MazG nucleoside triphosphate pyrophosphohydrolase (NTP-PPaseMycobacterium tuberculosis H37Ra is an avirulent relative of the typical laboratory strain H37Rv, both of which are derived from the parent strain H37 that was originally isolated from a 19-year-old patient with chronic pulmonary tuberculosis by Edward R. Baldwin in 1905 (1). Our systematic comparison of the complete genomic sequences of H37Rv and H37Ra identified multiple H37Ra-specific single nucleotide polymorphisms (SNPs) affecting about 30 genes including the virulence determinants phoP, nrp, and pks12 (2), consistent with the widely accepted notion (3-7) that the virulence attenuation of H37Ra is due to multiple mutations. One of these mutations, the A219E substitution located in a highly conserved sequence of H37Ra MazG, prompted the current study aiming to reveal the function of the corresponding prototype enzyme in M. tuberculosis (MtMazG) and compare its role with that of
BackgroundTrichinellosis, a widespread zoonosis, is regarded as an emerging or reemerging disease. Effective treatment and prognosis of trichinellosis depends on early diagnosis of the infection. The objective of this study was to identify sensitive and specific antigens for early diagnosis or effective vaccine antigens for preventing infection.MethodsThe somatic proteins of T. spiralis adult worms were separated by two-dimensional gel electrophoresis (2-DE). The separated proteins were probed with early infection sera from swine or mice infected with T. spiralis for 7 days. The primary immunoreactive spots were characterized by MALDI-TOF/TOF-MS analysis in combination with bioinformatics. The identified proteins were annotated using WEGO based on their functions. The immunodominant protein was chosen for expression as recombinant protein in E. coli and the purified recombinant protein was used to confirm its antigenicity by Western blot with the same infection sera.ResultsApproximately 300 spots were separated by 2-DE, with molecular weights ranging from 10 to 130 kDa, and pI values ranging from pH 4 to 10. The sera from swine and mice infected with T. spiralis for 7 days recognized 64 proteins. MALDI-TOF/TOF-MS analysis identified 55 proteins, some with different isoforms. Finally, 40 individual immunoreactive proteins were obtained with a wide range of biological functions. Several proteins, such as heat shock protein 70, 14-3-3 protein, and cysteine protease could be used as immunodiagnostic or vaccine antigens. Among these identified proteins, the highly immunodominant Ts14-3-3 was chosen for expression in E. coli and purified recombinant Ts14-3-3 was able to be strongly recognized by the same T. spiralis infected sera used for identifying these antigens, therefore the most promising antigen for early immunodiagnosis of Trichinella infection.ConclusionsA total of 64 proteins from the adult worm were recognized by early infection sera from swine and mice infected with T. spiralis for 7 days. Several proteins, are of particular interest as immunodiagnostic or vaccine antigens, especially with Ts14-3-3 as most promising due to its highly immunogenicity during early infection, expressed protein can be recognized by Trichinella early infection sera and the native Ts14-3-3 expression in both adult and larval stages.
e Ethambutol (EMB) plays a pivotal role in the chemotherapy of drug-resistant tuberculosis (TB), including multidrug-resistant tuberculosis (MDR-TB). Resistance to EMB is considered to be caused by mutations in the embCAB operon (embC, embA, and embB). In this study, we analyzed the embCAB mutations among 139 MDR-TB isolates from China and found a possible association between embCAB operon mutation and EMB resistance. Our data indicate that 56.8% of MDR-TB isolates are resistant to EMB, and 82.2% of EMB-resistant isolates belong to the Beijing family. Overall, 110 (79.1%) MDR-TB isolates had at least one mutation in the embCAB operon. The majority of mutations were present in the embB gene and the embA upstream region, which also displayed significant correlations with EMB resistance. The most common mutations occurred at codon 306 in embB (embB306), followed by embB406, embA(؊16), and embB497. Mutations at embB306 were associated with EMB resistance. DNA sequencing of embB306 -497 was the best strategy for detecting EMB resistance, with 89.9% sensitivity, 58.3% specificity, and 76.3% accuracy. Additionally, embB306 had limited value as a candidate predictor for EMB resistance among MDR-TB infections in China.M ultidrug-resistant tuberculosis (MDR-TB) is attributed to an estimated 3.7% new cases and 20.2% previously treated cases of TB annually worldwide and is becoming a major threat to global public health (1). In China, the significantly high prevalence (5.7% new cases and 25.6% previously treated cases) of MDR-TB makes TB control especially challenging (2). Ethambutol (EMB) is an important first-line anti-TB drug routinely recommended for therapy of drug-resistant TB, including MDR-TB. Disturbingly, in some regions of China, substantial proportions (51.3% to 66.7%) of MDR-TB isolates demonstrated EMB resistance (3-5). Development of new rapid and reliable molecular methods for detecting drug resistance is essential to optimize treatment regimens, prevent treatment failure, and thus reduce the further spread of drug-resistant isolates. However, these molecular assays require precise knowledge of the genetic mutations associated with drug resistance. Prior studies indicated that the characteristics of resistance-associated mutations vary in different regions (6, 7). EMB acts against TB by inhibiting membrane-associated arabinosyl transferases encoded by the embCAB operon (including embC, embA, and embB), which are involved in the synthesis of cell wall arabinogalactan (8,9). Approximately 50% to 70% of EMB-resistant TB isolates harbor mutations in a relatively short fragment (codons 306 -497) in embB genes, with mutations occurring most frequently at codon 306 in embB (embB306), embB406, and embB497 (5,8,(10)(11)(12)(13). Sequence analysis of this fragment has been a tool for the rapid detection of EMB resistance. However, approximately one third of EMB-resistant isolates do not carry changes in this region and therefore are not detectable by using DNA sequencing (12,14). Although other mutations in the embCAB...
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