Spectrum And Risk For HIV-1 Associated Mutations And The Efficacy Of Antiretroviral Therapy Among Infected Pediatric Patients MALHOTRA A, GAUR S, WHITLEY-WILLIAMS PN, PETROVA A
1Department of Pediatrics, Division of Infectious Disease, University of Medicine and Dentistry of New Jersey (UMDNJ) – Robert Wood Johnson Medical School New Brunswick, New Jersey, 08903 USA
Background: Introduction of combined therapy with reverse transcriptase and protease inhibitors has resulted in a considerable decrease in HIV related mortality, but it has also induced the development of multiple drug resistant HIV-1 variants. The few studies on HIV mutagenesis in infected children have not evaluated the impact of related mutations on pediatric HIV disease.
Methods: 42 HIV-1 infected children were enrolled and followed in the Robert Wood Johnson Pediatric Infectious Disease Clinic in New Brunswick, NJ, USA since 1999 to 2007. 41(97.6%) patients were assessed and demographic and treatment-related information was recorded as well as plasma viral load, CD4 T-lymphocyte counts and HIV genotype analysis. Between 2 and 5 measurements were obtained with 6-12 month intervals and a total of 119 measurements were assessed and analyzed for 40 patients (95%).
Results: 25 male and 16 female patients were enrolled. All participants were symptomatic and had preceding treatment history with combined ARV regimens including protease inhibitors (PI), nucleoside reverse transcriptase inhibitors (NRTI), and non-nucleoside reverse transcriptase inhibitors (NNRTI). Treatment regimens that included PIs were observed in 29/ 40 (72.5%) patients. Combined regimens did not significantly impact the incidence of NRTI and NNRTI associated mutations. Primary mutations in the protease gene increased the likelihood of plasma viral load (>10,000 copies/mL) irrespective of the child’s age, duration of therapy, or presence of NRTI and NNRTI mutations, P<0.008.
13 out of 40 (44.8%) were diagnosed with PI associated major mutations at study entry. Starting PI medication (n=4) among PI naïve patients (n=11) was associated with development of PI major mutations in 25% of cases. Anaysis of 119 HIV genotype measurements showed significant association between PI mutations and HIV viral load > 10,000 copies/mL (OR=2.84, 95% CI 1.21, 6.69).
Conclusions: Since primary PI mutations significantly increase the likelihood for high viral replication in HIV-infected pediatric patients, careful monitoring of PI resistant major mutations is important for maintaining low viral loads.
Bilirubin: An Endogenous Molecule With Antiviral Activity In Vitro Mancuso C1, Marchetti S2, Fadda G2, Santangelo R2 Institutes of 1Pharmacology and 2Microbiology, Catholic University School of Medicine, Roma, Italy
Background: In 1992, Nakagami et al. hypothesized an antiviral role for biliverdin, one of the by-products of the heme-degrading enzyme heme oxygenase, against the human herpes virus-6. However, biliverdin is rapidly reduced to bilirubin (BR), a molecule endowed with strong antioxidant and antinitrosative features. Aim of this work was to study if BR may reduce the replication of herpes simplex-1 (HSV-1), cytomegalovirus (CMV) and enterovirus (EV) in vitro.
Methods: Monolayers of Hep-2, Vero and MRC5 cells have been infected with clinical isolates of HSV-1, CMV and EV. To mimic the different pathophysiological situations occurring during viral infection, BR (1-10 µM) alone or in the presence of saturating human serum albumin (HSA, 10 µM) was given 2 hours (h) before, concomitantly and 2 h after viral infection. In selected experiments, BR and BR-HSA were pre-incubated with HSV-1 and EV and then given Hep-2 and Vero. After 24 and 48 h of incubation, infected cells were visualized by immunofluorescence and counted. CMV DNA copies were calculated by Real-Time PCR.
Results: When given before or together with HSV-1 and EV, BR (1-10 µM) and BR-HSA, significantly reduced viral replication after 24 and 48 h of incubation. A similar effect occurred when BR and BR-HSA, as above, were pre-incubated with HSV-1 and EV and then administered to cells. Finally, when Hep-2 and Vero cells were treated with BR (1-10 µM) following viral infection, the cytoprotective effect of the bile pigment against HSV-1 infection was evident only within the first 24 h of infection, whereas the effect on EV both at 24 and 48 h. When BR (5 µM) was given MRC5 fibroblasts before and together with CMV the viral load exhibited a significant decrease only after 5 and 7 days of incubation. In search for a mechanism to explain these cytoprotective effects of BR, the hypothesis of an increased cell stress response stress has been explored. Both Hep-2 and Vero cells incubated with BR (1-10 µM) exhibited a marked activation of the proto-oncogene Akt and the kinase JNK, two pathways involved in cell survival. Ultimately, BR (1-10 µM) dose-dependently increased nitric oxide production in Vero cells.
Conclusions: Bilirubin and BR-HSA, inhibited viral replication probably by increasing cellular stress response or cytotoxic endogenous molecules such as NO.
Isoniazid: It Was Or It Is A Magic Bullet? Preziosi P, Mancuso C Institute of Pharmacology, Catholic University School of Medicine, Roma, Italy
In 1952, Fox (US 2596069) reported the realization of the hydrazide of isonicotinic acid (isonicotinyl hydrazide or isoniazid, INH). The drug was an intermediate in the synthesis of the thiosemicarbazone of isonicotinoylaldehyde, a compound created by Fox himself during his search for novel thiosemicarbazone derivatives with antimycobacterial properties. These compounds had been studied by Domagk in the 1940s, and one, 4-acetylaminobenzaldehyde thiosemicarbazone (TB-1/698, aminothiozone, thiacetazone), had already been marketed as an antitubercular drug under the name of Conteben.
Previous studies on the antitubercular properties of pyridine bodies – in particular, nicotinamide – also contributed to the development of INH. Chorine (1945) had found the latter compound to possess in vivo activity against infections caused by mycobacteria in animals and, at very high doses an antitubercolar activity was found in humans as well (1951-52). Substitutions of its heterocyclic group rendered nicotinamide inactive as antimycobacterial drug [e.g., N-2-thiazolyl nicotinamide (1948-52)], unlike those of the amide group but all of substituted forms endowed with antitubercular activity as triamino isonicotinic acid and its methyl ester were lacking in vitamin properties. Relatively little attention had been focused on the hydrazine fraction of INH. The benzalbenzenic hydrazide had displayed in vitro activity at a concentration of 10-6 M. The hydrazides of nicotinic acid and its derivatives had very limited (but structurally interesting) in vitro but were inactive in vivo. The fundamental mechanism of action of isoniazid is the inhibition of mycophenolic acid synthesis.
INH proved to be an almost ideal antitubercular drug. In addition to being fully selective, the drug has never been exceeded by other anti-TB drugs. It displayed marked clinical activity in various forms of tuberculosis after a few days of treatment, and could be given during pregnancy. Both the frequency and severity of its adverse effects (hepatic, peripheral neuropathy that could be managed with vitamin B6 supplementation, CNS effects, immune, allergic, and hematologic disturbances) were more than acceptable. Given its activity at the CNS level, INH therapy has been proposed for certain neurologic disorders.
INH was (and is) a drug fully deserving of the appellative ‘magic bullet’.
A New Paradigm Of Endocrine Systems That Enables The Development Of Selective Hormonal Therapies MANDOKI JJ1, JIMÉNEZ-OROZCO A1, GARCÍA-MONDRAGÓN MJ1, MALDONA-DO-ESPINOZA A1, CASA-TIRAO B2 1Facultad de Medicina, 2Facultad de Filosofía y Letras, Universidad Nacional Autónoma de México
The concept of hormones as “chemical messengers” involves implicitly a paradigm which can be expressed briefly as follows: a hormone has a single physiological function; whether produced endogenously or administered exogenously, a hormone elicits always the same responses, which are independent of any other factors. Many observations by different groups of investigators do not support such notions; different endogenous macropulses of a hormone elicit different responses and thus have different physiological functions that are carried out independently of each other in a coherent manner. This contrasts with the diverse, incoherent effects produced by the exogenous administration of a hormone. As different endogenous macropulses of a hormone may be elicited by many different hormones, some of us have proposed earlier that hormone macropulses are components of multisignal messages which produce selective, coherent responses. By monitoring macropulses of a hormone together with their known diverse elicitors, as well as with their known diverse responses, the compositions and effects of the different multisignal messages of which the hormone is a part can be identified. Analytical methods recently developed by which numerous variables can be monitored simultaneously, make such goals realístic. The therapeutic production of such multisignal messages would elicit selective hormonal effects, and would avoid the production of undesirable adverse effects of exogenous single hormone administrations.
Searching For A Tool To Improve The Anti-Doping Action: The Project AR.I.E.T.T.A. (Artificial Intelligence Evoking Target Testing In Antidoping) MANFREDINI F1,2, MALAGONI AM2, LITMANEN H1,ZHUKOVSKAJA L1, JEANNIER P1, DAL FOLLO D2, BESSEBERG A1, GEISTLINGER M1, BAYER P1, FELISATTI M2 , CARRABRE JE1 1International Biathlon Union, Salzburg, Austria; 2Department of Biochemistry and Molecular Biology-Center Biomedical Studies applied to Sport–University of Ferrara, Ferrara, Italy
Background: Substances and methods used to increase the oxygen blood transport and the athlete's performance can be detected but the screening phase performed by International Federations remains a critical issue. The project AR.IE.T.T.A. aimed to develop a software able to analyze athletes’ haematological and performance profile and to point out those reflecting an abnormal pattern.
Methods: 120 Athletes belonging to the International Biathlon Union gave their written informed consent to the study. The haematological and performance data, previously collected were used to develop the AR.I.E.T.T.A. software.
Results: The software includes the following sections: 1) Log-in 2) Data-Entry: data can be loaded, stored and grouped 3) Analysis: data can be analysed, validated scores calculated, parameters displayed simultaneously as statistics, table/graphs, individual or subpopulation profiles 4) Screening: an immediate evaluation of the risk score of the present sample and/or the athlete under study can be obtained. The risk score is calculated combining different parameters, absolute values and inter-intra-individual variations considered concurrently with different weights.
Conclusions: AR.I.E.T.T.A. software enables a quick evaluation of blood results, favouring surveillance programs and timely target testing controls on athletes by the International Federations. Future studies aiming to validate the risk score and to improve the diagnostic phase will enable an upgrade of the system.
Albendazole In The Treatment Of Hydatidosis Of The Muscles MANOURAS A, LAGOUDIANAKIS E, PAPADIMA A, MARKOGIANNAKIS H, KATERGIANNAKIS V
First Department of Propaedeutic Surgery, Hippocrateion Hospital, Athens Medical School Athens, Greece
Background: Hydatid disease caused by the tapeworm Echinococcus granulosus is a worldwide problem especially in sheep and cattle raising countries. Muscle involvement is most commonly encountered as recurrence of previously treated disease or concurrently with primary lesions of the liver or lung. Furthermore, the rarity of muscle hydatidosis has unique implications in diagnosis and management.
Methods: We report here our observations on the usefulness of perioperative chemotherapy in surgical outcome in terms of morbidity and recurrence. We report on eight cases of primary echinococcus of the muscles presented in our clinic during a 10-year period.
Results: We have administered preoperative albendazole for one cycle of 28 days in 6 of our patients based on the size and appearance of the cyst. All patients underwent total pericystectomy without cyst rupture. We have not found any recurrences after minimum follow up of 12 months.
Conclusions: Muscle hydatidosis respond well to surgical intervention. Complete and intact removal of the cyst in muscular hydatidosis should be considered curative.
Enhanced Depot Vaccine Formulations, Vaccimax® And Depovax™, For Cancer And Pandemic Influenza Applications KARKADA, M, MACDONALD L, FUENTES-ORTEGA A, WEIR G, MANSOUR M Immunovaccine Technologies Inc., Halifax, Canada
Background: The development of a vaccine that can induce a robust cellular and humoral antigen specific immune response after a single dose would be ideal for pandemic flu and cancer immunotherapy. IVT has developed two novel depot vaccine formulations, VacciMax® and DepoVaxTM, which induce enhanced immune responses when compared to current common treatments.
Methods: To detect humoral responses induced by the recombinant H5 antigen (Vietnam/04), mice were immunized once with IVT’s vaccine formulations and once or twice with an appropriate control vaccine. Serum was collected at week 2, 3, 4 and monthly thereafter and titered using an H5 ELISA. The therapeutic efficacy of cacner vaccine formulations was tested in an established tumor challenge model. Mice bearing C3 tumors were vaccinated once with IVT’s depot formulations containing the immunodominant CTL epitope from HPV 16 E7. Tumor growth and survival was monitored for 6 weeks. Antigen specific CTL activity was detected by IFN-γ ELISPOT in the lymph nodes of HLA-A2/H2-D (AAD) transgenic mice immunized with HLA-A2 restricted CTL epitopes formulated in IVT’s depot vaccines.
Results: H5 formulated in our depot technology was able to raise a strong immune response within 18 days. At all time points tested, IVT depot formulation titers were superior to a single dose of the control alum vaccine and in the longer term superior or equal to the two dose alum vaccine. In the therapeutic C3 tumor challenge model, a depot formulation effectively eliminated C3 tumors after a single dose (100% tumor free mice) compared to non-vaccinated mice (0% tumor free, mean tumor size >2000 mm3). An HLA-A2 peptide-based therapeutic cancer vaccine has been designed for Breast/ Ovarian/ Prostate cancers and antigen-specific immune responses were detected in AAD mice after a single dose.
Conclusions: 1) Single dose capability and 100% response rate of IVT’s depot vaccines are significant in the context of a pandemic vaccine for which low initial responses and overall low individual response rates could lead to many deaths. 2) The multi-targeting strategy using tumor-specific peptides and potent cellular response induced by IVT’s depot vaccines indicates a promising immunotherapy for cancer.
Comprehensive HIV/AIDS Care And Treatment As A Need For Quality Provision Of Antiretroviral Therapy: A Case Study From Dar Es Salaam Region, Tanzania MAPUNJO SG1 AND URASSA DP2 1Ministry of Health and Social Welfare. DSM, Tanzania, 2Muhimbili University of Health and Allied Sciences, DSM, Tanzania
Background: The roll-out of Tanzania National HIV/AIDS care and treatment program began in October 2004, with a plan target to cover about 400,000 HIV infected Tanzanians in a period of five years. In delivering Antiretroviral Therapy (ART) a certain level of quality is recommended. The objective of this study was to determine the quality standards of health facilities in providing HIV/AIDS care and treatment in line with the Ministry of Health (MOH) stipulated guidelines.
Methods: A cross-sectional descriptive study was conducted to assess the quality standards in delivering ART in Dar es Salaam Region from May to July 2005. Ten health facilities (both public and private) already designated by MOH to provide ART, six of them since October 2004 (included purposively) and four since May 2005 (selected randomly). The other two facilities not designated were randomly picked and added. The checklist with the MOH required standards was used to assess the availability of equipments, staff, antiretrovirals, guidelines and adequacy of services provided.
Results: Regarding services provided, it was found that Comprehensive HIV/AIDS care and treatment was not fulfilled in all health facilities as recommended. More than half of the health facilities did not have Home Based Care (HBC) services. However, PASADA (Pastoral Activities and Services for people with AIDS in Dar es Salaam Archdiocese) with 14,000 patients, had a strong HBC with no patient lost to follow-up (The percentage of patient lost to follow- up ranged from 0% to 7.3%). Prevention of Mother to Child Transmission (PMTCT) services was found in two third of facilities. Although food support is included as an element of comprehensive HIV/AIDS care and treatment, only PASADA Voluntary Agency was providing nutrition services.
Figure: Percentage of health facilities with HIV care and treatment services
Conclusion: The advantage of comprehensive HIV/AIDS care and support services was shown by PASADA with the example of no patient lost to follow-up. The success shown by PASADA should be adopted by other health facilities for quality provision of ART.
