and critically involved in mammary gland development. Maspin is a cytosolic protein but is also localized to the nucleus and membrane, and is secreted. Aberrant methylation of the Maspin promoter is closely associated with Maspin gene silencing and is a common occurrence in cancer. Our laboratory has identified the aspartyl endopeptidase Cathepsin D (CatD) as a binding partner for Maspin. Unlike Maspin, which is downregulated in primary tumors and absent in metastatic breast cancer, CatD is excessively produced and aberrantly secreted by tumor cells. Studies depict a critical role for CatD in tumor growth (mitogenic effect), and invasion (proteolytic effect on matrix components) in breast cancer. Based on the importance of both Maspin and CatD, and specifically the reciprocity of their relationship in breast cancer, we embarked on identifying factors which might influence the Maspin and CatD partnership under normal conditions. Such an approach would shed light on how their alteration could lead to malignant growth and ultimately metastasis. Methods: We employed in vivo (mice model of mammary gland development), and in vitro (normal mammary epithelial and breast cancer cell lines grown on 3D matrices) models to decipher the Maspin and CatD partnership in the context of mammary gland development and during neoplastic breast cancer progression. Results: Our studies have illuminated a previously unidentified function for Maspin and its interaction with CatD in maintaining the differentiated secretory glandular phenotype of the mammary gland. In addition, the secretion of Maspin by mammary epithelial cells and its deposition into the extracelluar milieu plays an important role in matrix degradation by CatD. In this capacity Maspin could potentially regulate mammary tissue remodeling occurring under normal and pathological conditions. Conclusions: Studying this unique partnership has provided us with a critical view into several previously unidentified mechanisms of action for both of these proteins, and may contribute new strategies underlying Maspin- and CatD-based therapeutic approaches for combating breast cancer.
Anti-inflammatory, analgesic and antipyretic activities of Physalis minima Linn KHAN MA^1,2, KHAN H^3, KHAN S^4, MAHMOOD T^2, KHAN PM^4, JABAR A^5 1Department of Chemistry, Kohat University of Science and Technology, Kohat Pakistan, 2Pakistan Council of Scientific and Industrial Research Laboratories Complex, Peshawar, Pakistan, 3Gandhara College of Pharmacy, Gandhara University, Peshawar, Pakistan, 4Department of Chemistry, Jehanzeb Postgraduate College, Swat, Pakistan, 5Department of Chemistry, Islamia University, Bahawalpur, Pakistan Background: To evaluate anti-inflammatory, analgesic and antipyretic activities of Physalis minima Linn in order to discovery natural remedies for the management various painful and inflammatory conditions. Methods: NMRI mice (22–28 g) and Wistar rats (180–200 g) of either sex were obtained from the animal house facility of H.E.J. Research Institute of Chemistry, University of Karachi, Karachi, Pakistan. The carrageenan induced hind paw edema and Cotton pellet-induced granuloma tests were conducted for both crude extract and chloroform fraction. While the Acetic acid-induced abdominal constriction and Formalin test were performed to evaluate the analgesic potential of crude extract and chloroform fraction. The crude extract and chloroform fraction was also tested against Brewer’s yeast (Saccharomyces cerevisiae), induced fever. Results: The crude extract (58%) and chloroform fraction (62%) of Physalis minima significantly inhibited the carrageenan induced pay edema in rats at 400 mg/kg. In a dose dependent manner at 400 mg/kg, the crude methanol extract and chloroform fraction reduced granuloma (48%) and (62%) respectively. In a dose dependent manner at 400 mg/kg, the crude extract and chloroform fraction reduced the number of abdominal constriction (52%) and (38%) respectively. The crude methanol extract demonstrated (51%) and chloroform fraction (31%) activity in dose dependent way in the late phase in formalin induced pain. In case of antipyretic assay, the crude extract and chloroform fraction of Physalis minima expressed insignificant activity. Values of p < 0.05 were considered significant in all cases. Conclusions: Both the crude extract and chloroform fraction of the plant showed significant anti-nociceptiv and anti- inflammatory activity as compare to control, while the anti-pyretic response was insignificant.	Peptides Against Ageing