Decrease In Trypsin Activity Research Articles

Serum Activity of Proteolytic Enzyme Trypsin in Rats under Conditions of Water and Food Deprivation.

Trypsin is mainly regarded as a digestive enzyme, but there is evidence that activation of protease-activated receptor-2 (PAR-2) leads to behavioral changes. There are no data on trypsin activity in the serum of animals under conditions of thirst and starvation in the available literature. In our experiments, water deprivation led to a significant (p⩽0.05) increase in trypsin activity in rats, and food deprivation led to its decrease in comparison with controls (free access to water and food). After deprived rats received water and food, a decrease in trypsin activity was observed in both experimental groups. Changes in trypsin activity under conditions of water or food deprivation and after satiation were accompanied by shifts in some biochemical parameters of the bloods. Under conditions of metabolic stress (starvation and thirst), opposite changes in trypsin activity seem to indicate its participation in the mechanisms of adequate restructuring of metabolism and maintenance of vital processes in the body.

Double deficiency of cathepsin B and L in the mouse pancreas alters trypsin activity without affecting acute pancreatitis severity

BackgroundPremature intracellular trypsinogen activation has long been considered a key initiator of acute pancreatitis (AP). Cathepsin B (CTSB) activates trypsinogen, while cathepsin L (CTSL) inactivates trypsin(ogen), and both proteins play a role in the onset of AP. MethodsAP was induced by 7 hourly intraperitoneal injections of cerulein (50 μg/kg) in wild-type and pancreas-specific conditional Ctsb knockout (CtsbΔpan), Ctsl knockout (CtslΔpan), and Ctsb;Ctsl double-knockout (CtsbΔpan;CtslΔpan) mice. Pancreatic samples were collected and analyzed by histology, immunohistochemistry, real-time PCR, and immunoblots. Trypsin activity was measured in pancreatic homogenates. Peripheral blood was collected, and serum amylase activity was measured. ResultsDouble deletion of Ctsb and Cstl did not affect pancreatic development or mouse growth. After 7 times cerulein injections, double Ctsb and Ctsl deficiency in mouse pancreases increased trypsin activity to the same extent as that in Ctsl-deficient mice, while Ctsb deficiency decreased trypsin activity but did not affect the severity of AP. CtsbΔpan;CtslΔpan mice had comparable serum amylase activity and histopathological changes and displayed similar levels of proinflammatory cytokines, apoptosis, and autophagy activity compared with wild-type, CtsbΔpan, and CtslΔpan mice. ConclusionDouble deletion of Ctsb and Ctsl in the mouse pancreas altered intrapancreatic trypsin activity but did not affect disease severity and inflammatory response after cerulein-induced AP.

Study on the interaction of ethylene glycol with trypsin: Binding ability, activity, and stability

The study of the structural changes and thermal stabilities of proteins in the presence of various co-solutes is a critical element of basic research, drug discovery, and development. Ethylene glycol (EG) is typically known to be poisonous and teratogenic, causing skeletal and external abnormalities as well as other disorders. At pH 8, the influence of EG on the structure and thermal stability of trypsin was investigated using a variety of spectroscopic techniques. Spectroscopic techniques, fluorescence spectroscopy, circular dichroism (CD), and molecular dynamics simulation studies were used to evaluate the binding reaction between ethylene glycol and trypsin in the current study. Circular dichroism, Trp fluorescence, and absorption tests demonstrated that EG has an effect on secondary and tertiary structures of trypsin, and it boosts the protein's thermal stability, which increases with increasing EG concentration. This contact occurs through a static quenching process, according to fluorescence spectroscopy data, and van der Waals contributions and hydrogen bonds were also important in binding ethylene glycol to trypsin. The ethylene glycol binding constant (Kb) and the number of binding sites (n) were calculated using fluorescence quenching data. Kinetic technique analysis displayed a decrease in trypsin activity in the presence of ethylene glycol. Also, circular dichroism results confirmed the conformation changes resulting from the binding of ethylene glycol to trypsin. Molecular dynamics simulation using RMSD was used to test the stability of the trypsin-ethylene glycol complex in this study. Molecular docking research revealed that EG interacts with trypsin amino acid residues via hydrogen bonding and van der Waals interactions.

Effects of fluoranthene on digestive enzymes activity and relative growth rate of larvae of lepidopteran species, Lymantria dispar L. and Euproctis chrysorrhoea L.

Fluoranthene is one of the most abundant polycyclic aromatic hydrocarbon pollutants in the environment and it may accumulate in plant leaves which are the main food source for phytophagous insect species. The aim of this study was to establish the effects of dietary fluoranthene on specific activities of digestive enzymes and expression of their isoforms in the midgut, and the relative growth rates of Lymantria dispar and Euproctis chrysorrhoea larvae. Exposure to fluoranthene led to significantly decreased trypsin activity in the midgut of larvae of both species. Leucine aminopeptidase activity decreased significantly in the midgut of L. dispar larvae exposed to the lower concentration of fluoranthene, but that enzyme activity showed the opposite trend in E. chrysorrhoea larvae. There was no pollutant induced changes in lipase activity in L. dispar, while elevated enzyme activity was recorded in the midgut of E. chrysorrhoea larvae exposed to the lower concentration of fluoranthene. Different patterns of expression of enzyme isoforms were noticed. Relative growth rates of both species significantly decreased in fluoranthene treated larvae. These responses indicate to the significance of relationships between physiological changes and fitness-related traits in L. dispar and E. chrysorrhoea larvae affected by pollutant, and contribute to understanding the mechanisms of their adjustment to stressful conditions.

Mealworm larvae meal in diets for meagre juveniles: Growth, nutrient digestibility and digestive enzymes activity

This study aimed to evaluate the effect of including 10%, 20% and 30% of mealworm (Tenebrio molitor, TM) larvae meal (diets TM10, TM20 and TM30, respectively), replacing 25%, 50% and 75% of fishmeal (FM) in a isoproteic (50%) and isolipidic (19%) FM-based diet (CTR) for meagre (Argyrosomus regius). The diets were fed for 9 weeks to triplicate groups of fish with 18.0 g initial body weight and results were evaluated for growth performance; whole-body composition; amino acid (AA) catabolism and digestive enzymes activity; nutrients and chitin digestibility. A digestibility trial was also simultaneously conducted for 48 days. Growth and feed utilization parameters decreased linearly as dietary TM inclusion level increased, while mortality, hepatosomatic and visceral indexes, and AA catabolism enzymes activity were unaffected by the dietary treatments. Whole-body protein was reduced in fish fed TM20 and TM30 diets, with the latter group also having lower energy content than fish fed the CTR diet. The apparent digestibility coefficients (ADC) of dry matter, energy, protein, and AA (except leucine, valine and phenylalanine), decreased as dietary TM inclusion level increased. The lower ADC of protein was related to a linear decrease in trypsin activity. Lipase and total alkaline protease activities were also lower in fish fed the TM30 diet than in the other groups. A quadratic response of total alkaline protease with the increase of dietary TM meal was similar to that observed in the intensity of zymograms bands corresponding to proteases other than trypsin or chymotrypsin-like. Chitin ADC was null and no chitinolytic activity was found in the intestine. Overall, meagre presented a limited capacity to utilize TM, with a 10% TM dietary inclusion already resulting in significant impairment of fish digestive capacity and growth performance, while higher TM dietary levels further negatively affecting fish whole-body composition.

Ultraviolet irradiation of trypsin, lysozyme and β-galactosidase: how does UVC affect these enzymes when used as a secondary barrier against adventitious agents?

Trypsin is one of the essential raw materials used in the manufacturing of biopharmaceutical products. As an animal derived product, it can potentially carry a serious risk of contamination with adventitious agents that can result in production shut down and lost product. To mitigate these risks, several methods are currently being used in the industry to remove contamination including physical and chemical methods. Ultraviolet-C (UVC) light is known to inactivate adventitious agents that are resistant to physical and chemical methods and could be a secondary barrier strategy. In this study, we investigated the effect of UVC irradiation on the activity and structure of trypsin. Extreme doses of UVC light were applied to trypsin using a collimated beam apparatus. The effect of UVC light on trypsin enzymatic activity was measured using a colorimetric activity assay and the effect on structure was analyzed by spectrophotometry, gel electrophoresis, and mass spectrometry. To broaden the scope, the effect of UVC light on the activity of two additional enzymes, lysozyme and β-galactosidase, was also examined. At high doses of UVC light, changes to protein structure and protein fragmentation resulted in decreased trypsin activity. However, minimal damage was observed at doses applicable to inactivating adventitious agents, making UVC a feasible treatment for viral inactivation of trypsin products.

Trypsin and N-aminopeptidase (APN) activities in the hepatopancreas of an intertidal euryhaline crab: Biochemical characteristics and differential modulation by histamine and salinity

No studies are available about biochemical characteristics and modulation (i.e. by endogenous and/or environmental cues) of trypsin (a key digestive endoprotease) in hepatopancreas of intertidal euryhaline crabs neither on the possible concomitant modulation of key ectoproteases such as aminopeptidase-N (APN) involved in final steps of protein digestion. Furthermore, nothing is still known in decapods crustaceans about the role of histamine as primary chemical messenger for modulation of main components of digestive process (i.e. proteases). We determined biochemical characteristics and investigated the effect of histamine injections; of histamine in vitro and of acclimation of individuals to low and high salinity on trypsin and aminopeptidase-N (APN) activities in the hepatopancreas of the euryhaline crab Cyrtograpsus angulatus (Dana 1851). Trypsin activity was maximal at pH7.4 and at 45°C. APN activity increased from pH6.6 to 7.6–9.0 and was maintained high at 37–45°C. Both activities exhibited Michaelis-Menten kinetics (apparent Km: trypsin=0.36mM; APN=0.07mM). The injection of 10−4M histamine decreased trypsin activity (about 40%) in hepatopancreas while did not affect APN activity. Similarly, in vitro 10-4M histamine decreased trypsin activity (about 52%) in hepatopancreas but not APN activity. Trypsin activity in the hepatopancreas was not affected by acclimation of crabs to low (10psu) or high (40psu) environmental salinity while APN activity was increased (about 200%) in 10psu. The results show the differential modulation of trypsin and APN by distinct cues and point to histamine as modulator of intracellular trypsin by direct action on the hepatopancreas.

Binding characteristics of psoralen with trypsin: Insights from spectroscopic and molecular modeling studies

Psoralen (PSO) is a naturally occurring furanocoumarin with a variety of pharmacological activities, however very limited information on the interaction of PSO with trypsin is available. In this study, the binding characteristics between PSO and trypsin at physiological pH were investigated using a combination of fluorescence, UV–vis absorption, circular dichroism (CD), Fourier transform infrared (FT-IR) spectroscopic, chemometric and molecular modeling approaches. It was found that the fluorescence quenching of trypsin by PSO was a static quenching procedure, ascribing the formation of a PSO–trypsin complex. The binding of PSO to trypsin was driven mainly by hydrophobic forces as the positive enthalpy change and entropy change values. The molecular docking showed that PSO inserted into the active site pocket of trypsin to interact with the catalytic residues His57, Asp102 and Ser195 and may cause a decrease in trypsin activity. The results of CD and FT-IR spectra along with the temperature-induced denaturation studies indicated that the addition of PSO to trypsin led to the changes in the secondary structure of the enzyme. The concentration profiles and spectra of the three components (PSO, trypsin, and PSO–trypsin complex) obtained by multivariate curve resolution-alternating least squares analysis exhibited the kinetic processes of PSO–trypsin interaction. This study will be helpful to understand the mechanism of PSO that affects the conformation and activity of trypsin in biological processes.

Ontogenetic development of digestive enzymes in Sobaity sea bream Sparidentex hasta larvae under culture condition

Activities of the main enzymes involved in digestion (pepsin, trypsin, amylase, lipase and alkaline phosphatase) in Sobaity sea bream were studied from hatching up to the juvenile stages (40days after hatch, DAH). All enzyme activities except for pepsin were detected upon hatching, and their activity subsequently increased with age. Early presence of trypsin and amylase was attributed to a genetically programmed process. Trypsin exhibited fluctuating pattern (with tow peaks at 16 and 30DAH) triggered by food ingestion and was in response to the diet shifting during the rearing period. Both lipase and amylase activities were enhanced after the onset of the weaning depending upon the microbound diet components. Early presence and incremental trend of alkaline phosphatase from 5th DAH represented early maturation of enterocytes and consequently rapid growth of this species. Pepsin was first detected at 14DAH and showed a sharp increasing trend up to end of the experiment. Its progressive increase followed by a decrease in trypsin activity after accomplishing weaning exhibited a change in the digestive physiology to reach to an adult-type protein digestion. In conclusion, such early ontogenetic development of the above mentioned enzymes makes it possible to wean Sobaity sea bream larvae earlier than 25DAH in order to reduce live food usage and improve weaning efficiency. Statement of relevanceObtaining knowledge about digestive physiology makes it possible to quantify the capacity of larvae to digest and absorb different types of nutrients from live feed and/or microbound diets and to understand the right time to conduct weaning in Sobaity sea bream larvae earlier than what has been practiced in hatcheries in order to reduce live food usage and improve weaning efficiency.

Effects of exogenous protease supplementation on endogenous trypsin activity and gene expression in broilers.

Two hundred and forty one-day-old male broiler chickens (Ross-308) were fed corn-soybean basal diets with 0, 40, 80, or 160 mg/kg of an acid protease preparation from 1 to 42 days of age, in order to investigate the effects of an exogenous enzyme on growth performance, pancreatic trypsin activity and mRNA expression. Average daily gain (ADG) was greatest in the 160-mg/kg treatment group at 1-21 days, which was significantly higher than that in the control. After 42 days, average daily feed intake (ADFI) and ADG had significantly increased in the 80- and 160-mg/kg treatments (P < 0.05), with the 80-mg/kg treatment group having the highest ADFI and ADG values. The feed conversion ratio was not affected. Dietary acid protease at 80 and 160 mg/kg significantly decreased trypsin activity in the pancreas (relative to the control group) by 35.71 and 47.29%, respectively (P < 0.05). After 42 days, trypsin mRNA expression in the pancreas had significantly decreased by 19.5% in the 80-mg/kg treatment group relative to the control (P < 0.05). After 21 and 42 days, the diet supplemented with 160 mg/kg acid protease significantly decreased pancreatic trypsin mRNA by 19.6 and 37.7%, respectively, compared to the control. There were the 40-mg/kg treatment group and the control. Our results suggest that the amount of acid protease in the diet significantly affects trypsin activity and mRNA expression in broilers.

Mesenteric lymph drainage alleviates acute kidney injury induced by hemorrhagic shock without resuscitation.

This study aimed to investigate the effect of mesenteric lymph drainage on the acute kidney injury induced by hemorrhagic shock without resuscitation. Eighteen male Wistar rats were randomly divided into sham, shock, and drainage groups. The hemorrhagic shock model (40 mmHg, 3 h) was established in shock and drainage groups; mesenteric lymph drainage was performed from 1 h to 3 h of hypotension in the drainage group. The results showed that renal tissue damage occurred; the levels of urea, creatinine, and trypsin in the plasma as well as intercellular adhesion molecule-1 (ICAM-1), receptor of advanced glycation end-products (RAGE), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA), lactic acid (LA), and 2,3-DPG in the renal tissue were increased in the shock group after 3 h of hypotension. Mesenteric lymph drainage lessened the following: renal tissue damage; urea and trypsin concentrations in the plasma; ICAM-1, RAGE, TNF-α, MDA, and LA levels in the renal tissue. By contrast, mesenteric lymph drainage increased the 2,3-DPG level in the renal tissue. These findings indicated that mesenteric lymph drainage could relieve kidney injury caused by sustained hypotension, and its mechanisms involve the decrease in trypsin activity, suppression of inflammation, alleviation of free radical injury, and improvement of energy metabolism.

Interaction Between the Quorum Sensing and Stringent Response Regulation Systems in the Enterohemorrhagic Escherichia coli O157:H7 EDL933 Strain

In order to investigate the interaction of well-known regulation systems, quorum sensing and stringent response for the expression of virulence genes in enterohemorrhagic Escherichia coli (EHEC), E. coli strains with mutations in the two regulation systems, ΔluxS (ECM101) and ΔluxSΔrelAΔspoT (ECM201), and ΔluxS complemented strain to ECM201 (ECM202) were created from EHEC O157:H7 EDL933. Phenotypic characterization revealed that mutations in the stringent response system (ECM201 and ECM202) influenced the metabolic (defective utilization of arabinose and L-sorbose), enzymatic activities (decreased trypsin activity, and increased α-glucosidase activity) and motility. Our results suggest that quorum sensing interacts with stringent response regulation system.

Effects of curcumin on proinflammatory cytokines and tissue injury in the early and late phases of experimental acute pancreatitis

Background & aimsAcute pancreatitis (AP) varies from mild to severe necrotizing changes with high mortality. The objective of the current study was to investigate the effects of curcumin on tissue injury and proinflammatory cytokines in the early and late phases of AP. MethodsAP was induced by sodium taurocholate in rats (n = 140). First group was left untreated. Group II received 100 mg/kg curcumin daily starting 20 days before AP induction. The rats were allocated into 7 sub-groups (n:5) and were sacrificed at 2, 6, 12, 24, 72, 144 and 288 h following the induction of AP. Blood and pancreatic tissue samples were collected for biochemical and histopathologic evaluations and the assessment of protein and mRNA levels, as well. ResultsCurcumin decreased total histopathologic scores in comparison with those of the taurocholate group (P < 0.05). Curcumin increased Caspase-3 activity and decreased trypsin activity, while inhibited nuclear factor-κ (NF-κB) at all time points (P < 0.05) and moreover reduced activator protein-1 (AP-1). Curcumin decreased chemokine (except for 288 h), TNF-α (except for 2 and 24 h), IL-6 (except for 2, 6 and 288 h) and iNOS (except for 144 and 288 h) mRNA levels (P < 0.05). Curcumin serum nitric oxide (NO) (except for 144 and 288 h) levels were reduced, as well. ConclusionsIn conclusion, curcumin reduced tissue injury, trypsin activation and inhibited NF-κB and AP-1. However TNF-α, IL-6 and iNOS and NO were not inhibited at all time points. Therefore no direct correlation was detected in the subgroups between tissue injury, proinflammatory cytokines and oxidative enzymes.

Responses of digestive enzymes and body lipids to weaning times in yellowtail kingfishSeriola lalandi(Valenciennes, 1833) larvae

The optimal time to wean fish larvae from live feed to artificial feed was explored in yellowtail kingfish Seriola lalandi (YTK). The same weaning regime started at five different days post hatching (DPH), namely 10 DPH (W10), 13 DPH (W13), 16 DPH (W16), 19 DPH (W19) and 22 DPH (W22). The activities of trypsin, lipase and alkaline phosphatase were detected in fish from 8 DPH throughout the experiment, but pepsin activity was first detected in fish on 15 DPH in the W10 and W13 treatments. The increase in pepsin activity was concomitant with the decrease in trypsin activity. Total fish lipids after weaning reduced by 40% in the W10 and W13 treatments, and increased by 20% in the W19 and W22. Fish survival rate in the W22 treatment was significantly higher than that in the W10 and W13 treatments. The results suggest that 16 DPH is the earliest day to wean and the best weaning window for YTK larvae should be 19–22 DPH. This study provides enzymatic evidence to guide the weaning process for YTK larvae, and offers a useful approach to explore optimal weaning time for fish larvae.

Effect of disaccharides on the stabilization of bovine trypsin against detergent and autolysis

Osmolytes have been reported to stabilize biomolecules and even whole organisms against exposure to adverse environmental conditions. In this work, we report for the first time the use of some of these osmolytes, viz., the disaccharides trehalose and sucrose, in the stabilization of bovine trypsin against exposure to the anionic detergent sodium dodecyl sulfate and autolysis. Exposure of trypsin to SDS at a molar ratio of 1:45 led to decrease in trypsin activity by 61%. In the presence of 1 M sucrose and 1 M trehalose, the residual trypsin activity was found to increase to that of original enzyme activity. These two disaccharides were also found to slow down the rate of autolysis, resulting in residual activities of 80 and 88%, respectively, after incubation for 24 h. Active site titration showed retention of the fraction of active sites in the presence of trehalose. Fluorescence and CD spectroscopies were used to decipher the probable mechanism of this protective role of the disaccharides. Although complete resumption of secondary structure was not seen in the presence of the two disaccharides, the spectra of trypsin in the presence of stabilizers resembled the spectrum of native trypsin and were significantly different from the spectrum of detergent-denatured enzyme. Correlating the data obtained from spectroscopy with those obtained from activity assay, we propose that the retention of secondary structure of the enzyme is largely responsible for the retention of the functionally active form of trypsin.

Oral trypsin inhibitor can improve reflux esophagitis after distal gastrectomy concomitant with decreased trypsin activity

Background The pathogenesis of reflux esophagitis is not well understood and remains controversial. Distal gastrectomy serves as a model to assess the role of duodenal reflux with low gastric acidity in the development of reflux esophagitis. We investigated the clinical usefulness and antitrypsin activity after treatment with a trypsin inhibitor, camostat mesilate, against the reflux esophagitis after distal gastrectomy reconstructed with Billroth-I anastomosis. Methods Twenty-eight patients with gastroesophageal reflux disease after distal gastrectomy were prescreened according to esophageal pH level and trypsin activity, and consequently 11 patients were enrolled in the present clinical study. Esophageal and duodenal washings were aspirated for the evaluation of the pretreatment trypsin activity. Then 100 mg of camostat mesilate was administered orally. At 30 and 120 minutes after the administration, duodenal washings were aspirated for the evaluation of posttreatment trypsin activity. Thereafter, 300 mg of camostat mesilate was administered orally 3 times daily for a 4-week period. On the 28th day of administration, the grade of reflux esophagitis (Los Angeles classification) was re-evaluated under endoscopy and the esophageal washings were aspirated for the evaluation of trypsin activity. Results The trypsin activities in the duodenum both at 30 and 120 minutes after oral ingestion of camostat mesilate were decreased significantly in comparison with those in the pretreatment period in each patient ( P < .001). In 6 of 7 patients with detectable trypsin activity in the esophagus, the activities after the 28th day of treatment were lower than those in the pretreatment period, and the symptoms were milder than those before treatment ( P < .05). Furthermore, endoscopic evaluation showed that 40% of patients were grade B, C, and D after treatment (28th day), whereas 70% of patients were grade B, C, and D before the treatment. Conclusions Oral administration of trypsin inhibitor can improve reflux esophagitis after distal gastrectomy concomitant with decreased trypsin activity.

GSH Inhibits Trypsinization of the C-terminal Half of Human MRP1

MRP1 is a 190-kDa membrane glycoprotein that confers multidrug resistance to tumor cells. The accumulated evidence has proved that GSH interacts with MRP1 and stimulates drug transport. However, the mechanism of GSH-dependent drug transport by MRP1 remains unclear. In this study, we used limited tryptic digestion of MRP1 in isolated membrane vesicles, in the presence and absence of GSH, to investigate the influence of GSH on MRP1 conformation. We found that GSH inhibited the generation of an approximately 35-kDa C-terminal tryptic fragment (including a C-terminal His tag) termed C2 from MRP1. This effect of GSH was not because of direct inhibition of trypsin activity, and agosterol A enhanced the inhibitory effect of GSH. The main cleavage site in MRP1 for the generation of the C2 fragment by trypsin resided between TMD2 and NBD2 of MRP1. Limited tryptic digestion of membrane vesicles expressing various truncated and co-expressed MRP1 fragments in the presence and absence of GSH revealed that GSH inhibited the production of the C2 fragment only in the presence of the L(0) region of MRP1. Thus the L(0) region is required for the inhibition of trypsinization of the C-terminal half of MRP1 by GSH. These findings, together with previous reports, suggest that GSH induces a conformational change at a site within the MRP1 that is indispensable for the interaction of MRP1 with its substrates.

Maintenance ration, protein synthesis capacity, plasma insulin and growth of Atlantic salmon (Salmo salar L.) with genetically different trypsin isozymes

Growth was found to be associated with the changes of trypsin activity in the pyloric caecal tissues and the level of plasma insulin in Atlantic salmon (Salmo salar L.). A decrease in trypsin activity accompanied by an increase in plasma insulin was detected one month before an enhanced growth was observed. There were significant relationships between weight specific consumption rate, plasma insulin levels and fish growth. The correlation of weight specific consumption rate was higher with growth rate (R 2 D 0:7, p < 0:0001) than with plasma insulin concentration (R 2 D 0:4, p< 0:0001). When the comparison was made between Atlantic salmon carrying and lacking the trypsin variant TRP-292, the fish with the variant had lower maintenance ration (p < 0:05), higher capacity for protein synthesis in the white muscle (p< 0:02), and a greater ability to utilize the feed at a restricted ration than the fish without the variant. In Atlantic salmon lacking the variant, both plasma insulin concentrations and growth rates were significantly lower (p < 0:05) in the fish fed 0.5% bw day 1 than those fed 1% bw day 1 . Whilst the growth rates of TRP-292 salmon fed the different rations became similar one month after similar levels of plasma insulin were observed between them. The TRP-292 salmon may be defined as a high protein growth efficiency fish with low protein turnover rate. Genetic variation in trypsin isozyme pattern affects feed utilization, plasma insulin levels and growth in Atlantic salmon.

In vivo and in vitro effects of wheat germ agglutinin and Bowman-Birk soybean trypsin inhibitor, two potential transgene products, on midgut esterase and protease activities from Apis mellifera

Wheat germ agglutinin (WGA) and Bowman-Birk soybean trypsin inhibitor represent potential transgene products for inducing pest resistance in plants. The effects of these molecules were studied on midgut esterase and protease activities from Apis mellifera L., a major insect pollinator. Trypsin inhibitor and WGA did not exhibit an acute toxicity in A. mellifera. In vivo, trypsin inhibitor caused a decrease in the amount of trypsin activity and did not have a significant effect on esterase activity. In vitro, trypsin inhibitor inhibited about 80% of non-specific protease activity and 100% of trypsin activity. In vivo, WGA at high concentration in food (1 mg/ml) elicited a large decrease in trypsin activity and did not have a significant effect on esterase activity. In vitro, WGA did not have any significant effect on trypsin and non-specific protease activities but slightly activated esterase activity.

Proteinase inhibitory activity in the plasma of a mollusc: Evidence for the presence of α-macroglobulin in Biomphalaria glabrata

1. 1. A methylamine-sensitive inhibitor was present in the plasma of B. glabrata. 2. 2. This inhibitor decreased trypsin activity against a protein substrate, however trypsin retained activity against a low molecular weight substrate in the presence of the inhibitor. 3. 3. Snail plasma protected trypsin from inhbition by soybean trypsin inhibitor. 4. 4. The results give evidence for an α-macroglobulin proteinase inhibitor in the plasma of this gastropod mollusc.