Rice Coleoptile Growth under Water and in Air-Possible Effect of Buoyancy on Growth and Cell Walls

Maximum growth was achieved in rice coleoptiles (Oryza sativa L. cv. Sasanishiki) grown under water; they reached maximum length of 81.2 mm on day 5. The maximum length of coleoptiles grown in air or under water with air bubbling was 12.4 mm and 23.5 mm in day 5,respectively. Differences in coleoptile growth between air bubbling and air conditions, namely approximately 11 mm at day 5,could be due to buoyancy effect under water. Promoted growth under water was due to a decrease in cell wall extensibility. The decrease in cell wall extensibility could be related to the inhibition of the formation of diferulic acid-bridges among arabinoxylans in cell walls under water

Functional modifications associated with gastrointestinal tract organogenesis during metamorphosis in Atlantic halibut (Hippoglossus hippoglossus)

Background: Flatfish metamorphosis is a hormone regulated post-embryonic developmental event that transforms a symmetric larva into an asymmetric juvenile. In altricial-gastric teleost fish, differentiation of the stomach takes place after the onset of first feeding, and during metamorphosis dramatic molecular and morphological modifications of the gastrointestinal (GI-) tract occur. Here we present the functional ontogeny of the developing GI-tract from an integrative perspective in the pleuronectiforme Atlantic halibut, and test the hypothesis that the multiple functions of the teleost stomach develop synchronously during metamorphosis. Results: Onset of gastric function was determined with several approaches (anatomical, biochemical, molecular and in vivo observations). In vivo pH analysis in the GI-tract lumen combined with quantitative PCR (qPCR) of α and β subunits of the gastric proton pump (H+/K+-ATPase) and pepsinogen A2 indicated that gastric proteolytic capacity is established during the climax of metamorphosis. Transcript abundance of ghrelin, a putative orexigenic signalling molecule produced in the developing stomach, correlated (p < 0.05) with the emergence of gastric proteolytic activity, suggesting that the stomach’s role in appetite regulation occurs simultaneously with the establishment of proteolytic function. A 3D models series of the GI-tract development indicated a functional pyloric sphincter prior to first feeding. Observations of fed larvae in vivo confirmed that stomach reservoir function was established before metamorphosis, and was thus independent of this event. Mechanical breakdown of food and transportation of chyme through the GI-tract was observed in vivo and resulted from phasic and propagating contractions established well before metamorphosis. The number of contractions in the midgut decreased at metamorphic climax synchronously with establishment of the stomach’s proteolytic capacity and its increased peristaltic activity. Putative osmoregulatory competence of the GI-tract, inferred by abundance of Na+/K+-ATPase α transcripts, was already established at the onset of exogenous feeding and was unmodified by metamorphosis.Conclusions: The functional specialization of the GI-tract was not exclusive to metamorphosis, and its osmoregulatory capacity and reservoir function were established before first feeding. Nonetheless, acid production and the proteolytic capacity of the stomach coincided with metamorphic climax, and also marked the onset of the stomach’s involvement in appetite regulation via ghrelin.Peer Reviewe

Corticosteroid Activation of Atlantic Sea Lamprey Corticoid Receptor: Allosteric Regulation by the N-terminal Domain

Lampreys are jawless fish that evolved about 550 million years ago at the base of the vertebrate line. Modern lampreys contain a corticoid receptor (CR), the common ancestor of the glucocorticoid receptor (GR) and mineralocorticoid receptor (MR), which first appear in cartilaginous fish, such as sharks. Until recently, 344 amino acids at the amino terminus of adult lamprey CR were not present in the lamprey CR sequence in GenBank. A search of the recently sequenced lamprey germline genome identified two CR sequences, CR1 and CR2, containing the 344 previously un-identified amino acids at the amino terminus. CR1 also contains a novel four amino acid insertion in the DNA-binding domain (DBD). We studied corticosteroid activation of CR1 and CR2 and found their strongest response was to 11-deoxycorticosterone and 11-deoxycortisol, the two circulating corticosteroids in lamprey. Based on steroid specificity, both CRs are close to elephant shark MR and distant from elephant shark GR. HEK293 cells transfected with full-length CR1 or CR2 and the MMTV promoter have about 3-fold higher steroid-mediated activation compared to HEK293 cells transfected with these CRs and the TAT3 promoter. Deletion of the amino-terminal domain (NTD) of lamprey CR1 and CR2 to form truncated CRs decreased transcriptional activation by about 70% in HEK293 cells transfected with MMTV, but increased transcription by about 6-fold in cells transfected with TAT3, indicating that the promoter has an important effect on NTD regulation of CR transcription by corticosteroids.Comment: 27 pages, 6 figure

Bilirubin and biliverdin binding to raty protein (ligandin)

The binding of bilirubin to poly(-lysine) produces an optically active complex at pH 10.1. Circular dichroism spectra of these complexes are distinguishable from those generated by binding of bilirubin to the high affinity sites on albumin. Comparison of the circular dichroism spectra of bilirubin bound to the hepatic protein ligandin with those of bilirubin complexed with albumin or polylysine indicates that binding of bilirubin to ligandin occurs at two types of sites. These are distinguishable on the basis of their spectral properties, one resembling the high affinity site of bovine serum albumin and the other resembling polylysine. Complexes of biliverdin with albumin and ligandin bear similarities to the bilirubin-protein complexes. The native protein itself has an ordered structure which consists of 41% [alpha]-helix and is not altered by the binding of bilirubin.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/21676/1/0000064.pd

Hormonal control of tryptic enzyme activity in Atlantic cod larvae (Gadus morhua): Involvement of cholecystokinin during ontogeny and diurnal rhythm

Highlights • We describe the ontogenetic development of CCK and tryptic enzyme activity in larval cod • CCK is known to play a key role in regulating digestive processes • CCK concentrations increased during ontogeny suggesting a growing role in regulating digestive processes • A short-term experiment reveals a feedback mechanism between CCK and tryptic enzyme activity • Cod larvae have limited regulatory and digestive capacity to handle several meals in a short period The ontogenetic development of the gut hormone cholecystokinin (CCK) and the key proteolytic enzyme trypsin was described in Atlantic cod larvae (Gadus morhua) from first-feeding until 38 days post first-feeding (dpff). CCK is known to play a major role in the endocrine control of digestive processes in mammals and adult fish, but its regulatory role in the larval stages of marine fish is largely unknown. Only small amounts of CCK were found in the body (excluding head) in cod larvae at first-feeding, but CCK levels increased exponentially with development, suggesting a more pronounced role of CCK during ontogeny. Tryptic enzyme activity increased slightly until a standard length of ca. 8 mm (approx. 33 days dpff) with a significant increase in larvae larger than 8 mm standard length, indicating limited digestive capacity in the early stages. To entangle the short-term feedback mechanism between CCK and tryptic enzyme activity, we conducted a 12 hour feeding experiment at 21 dpff. Cod larvae receiving only algae revealed a noticeable response in tryptic enzyme activity within two hours in the morning, whereas larvae fed algae and rotifers at the same time showed a slightly delayed response up to four hours. Tryptic enzyme activity remained low in the group receiving only algae as well as the two fed groups in the afternoon. No reaction in tryptic enzyme activity was observed in larvae that received a second meal of rotifers in the afternoon, indicating limited regulatory and digestive capacity to handle several meals in a short period. CCK levels remained relatively constant throughout the day but increased in the afternoon in all three groups when tryptic enzyme activity was low, suggesting that a negative feedback mechanism between CCK and tryptic enzyme activity is present in larval cod at least from 21 dpff

Structure-Function Analysis of Diacylglycerol Acyltransferase Sequences from 70 Organisms

<p>Abstract</p> <p>Background</p> <p>Diacylglycerol acyltransferase families (DGATs) catalyze the final and rate-limiting step of triacylglycerol (TAG) biosynthesis in eukaryotic organisms. Understanding the roles of DGATs will help to create transgenic plants with value-added properties and provide clues for therapeutic intervention for obesity and related diseases. The objective of this analysis was to identify conserved sequence motifs and amino acid residues for better understanding of the structure-function relationship of these important enzymes.</p> <p>Results</p> <p>117 DGAT sequences from 70 organisms including plants, animals, fungi and human are obtained from database search using tung tree DGATs. Phylogenetic analysis separates these proteins into DGAT1 and DGAT2 subfamilies. These DGATs are integral membrane proteins with more than 40% of the total amino acid residues being hydrophobic. They have similar properties and amino acid composition except that DGAT1s are approximately 20 kDa larger than DGAT2s. DGAT1s and DGAT2s have 41 and 16 completely conserved amino acid residues, respectively, although only two of them are shared by all DGATs. These residues are distributed in 7 and 6 sequence blocks for DGAT1s and DGAT2s, respectively, and located at the carboxyl termini, suggesting the location of the catalytic domains. These conserved sequence blocks do not contain the putative neutral lipid-binding domain, mitochondrial targeting signal, or ER retrieval motif. The importance of conserved residues has been demonstrated by site-directed and natural mutants.</p> <p>Conclusions</p> <p>This study has identified conserved sequence motifs and amino acid residues in all 117 DGATs and the two subfamilies. None of the completely conserved residues in DGAT1s and DGAT2s is present in recently reported isoforms in the multiple sequences alignment, raising an important question how proteins with completely different amino acid sequences could perform the same biochemical reaction. The sequence analysis should facilitate studying the structure-function relationship of DGATs with the ultimate goal to identify critical amino acid residues for engineering superb enzymes in metabolic engineering and selecting enzyme inhibitors in therapeutic application for obesity and related diseases.</p

Feeding behaviour and digestion physiology in larval fish – current knowledge and gaps and bottlenecks in research

Food uptake follows rules defined by feeding behaviour that determines the kind and quantity of food ingested by fish larvae as well as how live prey and food particles are detected, captured and ingested. Feeding success depends on the progressive development of anatomical characteristics and physiological functions and on the availability of suitable food items throughout larval development. The fish larval stages present eco-morpho-physiological features very different from adults and differ from one species to another. The organoleptic properties, dimensions, detectability, movements characteristics and buoyancy of food items are all crucial features that should be considered, but is often ignored, in feeding regimes. Ontogenetic changes in digestive function lead to limitations in the ability to process certain feedstuffs. There is still a lack of knowledge about the digestion and absorption of various nutrients and about the ontogeny of basic physiological mechanisms in fish larvae, including how they are affected by genetic, dietary and environmental factors. The neural and hormonal regulation of the digestive process and of appetite is critical for optimizing digestion. These processes are still poorly described in fish larvae and attempts to develop optimal feeding regimes are often still on a ‘trial and error’ basis. A holistic understanding of feeding ecology and digestive functions is important for designing diets for fish larvae and the adaptation of rearing conditions to meet requirements for the best presentation of prey and microdiets, and their optimal ingestion, digestion and absorption. More research that targets gaps in our knowledge should advance larval rearing

A rapid, novel High Performance Liquid Chromatography method for the purification of Glutathione S-transferase: An application to the human placental enzyme

A simple High Performance Liquid Chromatography procedure is detailed for the purification of Glutathione S-transferase. The human placental transferase was used to assess its potential. Unlike conventional methods of purification, the procedure is rapid and resolution of the various forms is achieved in less than 20 min. Since recovery is essentially complete, it is possible to isolate different minor forms. Three forms, one major and two minor, were separated. The major form represented about 97% of the total recovered activity and exhibited a specific activity of 254.94 [mu]moles/min/mg protein with a purification of 1342-fold. Electrophoresis of the major form revealed the presence of a single band, suggesting homogeneity.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25703/1/0000257.pd