Expression, purification and structural analysis of the Pyrococcus abyssi RNA binding protein PAB1135

<p>Abstract</p> <p>Background</p> <p>The gene coding for the uncharacterized protein PAB1135 in the archaeon <it>Pyrococcus abyssi </it>is in the same operon as the ribonuclease P (RNase P) subunit Rpp30.</p> <p>Findings</p> <p>Here we report the expression, purification and structural analysis of PAB1135. We analyzed the interaction of PAB1135 with RNA and show that it binds efficiently double-stranded RNAs in a non-sequence specific manner. We also performed molecular modeling of the PAB1135 structure using the crystal structure of the protein Af2318 from <it>Archaeoglobus fulgidus </it>(<ext-link ext-link-id="2OGK" ext-link-type="pdb">2OGK</ext-link>) as the template.</p> <p>Conclusions</p> <p>Comparison of this model has lead to the identification of a region in PAB1135 that could be involved in recognizing double-stranded RNA.</p

Influencia del procesado industrial sobre las características químico-físicas y contenido en lípidos y antioxidantes del salvado de arroz

Se investigaron las características químico-físicas de los salvados de arroz en distintos momentos de su procesado industrial, incluyendo materia prima (salvados de arroz blanco y vaporizado), salvado peletizado y salvado desgrasado. Se determinó la humedad, la acidez y el índice de peróxidos. Utilizando extractos en éter de petróleo y cromatografía de gases, se estableció el contenido lipídico total y los perfiles de los ácidos grasos. Mediante HPLC se determinaron los tocoferoles y el γ-orizanol. El salvado de arroz vaporizado mostró la mayor concentración de lípidos y la menor acidez, lo que indica que el vaporizado es el proceso más eficaz para estabilizar el salvado. Los ácidos oleico, linoleico y palmítico fueron los mayoritarios en todas las muestras. El peletizado no produjo pérdidas de lípidos, ni de tocoferoles y γ-orizanol. Todas las muestras de salvado mostraron contenidos altos de α-tocoferol, intermedios de γ-tocoferol y pequeños de δ-tocoferol. Se detectaron nueve componentes del γ-orizanol, con una mayor proporción del componente eluido en cuarta posición, probablemente, ferulato de 24- metilén cicloartenilo. El salvado peletizado presentó el mayor contenido de tocoferoles, mientras que el de arroz vaporizado mostró el mayor contenido de γ-orizanol.A comparative study of the physico-chemical characteristics of rice bran during the successive steps of its industrial processing was carried out and included white and parboiled rice brans and pelletized and defatted rice brans. Moisture, acidity and peroxide index were determined. Using extracts in petroleum ether and gas chromatography, the total fat contents and the profiles of the fatty acids were established. The tocopherols and γ-oryzanol were determined using high performance liquid chromatography. The bran of parboiled rice showed the largest fat content and the lowest acidity, indicating that parboiling is the most effective process for bran stabilization. Oleic, linoleic and palmitic acids predominated in all the samples. Pelletization did not produce a loss in lipids, tocopherols or γ-oryzanol. All the samples showed higher contents of α-tocopherol, intermediate contents of γ-tocopherol and much lower concentrations of δ-tocopherol. Nine components of γ- oryzanol were detected, with a major proportion of the component that eluted in the fourth position, probably ferulate of 24-methylene cycloartenol. Pelletized rice bran showed the highest tocopherol content, whereas parboiled rice bran yielded the largest γ-oryzanol [email protected]

Different carbon nanostructured materials obtained in catalytic chemical vapor deposition

Different carbon nanostructured materials, such as nanotubes, nanofibers, nanosprings and nanooctopus, were grown by changing the metal catalyst and experimental parameters of the thermal chemical vapor deposition process. These experiments were performed using a tubular furnace and methane or acetylene as carbon feedstock gases. Thin films of Ni or Cu were deposited onto a SiO2/Si substrate and employed as catalysts. The effect of the growth temperature, metal catalyst and carbon gas precursor (methane or acetylene) on the final carbon nanoestructured material was studied by scanning electron microscopy, Raman spectroscopy and grazing incidence X-ray diffraction. Growth of multiwall carbon nanotubes (MWCNTs) was observed using both metal films and carbon precursor gases, whereas partially oxidized Ni films promoted formation of nanosprings. Experiments with reduced supply of methane resulted in octopus-like carbon nanostructures when a Cu film was used as a catalyst.Diferentes materiais nanoestruturados à base de carbono, tais como nanotubos, nanofibras, nanomolas e nanooctopus, foram obtidos através do processo de deposição química de vapor. Tais experimentos foram realizados em um forno tubular e variações nos parâmetros experimentais permitiram a obtenção das diferentes nanoestruturas de carbono. Filmes finos de Ni e Cu foram depositados sobre substratos de SiO2/Si e empregados como catalisadores. O efeito de diferentes gases precursores de carbono, da temperatura de crescimento e do metal catalítico sobre as características do material final foi investigado por microscopia eletrônica de varredura, espectroscopia Raman e difratometria de raios X com ângulo rasante. O uso dos gases metano e acetileno levaram à formação de nanotubos de carbono para ambos os filmes metálicos, enquanto Ni parcialmente oxidado promoveu o crescimento de nanomolas. Estruturas do tipo octopus foram obtidas a partir do uso de cobre associado a um fornecimento relativamente restrito de metano.11241132Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Unravelling the biological potential of Pinus pinaster bark extracts

Natural compounds from agro-food by-products have fostered interest in food industries. The aim of this study was to unravel potential uses for Pinus pinaster bark extracts (PBE). As functional features of this type of extracts are usually attributed to phenolic compounds, the extraction process was studied. Different PBEs were achieved, with high content in phenolic compounds, using different water/ethanol combinations as a solvent. These PBEs were chemically characterized, and their bioactivity and in vitro cell viability were evaluated. Extracts obtained with hydroethanolic solvents had higher content in phenolic and flavonoid compounds. All the PBEs presented high antioxidant, antibacterial and antihyperglycemic activities. Moreover, PBEs have low cytotoxicity and a selective activity against cancer cells as these were negatively affected. These features may allow the extracts to be used in food formulation and processing (as preservatives, antioxidants or bioactive ingredients), but they showed also potential for the pharmaceutical or nutraceutical sectors.This research was funded by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund (FEDER) under the scope of Norte2020—Programa Operacional Regional do Norte, by program INTERREG V-B Sudoe (REDVALUE, SOE1/P1/E0123) and by project MOBFOOD (POCI-01-0247-FEDER-024524), cofounded by PORTUGAL2020, Lisb@a2020, COMPETE 2020 and the European Union. Zlatina Genisheva is supported by the project OH2O (POCI-01-0145-FEDER-029145) funded by FCT and FEDER under the scope of Programa Operacional de Competividade e Internacionalizaçao (POCI)-COMPETE 2020 and PORTUGAL2020. Pedro Santos is recipient of a fellowship supported by a doctoral advanced training (call NORTE-69-2015-15), funded by the European Social Fund under the scope of Norte2020 (NORTE-08-5369-FSE-000036).info:eu-repo/semantics/publishedVersio

Design of green routes for cellulose extraction from a sugarcane by-product

info:eu-repo/semantics/publishedVersio

Risk assessment of exposure to multiple mycotoxins in food

Moulds may produce a diversity of toxins such as aflatoxins, ochratoxins, trichothecenes, zearalenone, fumonisins and others. Although toxicological, environmental and epidemiological studies have addressed the problem of these toxins one by one, more than one mycotoxin are found usually in the same contaminated food. Risk assessment for humans potentially exposed to multimycotoxins suffers very much from the lack of adequate food consumption data. Furthermore, for a given mycotoxin, synergism and antagonism with other mycotoxins, found in the same food commodities, are not taken into account. Aflatoxin B1 and ochratoxin A belong to the most frequently occurring mycotoxins. This has repeatedly been demonstrated, however, normally, the risk resulting from their simultaneous occurrence is not considered. A descriptive study was developed to monitor air fungal contamination in one hospital food unit

Identification of archaeal proteins that affect the exosome function in vitro

<p>Abstract</p> <p>Background</p> <p>The archaeal exosome is formed by a hexameric RNase PH ring and three RNA binding subunits and has been shown to bind and degrade RNA <it>in vitro</it>. Despite extensive studies on the eukaryotic exosome and on the proteins interacting with this complex, little information is yet available on the identification and function of archaeal exosome regulatory factors.</p> <p>Results</p> <p>Here, we show that the proteins PaSBDS and PaNip7, which bind preferentially to poly-A and AU-rich RNAs, respectively, affect the <it>Pyrococcus abyssi </it>exosome activity <it>in vitro</it>. PaSBDS inhibits slightly degradation of a poly-rA substrate, while PaNip7 strongly inhibits the degradation of poly-A and poly-AU by the exosome. The exosome inhibition by PaNip7 appears to depend at least partially on its interaction with RNA, since mutants of PaNip7 that no longer bind RNA, inhibit the exosome less strongly. We also show that FITC-labeled PaNip7 associates with the exosome in the absence of substrate RNA.</p> <p>Conclusions</p> <p>Given the high structural homology between the archaeal and eukaryotic proteins, the effect of archaeal Nip7 and SBDS on the exosome provides a model for an evolutionarily conserved exosome control mechanism.</p

Design of innovative biocompatible cellulose nanostructures for the delivery and sustained release of curcumin

Poor aqueous solubility, stability and bioavailability of interesting bioactive compounds is a challenge in the development of bioactive formulations. Cellulose nanostructures are promising and sustainable carriers with unique features that may be used in enabling delivery strategies. In this work, cellulose nanocrystals (CNC) and cellulose nanofibers were investigated as carriers for the delivery of curcumin, a model liposoluble compound. Nanocellulose modification with the surfactant cetyltrimethylammonium bromide (CTAB), tannic acid and decylamine (TADA), and by TEMPO-mediated oxidation were also tested and compared. The carrier materials were characterized in terms of structural properties and surface charge, while the delivery systems were evaluated for their encapsulation and release properties. The release profile was assessed in conditions that mimic the gastric and intestinal fluids, and cytotoxicity studies were performed in intestinal cells to confirm safe application. Modification with CTAB and TADA resulted in high curcumin encapsulation efficiencies of 90 and 99%, respectively. While no curcumin was released from TADA-modified nanocellulose in simulated gastrointestinal conditions, CNC-CTAB allowed for a curcumin-sustained release of ca. 50% over 8 h. Furthermore, the CNC-CTAB delivery system showed no cytotoxic effects on Caco-2 intestinal cells up to 0.125 g/L, meaning that up to this concentration the system is safe to use. Overall, the use of the delivery systems allowed for the reduction in the cytotoxicity associated with higher curcumin concentrations, highlighting the potential of nanocellulose encapsulation systems.info:eu-repo/semantics/publishedVersio

Amentadione is a new modulating agent for osteoarthritis in an ex-vivo co-culture preclinical assay

Osteoarthritis (OA) is a whole-joint disease where inflammation interplays with extracellular matrix mineralization in a cycle that leads to its degradation. The lack of effective preventing treatments and disease modifying agents, demands new therapeutic targets and development of effective drugs.FCT: SFRH/BD/111824/2015info:eu-repo/semantics/publishedVersio

Translating COGNISTAT and the Use of the Cognitive Interview Approach: Observations and Challenges

Cognitive functioning is a salient issue among people in the late adulthood stage where mental health declines with age. A common cognitive disability among elders is Dementia including Alzheimer’s disease. Cognitive screening tools such as the Mini- Mental Status Examination and the Montreal Cognitive Assessment are most commonly used to measure the cognitive ability areas leading to a diagnostic evaluation. COGNISTAT as a neuropsychological instrument is a recent screener being introduced to a few outpatient clinics. In this study; COGNISTAT is translated to Filipino for the first time using a rigorous procedure and pilot tested on elderly volunteers in local settings. The translation was done by an interdisciplinary team of a Geriatric Physician; a Speech Pathologist; and a Clinical Psychologist. The Filipino-translated COGNISTAT was administered to 22 elders in two batches using the cognitive interview method. Challenges were identified in the initial testing phase which has implications for future adaptations of foreign instruments