60,080 research outputs found

    Effect of enzymatic hydrolysis on bioactive properties of the mussel Mytilus galloprovincialis

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    Disserta√ß√£o de mestrado em Tecnologia e Ci√™ncia AlimentarOs oceanos e a diversidade de ambientes marinhos constituem uma fonte de diversos compostos qu√≠micos e biol√≥gicos de valor acrescentado. A valoriza√ß√£o destes recursos e a otimiza√ß√£o dos processos de extra√ß√£o recorrendo a tecnologias sustent√°veis e eficientes permite a s√≠ntese compostos biol√≥gicos que podem ser usados no desenvolvimento de alimentos funcionais, nutrac√™uticos, cosm√©ticos e medicamentos. O presente trabalho teve como objetivo estudar a otimiza√ß√£o do processo de hidr√≥lise enzim√°tica do mexilh√£o Mytilus galloprovincialis com a enzima Alcalase, para produ√ß√£o de p√©ptidos com propriedades antioxidante, anti-hipertensiva e antimicrobiana. O mexilh√£o foi obtido como subproduto de aquacultura da costa portuguesa, do qual foi aproveitada apenas a carne. A composi√ß√£o qu√≠mica aproximada determinada foi de 81,4 g de humidade, 12,6 g de prote√≠na, 1,7 g de l√≠pidos, 1,4 g de minerais e 2,8 g de hidratos de carbono. Foi usada a Metodologia de Superf√≠cie de Resposta atrav√©s de um desenho experimental composto central rotacional 23 com dezasseis ensaios para avaliar os efeitos das vari√°veis concentra√ß√£o de substrato (25, 50 e 75 g/150 g), concentra√ß√£o de enzima (1, 2 e 3 % v/m) e tempo (1, 2 e 3 h) tendo como resposta o grau de hidr√≥lise (GH %). Os resultados obtidos foram analisados atrav√©s do software Statistica vers√£o 10 e a signific√Ęncia estat√≠stica atrav√©s da an√°lise da vari√Ęncia (ANOVA). As condi√ß√Ķes √≥timas do ponto de vista industrial s√£o 25 g de substrato, 2 % de concentra√ß√£o de enzima e tempo de rea√ß√£o de 1 h. Os hidrolisados obtidos foram avaliados quanto √† sua capacidade antioxidante atrav√©s do m√©todo da atividade sequestradora do radical livre DPPH. O hidrolisado com maiores valores atividade antioxidante foi com concentra√ß√£o de substrato de 75 g/150 g, concentra√ß√£o de enzima de 2 % e tempo de rea√ß√£o de 2 h. O aumento concentra√ß√£o de substrato demonstrou influenciar positivamente a atividade antioxidante. Os ensaios de atividade anti-hipertensiva n√£o foram conclusivos para as amostras testadas. Os hidrolisados manifestaram atividade antimicrobiana nos microrganismos S. aureus e B. cereus. A bact√©ria E. coli n√£o sofreu qualquer inibi√ß√£o de crescimento. O ensaio com maior halo de inibi√ß√£o foi de concentra√ß√£o de substrato de 25 g, concentra√ß√£o de enzima de 1 % e tempo de rea√ß√£o de 1 h.Oceans and the diversity of marine environments are a source of diverse value-added chemical and biological compounds. The valorisation of these resources and the optimization of extraction processes using sustainable and efficient technologies allows the synthesis of biological compounds that can be used in the development of functional foods, nutraceuticals, cosmetics and medicines. The present work aimed to study the optimization of the enzymatic hydrolysis process of the mussel Mytilus galloprovincialis with the enzyme Alcalase, for the production of peptides with antioxidant, antihypertensive and antimicrobial properties. The mussel was obtained as an aquaculture byproduct from the Portuguese coast, which was used only for meat. The approximate chemical composition determined was 81.4 g moisture, 12.6 g protein, 1.7 g lipids, 1.4 g minerals and 2.8 g carbohydrates. Response Surface Methodology through a rotational central composite experimental design 23 with sixteen assays was used to evaluate the effects of substrate concentration (25, 50 and 75 g/150 g), enzyme concentration (1, 2 and 3 %) and time (1, 2 and 3 h) having as response the degree of hydrolysis (GH %). The results were analyzed using Statistica software version 10 and statistical significance through analysis of variance (ANOVA). The optimum conditions studied were 25 g of substrate, 3 % of enzyme concentration and 3 h reaction time, where a hydrolysis degree of 35.1 % was achieved. The hydrolysates obtained were evaluated for their antioxidant capacity by the DPPH free radical scavenging method. The hydrolyzate with the highest antioxidant activity was with 75 g/150 g substrate concentration, 2 % enzyme concentration and 2 h reaction time. Increased substrate concentration has been shown to positively influence antioxidant activity, however this does not apply to enzyme concentration and reaction time. No relationship was observed between the degree of hydrolysis and antioxidant capacity. Antihypertensive activity assays were not conclusive for the samples tested. Hydrolysates showed antimicrobial activity in S. aureus and B. cereus. The bacterium E. coli did not suffer any growth inhibition. The assay with the largest inhibition halo was 25 g/150 g substrate concentration, 1 % enzyme concentration and 1 h reaction time

    Monoolein cubosome formation using solvent-exchange in bulk and microfluidics: the effect of DODAB and DDAB addition

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    Disserta√ß√£o de mestrado em Biophysics and BionanosystemsMonoolein is an ubiquitous lipid in the production of drug delivery formulations mostly because of its propensity to form highly attractive cubosome nanoparticles. Cubosomes consist of dispersions of lipid bicontinuous cubic phases in water typically, stabilized with an amphiphilic polymer. Solvent-exchange is an energetically efficient and scalable process where the lipid is dissolved in a solvent miscible with water, like ethanol, and rapidly added to an aqueous solution. However, this process has issues mainly related to the poor size control over the particles produced. Previously, it was observed that by employing a solvent exchange approach in a microfluidic device, one could have control over the size of monoolein nanoparticles by manipulating the flow rate ratio between the ethanolic lipid solution and the two aqueous side-streams. It was shown that the particle size decreased with the increase of the flow rate ratio. In this work, we explore the effect of the addition of cationic lipid (DDAB or DODAB) to monoolein nanoparticles with the goal of embedding cationic charge on such particles to make them suitable for the encapsulation of nucleic acids. Contrary to neat monoolein, the presence of cationic lipids makes the final particle size independent of the flow rate. Additionally, the particles are markedly smaller, with the size of cationic lipid-monoolein nanoparticles dependent on total lipid concentration and the proportion of cationic lipid to monoolein. The size of the nanoparticles produced decrease with the increase of the ratio of cationic lipid to monoolein or by decreasing the total lipid concentration. In respect to the particles' structure, SAXS measurements did not provide evidence of cubic structures. Nonetheless, when samples are observed with Transmission Electron Microscopy with negative staining, hexagonal patterns are visible. This observation may indicate the presence of a cubic phase, but alternative scenarios should still be considered.Na produ√ß√£o de formula√ß√Ķes para entrega de f√°rmacos a monole√≠na √© considerado um l√≠pido universal com base na sua propens√£o para formar cubossomas. Estas nanopart√≠culas consistem em dispers√Ķes de fases c√ļbicas bicont√≠nuas em √°gua, tipicamente estabilizadas com um pol√≠mero anfif√≠lico. O m√©todo de troca de solventes √© um processo energeticamente eficiente e reproduz√≠vel onde o l√≠pido √© dissolvido num solvente misc√≠vel com a √°gua, como o etanol, e rapidamente adicionado a uma solu√ß√£o aquosa. No entanto, este processo apresenta alguns entraves como a aus√™ncia de controlo sobre o tamanho das part√≠culas produzidas. Anteriormente foi observado que a aplica√ß√£o do m√©todo de troca de solventes em microflu√≠dica possibilitou o controlo do tamanho das nanopart√≠culas de monole√≠na atrav√©s da manipula√ß√£o do r√°cio de fluxo entre a solu√ß√£o de l√≠pido dissolvido em etanol e as solu√ß√Ķes aquosas laterias, mais especificamente, pelo aumento do r√°cio de fluxo, o tamanho das part√≠culas diminu√≠a. Neste trabalho exploramos o efeito da adi√ß√£o de l√≠pido cati√≥nico (DDAB ou DODAB) nas nanopart√≠culas de monole√≠na com o objetivo de incorporar carga positiva nas part√≠culas, capacitando-as a encapsular √°cidos nucleicos. Contrariamente ao que acontecia num sistema de monole√≠na puro, na presen√ßa de l√≠pidos cati√≥nicos o tamanho final das part√≠culas torna-se independente das condi√ß√Ķes de fluxo. Al√©m disso, as part√≠culas que se formam s√£o significativamente mais pequenas, sendo que o tamanho das nanopart√≠culas de l√≠pido cati√≥nico e monole√≠na √© dependente da concentra√ß√£o de l√≠pido total a da propor√ß√£o de l√≠pido cati√≥nico para monole√≠na. Essencialmente, o tamanho das nanopart√≠culas produzidas diminui com o aumento do r√°cio de l√≠pido cati√≥nico para monole√≠na e pela diminui√ß√£o da concentra√ß√£o de l√≠pido total da formula√ß√£o. De acordo com a estrutura das part√≠culas, quando realizamos testes em SAXS n√£o h√° evid√™ncia de estruturas c√ļbicas, no entanto, quando as amostras s√£o submetidas a an√°lises em TEM com marca√ß√£o negativa, estruturas com padr√Ķes hexagonais s√£o vis√≠veis. Esta observa√ß√£o poder√° indicar a presen√ßa de uma fase c√ļbica, mas cen√°rios alternativos ainda t√™m que ser considerados

    Application of zebrafish and murine models in lipoprotein metabolism and atherosclerosis research

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    Cardiovascular diseases are still a major concern for the global health. The main underlying pathology of this disease is atherosclerosis which is characterized by the accumulation of lipids and immune cells in the arterial wall leading to a chronic local inflammation and lesion formation. In this thesis, we aimed to (1) validate the use of zebrafish in cholesterol metabolism and atherosclerosis research, (2) study the role of certain classes of scavenger receptors in lipoprotein uptake and cholesterol-based functions, and (3) validated two immune-based potential targets for atherosclerosis.</p

    Marker gene mRNA expressions in C2C12 cells grown in adipocyte differentiation medium.

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    (A) C2C12 cells were grown in adipocyte differentiation medium for a total of 5 days as described in the Materials and Methods. The concentrations of accumulated lipids were measured by the Oil Red O staining method. (B-G) Marker gene mRNA expressions in the C2C12 cells grown in adipocyte differentiation medium were monitored by real-time RT-PCR analysis using the specific primer set for each of the genes. The data indicate relative expressions compared to cells grown in growth medium after normalization with GAPDH mRNA expression. Error bars indicate standard deviations (n = 3). Asterisks (*: p p < 0.001) indicate that the difference is statistically significant between two groups.</p

    Metabolomic evaluation of PGPR defence priming in wheat (Triticum aestivum L.) cultivars infected with Puccinia striiformis f. sp. tritici (stripe rust)

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    Plant-microbe interactions are a phenomenal display of symbiotic/parasitic relationships between living organisms. Plant growth-promoting rhizobacteria (PGPR) are some of the most widely investigated plant-beneficial microbes due to their capabilities in stimulating plant growth and development and conferring protection to plants against biotic and abiotic stresses. As such, PGPR-mediated plant priming/induced systemic resistance (ISR) has become a hot topic among researchers, particularly with prospects of applications in sustainable agriculture. The current study applies untargeted ultra-high performance liquid chromatography-high-definition mass spectrometry (UHPLC-HDMS) to investigate PGPR-based metabolic reconfigurations in the metabolome of primed wheat plants against Puccinia striiformis f. sp. tricti (Pst). A seed bio-priming approach was adopted, where seeds were coated with two PGPR strains namely Bacillus subtilis and Paenibacillus alvei (T22) and grown under controlled conditions in a glasshouse. The plants were infected with Pst one-week post-germination, followed by weekly harvesting of leaf material. Subsequent metabolite extraction was carried out for analysis on a UHPLC-HDMS system for data acquisition. The data was chemometrically processed to reveal the underlying trends and data structures as well as potential signatory biomarkers for priming against Pst. Results showed notable metabolic reprogramming in primary and secondary metabolism, where the amino acid and organic acid content of primed-control, primed-challenged and non-primed-challenged plants were differentially reprogrammed. Similar trends were observed from the secondary metabolism, in which primed plants (particularly primed-challenged) showed an up-regulation of phenolic compounds (flavonoids, hydroxycinnamic acids-HCAs- and HCA amides) compared to the non-primed plants. The metabolomics-based semi-quantitative and qualitative assessment of the plant metabolomes revealed a time-dependent metabolic reprogramming in primed-challenged and primed-unchallenged plants, indicating the metabolic adaptations of the plants to stripe rust infection over time

    Divergence of Liver Lipidomes in Tibetan and Yorkshire Pigs Living at Different Altitudes

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    The Tibetan pig is a characteristic breed of the Qinghai-Tibet Plateau with distinct physiological and meat quality attributes. The liver lipid profile can offer an important perspective to explore the uniqueness of Tibetan pigs. A quantitative comparison of liver lipidomes revealed significant differences in the lipid profiles between Tibetan and Yorkshire pigs raised at different altitudes. The abundance of lipids in the livers of pigs raised at a high altitude was higher than that of pigs raised at a lower altitude, whereas the abundance of lipids in the livers of Yorkshire pigs was higher than that of Tibetan pigs raised at the same altitude. Of the 1101 lipids identified, 323 and 193 differentially abundant lipids (DALs) were identified in the pairwise comparisons of Tibetan and Yorkshire pigs raised at different altitudes, respectively. The DALs of Tibetan pigs consisted mainly of 161 triglycerides, along with several acylcarnitines, represented by carnitine C2:0, and significant changes in the abundance of some phospholipids. The DALs of Yorkshire pigs were more complex, with significant increases in the abundance of triglycerides, cholesteryl esters, and free fatty acids, and decreases in the abundance of some phospholipids. This research provides strong theoretical and data support for the high-quality development of the highland livestock industry

    –źmelioration by phytoadaptogene of effects of balneofactors of Truskavets‚Äô Spa on patients with post-radiation encephalopathy

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    Background.¬†We have previously explored effects of Ukrainian¬†phytocomposition ‚ÄúBalm Truskavets‚Äô‚Ä̬†on parameters of neuro-endocrine-immune complex and biophotonics in humans with maladaptation.¬†It is known that in patients with post-radiation encephalopathy the reaction to some stimuli is significantly changed, therefore it needs correction.¬†The purpose of this study is to test the ability of this phytocomposition to amelioration the effects of standard balneotherapeutic complex in patients with post-radiation encephalopathy.¬†Material and methods. The research was carried out through a retrospective analysis of the database of the Truskavetsian Scientific School of Balneology, which remained unpublished. The object of observation in 1997 were 19 men and 3 women with urate¬†urolithiasis and chronic pyelonephritis¬†who were exposed to pathogenic factors of the accident at the Ch–ĺrnobylian nuclear power plant during the liquidation of its consequences in 1986-87. The survey was conducted twice: on admission and after two weeks of rehabilitation in sanatorium ‚ÄúPerlyna Prykarpattya‚Ä̬†(Truskavets‚Äô Spa). 11 patients received standard balneotherapy¬†while¬†the other 11 patients additionally received the phytocomposition¬†‚ÄúBalm Truskavets‚Äô‚ÄĚ. According to the protocol, blood pressure,¬†routine hematological and biochemical blood parameters were determined. In addition, physical working capacity (PWC150)¬†as well as EEG, heart rate variability (HRV) and immunity parameters were determined.¬†Results. Standard balneotherapy increases the decreased level of T-helper lymphocytes, but further decreases the level of B-lymphocytes, glomerular filtration rate and PWC150, in combination with increased normal levels of blood creatinine and urea, as well as decreased levels of diastolic BP and heart rate.¬†This is accompanied by a further increase in the sympathetic tone and the leveling of the increased of ULF band HRV as marker of level in the plasma of catecholamines and glucocorticoids.¬†Additional use of phytocomposition limits the adverse effects of standard balneotherapy by modulating EEG and HRV parameters. Conclusion. Phytocomposition¬†"Balm¬†Truskavets'" by¬†modulating¬†the¬†parameters¬†of¬†the¬†nervous¬†system¬†limits¬†the¬†adverse¬†effects¬†of¬†standard¬†balneotherapy¬†at¬†the¬†Truskavets‚Äô¬†Spa¬†in¬†patients¬†with¬†post-radiation¬†encephalopathy

    Morphological and Molecular Investigations of Aquaporin-7 (AQP-7) in Male <i>Camelus dromedarius</i> Reproductive Organs

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    Aquaporins (AQP) are involved in bidirectional transfers of water and small solutes across cell membranes. They are present in all tissues. However, the expression of AQP-7 has not yet been demonstrated in the reproductive tract of the camelid Camelus dromedarius. The study presented here concerns the immunohistochemical evidence of aquaporin-7 (AQP-7) in different parts of the male genital tract of Camelus dromedarius. To check the immune reactivity levels of anti-AQP-7 antibody in the male genital tract of Camelus dromedarius, the testes (proximal part, distal part and rete testis), epididymis (head, body and tail), ductus deferens (initial, middle and ampullary part) and prostate gland (compact and disseminated part) were collected from 12 male camels during the rutting and non-rutting seasons and subjected to immunohistochemistry. The result showed that the highest level of AQP-7 mRNA expression was in the testis of rutting and non-rutting males compared to the ductus deferens, epididymis and prostate. In addition, the highest mRNA gene expression of AQP-7 was in rutting males compared to non-rutting males. AQP-7 mRNA expression was higher in the ret testis, the body of the epididymis, the ampullary part of the ductus deferens and the compact part of the prostate. The immune reactivity levels of AQP-7 in rutting males showed strong reactivity in the testis and prostate compared to the epididymis and ductus deferens. On the basis of the results, it can be concluded that the distribution of the AQP-7 transcript and protein varied among rutting and non-rutting seasons and that the physiological roles of AQP-7 in the transportation of lipids, energy and water should be considered the main challenge in the activity and establishment of male Camelus dromedarius fertility during the rutting and non-rutting seasons. Moreover, AQP-7 detection is critical in assessing regulation and screening for new modulators that can prompt the development of effective medication to enhance fertility during rutting and non-rutting seasons

    Untargeted lipidomics in overweight males with cardiovascular diseases - influence of smoking status : diploma thesis

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    PuŇ°enje duhana odgovorno je za prerani razvoj kardiovaskularnih bolesti razlińćitim mehanizmima, a abnormalne razine lipidnih seruma i razina lipoproteina jedna su od posljedica. Lipidi su biomolekule koje igraju vitalnu ulogu u raznim fiziopatologijama. Kako bi se otkrila funkcija lipida, od iznimne je vaŇĺnosti identificirati i kvantificirati pojedinańćne molekularne vrste lipida u sloŇĺenim bioloŇ°kim sustavima. Lipidomika je brzo razvijajuńáa analitińćka tehnika sposobna mjeriti stotine lipida i trenutno je na ńćelu znanstvenih istraŇĺivanja zbog vaŇĺnosti lipida u zdravlju i bolesti. Cilj ove studije bio je procijeniti uńćinak puŇ°enja cigareta na profil lipida u pacijenata s prekomjernom tjelesnom teŇĺinom s veńá razvijenom kardiovaskularnom boleŇ°ńáu pomońáu tekuńáe kromatografije (LC) spojene online s masenom spektrometrijom (MS). Sveukupno zapaŇĺanje ove studije bilo je da je doŇ°lo do poveńáanja koncentracije masnih kiselina i nekih specifińćnih triglicerida (TG) i diglicerida (DG) te do smanjenja koncentracije kolesterol estera (i zasińáene i nezasińáene) , fosfatidilkolina (osobito nezasińáene) i lizofosfatidikolina (osobito zasińáene), i TG zasińáen u puŇ°ańća u usporedbi s nepuŇ°ańćima. Stoga se na temelju ove studije moŇĺe reńái da puŇ°enje utjeńće i naruŇ°ava profil lipida, ali kod pacijenata s veńá postojeńáim kardiovaskularnim bolestima i mnogim dodatnim ńćimbenicima puŇ°enje moŇĺda nema tako znańćajan utjecaj kao prije razvoja bolestiTobacco smoking is responsible for the premature development of cardiovascular disease by various mechanisms, and abnormal serum lipid profile and lipoprotein levels are one of the consequences. Lipids are crucial small biomolecules and play vital roles in a variety of physio-pathological events. In order to unravel lipid function, it is of utmost importance to identify and quantify single lipid molecular species in complex biological systems. Lipidomics is a rapidly evolving analytical technique capable of measuring hundreds of lipids and is currently at the forefront of scientific research due to the importance of lipids in health and disease. The aim of this study was to evaluate the effect of cigarette smoking on lipid profile in overweight patients with already developed cardiovascular disease by liquid chromatography (LC) coupled online to mass spectrometry (MS). The overall observation of the present study was that, there was an increase in concentration of fatty acids, and some specific triglycerides (TG) and diglycerides (DG) and decrease in the concentration of colesteryl esters (both saturated and unsaturated) phosphatidylcholines (especially unsaturated) and lysophosphatidycolines (especially saturated), and triglycerides saturated in smokers compared to nonsmokers. Thus, it can be said, based on the present study, that smoking affects and deranges the lipid profile, but in patients with already existing cardiovascular diseases and many confounding factors smoking may not have such significant influence as before disease development
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