ABSTRACT
Tuberculosis continues to be a great source of concern in global health because of the large reservoir of humans infected with the bacilli and the appearance of clinical isolates resistant to a wide array of anti-tuberculosis drugs. New drugs with novel mechanisms of action on new targets are urgently required to reduce global tuberculosis burden. Mycobacterium tuberculosis nucleoid associated protein (NAP) HU has been shown to be druggable and essential for the organism's survival. In this study, four diarylethenes were synthesized using a one-pot decarboxylated Heck-coupling of coumaric acids with iodoanisoles. The prepared compounds 1-4 were tested for their in vitro growth inhibition of M. tuberculosis H37Rv using the spot culture growth inhibition assay, displaying minimum inhibitory concentrations between 9 and 22 µM. Their cytotoxicity against BHK-21 cell line showed half inhibition at concentrations between 98 and 729 µM. The most selective hit (SI = 81), demonstrated inhibition of M. tuberculosis HU protein involved in maintaining bacterial genome architecture.
Subject(s)
Antitubercular Agents , Bacterial Proteins/antagonists & inhibitors , Bacterial Proteins/metabolism , DNA-Binding Proteins/antagonists & inhibitors , DNA-Binding Proteins/metabolism , Mycobacterium tuberculosis/metabolism , Tuberculosis/drug therapy , Animals , Antitubercular Agents/chemical synthesis , Antitubercular Agents/chemistry , Antitubercular Agents/pharmacology , Cell Line , Cricetinae , HumansABSTRACT
BACKGROUND: The quality of multiplex polymerase chain reaction (PCR) assays depends on several factors. Therefore, it is important to establish the optimal conditions to achieve efficient amplification. The objective of this study was to implement a 5 × 4 factorial design combined with image analysis using agarose gels and an efficiency calculation to optimize a multiplex PCR assays for the detection of Salmonella enterica serovar typhimurium. METHODS: We used 12 ng of Salmonella DNA obtained from pure cultures and applied different annealing temperatures (65°C, 64.5°C, 63.3°C, 61.4°C, or 59°C) and different MgCl2 concentrations (1 mM, 1.5 mM, 2 mM, or 2.5 mM) to amplify regions of the fliC, rfbJ, and fljB genes. The 5 × 4 factorial design was performed using Statgraphics Plus software version 5.1, and the images were analyzed using Image Lab(TM) software. RESULTS: Superior amplification was obtained using an annealing temperature of 65°C and 2 mM MgCl2 . This finding was confirmed by calculating the efficiency of multiplex PCR assays (6.1%) at these conditions. CONCLUSION: We propose the application of factorial design and image analysis to determine the most suitable conditions for multiplex PCR optimization.
Subject(s)
Bacterial Typing Techniques/methods , Image Processing, Computer-Assisted/methods , Multiplex Polymerase Chain Reaction/methods , Research Design , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Electrophoresis, Agar Gel , Salmonella typhi/genetics , Salmonella typhi/isolation & purification , TemperatureABSTRACT
El infarto agudo del miocardio (IAM) es una de las enfermedades más comunes y su mortalidad es mayor después de las primeras horas de inicio. Es por ende importante hacer un rápido diagnóstico para acelerar la admisión del paciente y proveer terapia adecuada. El objetivo principal se basó en establecer diferencias de manejo entre el Jackson Memorial Hospital (JMH) de Miami y el Hospital Luis Vernaza (HLV) de Guayaquil. El siguiente es un estudio prospectivo. Se incluyeron 30 pacientes en Miami y 15 en Guayaquil que cumplieron con criterios diagnósticos para IAM. En el HLV se diagnosticó prácticamente por manifestaciones de isquemia y un electrocardiograma, no se utilizaron trombolíticos, ni inhidores de la gicloproteina...
