Dose Determination of Activated Charcoal in Management of Amitriptyline-Induced Poisoning by Reversed-Phase High-Performance Liquid Chromatography

Purpose: To assess the doses of activated charcoal currently used in the management of acute amitriptyline-induced drug poisoning and explore the possibility of using lower doses.Methods: Albino male Wistar rats, weighing 200 ± 20 g, were used for the study. The animals were divided into four groups of eight animals each. The concentration of amitriptyline in rat plasma was measured by high performance liquid chromatography (HPLC) for dose determination of activated charcoal. Chromatograms were established with acetonitrile: 70 mM KH2PO4 buffer (60: 40, v/v) solvent system on an Xterna® ms C18 SUM column (5 μm, 3.9 × 150 mm) and pH was adjusted to 4.5 with ortho-phosphoric acid. Mobile phase flow rate was 1 ml/min and ultraviolet (UV) detection was at 293 nm. Validation of the method was performed to determine its selectivity, linearity, precision, as well as limits of detection (LOD) and of quantification (LOQ).Results: Standard curves were linear, r2 = 0.996, for amitriptyline over the concentration range 10 - 60 ng/ml. Recovery (98.3 to 100.85 %) was in the selected concentration range of 10 - 60 ng/ml. The LOD and LOQ of the method for amitriptyline were 0.109 and 0.332 μg/ml, respectively. The validated method was successfully applied to measure plasma concentrations of amitriptyline and to measure the doses of activated charcoal currently used in the management of acute amitriptyline drug poisoning.Conclusion: The proposed RP-HPLC method enables determination of amitriptyline with good separation and resolution of the chromatographic peaks. Validation revealed that the method is sensitive, accurate and selective. Using half of the standard dose of the activated charcoal gave a comparable effect to the standard dose in reducing drug concentration in the blood. While, using quarter of the standard dose of activated charcoal does not have a cleared effect.Keywords: Amitriptyline, Activated charcoal, Drug poisoning, Reversed-phase high-performance liquid chromatograph

Active and Passive Immunization Protects against Lethal, Extreme Drug Resistant-Acinetobacter baumannii Infection

Extreme-drug-resistant (XDR) Acinetobacter baumannii is a rapidly emerging pathogen causing infections with unacceptably high mortality rates due to inadequate available treatment. New methods to prevent and treat such infections are a critical unmet medical need. To conduct a rational vaccine discovery program, OmpA was identified as the primary target of humoral immune response after intravenous infection by A. baumannii in mice. OmpA was >99% conserved at the amino acid level across clinical isolates harvested between 1951 and 2009 from cerebrospinal fluid, blood, lung, and wound infections, including carbapenem-resistant isolates, and was ≥89% conserved among other sequenced strains, but had minimal homology to the human proteome. Vaccination of diabetic mice with recombinant OmpA (rOmpA) with aluminum hydroxide adjuvant markedly improved survival and reduced tissue bacterial burden in mice infected intravenously. Vaccination induced high titers of anti-OmpA antibodies, the levels of which correlated with survival in mice. Passive transfer with immune sera recapitulated protection. Immune sera did not enhance complement-mediated killing but did enhance opsonophagocytic killing of A. baumannii. These results define active and passive immunization strategies to prevent and treat highly lethal, XDR A. baumannii infections

GWAS for quantitative resistance phenotypes in Mycobacterium tuberculosis reveals resistance genes and regulatory regions

YesDrug resistance diagnostics that rely on the detection of resistance-related mutations could expedite patient care and TB eradication. We perform minimum inhibitory concentration testing for 12 anti-TB drugs together with Illumina whole-genome sequencing on 1452 clinical Mycobacterium tuberculosis (MTB) isolates. We evaluate genome-wide associations between mutations in MTB genes or non-coding regions and resistance, followed by validation in an independent data set of 792 patient isolates. We confirm associations at 13 non-canonical loci, with two involving non-coding regions. Promoter mutations are measured to have smaller average effects on resistance than gene body mutations. We estimate the heritability of the resistance phenotype to 11 anti-TB drugs and identify a lower than expected contribution from known resistance genes. This study highlights the complexity of the genomic mechanisms associated with the MTB resistance phenotype, including the relatively large number of potentially causal loci, and emphasizes the contribution of the non-coding portion of the genome.Biomedical research grant from the American Lung Association (PI MF, RG-270912-N), a K01 award from the BD2K initiative (PI MF, ES026835), and an NIAID U19 CETR grant (P.I. M.M., AI109755), the Belgian Science Policy (Belspo) (L.R., C.J.M.)

In a search for efficient treatment for amyotrophic lateral sclerosis: Old drugs for new approaches

Recent progress in understanding the pathological changes in the nervous system and in certain other body systems (e.g., immune system) that lead to the development and progression of amyotrophic lateral sclerosis (ALS) revealed a number of molecular and cellular processes that can potentially be used as therapeutic targets. Many of these processes are compromised not only in ALS but also in other diseases and a repertoire of drugs able to restore, at least partially, their functionality has been developed. In this review, we briefly describe current approaches to the repurposing of such “old” drugs for treatment of patients with ALS