Yeast as a model organism for studying the evolution of non-standard genetic codes

During the last 30 years, a number of alterations to the standard genetic code have been uncovered both in prokaryotes and eukaryotic nuclear and mitochondrial genomes. But, the study of the evolutionary pathways and molecular mechanisms of codon identity redefinition has been largely ignored due to the assumption that non-standard genetic codes can only evolve through neutral evolutionary mechanisms and that they have no functional significance. The recent discovery of a genetic code change in the genus Candida that evolved through an ambiguous messenger RNA decoding mechanism is bringing that naive assumption to an abrupt end by showing, in a rather dramatic way, that genetic code changes have profound physiological and evolutionary consequences for the species that redefine codon identity. In this paper, the recent data on the evolution of the Candida genetic code are reviewed and an experimental framework based on forced evolution, molecular genetics and comparative and functional genomics methodologies is put forward for the study of non-standard genetic codes and genetic code ambiguity in general. Additionally, the importance of using Saccharomyces cerevisiae as a model organism for elucidating the evolutionary pathway of the Candida and other genetic code changes is emphasised.publishe

Steroid–quinoline hybrids for disruption and reversion of protein aggregation processes

Reversing protein aggregation within cells may be an important tool to fight protein-misfolding disorders such as Alzheimer’s, Parkinson’s, and cardiovascular diseases. Here we report the design and synthesis of a family of steroid−quinoline hybrid compounds based on the framework combination approach. This set of hybrid compounds effectively inhibited Aβ1−42 self-aggregation in vitro by delaying the exponential growth phase and/or reducing the quantity of fibrils in the steady state. Their disaggregation efficacy was further demonstrated against preaggregated Aβ1−42 peptides in cellular assays upon their endocytosis by neuroblastoma cells, as they reverted both the number and the average area of fibrils back to basal levels. The antiaggregation effect of these hybrids was further tested and demonstrated in a cellular model of general protein aggregation expressing a protein aggregation fluorescent sensor. Together, our results show that the new cholesterol−quinoline hybrids possess wide and marked disaggregation capacities and are therefore promising templates for the development of new drugs to deal with conformational disorders.Thanks are due to the University of Aveiro, FCT/MEC, Centro 2020 and Portugal2020, the COMPETE Program, and the European Union (FEDER Program) via the financial support to the research units LAQV-REQUIMTE (UIDB/50006/2020), IBiMED (UID/BIM/04501/2019) and CICECO- Aveiro Institute of Materials (UID/CTM/50011/2019), financed by national funds through the FCT/MCTES, to the Portuguese NMR Network, to the ThiMES Project (POCI-01- 0145-FEDER-016630), and to the PAGE Project “Protein Aggregation Across the Lifespan” (CENTRO-01-0145- FEDER-000003), including postdoctoral grants to H.M.T.A. (BPD/UI98/4861/2017) and R.N.d.S. (BPD/UI98/6327/2018). M.P. was supported by Ph.D. Grant SFRH/BD/135655/2018. A.R.S. and S.G. were supported by national funds (OE) through FCT, I.P., in the scope of the framework contract foreseen in numbers 4, 5, and 6 of Article 23 of the Decree-Law 57/2016 of August 29, changed by Law 57/2017 of July 19. Microphotographs were acquired in the LiM facility of iBiMED/UA, a member of the Portuguese Platform of BioImaging (PPBI) (POCI-01-0145-FEDER-022122).info:eu-repo/semantics/publishedVersio

Multiplex immunoassay for inflammatory proteins quantification in saliva – a methodologic approach

info:eu-repo/semantics/publishedVersio

A genetic code alteration generates a proteome of high diversity in the human pathogen Candida albicans

Background - Genetic code alterations have been reported in mitochondrial, prokaryotic, and eukaryotic cytoplasmic translation systems, but their evolution and how organisms cope and survive such dramatic genetic events are not understood. Results - Here we used an unusual decoding of leucine CUG codons as serine in the main human fungal pathogen Candida albicans to elucidate the global impact of genetic code alterations on the proteome. We show that C. albicans decodes CUG codons ambiguously and tolerates partial reversion of their identity from serine back to leucine on a genome-wide scale. Conclusion - Such codon ambiguity expands the proteome of this human pathogen exponentially and is used to generate important phenotypic diversity. This study highlights novel features of C. albicans biology and unanticipated roles for codon ambiguity in the evolution of the genetic code.publishe

Dynamic aspects of oral processing of curcumin-loaded solid lipid nanoparticles yoghurts by rheology and soft tribology assessment

Mouthfeel and consistency originating from the consumption of yoghurts represent decisive factors for the consumers’ acceptability. The present work aims to identify potential differences in the sensory perceptions of yoghurts with curcumin-loaded solid lipid nanoparticles (SLN) incorporation. Thus, rheology coupled with tribology were considered to simulate the dynamic aspects of oral processing. Similar values for the elastic (G’) and viscous (G’’) parameters were obtained for both control yoghurt and yoghurt with curcuminloaded SLN, as well as flow index (n) and consistency index (k) around 0.56 and 1.50 Pa.sn, respectively. However, the friction behaviour was characterised by significant differences in all tribological regimes, suggesting that the incorporation of nanoparticles in yoghurts could potentially affect the after-feel upon swallowing, whilst not directly impacting on yoghurt’s structure

Chemical composition, antibacterial and antioxidant activities of the essential oil from Vismia guianensis fruits

In recent decades, the essential oils of plants have drawn great interest as sources of natural products. Essential oil from the fruits of Vismia guianensis was tested for its chemical constituents and antimicrobial and antioxidant activities. Gas chromatography/mass spectrometry (GC-MS) analysis of the essential oil revealed the presence of 38 sesquiterpenoids. The major components were β-caryophyllene (25.8%), α-copaene (13.1%), and δ-cadinene (11.6%). Antimicrobial activities were measured against six species of Gram negative and seven species of Gram positive bacteria and showed antibacterial activity against the human pathogenic Gram-positive bacteria Staphylococcus lentus with minimum inhibitory concentration (MIC) values of 78 μg/ml. The antioxidant activity of the essential oil was evaluated using the beta carotene/linoleic acid assay and showed antioxidant activity.Key words: Vismia guianensis, chemical composition, antibacterial, antioxidant, fruits, essential oil

Non-transferrin-bound iron determination in blood serum using microsequential injection solid phase spectrometry – proof of concept

Non-transferrin-bound iron (NTBI) is a group of circulating toxic iron forms, which occur in iron overload or health conditions with dysregulation of iron metabolism. NTBI is responsible for increased oxidative stress and tissue iron loading. Despite its relevance as a biochemical marker in several diseases, a standardized assay is still lacking. Several methods were developed to quantify NTBI, but results show high inter-method and even inter-laboratory variability. Thus, the development of a consistent NTBI assay is a major goal in the management of iron overload and related clinical conditions. In this work, a micro sequential injection lab-on-valve (μSI-LOV) method in a solid phase spectrophotometry (SPS) mode was developed for the quantification of NTBI, using a bidentate 3,4–hydroxypyridinone (3,4-HPO) ligand anchored to sepharose beads as a chromogenic reagent. To attain SPS, the functionalized beads were packed into a column in the flow cell, and the analyte, NTBI retained as iron (III), formed a colored complex at the beads while eliminating the sample matrix. The dynamic concentration range was 1.62–7.16 μmol L−1 of iron (III), with a limit of detection of 0.49 μmol L−1 and a limit of quantification of 1.62 μmol L−1. The proposed μSI-LOV-SPS method is a contribution to the development of an automatic method for the quantification of the NTBI in serum samples.info:eu-repo/semantics/publishedVersio

Methane Emissions from Ruminants on Integrated Crop-Livestock Systems

Ruminant livestock produce ~80 million tonnes of methane (CH4) annually, accounting for ~33% of global anthropogenic emissions of CH4 (Beauchemin et al. 2008). CH4 is a powerful greenhouse gas, with global warming potential of 25 (Eckard et al. 2010), and represents a significant loss of dietary energy (2 to 12% of gross energy of feeds; Patra 2012) in the ruminant production system. Despite greenhouse gas (GHG) emissions have become an increasingly important topic worldwide, there is still a high variability around the estimated values of these emissions, mainly about emissions attributable to livestock (range from 8 to 51%; Herrero et al. 2011). This variability creates confusion among researchers, policy makers and the public, particularly in tropical/sub-tropical regions due substantial uncertainties. Therefore, using rigorous and internationally accepted protocols, a Brazilian national project was established in order to contribute for the estimates of GHG emissions attributable to livestock in Brazilian ruminant production systems. Moreover, enteric CH4 emissions are a major challenge for research, in order to develop technologies and strategies for sustainable ruminant production systems in the future (Eckard et al. 2010). In recent years, integrated crop-livestock systems (ICLS) have gained interest due to, for example, the abatement of methane from livestock production: directly through a reduction in CH4 per unit of animal products resulting from the increase on feed quality and animal welfare (i.e. improved environmental temperature for ICLS with trees), and indirectly through reduction of area submitted to land use changes (i.e. leading to a loss of soil C stocks). This paper deals with the preliminary results from CH4 emissions by beef heifers grazing in two ICLS (i.e. production system that integrates corn or soybeans crops, during the warm season, and cattle grazing on a cool season pasture, on the same area and in the same cropping year, with or without trees), how these findings contributes to determine the soil C balance and mitigation measures

Cannabidiol and cannabigerol exert antimicrobial activity without compromising skin microbiota

Cannabidiol (CBD) and cannabigerol (CBG) are two pharmacologically active phytocannabinoids of Cannabis sativa L. Their antimicrobial activity needs further elucidation, particularly for CBG, as reports on this cannabinoid are scarce. We investigated CBD and CBG’s antimicrobial potential, including their ability to inhibit the formation and cause the removal of biofilms. Our results demonstrate that both molecules present activity against planktonic bacteria and biofilms, with both cannabinoids removing mature biofilms at concentrations below the determined minimum inhibitory concentrations. We report for the first time minimum inhibitory and lethal concentrations for Pseudomonas aeruginosa and Escherichia coli (ranging from 400 to 3180 µM), as well as the ability of cannabinoids to inhibit Staphylococci adhesion to keratinocytes, with CBG demonstrating higher activity than CBD. The value of these molecules as preservative ingredients for cosmetics was also assayed, with CBG meeting the USP 51 challenge test criteria for antimicrobial effectiveness. Further, the exact formulation showed no negative impact on skin microbiota. Our results suggest that phytocannabinoids can be promising topical antimicrobial agents when searching for novel therapeutic candidates for different skin conditions. Additional research is needed to clarify phytocannabinoids’ mechanisms of action, aiming to develop practical applications in dermatological use.info:eu-repo/semantics/publishedVersio

Microbial DNA extraction methods for microbial screening - saliva vs biofilm comparison

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