Gene expression profiles in Atlantic salmon adipose-derived stromo-vascular fraction during differentiation into adipocytes

<p>Abstract</p> <p>Background</p> <p>Excessive fat deposition is one of the largest problems faced by salmon aquaculture industries, leading to production losses due to high volume of adipose tissue offal. In addition, increased lipid accumulation may impose considerable stress on adipocytes leading to adipocyte activation and production and secretion of inflammatory mediators, as observed in mammals.</p> <p>Results</p> <p>Microarray and qPCR analyses were performed to follow transcriptome changes during adipogenesis in the primary culture of adipose stromo-vascular fraction (aSVF) of Atlantic salmon. Cellular heterogeneity decreased by confluence as evidenced by the down-regulation of markers of osteo/chondrogenic, myogenic, immune and vasculature lineages. Transgelin (TAGLN), a marker of the multipotent pericyte, was prominently expressed around confluence while adipogenic PPARγ was up-regulated already in subconfluent cells. Proliferative activity and subsequent cell cycle arrest were reflected in the fluctuations of pro- and anti-mitotic regulators. Marked regulation of genes involved in lipid and glucose metabolism and pathways producing NADPH and glycerol-3-phosphate (G3P) was seen during the terminal differentiation, also characterised by diverse stress responses. Activation of the glutathione and thioredoxin antioxidant systems and changes in the iron metabolism suggested the need for protection against oxidative stress. Signs of endoplasmic reticulum (ER) stress and unfolded protein response (UPR) occured in parallel with the increased lipid droplet (LD) formation and production of secretory proteins (adipsin, visfatin). The UPR markers XBP1 and ATF6 were induced together with genes involved in ubiquitin-proteasome and lysosomal proteolysis. Concurrently, translation was suppressed as evidenced by the down-regulation of genes encoding elongation factors and components of the ribosomal machinery. Notably, expression changes of a panel of genes that belong to different immune pathways were seen throughout adipogenesis. The induction of AP1 (Jun, Fos), which is a master regulator of stress responses, culminated by the end of adipogenesis, concurrent with the maximal observed lipid deposition.</p> <p>Conclusions</p> <p>Our data point to an intimate relationship between metabolic regulation and immune responses in white adipocytes of a cold-blooded vertebrate. Stress imposed on adipocytes by LD formation and expansion is prominently reflected in the ER compartment and the activated UPR response could have an important role at visceral obesity in fish.</p

Korišćenje in vitro model sistema za proučavanje osnovnih bioloških procesa kod riba

With the increasing replacement of fish meal and fish oil with new ingredients in aquaculture diets, imbalances in amino acids, fatty acids, vitamins and minerals can occur. The metabolic and regulatory processes underlying these nutrition-induced imbalances in fish are still not fully understood. At the cellular level, essential dietary compounds and micro-nutrients have been shown to influence lineage determination, differentiation and proliferation of certain cell types, and hence the development of tissue structures and organogenesis. An improved understanding of cellular and molecular events occurring during development in teleosts will enable us to better characterize and define particular requirements, customize feed components, and thus enable development of sustainable feeds, minimize the occurrence of disorders as well as maintain the continuous growth of the fish. In vitro techniques have a great potential in experiments involving mechanism based hypothesis testing, where there is a significant need for a complete understanding of basic biological processes. Cell cultures provide means to study single-factor effects and the combinations thereof in detail, and further, to investigate the role of particular nutrients and their specific gene interactions, which are not possible when working at the organism level. In addition, whole organisms are complex and vary individually, depending on age, sex, health status, type of meal, genetics etc., which makes it difficult to accurately simulate nutritional processes. In vitro experiments offer the unique opportunity to develop standardized methods to study quality of novel and fortified feed products by studying the cellular and molecular effect of different types of food products, ranging from proteins to n-3 fatty acids (FAs) and from fat-soluble vitamins to minerals and trace elements. The system may also be used for studies on the development of functional feeds such as probiotics, prebiotics, bioactive peptides, lipase inhibitors, fat and cholesterol binders and antioxidants. Results from selected in vitro trials, showing how different nutrients may influence development of fat, bone and muscle cells and lipid metabolism in liver cells, will be presented.Zamena ribljeg brašna i ulja sa novim, alternativnim izvorima proteina i masti u hrani za ribe može dovesti do narušavanja odnosa amino kiselina, masnih kiselina i minerala kod ovih organizama. Promene u metaboličkim i regulatornim procesima koje mogu da nastanu loše balansiranom ishranom još uvek nisu dobro proučene kod riba. Esencijalne hranljive materije i mikronutrijenti mogu da utiču na razvoj pojedinih ćelijskih linija, njihovu proliferaciju, a samim tim i na pravilan razvoj tkiva i organa. Bolje razumevanje ćelijskih i molekularnih procesa, koji se dešavaju tokom razvića košljoriba će nam omogućiti da bolje razumemo i definišemo određene zahteve za ishranom na svakom stupnju razvoja riba, prilagodimo komponente riblje hrane, a samim tim i omogućimo normalan razvoj i rast riba, kao i da sprečimo pojavu različitih poremećaja. Korišćenje in vitro sistema predstavlja veliki potencijal za testiranje novih komponenata hrane i daje nam mogućnost za razumevanje osnovnih bioloških procesa kod riba. Prednost korišćenja ćelijskih kultura je što nam one omogućavaju da proučavamo uticaj pojedinačnih faktora, ali i kombinaciju dva ili više različitih faktora i njihov uticaj na gene, što nije moguće istraživati na nivou organizma. Pored toga proučavanje organizma kao celine je jako složeno, jer zavisi od pola, zdravstvenog stanja jedinke, ishrane, genetike, itd., te je na ovaj način lakše simulirati procese ishrane. In vitro eksperimenti nude jedinstvenu priliku da razvijemo standardizovane metode za proučavanje kvaliteta i efekata novih hranljivih materija: od proteina, preko masnih kiselina i minerala, do elemenata, koji se u hrani nalaze u tragovima. Ovi sistemi mogu se koristiti i za proučavanje novih, funkcionalnih hraniva, kao što su: probiotici, prebiotici, bioaktivni peptidi, masti, inhibitori lipaze, antioksidansi. Biće prikazani rezultati odabranih in vitro ispitivanja uticaja različitih hraniva na metabolizam masnog tkiva, kostiju i mišića kao i na sam metabolizam masti

Low Omega-3 Levels in the Diet Disturbs Intestinal Barrier and Transporting Functions of Atlantic Salmon Freshwater and Seawater Smolts

Due to a limited access to marine raw materials from capture fisheries, Atlantic salmon feeds are currently based on mainly plant ingredients (75%) while only 25% come from traditional marine ingredients including marine fish meal and fish oil. Thus, current feeds contain less of the essential omega-3 fatty acids. The aim of the study was to assess the impact of different omega-3 levels in fish feed on intestinal barrier and transporting functions of Atlantic salmon freshwater and seawater smolts. Atlantic salmon were fed three levels of omega-3 (2, 1 and 0.5%) and fish performance was followed through smoltification and the subsequent seawater acclimation. Intestinal barrier and transporting functions were assessed using Ussing chamber methodology and combined with transcript analysis of tight junction related proteins and ion transporters. A linear decrease in growth was observed with decreasing omega-3 levels. Low (0.5%) inclusion of omega-3 impaired the barrier function of the proximal intestine compared to 2% inclusion. Further, low levels of omega-3 decrease the transepithelial electrical potential across the epithelium indicating disturbed ion transport. It can be concluded that low dietary levels of omega-3 impair somatic growth and intestinal function of Atlantic salmon.publishedVersio

Dietary Level of the Omega-3 Fatty Acids EPA and DHA Influence the Flesh Pigmentation in Atlantic Salmon

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Improved fillet quality in harvest-size Atlantic salmon fed high n-3 canola oil as a DHA-source

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PENINGKATAN KETERAMPILAN BERPIKIR KREATIF DAN KETERAMPILAN PROSES SAINS SISWA SEKOLAH DASAR MELALUI MODEL PEMBELAJARAN BERBASIS MASALAH

Penelitian ini didasarkan pada masih rendahnya kualitas pembelajaran di SD dalam mengimplementasikan pembelajaran yang dapat meningkatkan keterampilan berpikir kreatif dan keterampilan proses sains. Berdasarkan alasan tersebut tujuan penelitian ini yaitu untuk mendeskripsikan apakah data peningkatan keterampilan berpikir kreatif dan keterampilan proses sains siswa yang mendapatkan pembelajaran berbasis masalah (PBM) lebih baik dibandingkan siswa yang mendapatkan pembelajaran bukan PBM. Metode penelitian yang digunakan yaitu quasi eksperiment dengan desain pre- and post test design. Penelitian ini dilaksanakan di kelas V salah satu sekolah dasar negeri di Kabupaten Majalengka pada tahun ajaran 2014/2015 dengan subjek penelitian sebanyak 24 siswa kelas eksperimen dan 24 siswa kelas kontrol. Kelas ekperimen diberi perlakuan PBM, sedangkan kelas kontrol dengan bukan PBM. Kedua kelompok diberikan pre test dan post test dengan menggunakan instrumen tes yang sama. Instrumen yang digunakan terdiri atas butir soal uraian untuk mengukur keterampilan berpikir kreatif, soal pilihan ganda untuk mengukur keterampilan proses sains, dan lembar observasi untuk melihat keterlaksanaan pembelajaran. Data pre test dan post test diolah menggunakan bantuan program SPSS 20 for Windows. Hasil analisis data menunjukan bahwa keterampilan berpikir kreatif dan keterampilan proses sains pada kelas eksperimen baik secara keseluruhan maupun pada setiap indikatornya mengalami peningkatan yang lebih baik dibanding siswa kelas kontrol. Adapun peningkatan secara keseluruhan untuk keterampilan berpikir kreatif yaitu sebesar 0,58 sedangkan peningkatan keterampilan proses sains yaitu sebesar 0,56 dimana keduanya berada pada kategori sedang. Sehingga dapat disimpulkan bahwa peningkatan keterampilan berpikir kreatif dan keterampilan proses siswa yang mendapatkan pembelajaran berbasis masalah (PBM) lebih baik dibandingkan siswa yang mendapatkan pembelajaran bukan PBM. ---------- This study based on education in elementary school is still low in implementing creative thinking ability and science process ability. Based on these reason, the purpose of this research to describe an overview of the impact of problem based learning on creative thinking ability and science process ability. To a chieve these objektive the study used quasi exsperimental method with pre and post test design. The subjek were students of class V in Majalengka one vocational school year 2014/2015. Which 24 student in experimental class and 24 student in control class. In this design, the exsperimental class treated with problem based learning, whereas the untreated control class non problem based learning. Both groups were given a pre-test and post-test using the same test instruments. The instrument used consisted of items description for measuring creative thinking skills, multiple choice questions to measure the science process skills, and the observation sheet to see the feasibility study. Pre-test and post test is processed using SPSS 20 for Windows. The results showed the impact of problem based learning can improve creative thinking ability (N-gain 0,58) and science process ability (N-gain 0,56). Results of this study can be concluded that an increase in creative thinking skills ability and science process ability of the students who received problem-based learning was better than the students who get non problem based learning

Different Dietary Ratios of Camelina Oil to Sandeel Oil Influence the Capacity to Synthesise and Deposit EPA and DHA in Zucker Fa/Fa Rats

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Long-term feeding of Atlantic salmon with varying levels of dietary EPA+DHA alters the mineral status, but does not affect the stress responses after mechanical delousing stress

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Dietary Fish Oil Alters DNA Methylation of Genes Involved in Polyunsaturated Fatty Acid Biosynthesis in Muscle and Liver of Atlantic Salmon (Salmo salar)

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