MALDI-TOF MS Assessment to Identify Environmental Mycobacteria

Over the past few decades, there has been a significant increase in the number of mycobacterial species described. Currently, the genus?Mycobacterium?consists of 170 species. Most species are called nontuberculous mycobacteria (NTM) and are potentially or rarely pathogenic and ubiquitous. One of the main challenges in mycobacteriology is the rapid and precise identification of these microorganisms. In this work, we compared two protein extraction protocols for the identification of 38 reference strains and clinical isolates, representing 27 species, by mass spectrometry (MALDI-TOF MS) to subsequently use the best method for identifying environmental mycobacteria. The results obtained with reference strains and clinical isolates showed that protocol A was effective in identifying 92.1% of mycobacterial specimens at the species level and protocol B, 50%. Therefore, protocol A was evaluated for the rapid identification of 27 environmental mycobacterial isolates. These isolates were subjected to PCR-restriction enzyme analysis (PRA-hsp65). Two isolates were misidentified by PRA-hsp65, whereas MALDI-TOF MS was able to identify them correctly. The results were confirmed by?hsp65 and 16S rRNA gene sequencing. Mass spectrometry has the advantage of being a simpler and faster technique than PRA-hsp65, and our results showed that MALDI-TOF MS is a valuable tool for the identification of environmental mycobacterial isolates.

Triterpenes from Cabralea canjerana as in Vitro Inhibitors to Light Reactions of Photosynthesis

Aims: Cabraleadiol (1), Ocotilone (2) and Odoratone (3) are three triterpenes isolated from Cabralea canjerana (Vell.) Mart. (Meliaceae). They were chemically characterized, and their effect was tested on the light reaction of photosynthesis. Study Design: Natural products were used as models to find new models for inhibitors of photosynthesis. Place and Duration of Study: Departmento de Química, Universidade Federal de S&atildeo Carlos and Departmento de Bioquímica, Facultad de Química, Universidad Nacional Autonoma de México, between May 2013 and January 2014. Methodology: The natural products had their effect on the light reaction of photosynthesis studied by pollarography and Chlorophyll a (Chl a) fluorescence transients. Results: The compounds inhibited ATP synthesis and electron transport rate (basal, phosphorylating and uncoupled). Therefore, they act as Hill reactions inhibitors. Their inhibition site were located in the range of electron flow from OEC complex and between P680 to QA of PS II, and inhibited the photosystem II (PS II) by inducing the appearance of a K-band which is an indicative that the photochemical apparatus is failing at the donor side of PS II interacting at the OEC complex and by transforming active reaction centers to “heat sinks” or the formation of silent reaction centers unable to reduce QA. Conclusion: Furthermore, these triterpenes inhibit PS II and induce the appearance of small G band which is related with the decreased plastoquinone (PQ) pool reduction.