No abstract

No abstract

Mycobacterium tuberculosis is the cause of one of the diseases with the highest mortality and morbidity rate in the Americas and in the world. In developing countries, the diagnosis of tuberculosis (TB) is based on baciloscopy and bacteriological cultures. The first method has a low sensitivity, and the second can take several weeks to reach a confirmatory diagnosis. The lack of a rapid diagnosis compromises the efforts to control this disease and favors the transmission of tuberculosis to the susceptible population. In this work, we present the synthesis, amine-silanization, characterization and bio-functionalization of magnetic nanoparticles (MNPs) to develop a sandwich ELISA to detect and concentrate antigens from M. tuberculosis. For this purpose, a recombinant mycobacterial heat shock protein Hsp16.3, which contributes to the persistence of TB, was cloned and expressed in the E. coli system. Polyclonal antibodies anti-Hsp16.3 were produced in a rabbit and in mice. Magnetic nanoparticles were synthesized by co-precipitation, amine-functionalized and characterized by several physical-chemical methods. The XRD, Mossbauer spectroscopy, zeta potential, TEM, and FTIR all proved the successful preparation of the MNPs showing a diffraction crystal diameter of 10.48 ± 2.56 nm, superficial net charge of : +23.57 ± 2.87 mV, characteristic patterns of magnetite and a structure similar to a sphere. Additionally, it showed a magnetization saturation of 37.06 emu.g−1. For the functionalization of nanoparticle surfaces with anti-Hsp16.3, the active ester method was used for bond formation, and parameters such as time of incubation, coupling agents ratio (EDC/NHS) and concentration as well as surface saturation level of amine-silanized MNPs (MNP@Si@NH2) were standardized. Finally, bio-functionalized MNPs were used to detect, fix and concentrate the recombinant antigen Hsp16.3 from M. tuberculosis in a sandwich ELISA-MNP assay.

No abstract

En el presente trabajo, se reporta la síntesis y caracterización de nanopartículas magnéticas de magnetita/plata (Fe3O4/Ag) para el estudio de sus propiedades antibacterianas frente a las bacterias Enterobacter aerogenes (Gramnegativa) y Enterococcus faecalis (Gram-positiva). Las nanopartículas magnéticas de magnetita (MNPs) se sintetizaron por el método de solgel, usando bromuro de cetiltrimetilamonio (CTAB) como surfactante. Posteriormente, en la dispersión coloidal de magnetita, se llevó a cabo la reducción química in situ de iones de plata, usando glucosa como agente reductor y polivinilpirrolidona (PVP) como agente dispersante, para obtener un nanocompuesto magnético Fe3O4/Ag. El análisis morfológico y espectroscópico de las nanopartículas de Fe3O4 y del nanocompuesto de Fe3O4/Ag fue realizado mediante la espectroscopia infrarroja con transformada de Fourier (FTIR), Raman, y Mössbauer (MS), además de la técnica de difracción de rayos X (DRX), la microscopía electrónica de barrido (SEM) y espectroscopia de energía dispersiva de rayos X (EDS). Las nanopartículas de Fe3O4 resultaron esféricas con un diámetro medio de 10 nm y el nanocompuesto de Fe3O4/Ag con un tamaño medio de 28 nm, el test antibacteriano indicó que el uso del nanocompuesto de Fe3O4/Ag a una concentración de 5 mg·mL-1 permite una inhibición total del crecimiento de los microorganismos estudiados a partir de una concentración inicial 108 bacterias mL-1.

The fortification of dairy beverages is a widely developed strategy using non-heme or heme iron. Heme iron has a higher bioavailability. The investigation aimed to elaborate pasteurized milk with fortified chocolate flavor with heme iron that has good sensory acceptability. The preparation of the flavored milk was carried out based on the regulations and heme iron, obtained from a commercial source of whole blood of porcine origin, was added before the pasteurization process to achieve its complete dilution. The concentration of iron and chocolate flavoring was established as variables in order to evaluate the optimal formulation based on Sensory Acceptability (SA). The experimental design was a 32 factorial design in which eight formulations were established, which were sensory acceptability evaluated by a total of 35 school-age children, aged between 8 and 11 years using a five-point facial hedonic scale. The results of the analysis of variance and optimization of the response showed that SA was 4.71 (on a scale of 1 to 5) for a fortification of 6.76 mg Fe kg-1 sample and a chocolate concentration of 2.0 g kg-1 sample. The physicochemical characterization indicated a higher percentage of carbohydrates, a higher concentration of iron (9.3 mg Fe kg-1 sample) and vitamin C (349.0 mg kg-1 sample) with respect to fresh milk. According to the physicochemical and microbiological results, the approximate life time of the beverage was 5 days, which is in accordance with Peruvian regulations. These results showed a method of fortification of flavored milk that allowed the use of heme iron, whose content could contribute to the daily requirement of this mineral in children aged between 8 and 11 years old (8 mg of iron per day).