Uso de imunocaptura para o isolamento de bactérias do ambiente.

Obtenção do soro. Imunocaptura. Procedimentos para imunocaptura. Preparo das amostras. Amostras de plantas.bitstream/CNPAB-2010/32965/1/doc198.pd

The open pan-genome architecture and virulence landscape of Mycobacterium bovis

Animal tuberculosis (TB) is an emergent disease caused by Mycobacterium bovis, one of the animal-adapted ecotypes of the Mycobacterium tuberculosis complex (MTC). In this work, whole-genome comparative analyses of 70 M. bovis were performed to gain insights into the pan-genome architecture. The comparison across M. bovis predicted genome composition enabled clustering into the core- and accessory-genome components, with 2736 CDS for the former, while the accessory moiety included 3897 CDS, of which 2656 are restricted to one/two genomes only. These analyses predicted an open pan-genome architecture, with an average of 32 CDS added by each genome and show the diversification of discrete M. bovis subpopulations supported by both core- and accessory-genome components. The functional annotation of the pan-genome classified each CDS into one or several COG (Clusters of Orthologous Groups) categories, revealing ‘transcription’ (total average CDSs, n=258), ‘lipid metabolism and transport’ (n=242), ‘energy production and conversion’ (n=214) and ‘unknown function’ (n=876) as the most represented. The closer analysis of polymorphisms in virulence-related genes in a restrict group of M. bovis from a multi-host system enabled the identification of clade-monomorphic non-synonymous SNPs, illustrating clade-specific virulence landscapes and correlating with disease severity. This first comparative pan-genome study of a diverse collection of M. bovis encompassing all clonal complexes indicates a high percentage of accessory genes and denotes an open, dynamic non-conservative pan-genome structure, with high evolutionary potential, defying the canons of MTC biology. Furthermore, it shows that M. bovis can shape its virulence repertoire, either by acquisition and loss of genes or by SNP-based diversification, likely towards host immune evasion, adaptation and persistence.info:eu-repo/semantics/publishedVersio

Genome-wide estimation of recombination, mutation and positive selection enlightens diversification drivers of Mycobacterium bovis

Genome sequencing has reinvigorated the infectious disease research feld, shedding light on disease epidemiology, pathogenesis, host–pathogen interactions and also evolutionary processes exerted upon pathogens. Mycobacterium tuberculosis complex (MTBC), enclosing M. bovis as one of its animal-adapted members causing tuberculosis (TB) in terrestrial mammals, is a paradigmatic model of bacterial evolution. As other MTBC members, M. bovis is postulated as a strictly clonal, slowly evolving pathogen, with apparently no signs of recombination or horizontal gene transfer. In this work, we applied comparative genomics to a whole genome sequence (WGS) dataset composed by 70 M. bovis from diferent lineages (European and African) to gain insights into the evolutionary forces that shape genetic diversifcation in M. bovis. Three distinct approaches were used to estimate signs of recombination. Globally, a small number of recombinant events was identifed and confrmed by two independent methods with solid support. Still, recombination reveals a weaker efect on M. bovis diversity compared with mutation (overall r/m= 0.037). The diferential r/m average values obtained across the clonal complexes of M. bovis in our dataset are consistent with the general notion that the extent of recombination may vary widely among lineages assigned to the same taxonomical species. Based on this work, recombination in M. bovis cannot be excluded and should thus be a topic of further efort in future comparative genomics studies for which WGS of large datasets from diferent epidemiological scenarios across the world is crucial. A smaller M. bovis dataset (n= 42) from a multi-host TB endemic scenario was then subjected to additional analyses, with the identifcation of more than 1,800 sites wherein at least one strain showed a single nucleotide polymorphism (SNP). The majority (87.1%) was located in coding regions, with the global ratio of non-synonymous upon synonymous alterations (dN/dS) exceeding 1.5, suggesting that positive selection is an important evolutionary force exerted upon M. bovis. A higher percentage of SNPs was detected in genes enriched into “lipid metabolism”, “cell wall and cell processes” and “intermediary metabolism and respiration” functional categories, revealing their underlying importance in M. bovis biology and evolution. A closer look on genes prone to horizontal gene transfer in the MTBC ancestor and included in the 3R (DNA repair, replication and recombination) system revealed a global average negative value for Taijima’s D neutrality test, suggesting that past selective sweeps and population expansion after a recent bottleneck remain as major evolutionary drivers of the obligatory pathogen M. bovis in its struggle with the host.info:eu-repo/semantics/publishedVersio

A Complex Case of Cholestasis in a Patient with ABCB4 and ABCB11 Mutations

The low-phospholipid-associated cholelithiasis (LPAC) syndrome is a form of symptomatic cholelithiasis occurring in young adults, characterized by recurrence of symptoms after cholecystectomy and presence of hepatolithiasis. The case refers to a healthy 39-year-old Caucasian male who presented with abdominal pain and jaundice. His blood tests showed conjugated hyperbilirubinemia and elevated liver enzymes (total bilirubin 6.65 mg/dL, γ-glutamyltransferase 699 IU/L) and abdominal computed tomography revealed dilation of common bile duct and left intrahepatic ducts. Magnetic resonance cholangiopancreatography identified choledocholithiasis, retrieved by endoscopic retrograde cholangiopancreatography, after which there was a worsening of jaundice (total bilirubin 23 mg/dL), which persisted for several weeks, possibly due to ciprofloxacin toxicity. After an extensive workup including liver biopsy, the identification of two foci of hepatolithiasis on reevaluation abdominal ultrasound raised the hypothesis of LPAC syndrome and the patient was started on ursodeoxycholic acid, with remarkable improvement. Genetic testing identified the mutation c.1954A>G (p.Arg652Gly) in ABCB4 gene (homozygous) and c.1331T>C (p.Val444Ala) in ABCB11 gene (heterozygous). In conclusion, we describe the unique case of an adult male with choledocholithiasis, hepatolithiasis, and persistent conjugated hyperbilirubinemia after retrieval of stones, fulfilling the criteria for LPAC syndrome and with possible superimposed drug-induced liver injury, in whom ABCB4 and ABCB11 mutations were found, both of which had not been previously described in association with LPAC.info:eu-repo/semantics/publishedVersio

Traditional free-weight vs. variable resistance training applied to elite young soccer players during a short preseason: effects on strength, speed, and power performance

Maximizing the neuromuscular capacities of players is a critical challenge during short soccer preseasons. This study compared the effects of two strength-power training regimes, on the strength, speed, and power performance of elite young soccer players during a 4-week preseason. Twenty-five under-20 players from the same club were pair-matched in two training groups as follows: traditional training group (TTG) (n=13), athletes performed half-squat (HS) and jump-squat (JS) exercises as traditionally prescribed; and EB group (EBG) (n=12), athletes performed HS and JS with EB attached to the barbell. Vertical jump height, 20-m sprint velocity, change-of-direction (COD) speed, HS and JS power, and one-repetition maximum (1RM) in the HS were assessed pre, post 2-week, and post 4-week of training. An ANOVA two-way with repeated measures was used to assess the effects of both training protocols over the experimental period. Both strategies were effective for significantly improving HS and JS power (effect sizes [ES] 1.00 - 1.77), HS 1RM (ES = 1.68 and 1.51 for TTG and EBG, respectively), vertical jumping ability (ES 0.37 - 0.65), and COD speed (ES = 0.81 and 0.39 for TTG and EBG, respectively), when comparing pre- and post-measures. In contrast, both TTG and EBG failed to increase 20-m sprint velocity (ES ranging between -0.54 and 0.23). In conclusion, both training schemes were able to improve the strength and power performance, but not the sprint capacity of young soccer players. To accelerate strength gains over very-short time periods (i.e., 2-week), variable resistance training may be advantageous. Conversely, to optimize power adaptations in ballistic exercises across a similar time period, traditional FW training may be preferred