Standardization of natural mycolic acid antigen composition and production for use in biomarker antibody detection to diagnose active tuberculosis

This research article published by Elsevier B.V., 2016Mycobacterium tuberculosis, the causative agent of tuberculosis, is characterized by the abundance of species specific, antigenic cell wall lipids called mycolic acids. These wax-like molecules all share an identical, amphiphilic mycolic motif, but have different functional groups in a long hydrophobic hydrocarbon mero-chain that divide them into three main classes: alpha-, keto- and methoxy-mycolic acids. Whereas alpha-mycolic acids constitutively maintain an abundance of around 50%, the ratio of methoxy- to keto-mycolic acid types may vary depending on, among other things, the growth stage of M. tuberculosis. In human patients, antibodies to mycolic acids have shown potential as diagnostic serum biomarkers for active TB. Variations in mycolic acid composition affect the antigenic properties and can potentially compromise the precision of detection of anti-mycolic acids antibodies in patient sera to natural mixtures. We demonstrate this here with combinations of synthetic mycolic acid antigens, tested against TB patient and control sera. Combinations of methoxy- and α-mycolic acids are more antigenic than combinations of keto- and α-mycolic acids, showing the former to give a more sensitive test for TB biomarker antibodies. Natural mixtures of mycolic acids isolated from mature cultures of M. tuberculosis H37Rv give the same sensitivity as that with synthetic methoxy- and α-mycolic acids in combination, in a surface plasmon resonance inhibition biosensor test. To ensure that the antigenic activity of isolates of natural mycolic acids is reproducible, we cultured M. tuberculosis H37Rv on Middlebrook 7H10 solid agar plates to stationary growth phase in a standardized, optimal way. The proportions of mycolic acid classes in various batches of the isolates prepared from these cultures were compared to a commercially available natural mycolic acid isolate. LC-MS/MS and NMR data for quantitation of mycolic acids class compositions show that the variation in batches is small, suggesting that the quality of the results for anti-mycolic acid antibody detection in the TB patients should not be affected by different batches of natural mycolic acid antigens if prepared in a standard way

Wat is de beste plek voor kinderen met externaliserende gedragsproblemen in het reguliere onderwijs? Uitkomsten van een klasindelinginterventie

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INFLUÊNCIA DO ANELAMENTO E ESTIOLAMENTO DE RAMOS NA PROPAGAÇÃO DA LARANJEIRA VALÊNCIA (Citrus sinensis Osbeck) ATRAVÉS DE ESTACAS

Objetivando estudar a influência do condicionamento de ramos, através do anelamento e estiolamento parcial, sobre o enraizamento de estacas de laranjeira cv. Valência, conduziu-se o experimento no período de fevereiro a dezembro de 1993. Para o anelamento foi utilizado arame fino de cobre com o qual provocou-se um estrangulamento na base do ramo. No estiolamento, utilizou-se fita preta de polietileno, que foi enrolada na base das brotações. De cada ramo condicionado, obteve-se apenas uma estaca com 15 cm de comprimento, diâmetro médio de 0,5 cm e um par de folhas apicais, cortadas pela metade. O plantio das estacas foi realizado em 12.04.93, em substrato de areia lavada e peneirada. Adotou-se o delineamento inteiramente casualizado, em esquema fatorial 3x3, resultante das combinações entre os períodos de zero, 30 e 60 dias dos fatores anelamento e estiolamento. Verificou-se que não ocorreu enraizamento nas estacas que permaneceram três meses no substrato e variou de 0 até 26,04% para aquelas que permaneceram oito meses. O período de estiolamento influenciou significativamente o percentual de estacas enraizadas.<br>With the aim of studying the effect of branch conditioning through girdling and banding on Valência orange cuttings a rooting trial was carried out from February to December of 1993. The girdling was performed with a thin cooper wire tightly tied at the branch base reaching the log, and the banding with black poliethylene tape, covering the branch at the base of the sprouts. Each conditioned branch gave one cutting of 15 cm length, 0.5 cm diameter and a pair of half cut apical leaves. Cuttings were planted on Dec.04, 93, in washed and sieved sand substrate. The experimental design was completely randomized, with a factorial 3 x 3 from 0, 30 and 60 day periods, girdling and banding. There was no rooting in cuttings that remained three months in the substrate and there was up to 26.04% rooting for those ones that lasted for eight months in the substrate. The banding period significantly affected the cuttings rooting percentage