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Digestive Enzymes (digestive + enzyme)
Terms modified by Digestive Enzymes Selected AbstractsCommunity heterogeneity and single-cell digestive activity of estuarine heterotrophic nanoflagellates assessed using lysotracker and flow cytometryENVIRONMENTAL MICROBIOLOGY, Issue 7 2010Eva Sintes Summary Heterotrophic nanoflagellates (HNFs) are an essential component of all aquatic microbial food webs, and yet the exploration of the numerical and single-cell responses of these organisms in mixed assemblages still represents a major technical challenge. LysoTracker Green staining combined with flow cytometry was recently proposed for the enumeration of aquatic HNFs. Here we show that LysoTracker Green not only allows the enumeration of HNFs in estuarine samples with a wide range of HNF abundances, but also allows the discrimination of distinct HNF populations in mixed assemblages. In addition, the resulting cytometric parameters can be used to characterize cell size and the level of activity of the cells in the different populations that are detected. LysoTracker Green accumulates preferentially in lysosomes, and we demonstrate that the green fluorescence emission from HNF cells stained with LysoTracker strongly correlates with cell-specific ,-glucosaminidase (,-Gam) activity, a key digestive enzyme of lysosomal origin in eukaryotic cells. Our results further show that different populations that develop in estuarine regrowth cultures are characterized by different intrinsic ranges of size and of feeding activity, and that there is a wide range of single-cell responses within these HNF populations. We found a large degree of uncoupling between cell size and feeding activity, both between and within HNF populations, and there appears to be no clear allometric scaling of feeding activity. We were able to reconstruct the succession of distinct HNF populations that developed during the regrowth experiments, and explore the complex interactions that occurred between numerical (change in abundance of the cytometric populations) and single-cell HNF responses. [source] Effects of dietary lipid levels on the growth, digestive enzyme, feed utilization and fatty acid composition of Japanese sea bass (Lateolabrax japonicus L.) reared in freshwaterAQUACULTURE RESEARCH, Issue 2 2010Gang Luo Abstract Triplicate groups of 40 Japanese sea bass Lateolabrax japonicus reared in freshwater (average weight, 9.52±0.47 g) were fed with six isonitrogenous (,46% crude protein) diets containing 6%, 8%, 10%, 12%, 14% or 16% lipid for 10 weeks respectively. The results showed that the maximum weight gain (WG), specific growth rate (SGR), feed intake (FI) and protein efficiency ratio (PER) all occurred at the 10% lipid level (P<0.05) and growth depression occurred when the dietary lipid level was over 12%. Whole body and liver lipid concentrations were enhanced with the increase in the dietary lipid levels, but the muscle lipid content did not significantly change with the increase in the dietary lipid levels. Both liver pepsin and trypsin activities increased with dietary lipid levels ranging from 6% to 10%, and then decreased with a further increase in the dietary lipid content. Liver lipase activities showed a positive correlation with dietary lipid levels, but amylase activities were not markedly influenced by dietary lipid levels. High proportions of 18:1n-9, 20:1n-9, eicosapentaenoic acid (20:5n-3; EPA), 22:1n-11 and docosahexaenoic acid (22:6n-3; DHA), and low concentrations of n-6 fatty acids, particularly 18:2n-6 occurring in the liver and muscle, to some extent, reflected the fatty acid composition in experimental diets. [source] Effect of water temperature on the growth performance and digestive enzyme activities of Chinese longsnout catfish (Leiocassis longirostris Günther)AQUACULTURE RESEARCH, Issue 16 2009Hongyue Zhao Abstract The present study was carried out to investigate the influence of water temperature on the growth performance and digestive enzyme (pepsin, trypsin and lipase) activities of Chinese longsnout catfish. Triplicate groups of Chinese longsnout catfish (35.6±0.48 g, mean±SE) were reared at different water temperatures (20, 24, 28 and 32 °C). The feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) were significantly affected by water temperatures and regression relationships between water temperature and FI, SGR as well as FER were expressed as FR=,0.016T2+0.91T,10.88 (n=12, R2=0.8752), SGR=,0.026T2+1.39T,17.29 (n=12, R2=0.7599) and FER=,0.013T2+0.70T,8.43 (n=12, R2=0.7272). Based on these, the optimum temperatures for FR, SGR and FER were 27.66, 26.69 and 26.44 °C respectively. The specific activities of digestive enzymes at 24 or 28 °C were significantly higher than that at 20 or 32 °C. In addition, there was a significant linear regression between FR or SGR and specific activities of pepsin and lipase, which indicated that pepsin and lipase played important roles in regulating growth through nutrient digestion in Chinese longsnout catfish. [source] Growth, digestive capacity and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of dietary inositolAQUACULTURE RESEARCH, Issue 8 2009Wei-Dan Jiang Abstract A 60-day feeding trial was carried out with juvenile Jian carp (Cyprinus carpio var. Jian) to study the effects of myo -inositol (MI) on the growth, digestive enzyme and intestinal microbial population. Diets with seven levels of inositol (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg MI kg,1 diet) were fed to Jian carp (initial weight 22.28±0.07 g). Per cent weight gain (PWG) was improved with increasing inositol levels up to 535.8 mg MI kg,1 diet (P<0.05), and plateaued (P>0.05). The protein production value, lipid production value and ash production value were increased with increasing dietary inositol levels up to 384.2, 838.8 and 838.8 mg MI kg,1 diet respectively (P<0.05). Although intestinal protein content and trypsin activity were not affected by inositol levels (P>0.05), chymotrypsin, lipase and amylase activities in intestine were the lowest for fish fed the MI-unsupplemented diet (P<0.05). Alkaline phosphatase, Na+, K+ -ATPase, ,-glutamyl transpeptidase and creatinkinase activities in the intestine were increased with an increase in the inositol levels up to 384.2,687.3 mg MI kg,1 diet (P<0.05). Intestinal Aeromonas hydrophila and Escherichia coli decreased with an increase in the levels of dietary inositol up to 232.7 and 687.3 mg MI kg,1 diet respectively (P<0.05), while Lactobacillus in the intestine increased with an increase in inositol levels up to 990.3 mg MI kg,1 diet (P<0.05). In conclusion, inositol improved growth, digestive capacity and intestinal microbial population of juvenile Jian carp, and the dietary inositol requirement for PWG of juvenile Jian carp is 518.0 mg MI kg,1 diet. [source] The feeding behavior of Trichogramma brassicae: new evidence for selective ingestion of solid foodENTOMOLOGIA EXPERIMENTALIS ET APPLICATA, Issue 1 2000Z.X. Wu Abstract A descriptive study of the feeding behavior and structures of Trichogramma brassicae Bezdenko (Hymenoptera: Trichogrammatidae) was conducted. Based on direct observational and biochemical evidence, larvae feed predominantly on particulate materials, starting ca. 25 h post-oviposition. Feeding lasted for ca. 9 h, at 25±1 °C. During this feeding period the shape of the larvae changed from vermiform to pyriform and then to sacciform, resulting in a ca. 40-fold increase in body size. Larvae used elaborate feeding behaviors as they pulled solid food particles to their oral opening, broke small particles from larger ones, and took the particles into the stomodaeum, which is a powerful pump. In the stomodaeum, peristaltic movement further macerated the particles, which eventually passed through the cardiac valve into the midgut. As indicated by changes in fluorescently labeled casein, digestive enzymes aid in the extra-oral chemical digestion of food. The contents of the gut, during and shortly after feeding, were almost entirely closely packed solid particles. The behavioral activity of feeding larvae centered almost exclusively on processing and ingesting solid food particles. The rapid larval growth is much more plausibly explained by their feeding on the highly concentrated nutrients found in solid foods, rather than the extensive concentration required if dilute liquids were the principal source of nutrients. The implications of these findings for the development of practical artificial diets are discussed. [source] Digestive peptidases in Tenebrio molitor and possibility of use to treat celiac diseaseENTOMOLOGICAL RESEARCH, Issue 3 2007Elena N. ELPIDINA Abstract Digestion in Tenebrio molitor larvae occurs in the midgut, where there is a sharp pH gradient from 5.6 in the anterior midgut (AM) to 7.9 in the posterior midgut (PM). Accordingly, digestive enzymes are compartmentalized to the AM or PM. Enzymes in the AM are soluble and have acidic or neutral pH optima, while PM enzymes have alkaline pH optima. The main peptidases in the AM are cysteine endopeptidases presented by two to six subfractions of anionic proteins. The major activity belongs to cathepsin L, which has been purified and characterized. Serine post-proline cleaving peptidase with pH optimum 5.3 was also found in the AM. Typical serine digestive endopeptidases, trypsin-like and chymotrypsin-like, are compartmentalized to the PM. Trypsin-like activity is due to one cationic and three anionic proteinases. Chymotrypsin-like activity consists of one cationic and four anionic proteinases, four with an extended binding site. The major cationic trypsin and chymotrypsin have been purified and thoroughly characterized. The predicted amino acid sequences are available for purified cathepsin L, trypsin and chymotrypsin. Additional sequences for putative digestive cathepsins L, trypsins and chymotrypsins are available, implying multigene families for these enzymes. Exopeptidases are found in the PM and are presented by a single membrane aminopeptidase N-like peptidase and carboxypeptidase A, although multiple cDNAs for carboxypeptidase A were found in the AM, but not in the PM. The possibility of the use of two endopeptidases from the AM , cathepsin L and post-proline cleaving peptidase , in the treatment of celiac disease is discussed. [source] A multivariate biomarker-based model predicting population-level responses of Daphnia magnaENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 9 2003Wim M. De Coen Abstract A multivariate model is proposed relating short-term biomarker measurements in Daphnia magna to chronic effects (21-d exposure) occurring at the population level (time to death, mean brood size, mean total young per female, intrinsic rate of natural increase, net reproductive rate, and growth). The results of the short-term exposure (48h-96 h) to eight model toxicants (cadmium, chromium, mercury, tributyl tin, linear alkylsulfonic acid, sodium pentachlorophenolate, lindane, and 2,4-dichloro-phenoxyacetic acid) on the following biomarkers were used for the multivariate model: digestive enzymes (amylase, cellulase, ,-galactosidase, trypsin, and esterase), enzymes of the intermediary metabolism (glycogen phosphorylase, glucose-6-phosphate de-hydrogenase, pyruvate kinase, lactate dehydrogenase, and isocitrate dehydrogenase), cellular energy allocation (CEA) (protein, carbohydrate, and lipid content and electron transport activity), and DNA damage and antioxidative stress activity. Using partial least squares to latent structures (PLS), a two-component model was obtained with R2 of 0.68 and a Q2 value of 0.60 based on the combined analysis of a limited number of the 48- and 96-h biomarker responses. For the individual population-level responses, the R2 values varied from 0.66 to 0.77 and the Q2 values from 0.52 to 0.69. Energy-related biomarkers (cellular energy allocation, lipid contents, anaerobic metabolic activity,pyruvate kinase, and lactate dehydrogenase), combined with parameters related to oxidative stress (catalase) and DNA damage measured after 48 and 96 h of exposure, were able to predict long-term effects at higher levels of biological organization. [source] X-ray crystallography and structural stability of digestive lysozyme from cow stomachFEBS JOURNAL, Issue 8 2009Yasuhiro Nonaka In ruminants, some leaf-eating animals, and some insects, defensive lysozymes have been adapted to become digestive enzymes, in order to digest bacteria in the stomach. Digestive lysozyme has been reported to be resistant to protease and to have optimal activity at acidic pH. The structural basis of the adaptation providing persistence of lytic activity under severe gastric conditions remains unclear. In this investigation, we obtained the crystallographic structure of recombinant bovine stomach lysozyme 2 (BSL2). Our denaturant and thermal unfolding experiments revealed that BSL2 has high conformational stability at acidic pH. The high stability in acidic solution could be related to pepsin resistance, which has been previously reported for BSL2. The crystal structure of BSL2 suggested that negatively charged surfaces, a shortened loop and salt bridges could provide structural stability, and thus resistance to pepsin. It is likely that BSL2 loses lytic activity at neutral pH because of adaptations to resist pepsin. [source] Transcriptional signatures in response to wheat germ agglutinin and starvation in Drosophila melanogaster larval midgutINSECT MOLECULAR BIOLOGY, Issue 1 2009H.-M. Li Abstract One function of plant lectins such as wheat germ agglutinin is to serve as defences against herbivorous insects. The midgut is one critical site affected by dietary lectins. We observed marked cellular, structural and gene expression changes in the midguts of Drosophila melanogaster third instar larvae that were fed wheat germ agglutinin. Some of these changes were similar to those observed in the midguts of starved D. melanogaster. Dietary wheat germ agglutinin caused shortening, branching, swelling, distortion and in some cases disintegration of the midgut microvilli. Starvation was accompanied primarily by shortening of the microvilli. Microarray analyses revealed that dietary wheat germ agglutinin evoked differential expression of 61 transcripts; seven of these were also differentially expressed in starved D. melanogaster. The differentially transcribed gene clusters in wheat germ agglutinin-fed larvae were associated with (1) cytoskeleton organization; (2) digestive enzymes; (3) detoxification reactions; and (4) energy metabolism. Four possible transcription factor binding motifs were associated with the differentially expressed genes. One of these exhibited substantial similarity to MyoD, a transcription factor binding motif associated with cellular structures in mammals. These results are consistent with the hypothesis that wheat germ agglutinin caused a starvation-like effect and structural changes of midgut cells of D. melanogaster third-instar larvae. [source] Genes encoding a group of related small secreted proteins from the gut of Hessian fly larvae [Mayetiola destructor (Say)]INSECT SCIENCE, Issue 5 2006MING-SHUN CHEN Abstract A group of related genes has been isolated and characterized from the gut of Hessian fly larvae [Mayetiola destructor (Say)]. Members in this group appear to encode proteins with secretary signal pep tides at the N-terminals. The mature putative proteins are small, acidic proteins with calculated molecular masses of 14.5 to 15.3 kDa, and isoelectric points from 4.56 to 4.88. Northern blot analysis revealed that these genes are expressed predominantly in the gut of Hessian fly larvae and pupae. Two related genes, G10K1 and G10K2, were isolated as tandem repeats. Both genes contain three exons and two introns. The intron/exon boundaries were conserved in terms of amino acid encoding, suggesting that they arose by gene duplication. The fact that the frequency of this group of clones in a gut cDNA library higher than that of total cDNA clones encoding digestive enzymes suggested that this group of proteins may perform an important function in the gut physiology of this insect. However, the exact functions of these proteins are as yet known since no sequence similarity could be identified between these proteins and any known sequences in public databases using standard methods. [source] Encapsulation efficiency and release behaviors of bovine serum albumin loaded in alginate microspheres prepared by sprayingJOURNAL OF APPLIED POLYMER SCIENCE, Issue 4 2008Jie Zhang Abstract Spraying and spraying with an electrostatic field (SEF) were employed to prepare alginate microspheres for delivering proteins, especially for intestinal digestive enzymes and cytokines. The encapsulation efficiency (EE) of a model protein [bovine serum albumin (BSA)] at a pH value lower than the isoelectric point was 20% higher than that at a natural pH. Moreover, for the microspheres prepared by SEF, EE improved significantly with increasing electric voltage. The interactions between BSA and the alginate microspheres were identified with Fourier transform infrared spectroscopy. The release profiles in vitro showed a controlled and pH-responsive release manner for the encapsulated BSA. A first-order release equation was postulated and modified to describe the release kinetics with an obviously initial burst release related to the eroded porous matrix. The equation fit the release data well when the pH value and composition of the release media were changed. The analysis of the release kinetics indicated that the drug release rate was in an inverse ratio to the diameter of the microspheres. Increasing the gas flow rate or electric voltage decreased both the mean diameter and size distribution of the microspheres significantly and enhanced the release rate of loaded drugs from alginate microspheres. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis analysis revealed that BSA kept its structural integrity during the encapsulation and release process. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2008 [source] Molecular mechanisms of pancreatitis: Current opinionJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 9 2008Alain Vonlaufen Abstract Pancreatitis (necroinflammation of the pancreas) has both acute and chronic manifestations. Gallstones are the major cause of acute pancreatitis, whereas alcohol is associated with acute as well as chronic forms of the disease. Cases of true idiopathic pancreatitis are steadily diminishing as more genetic causes of the disease are discovered. The pathogenesis of acute pancreatitis has been extensively investigated over the past four decades; the general current consensus is that the injury is initiated within pancreatic acinar cells subsequent to premature intracellular activation of digestive enzymes. Repeated attacks of acute pancreatitis have the potential to evolve into chronic disease characterized by fibrosis and loss of pancreatic function. Our knowledge of the process of scarring has advanced considerably with the isolation and study of pancreatic stellate cells, now established as the key cells in pancreatic fibrogenesis. The present review summarizes recent developments in the field particularly with respect to the progress made in unraveling the molecular mechanisms of acute and chronic pancreatic injury secondary to gallstones, alcohol and genetic factors. It is anticipated that continued research in the area will lead to the identification and characterization of molecular pathways that may be therapeutically targeted to prevent/inhibit the initiation and progression of the disease. [source] Intestinal function and body growth of broiler chickens on maize-based diets supplemented with mimosa tannins and a microbial enzymeJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 12 2004Paul A Iji Abstract A study was conducted to evaluate the effects of tannin (0, 5, 15, 20 and 25 g kg,1 diet) and a microbial enzyme supplement (MES) on the feed consumption, body growth and digestive physiology of broiler chickens between hatch and 22 days of age. Feed intake, body weight and body weight gain declined (p < 0.001) with an increase in dietary tannin content. Feed conversion efficiency was increased (p < 0.001) in line with dietary tannin level, up to 15 g kg,1 diet. There were no significant effects of dietary treatment on the protein content of pancreatic tissue or activities of pancreatic and jejunal enzymes. The ileal digestibilities of energy, protein, arginine, alanine and leucine were reduced (p < 0.001) as dietary tannin level rose to 20 g kg,1 diet and beyond. The digestibilities of methionine and phenylalanine were also negatively affected (p < 0.01) at the highest level of dietary tannins, while phosphorus digestion was improved (p < 0.05) on diets containing tannin. Apart from an increase (p < 0.01) in the protein content of the jejunal mucosa of birds on the diet with 20 g tannin kg,1 diet, there were no significant effects of the MES on most of the variables assessed. The results demonstrate the negative effects of tannin, especially at high levels of inclusion in the diet. However, neither tannins nor MES influenced the activities of digestive enzymes assessed, suggesting that a wider range of factors may be involved in regulating the effects of tannins on poultry. Copyright © 2004 Society of Chemical Industry [source] Digestive system and feeding mode in Cambrian naraoiid arthropodsLETHAIA, Issue 2 2002JEAN VANNIER The function of the digestive system of naraoiid arthropods is interpreted in the light of new observations on Early Cambrian specimens from China and detailed comparisons with Recent crustaceans and other arthropods. In naraoiids, paired tubular diverticulae ending as blind caeca are present along the entire midgut, and are interpreted as sites for the secretion of digestive enzymes. Naraoia bears one pair of long, ramifying, distensible diverticulae, possibly used for both food storage and digestion as suggested by Recent analogues (e.g. branchiuran and isopod crustaceans and limulids). Naraoiids were probably epibenthic scavengers/predators rather than mud-eaters. They were either opportunistic intermittent feeders (Naraoia) or more regular feeders (Misszhouia). The mud-fills of the alimentary canals are likely to be artefacts due to taphonomic and weathering processes or, less likely, to sediment ingestion by animals trapped alive in turbiditic flows. The case study of naraoiid arthropods adds to other fossil evidence supporting the idea that predation played a key role in the Early Cambrian food-webs and that organs adapted for this purpose had already reached a high level of diversity and anatomical sophistication. [source] Review article: proteinase-activated receptors , novel signals for gastrointestinal pathophysiologyALIMENTARY PHARMACOLOGY & THERAPEUTICS, Issue 3 2000Vergnolle Proteinase-activated receptors (PARs) have the common property of being activated by the proteolytic cleavage of their extracellular N-terminal domain. The new NH2 -terminus acts as a ,tethered ligand' binding and activating the receptor itself. Four members of this family have been cloned, three of which are activated by thrombin (PAR-1, PAR-3 and PAR-4) while the fourth (PAR-2) is activated by trypsin or mast cell tryptase. In physiological or pathophysiological conditions, the gastrointestinal tract is exposed more than other tissues to proteinases (digestive enzymes, proteinases from pathogens or proteinases from inflammatory cells) that can activate PARs. Since PARs are highly expressed throughout the gastrointestinal tract, the study of the role of PARs in these tissues appears to be particularly important. It has already been shown that PAR-2 activation induces calcium mobilization and eicosanoid production in enterocytes as well as changes in ion transport in jejunal tissue segments. PAR-2 activation also causes calcium mobilization and stimulates amylase release from pancreatic acini. Moreover, both PAR-1 and PAR-2 activation can alter the gastrointestinal motility. In inflammatory or allergic conditions, the proteinases that constitute the major agonists for PARs (thrombin, trypsin and mast cell tryptase) are usually released. The activation of PARs by these proteinases might contribute to the gastrointestinal disorders associated with these pathologies. A complete understanding of the role of PARs in the gastrointestinal tract will require the development of selective receptor antagonists that are not yet available. Nonetheless, the use of PAR agonists has already highlighted new potential functions for proteinases in the gastrointestinal tract, thus the control of PAR activation might represent a promising therapeutic target. [source] A New Method for the Treatment of Sperm Samples for Ultrastructural Study Based on the Use of Animal Tissues as Biological ContainersMICROSCOPY RESEARCH AND TECHNIQUE, Issue 9 2007Concepción Junquera Abstract The study of the ultrastructure of spematozoa by means of transmission electron microscopy often presents with problems of interpretation according to the method employed, depending on whether samples are either centrifuged previously to the fixation or immersed in viscous gels. The major problems of interpretation are: changes in the location of vesicles originated during the maturation process and modifications in the adsorption of seminal plasma proteins to the sperm membrane surface. The aim of our study is to communicate an original new method for the treatment of spermatozoa for ultrastructural study. Our method is based on the use of animal tissues as biological containers, inside which the spermatic suspensions are included. We developed this method using fresh sperm samples taken from mature Rasa aragonesa rams. As biological container, we used 2.5-cm long segments of the intestine of 1-week-old chickens (Gallus gallus) (diameter around 4 mm). To avoid any influence of digestive enzymes of the mucosa on the sperm surface, we put each intestine fragment inside out by means of microdissection forceps under bifocal optical microscope and cold light. One of the edges was tied with thin suture silk. The sperm suspension was injected in the optimal experimental condition and amount. Finally, the still open edge of the intestine segment was tied with silk in the same way as the other segment edge. By using this technique, we can perform a suitable morphological study at an ultrastructural level. In addition, the functional relationship of the ultrastructural components of the target cells is correctly preserved. Microsc. Res. Tech., 2007. © 2007 Wiley-Liss, Inc. [source] Immune-compromised state in the rat pancreas after chronic alcohol exposure: the role of peroxisome proliferator-activated receptor ,,THE JOURNAL OF PATHOLOGY, Issue 4 2007F Fortunato Abstract Alcohol exposure is known to sensitize acinar cells to various insults but the pathophysiological mechanisms of alcoholic pancreatitis remain unknown. Alcohol abuse has been shown to mediate an anti-inflammatory response and periods of immune suppression seem to be associated with organ injury and mortality. The purpose of this study was to determine the mechanisms by which alcohol exerts transcriptional activities in the rat pancreas and how alcohol alters the inflammatory response. Using the Lieber,DeCarli alcohol/control diet, rats that were fed with alcohol over 14 weeks demonstrated a decrease of inflammatory cells in pancreatic tissue compared to controls. The anti-inflammatory effects of alcohol were confirmed by decreased expression of pro-inflammatory cytokines including TNF,, IL-1,, IL-18, TGF,, and MCP-1. In addition, alcohol significantly increased the activity of PPAR,, which is a known anti-inflammatory transcription factor, while pro-inflammatory factors including AP-2 and EGR-1 were significantly suppressed. NF,B binding showed a tendency towards a reduction. Electron microscopy studies revealed enlarged and injured mitochondria and lysosomes, accompanied by peri-cellular fibrosis. Furthermore, alcohol exposure increased the activities of trypsin and cathepsin B, both known to be critical in initiating acinar cell injury and pancreatitis. Despite the known alcohol-mediated acinar cell and mitochondrial injury, the mitochondrial-mediated apoptotic pathway was attenuated. These data demonstrate that the pancreas exposed to alcohol maintains an anti-inflammatory state by activating PPAR,. Intracellular mitochondrial and lysosomal damage after chronic alcohol exposure induces premature activation of digestive enzymes and establishment of peri-cellular fibrosis in the absence of inflammation. Copyright © 2007 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [source] The Prenatal Development and Histochemistry of the Ileal Mucins in the Bovine FetusesANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 6 2009F. Beyaz Summary Few studies exist regarding the distribution of intestinal mucins in fetuses of mammalians such as cattle and sheep. In this study, we aimed to describe the changes in the mucin production by ileal epithelium of bovine fetuses during their prenatal development. The goblet cells showed heterogeneity in mucins and the apical cytoplasm of the enterocytes demonstrated Periodic acid Schiff-positive reaction which declined gradually towards the birth. Moreover, the number of the goblet cells containing acidic and mixed mucins augmented, whereas those containing neutral mucins decreased with advancing gestational age. After sixth month of gestation, with the initiation of the ileal Peyer patches and follicle-associated epithelium development, a gradual increase in the number of goblet cells containing sulfomucins was also noticed towards the birth. The presence of different mucins in the ileum of bovine fetuses throughout prenatal development might play a role in the protection of the intestinal mucosa against urinary waste products in swallowed amniotic fluid and bile. Furthermore, mucins can also contribute for the formation of meconium in intra-uterine life and building of strong intestinal barrier with predominating sulfomucins, protecting the intestine against potential pathogens and digestive enzymes after birth. [source] Effects of non-starch polysaccharides enzyme, phytase and citric acid on activities of endogenous digestive enzymes of tilapia (Oreochromis niloticus × Oreochromis aureus)AQUACULTURE NUTRITION, Issue 4 2009J.S. LI Abstract Non-starch polysaccharides (NSPs) enzyme, phytase and citric acid have been shown to increase digestibility in pigs, poultry and some fish. To examine their roles in digestion in tilapia (Oreochromis niloticus × Oreochromis aureus), the present study was designed to evaluate their effects on activities of a number of endogenous digestive enzymes in tilapia. Neither NSPs enzyme (1 g kg,1) nor phytase (1 g kg,1) affected the activities of protease in digestive organs. In contrast, NSPs enzyme increased the activities of amylase in the hepatopancreas and intestine by 11.4% and 49.5%, respectively, while phytase (1 g kg,1) increased the activities of amylase in the hepatopancreas and intestine by 14.0% and 24.1%, respectively. Citric acid (10 g kg,1) increased the activities of protease in stomach by 29.6%, but reduced the activities of protease in the intestine by 35.1%. Citric acid increased the activities of amylase in the hepatopancreas and intestine by 30.7% and 29.4%. Lipase activities were not affected by NSPs enzyme, phytase or citric acid. Above results suggest that NSPs enzyme, phytase and citric acid primarily influence the activities of amylase in digestive tract of tilapia. [source] Effect of microencapsulated diets supplemented with genetically modified bacteria on the growth and survival of Fenneropenaeus indicus postlarvaeAQUACULTURE NUTRITION, Issue 1 2007S. SIRVAS-CORNEJO Abstract Microencapsulated diets were prepared and supplemented with two genetically modified bacteria that produced digestive enzymes. One produced a protease (strain Escherichia coli XL1Bluep635), and the other a lipase and a protease (strain E. coli XL1Bluep7). Fenneropenaeus indicus at the postlarval 1 stage (PL1) were fed these diets for 16 days, and their total length and survival were recorded every 2 days. The results were analyzed by anova and sequential Turkey,Kramer analysis. Shrimp fed on the diet supplemented with strain E. coli XL1Bluep635 (diet 635) exhibited the fastest growth rate of 0.26 mm day,1, followed by shrimp fed on the control commercial unsupplemented diet CD2 , 0.21 mm day,1, and shrimp fed on diet supplemented with strain E. coli XL1Bluep7 (diet 7) , 0.20 mm day,1. The growth rates of shrimp fed on diet supplemented with control strain E. coli XL1BluepUC19 (diet XL1), and those fed on unsupplemented diet prepared in our laboratory (D2), were 0.15 and 0.14 mm day,1, respectively. The survival of shrimp fed on diets CD2 and 635 showed the same level of survival of 83.3%, followed by those fed on diet D2 with 76.6%. Shrimp fed on diet 7, showed 71.6% survival, and those fed on diet XL1, 55%. [source] Effect of water temperature on the growth performance and digestive enzyme activities of Chinese longsnout catfish (Leiocassis longirostris Günther)AQUACULTURE RESEARCH, Issue 16 2009Hongyue Zhao Abstract The present study was carried out to investigate the influence of water temperature on the growth performance and digestive enzyme (pepsin, trypsin and lipase) activities of Chinese longsnout catfish. Triplicate groups of Chinese longsnout catfish (35.6±0.48 g, mean±SE) were reared at different water temperatures (20, 24, 28 and 32 °C). The feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) were significantly affected by water temperatures and regression relationships between water temperature and FI, SGR as well as FER were expressed as FR=,0.016T2+0.91T,10.88 (n=12, R2=0.8752), SGR=,0.026T2+1.39T,17.29 (n=12, R2=0.7599) and FER=,0.013T2+0.70T,8.43 (n=12, R2=0.7272). Based on these, the optimum temperatures for FR, SGR and FER were 27.66, 26.69 and 26.44 °C respectively. The specific activities of digestive enzymes at 24 or 28 °C were significantly higher than that at 20 or 32 °C. In addition, there was a significant linear regression between FR or SGR and specific activities of pepsin and lipase, which indicated that pepsin and lipase played important roles in regulating growth through nutrient digestion in Chinese longsnout catfish. [source] Growth and digestive enzymes of Macrobrachium rosenbergii juveniles: effect of different stocktypes and dietary protein levels under a similar culture environmentAQUACULTURE RESEARCH, Issue 12 2009Vidya Sagar Abstract A feeding trial was conducted to study the effect of dietary protein (DP) levels on the growth and digestive enzyme activities of different wild stocks of Macrobrachium rosenbergii juveniles. Wild juveniles of M. rosenbergii were collected from the west coast of India, Gujarat (G), Maharashtra (M) and from the east coast of India, Andhra Pradesh (A), and raised in culture ponds of 200 m2 at 1 juvenile m,2. All the animals were tagged individually with Elastomer tags of a particular colour assigned to their respective stock and acclimatized for 7 days before being released into the pond at a ratio of 70:65:65 (A:M:G). Each of the two feeds, the first with 27% DP, termed the suboptimum level (S), and the second 32% DP, termed the optimum level (O), was fed in duplicate ponds at 6% of the body mass for the first 30 days and 4% for the last 30 days. The average weight of stocked prawn, respectively, in O DP and S DP fed ponds was 0.90 ± 0.04 and 1.06 ± 0.08 g for the G stock, 0.80 ± 0.07 and 1.01 ± 0.1 g for the M stock and 3.06 ± 0.13 and 3.10 ± 0.23 g for the A stock. Both the protein level and the stock type had a significant (P<0.05) effect on the weight gain% of the prawn. There was an approximate 95% change in weight gain with a DP change. Similarly, G and M stocks exhibited significantly higher (P<0.05) growth rates of approximately 90% than the A stock, although no difference was noted between the G and the M stocks. However, for protein × stock (interaction) levels, there was no significant difference (P>0.05) among the groups. Although insignificant, the survival rates among the different stocks varied from 56% to 77%. Optimum protein level showed a significant increase (P<0.05) in the specific growth rate (SGR). Feed conversion ratio, feed efficiency ratio, protein efficiency ratio and net protein utilization were not affected either due to DP, stock type or their interaction. The O × A group exhibited the maximum variation in body weight. Digestive enzyme activities were similar in all the groups, but enzymes for phospho-monoesterase were significantly higher (P<0.05) at O DP. Both the G and the M stock showed a significantly higher (P<0.05) alkaline phosphatase activity while acid phosphatase activity was significantly higher (P<0.05) in the M stock only. Overall, the G and M stocks showed higher growth responses compared with the A stock. [source] Activities of digestive enzymes during embryonic development in the crayfish Procambarus clarkii (Decapoda)AQUACULTURE RESEARCH, Issue 12 2009Ying Dai Abstract The red swamp crayfish, Procambarus clarkii (Girard), has become an important freshwater species for Chinese inland aquaculture because of its high commercial value and consumer demand. The aim of this study was to gather information about the activity of digestive enzymes at different embryonic stages of P. clarkii in order to increase our knowledge about digestive physiology and to guide technology for maternal culture so as to improve the hatching rate. Embryonic developmental stages were divided into six stages: I, fertilized egg; II, cleavage and blastula; III, gastrula; IV, egg nauplius; V, eye pigment forming; and VI, prepared for hatching. Pepsin-specific activity decreased significantly from stage I to stage IV. Although it increased at stage V, the activity level declined again before hatching. Both trypsin- and amylase-specific activity levels dramatically increased in the middle stages of embryogenesis, whereas at the other embryonic stages the activities of these two digestive enzymes were much lower. Lipase-specific activity exhibited a low level during all embryonic stages. The pattern of digestive enzyme activity was related to organogenesis and the utilization of yolk proteins at different embryonic stages. [source] A method for the histochemical localization of digestive enzymes in whole freeze-substituted larval fish embedded in glycol methacrylateAQUACULTURE RESEARCH, Issue 7 2009Grant W Vandenberg Abstract In order to study the localization of digestive enzymes in larval walleye (Sander vitreus vitreus), a novel method of low-temperature processing of whole, unfixed larvae and subsequent embedding in glycol methacrylate (GMA) was developed. Larval walleye aged 2,10 days post hatch were flash-frozen in liquid nitrogen and transferred into pre-chilled acetone for 12 h at a temperature of ,25 °C. Acetone was gradually replaced with increasing concentrations of GMA resin monomer over a 6-h period. The polymer (100%) was further allowed to infiltrate the larvae for 36 h. Resin-embedded larvae were transferred to embedding moulds and polymerized overnight at ,25 °C. Four micrometre sections were stained to identify either alkaline phosphatase, non-specific esterase, aminopeptidase M or dipeptidyl peptidase IV. All enzymes studied were readily detected and accurately localized, and no enzyme diffusion was observed. Therefore, freeze substitution combined with low-temperature GMA embedding allows the maintenance of excellent tissue morphology and accurate enzyme localization in whole larval walleye specimens from 2 to 10 days post hatch. It is recommended, however, that samples be frozen in pre-cooled fluorocarbon-based liquid coolants in order to assure optimal tissue preservation. [source] Digestive enzyme spectra in crustacean decapods (Paleomonidae, Portunidae and Penaeidae) feeding in the natural habitatAQUACULTURE RESEARCH, Issue 3 2009Maria Santos Reis Bonorino Figueiredo Abstract This work describes the profile of five proteases and four carbohydrases from the crustacean decapods Macrobrachium australiense (Holthuis), Scylla serrata (Forskal), Portunus pelagicus (Linnaeus), Penaeus esculentus, Penaeus plebejus (Hess) and Metapenaeus bennettae (Racck & Dall), feeding in the natural habitat, in order to provide an indication of their digestive capabilities. The results raised the following points. First, species from each family showed a particular suite of digestive enzymes. Second, the activity of cellulase from M. australiensis and S. serrata, using AZCL-HE cellulose as the substrate, was around 90% higher than that observed with AZO-CM cellulose. However, for P. pelagicus and P. esculentus, the enzyme activity was better with AZO-CM cellulose. Third, M. australiense displayed the highest ratio of amylase to protease activity. In contrast, Portunidae species, P. pelagicus and S. serrata showed the lowest ratios. Fourth, comparison of the laminarinase activity of M. bennettae and P. esculentus in October (Spring) and December (early Summer) showed a significant decrease in December. Finally, the wide distribution of digestive enzymes in these crustaceans may reflect different feeding habits and habitats. [source] Development and distribution of intestinal enzymatic activity in Paralabrax maculatofasciatus (Steindachner, 1868) larvae fed with live preyAQUACULTURE RESEARCH, Issue 2 2009Renato Peña Abstract We describe the development and distribution of intestinal aminopeptidase M, dipeptidyl aminopeptidase IV, non-specific esterase, alkaline phosphatase and acid phosphatase, using enzyme histochemistry techniques, in the spotted sand bass larvae (Paralabrax maculatofasciatus) under culture conditions. All digestive enzymes tested showed a positive reaction from first feeding (day 2) and throughout the study period (day 30). At first feeding, the main enzymatic activity was in the mucosa throughout the intestines. Later, enzymatic activity occurred in the liver, kidney and stomach. All enzymatic activities increased from days 15 to 20, remaining constant until the end of the study. This enzymatic activity suggests the onset of maturation of the digestive tract. After day 20, a positive reaction was recorded in the pyloric caeca for all tested enzymatic activities. Our study confirms the digestive and absorptive functions in the intestines in spotted sand bass larvae from first feeding. It also brings new insight to establish an early weaning strategy during cultivation of spotted sand bass larvae. [source] Digestive enzyme activity at different developmental stages of blackspot seabream, Pagellus bogaraveo (Brunnich 1768)AQUACULTURE RESEARCH, Issue 4 2008Laura Ribeiro Abstract Blackspot seabream, Pagellus bogaraveo (Brunnich), has been identified as a potential species to diversify European aquaculture production. Although rearing aspects have been widely investigated, little information exists on the nutritional requirements for this species. The aim of this study was to build up information on the activity of digestive enzymes at certain developmental stages of blackspot seabream in order to understand the nutritional needs of larvae and post larvae. Fish larvae were reared from hatching to 55 days after hatching (dah), and the feeding plan consisted in rotifers (5,35 dah), Artemia naupli (30,35 dah) metanaupli (35,45) and Gemma microdiet (45,55 dah). At 7, 11, 21, 45 and 55 days after hatching (dah), pooled samples of fish larvae were collected for analysis of trypsin, amylase, lipase, alkaline phosphatase and leucine,alanine peptidase activity. Up to 21 dah, the whole larvae body was used for enzymatic analysis, whereas in older larvae only the dissected abdominal cavity was used. Blackspot seabream body dry weight growth was exponential, increasing from 60 ,g at 5 dah to 30±9.7 mg at 55 dah. Amylase specific activity decreased significantly during development, exhibiting at 11 dah (0.6 U mg,1 protein) an average value 2.7 times lower than at 7 dah, and remaining stable between 45 and 55 dah (0.7 U mg protein,1). Trypsin specific activity remained constant until 21 dah (between 38 and 44 mU mg protein,1), which could be related to the larvae feeding regime. At later stages of development, lipase-specific activity exhibited a significant increase (P<0.05), being three times higher at 55 dah (8 U mg protein,1) than at 45 dah. The total activity of the studied digestive enzymes increased significantly during larval development (until 21 dah), whereas afterwards only lipase and leucine,alanine peptidase increased significantly between 45 and 55 dah. The pattern of digestive enzymes activity was related to organogenesis and the type of food used at different developmental stages. [source] Production and characterization of a recombinant beta-1,4-endoglucanase (glycohydrolase family 9) from the termite Reticulitermes flavipesARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 3 2010Xuguo Zhou Abstract Cell-1 is a host-derived beta-1,4-endoglucanase (Glycohydrolase Family 9 [GHF9]) from the lower termite Reticulitermes flavipes. Here, we report on the heterologous production of Cell-1 using eukaryotic (Baculovirus Expression Vector System; BEVS) and prokaryotic (E. coli) expression systems. The BEVS-expressed enzyme was more readily obtained in solubilized form and more active than the E. coli,expressed enzyme. Km and Vmax values for BEVS-expressed Cell-1 against the model substrate CMC were 0.993% w/v and 1.056,µmol/min/mg. Additional characterization studies on the BEVS-expressed enzyme revealed that it possesses activity comparable to the native enzyme, is optimally active around pH 6.5,7.5 and 50,60°C, is inhibited by EDTA, and displays enhanced activity up to 70°C in the presence of CaCl2. These findings provide a foundation on which to begin subsequent investigations of collaborative digestion by coevolved host and symbiont digestive enzymes from R. flavipes that include GHF7 exoglucanases, GHF1 beta glucosidases, phenol-oxidizing laccases, and others. © 2010 Wiley Periodicals, Inc. [source] Control of the release of digestive enzymes in the larvae of the fall armyworm, Spodoptera frugiperdaARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 1 2010Digali Lwalaba Abstract There is a basal level of enzyme activity for trypsin, aminopeptidase, amylase, and lipase in the gut of unfed larval (L6) Spodoptera frugiperda. Trypsin activity does not decrease with non-feeding, possibly because of the low protein levels in plants along with high amino acid requirements for growth and storage (for later reproduction in adults). Therefore, trypsin must always be present so that only a minimal protein loss via egestion occurs. Larvae, however, adjust amylase activity to carbohydrate ingestion, and indeed amylase activity is five-fold higher in fed larvae compared to unfed larvae. Gut lipase activity is low, typical of insects with a high carbohydrate diet. A flat-sheet preparation of the ventriculus was used to measure the release of enzymes in response to specific nutrients and known brain/gut hormones in S. frugiperda. Sugars greatly increase (>300%) amylase release, but starch has no effect. Proteins and amino acids have little or no effect on trypsin or aminopeptidase release. The control of enzyme release in response to food is likely mediated through neurohormones. Indeed, an allatostatin (Spofr-AS A5) inhibits amylase and trypsin, and allatotropin (Manse- AT) stimulates amylase and trypsin release. Spofr-AS A5 also inhibits ileum myoactivity and Manse-AT stimulates myoactivity. The epithelial secretion rate of amylase and trypsin was about 20% of the amount of enzyme present in the ventricular lumen, which, considering the efficient counter-current recycling of enzymes, suggests that the secretion rate is adequate to replace egested enzymes. © 2009 Wiley Periodicals, Inc. [source] Mammalian target of rapamycin signaling is crucial for joint destruction in experimental arthritis and is activated in osteoclasts from patients with rheumatoid arthritisARTHRITIS & RHEUMATISM, Issue 8 2010Daniel Cejka Objective Activation of the mammalian target of rapamycin (mTOR) pathway is important for immune cell activation and bone metabolism. To date, the contribution of mTOR signaling to joint inflammation and structural bone and cartilage damage is unknown. The aim of this study was to investigate the potential of inhibiting mTOR as a treatment of inflammatory arthritis. Methods Human tumor necrosis factor,transgenic mice in which inflammatory arthritis was developing were treated with 2 different mTOR inhibitors, sirolimus or everolimus. The effects of treatment on clinical disease activity, inflammation, and localized joint and cartilage destruction were studied. In addition, the effects of mTOR inhibition on osteoclast survival and expression of key molecules of osteoclast function were analyzed in vitro. Moreover, synovial tissue from patients with rheumatoid arthritis (RA) was assessed for activation of the mTOR pathway. Results Inhibition of mTOR by sirolimus or everolimus reduced synovial osteoclast formation and protected against local bone erosions and cartilage loss. Clinical signs of arthritis improved after mTOR inhibition, and histologic evaluation showed a decrease in synovitis. In vitro, mTOR inhibition down-regulated the expression of digestive enzymes and led to osteoclast apoptosis. Moreover, mTOR signaling was shown to be active in the synovial membrane of patients with RA, particularly in synovial osteoclasts. Conclusion Signaling through mTOR is an important link between synovitis and structural damage in inflammatory arthritis. Current pharmacologic inhibitors of mTOR could be effective in protecting joints against structural damage. [source] | ||||||||||||||||||||||