A Paradigm Shift from Emergency Response to Reconstruction and Rehabilitation: Creation of Peak National Body for Disaster Management in Pakistan

The earthquake of 8 October 2005, an unprecedented disaster in the history of Pakistan, led to an equally exceptional national response. Reconstruction and rehabilitation of affected areas was indeed a herculean task. The Earthquake Reconstruction and Rehabilitation Authority (ERRA) was immediately established as a peak national body with extraordinary powers and mandate to ensure coordinated actions for rescue, relief, reconstruction and rehabilitation. The national institutional set up was forced to readjust rapidly to convert this adversity into an opportunity to improve its capability to deal with disasters. This paper aims to provide an overview of the institutional strategy and measures undertaken in the wake of the 2005 earthquake. It looks at the strengths and weaknesses of installing an efficient entity largely adopting a command and control approach to efficiently and effectively deliver reconstruction projects on the ground. The paper seeks to derive lessons that can be useful for governments considering the setting up of comprehensive proactive disaster management systems

UV SPECTROPHOTOMETRIC ASSAY METHOD DEVELOPMENT AND VALIDATION OF DABIGATRAN ETEXILATE IN CAPSULES

Objective: To develop a simple and a cheap UV spectrophotometric method for the quantitative estimation of Dabigatran etexilate in capsules and validate as per ICH guidelines.Methods: The optimized method uses triethyl ammonium phosphate aqueous solution (pH 2.5) as a solvent for the estimation of assay of Dabigatran etexilate in capsules at a wavelength of 325 nm.Results: The developed method resulted in Dabigatran etexilate exhibiting linearity in the range 5-15μg/ml. The assay precision is exemplified by relative standard deviation of 1.07%. Percentage Mean recovery was found to be in the range of 98â€102, during accuracy studies. Method was found to be robust with respect to wavelength and pH of the solvent. Conclusion: A simple and a cheap UV spectrophotometric method was developed and validated for the quantitative estimation of Dabigatran etexilate in capsules as per ICH guidelines and hence it can be used for routine analysis in various pharmaceutical industries.Â

Anxiety, Depression, and Pain: Considerations in the Treatment of Patients with Uncontrolled Hypertension

PURPOSE OF REVIEW: The association between mental health, pain, and treatment-resistant hypertension is an important consideration for treating physicians. We review and discuss the connection between conditions of anxiety, depression, and chronic pain and their effect on uncontrolled hypertension. RECENT FINDINGS: There is significant co-occurrence of hypertension with anxiety, depression, and chronic pain which may lead to undertreatment of hypertension and undertreatment of the underlying mental health disorder. The association between mental health and hypertension is complex and is modulated by physiologic and environmental factors. Physicians treating patients with hypertension should be cognizant of the role anxiety, depression, and chronic pain play in treatment efficacy and compliance. Patients undergoing treatment should be screened for mental health disorders at treatment initiation and frequently thereafter to ensure optimal overall health and compliance

A reductive aminase from Aspergillus oryzae

Reductive amination is one of the most important methods for the synthesis of chiral amines. Here we report the discovery of an NADP(H)-dependent reductive aminase from Aspergillus oryzae (AspRedAm, Uniprot code Q2TW47) which can catalyse the reductive coupling of a broad set of carbonyl compounds with a variety of primary and secondary amines with up to >98% conversion and with up to >98% enantiomeric excess. In cases where both carbonyl and amine show high reactivity, it is possible to employ a 1:1 ratio of the substrates, forming amine products with up to 94% conversion. Steady-state kinetic studies establish that the enzyme is capable of catalysing imine formation as well as reduction. Crystal structures of AspRedAm in complex with NADP(H) and also with both NADP(H) and the pharmaceutical ingredient (R)-rasagiline are reported. We also demonstrate preparative scale reductive aminations with wild-type and Q240A variant biocatalysts displaying total turnover numbers of up to 32,000 and space time yields up to 3.73 g L-1 d-1

Stereoselectivity and Structural Characterisation of an Imine Reductase (IRED) from Amycolatopsis orientalis

The imine reductase AoIRED from Amycolatopsis orientalis (Uniprot R4SNK4) catalyzes the NADPH-dependent reduction of a wide range of prochiral imines and iminium ions, predominantly with (S)-selectivity and with e.e.s of up to >99%. AoIRED displays up to 100-fold greater catalytic efficiency for 2-methyl-1-pyrroline (2MPN) compared to other IREDs, such as the enzyme from Streptomyces sp. GF3546, which also exhibits (S)-selectivity, and thus AoIRED is an interesting candidate for preparative synthesis. AoIRED exhibits unusual catalytic properties, with inversion of stereoselectivity observed between structurally similar substrates, and also, in the case of 1-methyl-3,4-dihydroisoquinoline, for the same substrate, dependent on the age of the enzyme after purification. The structure of AoIRED has been determined in an ‘open’ apo-form, revealing a canonical dimeric IRED fold in which the active site is formed between the N- and C-terminal domains of participating monomers. Co-crystallisation with NADPH gave a ‘closed’ form in complex with the cofactor, in which a relative closure of domains, and associated loop movements, has resulted in a much smaller active site. A ternary complex was also obtained by co-crystallization with NADPH and 1-methyl-1,2,3,4-tetrahydroisoquinoline [(MTQ], and reveals a binding site for the (R)-amine product which places the chiral carbon within 4 Å of the putative location of the C4 atom of NADPH that delivers hydride to the C=N bond of the substrate. The ternary complex has permitted structure-informed mutation of the active site, resulting in mutants including Y179A, Y179F and N241A, of altered activity and stereoselectivity

Efficient clinical-grade γ-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing

γ-Retroviral vectors (γ-RV) are powerful tools for gene therapy applications. Current clinical vectors are produced from stable producer cell lines which require minimal further downstream processing, while purification schemes for γ-RV produced by transient transfection have not been thoroughly investigated. We aimed to develop a method to purify transiently produced γ-RV for early clinical studies. Here, we report a simple one-step purification method by high-speed centrifugation for γ-RV produced by transient transfection for clinical application. High-speed centrifugation enabled the concentration of viral titers in the range of 107-108 TU/mL with >80% overall recovery. Analysis of research-grade concentrated vector revealed sufficient reduction in product- and process-related impurities. Furthermore, product characterization of clinical-grade γ-RV by BioReliance demonstrated two-logs lower impurities per transducing unit compared with regulatory authority-approved stable producer cell line vector for clinical application. In terms of CAR T cell manufacturing, clinical-grade γ-RV produced by transient transfection and purified by high-speed centrifugation was similar to γ-RV produced from a clinical-grade stable producer cell line. This method will be of value for studies using γ-RV to bridge vector supply between early- and late-stage clinical trials

Synergistic Chemo/Biocatalytic Synthesis of Alkaloidal Tetrahydroquinolines

The power of complementary chemocatalytic and biocatalytic transformations is demonstrated in the asymmetric synthesis of 2-substituted tetrahydroquinolines. A series of racemic tetrahydroquinolines were synthesized through a convergent one-pot Rh­(I)-catalyzed addition/condensation sequence of alkyl vinyl ketones and aminophenylboronic acids. The resulting tetrahydroquinolines were thereafter shown to be substrates for the flavin-dependent enzyme cyclohexylamine oxidase, and preparative-scale deracemizations have been demonstrated on these high-value targets