243 resultados para DAG


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ABSTRACT The Baltic Sea is a vulnerable ecosystem currently undergoing a number of changes, both natural and human induced. The changes are likely to affect the species found on these shores, e.g. their distribution and interactions with other species. Blue mussels (Mytilus trossulus x Mytilus edulis) provide one of the main biogenic hard structures on the shallow shores of the Baltic Sea where they aggregate into dense beds and provide a number of resources for over 40 associated macrofaunal species, thus functioning as ecosystem engineers. The blue mussel, being a marine species, is highly likely to be affected by any changes in sea water salinity, circulation and/or water balance. These changes could trickle down also to affect the associated macrofaunal communities. The aims of this thesis were three-fold: first, I examined and described the macrofaunal communities found within blue mussel patches since the fauna associated with mussel patches had never been described in the study area prior to this thesis. Second, I explored how changes in mussel density, size as well as patch size and shape would affect the mussel communities. Finally, I tested how general landscape theories derived from terrestrial studies function in blue mussel systems. Theories included the structural heterogeneity hypothesis, species-area relationships, edge effects and patch isolation effects. The work shows that blue mussels in the northern Baltic Sea have an indisputable function as diversity hotspots and that the faunal assemblages found in mussel patches are extremely rich and unique. Further on, it shows that changes in mussel biomass, size, patch size and amount of edge have the potential to alter the faunal assemblages and diversity within patches. Finally, it shows that although some landscape theories, such as the structural heterogeneity hypothesis, seem to apply also in blue mussel communities, others cannot be directly applied due to the different prevailing conditions in the study system. This is a pioneering work looking at diversity shaping processes on the rocky shores of the Gulf of Finland, making up over 40% of the total water basin. A focus on niche construction, positive facilitation effects and ecosystem engineering could provide new insights and methods for conservation biology, but before this can be done, we need to fully understand the circumstances under which a species becomes an ecosystem engineer and recognize the systems in which it functions.

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A key step in the triacylglycerol (TAG) biosynthetic pathway is the final acylation of diacylglycerol (DAG) by DAG acyltransferase. In silico analysis has revealed that the DCR (defective in cuticular ridges) (At5g23940) gene has a typical HX4D acyltransferase motif at the N-terminal end and a lipid binding motif VX(2)GF at the middle of the sequence. To understand the biochemical function, the gene was overexpressed in Escherichia coli, and the purified recombinant protein was found to acylate DAG specifically in an acyl-CoA-dependent manner. Overexpression of At5g23940 in a Saccharomyces cerevisiae quadruple mutant deficient in DAG acyltransferases resulted in TAG accumulation. At5g23940 rescued the growth of this quadruple mutant in the oleate-containing medium, whereas empty vector control did not. Lipid particles were localized in the cytosol of At5g23940-transformed quadruple mutant cells, as observed by oil red O staining. There was an incorporation of 16-hydroxyhexadecanoic acid into TAG in At5g23940-transformed cells of quadruple mutant. Here we report a soluble acyl-CoA-dependent DAG acyltransferase from Arabidopsis thaliana. Taken together, these data suggest that a broad specific DAG acyltransferase may be involved in the cutin as well as in the TAG biosynthesis by supplying hydroxy fatty acid.

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Forskningen utreder skillnader mellan finsk- och svenskspråkiga skolor i en kommun i Nylands län utgående från elevvårdens synvinkel. Syftet är att kartlägga situationen i en kommun och synliggöra de skillnader och faktorer som påverkar det sociala kapitalets mängd i skolorna. Skillnaderna betraktas utgående från det sociala kapitalets inverkan på gemenskapen och från ekologisk synvinkel. Sambandet mellan det sociala stöd och den sociala kontroll som särskilt de vuxna i skolan producerar, har betydelse för skolelevers välmående. För att kunna bilda en socialt stödande och socialt kontrollerande atmosfär krävs det funktionella förändringar i skolan. Särskilt de vuxna i skolan skulle behöva mera gemenskap. Elevvårdsarbetet och skolarbetet riktar sig främst mot elever i dag, fastän de vuxna skulle behöva stärka sina sociala förhållanden. Bris i uppkomsten av socialt stöd och kontroll beror främst på problem i samarbete och kommunikation mellan de vuxna i skolan. Skolkuratorerna är de enda professionella inom skolan som i sitt arbete tar hela skolan som gemenskap i beaktande. Denna forskning är en abduktiv kvalitativ fallstudie som är tili sin karaktär beskrivande. Som data används intervjuer av elevvärdspersonalen i kommunen och enkäten Hälsa i skolan av Institutet för hälsa och välfärd från år 2008. Viktigaste källan för forskningen är Noora Ellone (2008) forskning "Kasvuyhteisö nuoren turvana".

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The developing seeds of Actinodaphne hookeri were investigated to delineate their ability to synthesize large amounts of trilaurin. Until 88 days after flowering the embryos contained 71% neutral lipids (NL) and 29% phospholipids (PL) and both these components contained C-16:0, C-18:0, C-18:2, and C-18:3 as the major fatty acids (FA). At 102 days after flowering the seeds began to accumulate triacylglycerols (TAG) and to synthesize lauric acid (C-12:0). By 165 days after flowering, when the seeds were mature, they contained about 99% NL and 1% FL. At this stage the TAG contained exclusively C-12:0, while the PL consisted of long-chain fatty acids (LCFA) only. Leaf lipids in contrast did not contain any C-12:0. Experiments on [1-C-14]acetate incorporation into developing seed slices showed that at 88 days after flowering only 4% of the label was in TAG, 1% in diacylglycerols (DAG), and 87% in FL. One hundred two days after flowering seeds incorporated only 2% of the label into TAG, 30% into DAG, and 64% into FL. In contrast at 114 days after flowering 71% of the label was incorporated into TAG, 25% into DAG, and only 2% into FL. Analysis of labeled FA revealed that up to 102 days after flowering it was incorporated only into LCFA, whereas at 114 days after flowering it was incorporated exclusively into C-12:0. Furthermore, 67% of the label in PL at 114 days after flowering was found to be dilaurylglycerophosphate. Analysis of the label in DAG at this stage showed that it was essentially in dilaurin species. These observations indicate the induction of enzymes of Kennedy pathway for the specific synthesis of trilaurin at about 114 days after flowering, Homogenates of seeds (114 days after flowering) incubated with labeled FA in the presence of glycerol-3-phosphate and coenzymes A and ATP incorporated 84% of C-12:0 and 61% of C-14:0, but not C-16:0, C-18:2, and C-18:3, into TAG. In contrast the LCFA were incorporated preferentially into FL. It is concluded that, between 102 and 114 days after flowering, a switch occurs in A. hookeri for the synthesis of C-12:0 and trilaurin which is tissue specific. Since the seed synthesizes exclusively C-12:0 at 114 days after flowering onwards and incorporates specifically into TAG, this system appears to be ideal for identifying the enzymes responsible for medium-chain fatty acid as well as trilaurin synthesis and for exploiting them for genetic engineering. (C) 1994 Academic Press, Inc.

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Background: Phosphorylation by protein kinases is central to cellular signal transduction. Abnormal functioning of kinases has been implicated in developmental disorders and malignancies. Their activity is regulated by second messengers and by the binding of associated domains, which are also influential in translocating the catalytic component to their substrate sites, in mediating interaction with other proteins and carrying out their biological roles. Results: Using sensitive profile-search methods and manual analysis, the human genome has been surveyed for protein kinases. A set of 448 sequences, which show significant similarity to protein kinases and contain the critical residues essential for kinase function, have been selected for an analysis of domain combinations after classifying the kinase domains into subfamilies. The unusual domain combinations in particular kinases suggest their involvement in ubiquitination pathways and alternative modes of regulation for mitogen-activated protein kinase kinases (MAPKKs) and cyclin-dependent kinase (CDK)-like kinases. Previously unexplored kinases have been implicated in osteoblast differentiation and embryonic development on the basis of homology with kinases of known functions from other organisms. Kinases potentially unique to vertebrates are involved in highly evolved processes such as apoptosis, protein translation and tyrosine kinase signaling. In addition to coevolution with the kinase domain, duplication and recruitment of non-catalytic domains is apparent in signaling domains such as the PH, DAG-PE, SH2 and SH3 domains. Conclusions: Expansion of the functional repertoire and possible existence of alternative modes of regulation of certain kinases is suggested by their uncommon domain combinations. Experimental verification of the predicted implications of these kinases could enhance our understanding of their biological roles.

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The biosynthesis of triacylglycerol (TAG) occurs in the microsomal membranes of eukaryotes. Here, we report the identification and functional characterization of diacylglycerol acyltransferase (DGAT), a member of the 10 S cytosolic TAG biosynthetic complex (TBC) in Rhodotorula glutinis. Both a full-length and an N-terminally truncated cDNA clone of a single gene were isolated from R. glutinis. The DGAT activity of the protein encoded by RgDGAT was confirmed in vivo by the heterologous expression of cDNA in a Saccharomyces cerevisiae quadruple mutant (H1246) that is defective in TAG synthesis. RgDGAT overexpression in yeast was found to be capable of acylating diacylglycerol (DAG) in an acyl-CoA-dependent manner. Quadruple mutant yeast cells exhibit growth defects in the presence of oleic acid, but wild-type yeast cells do not. In an in vivo fatty acid supplementation experiment, RgDGAT expression rescued quadruple mutant growth in an oleate-containing medium. We describe a soluble acyl-CoA-dependent DAG acyltransferase from R. glutinis that belongs to the DGAT3 class of enzymes. The study highlights the importance of an alternative TAG biosynthetic pathway in oleaginous yeasts.

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We compared the capture efficiency and catch dynamics of collapsible square and conical pots used in resource assessment and harvesting of red king crabs (Paralithodes camtschaticus [Tilesius, 1815]) in the Barents Sea. After two days of soaking, square pots caught three times as many crabs as conical pots, and their catches consisted of a higher proportion of male crabs and male crabs larger than 160 mm carapace length compared to the catches in the conical pots. Catches in the square pots did not increase as soak times were increased beyond two days, which indicates equilibrium between the rate of entries into and the rate of exits from the pots. Catches in conical pots, however, increased with increasing soak times up to eight days, the longest soak time examined in this study. These findings demonstrate the higher efficiency of square pots and the importance of understanding catch dynamics when making population assessments based on catchper-unit-of-effort data. The favorable catch characteristics and handling properties of the collapsible square pot may make this pot design suitable for other crab fisheries, as well.

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We have formulated a model for analyzing the measurement error in marine survey abundance estimates by using data from parallel surveys (trawl haul or acoustic measurement). The measurement error is defined as the component of the variability that cannot be explained by covariates such as temperature, depth, bottom type, etc. The method presented is general, but we concentrate on bottom trawl catches of cod (Gadus morhua). Catches of cod from 10 parallel trawling experiments in the Barents Sea with a total of 130 paired hauls were used to estimate the measurement error in trawl hauls. Based on the experimental data, the measurement error is fairly constant in size on the logarithmic scale and is independent of location, time, and fish density. Compared with the total variability of the winter and autumn surveys in the Barents Sea, the measurement error is small (approximately 2–5%, on the log scale, in terms of variance of catch per towed distance). Thus, the cod catch rate is a fairly precise measure of fish density at a given site at a given time.

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磷脂酰甘油(PG)是植物类囊体膜中唯一的磷脂,在它的sn-2位上总是连着一个棕榈酸(16:0)或反式十六碳烯酸(16:1 trans)。由于PG的分子结构独特,对它的功能已有了很多研究,目前认为PG在维持类囊体膜的结构与功能方面具有非常重要的作用。缺磷胁迫下,蓝藻、衣藻及拟南芥、大麦等物种中均检测到了PG含量的下降。对这一现象的常见解释是缺磷导致了PG生物合成受阻,从而引起了其含量的降低。但迄今为止尚没有试验证据支持。本研究比较了缺磷对不同叶龄的小麦与烟草叶片中PG含量与PG水解酶的活性的影响,同时对缺磷叶片酶粗提液水解外源PG后的主要产物、几种磷脂酶抑制剂对上述酶反应的影响等进行了研究,以阐明缺磷条件下叶片中PG含量下降的主要原 因。 缺磷小麦第一叶完全展开时,PG含量与PG水解酶活性均与对照相似;而第三叶完全展开时,尽管缺磷第三叶中PG水解酶活性也与对照相似,但其PG含量低于对照。这一结果表明,在小麦叶片完全展开之前,缺磷条件未影响叶片中的PG水解酶活性,第三叶中较低的PG含量应由PG的生物合成受阻引起。并且,由于缺磷植株第一叶完全展开时PG含量未受影响而第三叶中却表现出了轻微降低,可以推测叶片萌发越晚,PG生物合成受到的抑制就会越严重。 为了研究叶片衰老过程中PG含量下降的原因,我们比较了6,10,14与18日龄时缺磷与对照小麦植株第一叶中PG的相对含量与PG水解酶活性。研究发现:6日龄时,刚刚完全展开的缺磷和对照小麦第一叶中无论是PG含量还是PG水解酶活性都较为相似;而随着叶片的逐渐衰老,缺磷植株第一叶中PG含量大幅度下降,同时伴随着PG水解酶活性的急剧上升。18日龄时,缺磷小麦第一叶中的PG含量较对照降低了69.1%,其PG水解酶活性也远高于对照,37ºC下温育30min后,缺磷叶片的酶粗提液使外源PG含量降低了74.16%,而对照中只降低了13.7%。上述结果表明,缺磷条件下,小麦叶片中PG含量降低的程度与PG水解酶活性的强弱密切相关,PG水解加剧是导致老叶中PG含量降低的一个重要原因。 磷脂酶是水解磷脂的主要酶类。目前在植物体中发现的磷脂酶种类主要有磷脂酶D(PLD)、磷脂酶C(PLC)与磷脂酶A(PLA)。通过薄层层析(TLC),我们发现缺磷小麦叶片的酶粗提液水解外源PG后的主要产物是磷脂酸(PA)、二脂酰甘油(DAG)与游离脂肪酸(FFA)。将n-丁醇加入到缺磷小麦叶片的体外酶反应体系中后,观察到PA、DAG与FFA的生成量均表现出一定程度的降低。由于n-丁醇是PA经PLD途径生成的抑制剂,因此,上述结果表明PLD参与了缺磷条件下小麦叶片中PG的水解。硫酸新霉素是PLC的非特异性抑制剂,低浓度的硫酸新霉素(100μM 和 200μM )加入到缺磷小麦叶片的体外酶反应体系后,三种产物的生成受到了严重抑制,表明PLC也与缺磷叶片中PG的降解密切相关。 为了进一步分析缺磷导致PG含量降低的原因,我们以烟草为试验材料,检测了缺磷胁迫对烟草嫩叶和老叶中的PG含量、PG水解酶活性、与PG降解相关的酶的种类及PLC、PLDα、PLDβ与PAT-1基因在mRNA上表达水平的的影响。结果表明,缺磷烟草叶片中PG含量的降低由PG生物合成受阻与PG降解加剧共同导致,PLC和PLD活性与烟草叶片中PG的降解有关。缺磷植株老叶中PG水解酶活性及PLC、PLDα、PLDβ基因在mRNA水平上的表达量均高于对照,表明在磷胁迫条件下,老叶中PG水解酶活性可能受到转录水平上的调节, PLC、PLDα、PLDβ转录活性的增强导致了PLC、PLD活性加强,从而引起PG降解的加剧,最终导致了PG含量的降低。与PLC、PLDα和PLDβ不同,缺磷胁迫对patatin蛋白(表现PLA2活性)的编码基因PAT-1在转录水平上的表达无影响,TLC分析PG的水解产物也未检测到溶血磷脂酰甘油(LPG)的生成。由此可见,PLA活性可能与缺磷条件下PG的降解无关。

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磷脂酰甘油(phosphatidylglycerol, PG)是类囊体膜(也叫光合膜)中唯一的一种磷脂。在蓝藻中,PG的合成途径为:磷脂酸(phosphatidic acid, PA)胞嘧啶双磷酸-二酰基甘油 (cytidine diphosphate diacylglycerol, CDP-DAG) 磷酸磷脂酰甘油 (phosphatidylglycerol phosphate, PGP)PG。其中最后一步反应是由PGP去磷酸化而生成PG,催化该反应的是PGP磷酸酶。然而迄今为止,PGP磷酸酶还没有在蓝藻和高等植物中得到克隆和鉴定。本工作在鱼腥藻Anabaena sp. PCC7120中通过将一个可能编码PGP磷酸酶的基因(alr1715)进行突变,获得缺失PG的突变体。与野生型相比,该突变体PG的含量降低了30%左右。突变后的蓝藻藻丝发黄、生长缓慢,叶绿素含量降低。整体细胞的光合作用活性、光系统II(photosystem II,PSII)的放氧活性以及PSII反应中心的光能转化效率显著下降,传递给PSII的激发能减少。

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研究以昆明山海棠根部水抽提物(Tripterygium hypoglaucum (Level) Hutch,THH)处理中国仓鼠V_(79)细胞,通过检测V_(79)细胞C-M细胞频率以及二酰基甘油(1,2-diacylgcerol,DAG)的含量测定,分析了THH诱发非整倍体与细胞醇磷酯信号通路的关系。结果指出:THH能在1mg/ml、2mg/ml两个剂量上使V_(79)细胞的DAG含量显著升高(P<0.001),并明显的提高C-M细胞频率(P<0.05),提示肌醇酯信号通路是介导THH诱发非整倍体的途径之一。

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Extracellular polymeric substances (EPS) from four filamentous cyanobacteria Microcoleus vaginatus, Scytonema javanicum, Phormidium tenue and Nostoc sp. and a coccoid single-cell green alga Desmococcus olivaceus that had been separated from desert algal crusts of Tegger desert of China, were investigated for their chemical composition, structure,and physical properties. The EPS contained 7.5-50.3% protein (in polymers ranging from 14 to more than 200 kD, SDS-PAGE) and 16.2-46.5% carbohydrate (110-460 kD, GFC). 6-12 kinds of monosaccharides, including 2-O-methyl rhamnose, 2-O-methyl glucose, and N-acetyl glucosamine were found. The main carbohydrate chains from M. vaginatus and S. javanicum consisted mainly of equal proportion of Man, Gal and Glc, that from P. tenue consisted mainly of arabinose, glucose and rhamnose. Arabinose was present in pyranose form, mainly alpha-L 1 --> 3 linked, with branches on C4 of almost half of the units. Glucose was responsible for the terminal units, in addition of having some units as beta1 --> 3 and some as beta1 --> 4 linked. Rhamnose was mainly 1 --> 3 linked with branches on C2 on half of the units. The carbohydrate polymer from D. olivaceus was composed mainly of beta1 --> 4 linked xylose, galactose and glucose. The galactose part was present both in beta-pyranose and -furanose forms. Arabinose in alpha-L-furanose form was mainly present as 1 --> 2 and 1 --> 2, 5 linked units, rhamnose only as alpha 1 --> 3 and xylose as beta 1 --> 4. The backbone of the polysaccharide from Nostoc sp. was composed of beta-1 --> 4 linked xylose, galactose and glucose. Most of the glucose was branched on position C6, terminal glucose and 2-O-methyl glucose units are also present. The relationship between structure, physical properties and potential biological function is discussed. (C) 2003 Elsevier Ltd. All rights reserved.

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Hot water-soluble polysaccharides woe extracted from field colonies and suspension cultures of Nostoc commune Vaucher, Nostoc flagelliforme Berkeley et Curtis, and Nostoc sphaeroides Kutzing. Excreted extracellular polymeric substances (EPS) were isolated from the media in which the suspension cultures were grown. The main monosaccharides of the field colony polysaccharides from the three species were glucose, xylose, and galactose, with an approximate ratio of 2:1:1. Mannose was also present, but the levels varied among the species, and arabinose appeared only in N. flagelliforme. The compositions of the cellular polysaccharides and EPS from suspension cultures were more complicated than those of the field samples and varied among the different species. The polysaccharides from the cultures of N. flagelliforme had a relatively simple composition consisting of mannose, galactose, glucose, and glucuronic acid, but no xylose, as was found in the field colony polysaccharides. The polysaccharides from cultures of N. sphaeroides contained glucose (the major component), rhamnose, fucose, xylose, mannose, and galactose. These same sugars were present in the polysaccharides from cultures of N. commune, with xylose as the major component. Combined nitrogen in the media had no qualitative influence on the compositions of the cellular polysaccharides but affected those of the EPS of N. commune and N. flagelliforme. The EPS of N. sphaeroides had a very low fetal carbohydrate content and thus was not considered to be polysaccharide in nature. The field colony polysaccharides could be separated by anion exchange chromatography into neutral and acidic fractions having similar sugar compositions. Preliminary linkage analysis showed that 1) xylose, glucose, and galactose were 1-->4 linked, 2) mannose, galactose, and xylose occurred as terminal residues, and 3) branch points occurred in glucose as 1-->3,4 and 1-->3,6 linkages and in xylose as a 1-->3,4 linkage. The polymer preparations from field colonies had higher kinematic viscosities than those from corresponding suspension cultures. The high viscosities of the polymers suggested that they might DE suitable for industrial uses.

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This paper describes an attractive method to make biodiesel from soybean soapstock (SS). A novel recovery technology of acid oil (AO) from SS has been developed with only sulfuric acid solution under the ambient temperature (25 +/- 2 degrees C). After drying, AO contained 50.0% FFA, 15.5% TAG 6.9% DAG 3.1% MAG 0.8% water and other inert materials. The recovery yield of AO was about 97% (w/w) based on the total fatty acids of the SS. The acid oil could be directly converted into biodiesel at 95 degrees C in a pressurized reactor within 5 hours. Optimal esterification conditions were determined to be a weight ratio of 1 : 1.5 : 0.1 of AO/methanol/sulfuric acid. Higher reaction temperature helps to shorten the reaction time and requires less catalyst and methanol. Ester content of the biodiesel derived from AO through one-step acid catalyzed reaction is around 92%. After distillation, the purity of the biodiesel produced from AO is 97.6% which meets the Biodiesel Specification of Korea. The yield of purified biodiesel was 94% (w/w) based on the total fatty acids of the soapstock.