47 research outputs found

    The <i>N</i>-myristoylome of <i>Trypanosoma cruzi</i>

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    Protein N-myristoylation is catalysed by N-myristoyltransferase (NMT), an essential and druggable target in Trypanosoma cruzi, the causative agent of Chagas’ disease. Here we have employed whole cell labelling with azidomyristic acid and click chemistry to identify N-myristoylated proteins in different life cycle stages of the parasite. Only minor differences in fluorescent-labelling were observed between the dividing forms (the insect epimastigote and mammalian amastigote stages) and the non-dividing trypomastigote stage. Using a combination of label-free and stable isotope labelling of cells in culture (SILAC) based proteomic strategies in the presence and absence of the NMT inhibitor DDD85646, we identified 56 proteins enriched in at least two out of the three experimental approaches. Of these, 6 were likely to be false positives, with the remaining 50 commencing with amino acids MG at the N-terminus in one or more of the T. cruzi genomes. Most of these are proteins of unknown function (32), with the remainder (18) implicated in a diverse range of critical cellular and metabolic functions such as intracellular transport, cell signalling and protein turnover. In summary, we have established that 0.43–0.46% of the proteome is N-myristoylated in T. cruzi approaching that of other eukaryotic organisms (0.5–1.7%)

    Is There a Seamount Effect on Microbial Community Structure and Biomass? The Case Study of Seine and Sedlo Seamounts (Northeast Atlantic)

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    Seamounts are considered to be “hotspots” of marine life but, their role in oceans primary productivity is still under discussion. We have studied the microbial community structure and biomass of the epipelagic zone (0–150 m) at two northeast Atlantic seamounts (Seine and Sedlo) and compared those with the surrounding ocean. Results from two cruises to Sedlo and three to Seine are presented. Main results show large temporal and spatial microbial community variability on both seamounts. Both Seine and Sedlo heterotrophic community (abundance and biomass) dominate during winter and summer months, representing 75% (Sedlo, July) to 86% (Seine, November) of the total plankton biomass. In Seine, during springtime the contribution to total plankton biomass is similar (47% autotrophic and 53% heterotrophic). Both seamounts present an autotrophic community structure dominated by small cells (nano and picophytoplankton). It is also during spring that a relatively important contribution (26%) of large cells to total autotrophic biomass is found. In some cases, a “seamount effect” is observed on Seine and Sedlo microbial community structure and biomass. In Seine this is only observed during spring through enhancement of large autotrophic cells at the summit and seamount stations. In Sedlo, and despite the observed low biomasses, some clear peaks of picoplankton at the summit or at stations within the seamount area are also observed during summer. Our results suggest that the dominance of heterotrophs is presumably related to the trapping effect of organic matter by seamounts. Nevertheless, the complex circulation around both seamounts with the presence of different sources of mesoscale variability (e.g. presence of meddies, intrusion of African upwelling water) may have contributed to the different patterns of distribution, abundances and also changes observed in the microbial community

    The +4G Site in Kozak Consensus Is Not Related to the Efficiency of Translation Initiation

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    The optimal context for translation initiation in mammalian species is GCCRCCaugG (where R = purine and “aug” is the initiation codon), with the -3R and +4G being particularly important. The presence of +4G has been interpreted as necessary for efficient translation initiation. Accumulated experimental and bioinformatic evidence has suggested an alternative explanation based on amino acid constraint on the second codon, i.e., amino acid Ala or Gly are needed as the second amino acid in the nascent peptide for the cleavage of the initiator Met, and the consequent overuse of Ala and Gly codons (GCN and GGN) leads to the +4G consensus. I performed a critical test of these alternative hypotheses on +4G based on 34169 human protein-coding genes and published gene expression data. The result shows that the prevalence of +4G is not related to translation initiation. Among the five G-starting codons, only alanine codons (GCN), and glycine codons (GGN) to a much smaller extent, are overrepresented at the second codon, whereas the other three codons are not overrepresented. While highly expressed genes have more +4G than lowly expressed genes, the difference is caused by GCN and GGN codons at the second codon. These results are inconsistent with +4G being needed for efficient translation initiation, but consistent with the proposal of amino acid constraint hypothesis

    Morphometric study of the root anatomy in furcation area of mandibular first molars

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    Furcation involvement in periodontal disease has been a challenge for the dentist. OBJECTIVE: The aim of this study was to investigate root dimensions in the furcation area of 233 mandibular first molars. MATERIAL AND METHODS: Digital photomicrographs were used to obtain the following measurements on the buccal and lingual surfaces of each tooth: root trunk height (RT), horizontal interadicular distance obtained 1 mm (D1) and 2 mm (D2) below the fornix and interadicular angle (IA). RESULTS: Mean± standard deviation of buccal and lingual furcation measurements were, respectively, 1.37±0.78 mm and 2.04±0.89 mm for RT; 0.86±0.39 mm and 0.71±0.42 mm for D1; 1.50±0.48 mm and 1.38±0.48 mm for D2; 41.68±13.20º and 37.78±13.18º for IA. Statistically significant differences were found between all measured parameters for buccal and lingual sides (p<0.05, paired t test). CONCLUSIONS: In conclusion, the lingual furcation of mandibular first molars presented narrower entrance and longer root trunk than the buccal furcation, suggesting more limitation for instrumentation and worse prognosis to lingual furcation involvements in comparison to buccal lesions

    The Complex and Important Cellular and Metabolic Functions of Saturated Fatty Acids

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    This review summarizes recent findings on the metabolism and biological functions of saturated fatty acids (SFA). Some of these findings show that SFA may have important and specific roles in the cells. Elucidated biochemical mechanisms like protein acylation (N-myristoylation, S-palmitoylation) and regulation of gene transcription are presented. In terms of physiology, SFA are involved for instance in lipogenesis, fat deposition, polyunsaturated fatty acids bioavailability and apoptosis. The variety of their functions demonstrates that SFA should no longer be considered as a single group

    Qualidade do melão cantaloupe cv. Hy-Mark minimamente processado e armazenado sob refrigeração Quality of cantaloupe melon cv. Hy-Mark minimally processed and refrigerated

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    A maioria das pesquisas em melão minimamente processado é concentrada no tipo 'cantaloupe', devido à sua importância no mercado internacional. O objetivo deste trabalho foi avaliar a qualidade do melão cantaloupe cv. Hy-Mark minimamente processado. Os frutos foram lavados e sanitizados, sendo em seguida cortados na forma de cubos, imersos em solução de CaCl2, acondicionados em embalagens flexíveis PET (tereftalato de polietileno) e armazenados a 4ºC por 9 dias. O delineamento foi conduzido inteiramente casualizado, com três repetições. A intervalos de três dias, amostras foram coletadas e analisadas quanto a coliformes totais e fecais, e contagem de bactérias aeróbias mesófilas e bolores e leveduras, pH, sólidos solúveis totais, acidez, vitamina C, açúcares redutores totais, atividade de água, firmeza e análise sensorial através do atributo de aceitação global. O melão minimamente processado apresentou boa estabilidade física, físico-química, microbiológica e sensorial, estimando-se em 9 dias a vida útil deste produto a 4ºC.<br>Most of the research into minimally processed melon is focused on cantaloupe melon, due to its importance in the international market. The objective of this work was to evaluate the quality of cantaloupe melon cv. Hy-Mark, minimally processed and refrigerated. Fruits were washed, sanitized, cut and imbedded in a calcium chloride solution and packed in PET and stored at 4ºC during 9 days. At three-day intervals samples were collected and analyzed for total and fecal coliforms, counting of aerobic mesophilic bacterias, molds and yeast, SS (soluble solids), TTA (total titrable acidity), firmness, pH, TRS (total reducing sugar), Aw, vitamin C, color and acceptability. The minimally processed melon showed good overall physical, pyisico-chemical, microbiological and sensorial stability during the storage period
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