Phosphatidylcholine:diacylglycerol cholinephosphotransferase's unique regulation of castor bean oil quality

The enzyme phosphatidylcholine:diacylglycerol cholinephosphotransferase facilitates accumulation of seed oil with three hydroxylated acyl groups in Ricinus communis.Castor bean (Ricinus communis) seed oil (triacylglycerol [TAG]) is composed of similar to 90% of the industrially important ricinoleoyl (12-hydroxy-9-octadecenoyl) groups. Here, phosphatidylcholine (PC):diacylglycerol (DAG) cholinephosphotransferase (PDCT) from castor bean was biochemically characterized and compared with camelina (Camelina sativa) PDCT. DAGs with ricinoleoyl groups were poorly used by Camelina PDCT, and their presence inhibited the utilization of DAG with "common" acyl groups. In contrast, castor PDCT utilized DAG with ricinoleoyl groups similarly to DAG with common acyl groups and showed a 10-fold selectivity for DAG with one ricinoleoyl group over DAG with two ricinoleoyl groups. Castor DAG acyltransferase2 specificities and selectivities toward different DAG and acyl-CoA species were assessed and shown to not acylate DAG without ricinoleoyl groups in the presence of ricinoleoyl-containing DAG. Eighty-five percent of the DAG species in microsomal membranes prepared from developing castor endosperm lacked ricinoleoyl groups. Most of these species were predicted to be derived from PC, which had been formed by PDCT in exchange with DAG with one ricinoleoyl group. A scheme of the function of PDCT in castor endosperm is proposed where one ricinoleoyl group from de novo-synthesized DAG is selectivity transferred to PC. Nonricinoleate DAG is formed and ricinoleoyl groups entering PC are re-used either in de novo synthesis of DAG with two ricinoleoyl groups or in direct synthesis of triricinoleoyl TAG by PDAT. The PC-derived DAG is not used in TAG synthesis but is proposed to serve as a substrate in membrane lipid biosynthesis during oil deposition

Development of a fluorescent nanosensor for ribose

AbstractTo analyze ribose uptake and metabolism in living cells, nanosensors were engineered by flanking the Escherichia coli periplasmic ribose binding protein with two green fluorescent protein variants. Following binding of ribose, fluorescence resonance energy transfer decreased with increasing ribose concentration. Five affinity mutants were generated covering binding constants between 400 nM and 11.7 mM. Analysis of nanosensor response in COS-7 cells showed that free ribose accumulates in the cell and is slowly metabolized. Inhibitor studies suggest that uptake is mediated by a monosaccharide transporter of the GLUT family, however, ribose taken up into the cell was not or only slowly released, indicating irreversibility of uptake

Fluorescence resonance energy transfer sensors for quantitative monitoring of pentose and disaccharide accumulation in bacteria

<p>Abstract</p> <p>Background</p> <p>Engineering microorganisms to improve metabolite flux requires detailed knowledge of the concentrations and flux rates of metabolites and metabolic intermediates <it>in vivo</it>. Fluorescence resonance energy transfer sensors represent a promising technology for measuring metabolite levels and corresponding rate changes in live cells. These sensors have been applied successfully in mammalian and plant cells but potentially could also be used to monitor steady-state levels of metabolites in microorganisms using fluorimetric assays. Sensors for hexose and pentose carbohydrates could help in the development of fermentative microorganisms, for example, for biofuels applications. Arabinose is one of the carbohydrates to be monitored during biofuels production from lignocellulose, while maltose is an important degradation product of starch that is relevant for starch-derived biofuels production.</p> <p>Results</p> <p>An <it>Escherichia coli </it>expression vector compatible with phage λ recombination technology was constructed to facilitate sensor construction and was used to generate a novel fluorescence resonance energy transfer sensor for arabinose. In parallel, a strategy for improving the sensor signal was applied to construct an improved maltose sensor. Both sensors were expressed in the cytosol of <it>E. coli </it>and sugar accumulation was monitored using a simple fluorimetric assay of <it>E. coli </it>cultures in microtiter plates. In the case of both nanosensors, the addition of the respective ligand led to concentration-dependent fluorescence resonance energy transfer responses allowing quantitative analysis of the intracellular sugar levels at given extracellular supply levels as well as accumulation rates.</p> <p>Conclusion</p> <p>The nanosensor destination vector combined with the optimization strategy for sensor responses should help to accelerate the development of metabolite sensors. The new carbohydrate fluorescence resonance energy transfer sensors can be used for <it>in vivo </it>monitoring of sugar levels in prokaryotes, demonstrating the potential of such sensors as reporter tools in the development of metabolically engineered microbial strains or for real-time monitoring of intracellular metabolite during fermentation.</p

Downregulation of the INDEHISCENT Gene by RNAi Resulted in Desired Pod Shatter Reduction of Lepidium campestre in Subsequent Generations

Wild species field cress (Lepidium campestre) has favorable agronomic traits, making it a good candidate for future development as an oil and catch crop. However, the species is very prone to pod shatter, resulting in severe yield losses. This is one of the important agronomic traits that needs to be improved in order to make this species economically viable. In this study, we cloned the L. campestre INDEHISCENT (LcIND) gene and prepared two LcIND-RNAi constructs with the IND promoter (long 400 bp and short 200 bp) from Arabidopsis. A number of stable transgenic lines were developed and evaluated in terms of pod shatter resistance. The majority of the transgenic lines showed increased resistance to pod shatter compared to the wild type, and this resistance was maintained in four subsequent generations. The downregulation of the LcIND gene by RNAi in the transgenic lines was confirmed by qRT-PCR analysis on T-3 lines. Southern blot analysis showed that most of the analyzed lines had a single-copy integration of the transgene, which is desirable for further use. Our results show that it is possible to generate stable transgenic lines with desirable pod shatter resistance by downregulating the LcIND gene using RNAi in field cress, and thus speeding up the domestication process of this wild species

Mast Cell Accumulation in Glioblastoma with a Potential Role for Stem Cell Factor and Chemokine CXCL12

Glioblastoma multiforme (GBM) is the most common and malignant form of glioma with high mortality and no cure. Many human cancers maintain a complex inflammatory program triggering rapid recruitment of inflammatory cells, including mast cells (MCs), to the tumor site. However, the potential contribution of MCs in glioma has not been addressed previously. Here we report for the first time that MCs infiltrate KRas+Akt-induced gliomas, using the RCAS/TV-a system, where KRas and Akt are transduced by RCAS into the brains of neonatal Gtv-a- or Ntv-a transgenic mice lacking Ink4a or Arf. The most abundant MC infiltration was observed in high-grade gliomas of Arf−/− mice. MC accumulation could be localized to the vicinity of glioma-associated vessels but also within the tumor mass. Importantly, proliferating MCs were detected, suggesting that the MC accumulation was caused by local expansion of the MC population. In line with these findings, strong expression of stem cell factor (SCF), i.e. the main MC growth factor, was detected, in particular around tumor blood vessels. Further, glioma cells expressed the MC chemotaxin CXCL12 and MCs expressed the corresponding receptor, i.e. CXCR4, suggesting that MCs could be attracted to the tumor through the CXCL12/CXCR4 axis. Supporting a role for MCs in glioma, strong MC infiltration was detected in human glioma, where GBMs contained significantly higher MC numbers than grade II tumors did. Moreover, human GBMs were positive for CXCL12 and the infiltrating MCs were positive for CXCR4. In conclusion, we provide the first evidence for a role for MCs in glioma

Plant Acyl-CoA:Lysophosphatidylcholine Acyltransferases (LPCATs) Have Different Specificities in Their Forward and Reverse Reactions

Background: Acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT) enzymes have central roles in acyl editing of phosphatidylcholine. Results: Plant LPCATs were expressed in yeast and biochemically characterized. Conclusion: LPCATs can edit acyl composition of phosphatidylcholine through their combined forward and reverse reactions. Significance: Plant LPCATs play a role in editing both sn-positions of PC and remove ricinoleic acid with high selectivity from this lipid

Camelina sativa phosphatidylcholine:diacylglycerol cholinephosphotransferase-catalyzed interconversion does not discriminate between substrates

Phosphatidylcholine:diacylglycerol cholinephosphotransferases (PDCT) regulate the fatty acid composition of seed oil (triacylglycerol, TAG) by interconversion of diacylglycerols (DAG) and phosphatidylcholine (PtdCho). PtdCho is the substrate for polyunsaturated fatty acid biosynthesis, as well as for a number of unusual fatty acids. By the action of PDCT, these fatty acids can be transferred into the DAG pool to be utilized in TAG biosynthesis by the action of acyl-CoA:DAG and phospholipid:diacylglycerol acyltransferases. Despite its importance in regulating seed oil composition, biochemical characterization of PDCT enzymes has been lacking. We characterized Camelina sativa PDCT in microsomal preparations of a yeast strain expressing Camelina PDCT and lacking the capacity of producing TAG. Camelina PDCT was specific for PtdCho and the sn-1,2 enantiomer of DAG and could not utilize ceramide. The interconversion reaches equilibrium within 15 min of incubation, indicating that only distinct pools of DAG and PtdCho were available for exchange. However, the pool sizes of DAG and PtdCho involved in the exchange were not fixed but increased with the amount of exogenous DAG or PtdCho added. Camelina PDCT showed about the same selectivity for di-oleoyl, di-linoleoyl, and di-linolenoyl species in both PtdCho and DAG substrates, suggesting that no unidirectional transfer of particular unsaturated substrates occurred. Camelina PDCT had a good activity with erucoyl-DAG as a substrate despite low erucic acid levels in PtdCho in plant species accumulating a high amount of this fatty acid in the seed oil

Intellektuell gemenskap : En studie om fysisk aktivitet i grupp på daglig verksamhet

Syfte och frågeställning Syftet är att undersöka vilka förutsättningar som påverkar deltagande i fysisk aktivitet hos personer med intellektuell funktionsnedsättning som går på daglig verksamhet. Vilka faktorer främjar till deltagandet i fysisk aktivitet? Vilka faktorer hindrar till deltagandet i fysisk aktivitet? Vad är upplevelsen av organiserad gruppträning? Metod En kvalitativ design med intervju som valdes som undersökningsmetod. Urvalet består av 4 arbetstagare och 1 coach verksamma vid dagliga verksamheter. Datainsamlingen genomfördes genom semistrukturerade intervjuer, där två intervjuguider utformades. En anpassad till arbetstagarna och en till coachen. Intervjuguiden grundades på KASAMs tre komponenter begriplighet, hanterbarhet och meningsfullhet. Efter transkribering av intervjuerna analyserades dessa genom en innehållsanalys. Resultat Arbetstagarna ansåg att fysisk aktivitet har positiv inverkan på den fysiska och psykiska hälsan. Socialt stöd upplevs vara en avgörande faktor till deltagande i fysisk aktivitet enligt coach och arbetstagare. Hinder till deltagande anges vara psykisk ohälsa, kommunikationssvårigheter och negativa känslor kring transportering. Samtliga arbetstagare ansåg att social interaktion, delaktighet och gemenskap är de viktigaste faktorerna till deltagande i fysisk aktivitet. Dessa faktorer framkom i analysen mot känsla av sammanhang (KASAM) där samspelet mellan begriplighet, hanterbarhet och meningsfullhet påverkar deltagandet i fysisk aktivitet. Slutsats Socialt stöd, social interaktion och upplevd förbättrad ångesthantering är främjande faktorer som skapar en känsla av sammanhang kring fysisk aktivitet på daglig verksamhet för personer med IF. Hinder och faktorer som motverkar känslan av sammanhang och till deltagande i fysisk aktivitet är transport till och från aktiviteten, den psykiska ohälsan samt brist på initiativtagande. Organiserad gruppträning främjar fysisk aktivitet eftersom den organiseras av andra personer.Aim The aim for this study was to investigate the conditions that affect participation in physical activity for people with intellectual disabilities who go to daily centers. • What factors promote participation in physical activity? • What factors prevent participation in physical activity? • What is the experience of organized group training? Method A qualitative method was chosen with a total of 5 interviews: 4 people with intellectual disability (ID) and 1 coach working on a daily center. The data collection was conducted through semi-structured interviews and two interview guides were designed. One was adapted to the match the people with ID and one to the coach. The interview guide and analysis was based on the three components of the sense of coherence theory (SOC): comprehensibility, manageability and meaningfulness. After transcribing the interviews, these were analyzed by a content analysis. Results People with ID in this study felt that physical activity has a positive impact on physical and mental health. Social support is perceived to be an important factor in participating in physical activity according to coach and people with ID. Things that prevented participation was stated to be mental illness, communication difficulties and negative feelings about transportation. Every participated person with ID considered social interaction, participation and community was the most important factors for participation in physical activity. These factors emerged in the analysis with SOC, where the interaction between comprehensibility, manageability and meaningfulness affects participation in physical activity. Conclusion With sense of coherence it seems like the people with ID, in this study, enables participation in physical activity. Social support, social interaction and improved management of anxiety are promotional factors for participation. Factors that prevent participation in physical activity was transport to and from the activity, mental illness and lack of initiative. Organized group training promotes physical activity because it is organized by other people

Rokotusinfo lapsille : Internetsivut Terveysnettiin

Rokotettaessa elimistöön viedään tautia aiheuttavaa mikrobia tai sen osaa sellaisessa muodossa, että se saa elimistössä aikaan pitkäkestoisen suojan tätä taudinaiheuttajaa vastaan. Rokote voidaan antaa pistoksena tai suun kautta. Rokottaminen on sekä luonnollinen, että myös edullinen keino ehkäistä tartuntatauteja ja estää niiden leviäminen. Suurin osa lapsista rokotetaan yleisen rokotusohjelman mukaisesti. Rokotusohjelman tavoitteena on suojata väestöä mahdollisimman hyvin tarttuvilta taudeilta. Kansallinen rokotusohjelma uudistui viimeksi vuonna 2005, jolloin otettiin käyttöön uusia yhdistelmärokotteita ja pistosten määrää voitiin näin vähentää. (Terveyden ja hyvinvoinnin laitos 2011) Projektin tehtävänä oli luoda tietopaketti rokotuksista Terveysnetin juniorinettiin alakouluikäisille lapsille. Projektin tavoitteena on antaa tietoa alakouluikäisille lapsille rokotuksista. Terveysnetti on Turun ammattikorkeakoulun ylläpitämä sivusto. Projektin tuloksena syntyneillä internetsivuilla käsitellään rokotteita, rokotteilla ehkäistäviä tauteja, rokottamisen syitä ja rokotustilannetta. Internetsivut ovat yksinkertaiset, lapsille suunnatut ja helppolukuiset, jotta alakouluikäiset saavat niistä helposti ikätasolleen sopivaa tietoa. Sivuston toteutuksessa on käytetty tuoreita ja luotettavia lähteitä. Sivut ovat käytettävissä osoitteessa http://terveysnetti.turkuamk.fi/ juniorinetti osiossa. Tulevaisuuden haasteena on rokotuskattavuuden ylläpitäminen, sillä jo pienikin kattavuuden aleneminen voisi tuoda Suomeen takaisin tauteja, jotka on jo onnistuttu hävittämään.Vaccine contains a pathogenic microbe or part of it in such form that it causes a long-term resistance against this pathogen. Vaccine can be given by injection or orally. Vaccination is the natural and an inexpensive way to prevent infectious diseases and to stop the spreading. Vaccination coverage in Finland has been traditionally high. The goal of Finnish vaccination program is to protect the population against infectious diseases as well as possible. Finnish vaccination program was renewed in 2005. The number of injections decreased due to the new combination vaccines. (THL 2011.) The purpose of the project was to create information package for vaccinations on Terveysnetti (Health net) for children. The goal of the project is to give children information about vaccinations. Terveysnetti is a site maintained by Turku University of Applied Sciences. Web pages provide information on vaccines, vaccine-preventable diseases and causes of vaccination. Web pages are simple and easy to read, so children can understand the information easily. New and reliable sources were used of making the website. Pages can be found at: http://terveysnetti.turkuamk.fi/ The future challenge is to keep vaccination coverage high, because even a small decrease in coverage could bring diseases which has already been eradicated back to Finland