Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss     

Jamie S. Harmon  Kim G. Michelsen  Dr. Mark A. Sheridan 《Fish physiology and biochemistry》1991,9(4):361-368

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.  相似文献   

A review of antimicrobial peptides: defensins and related cationic peptides     

Evans EW  Harmon BG 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》1995,24(4):109-116

Cationic antimicrobial peptides are present throughout the plant and animal kingdoms and bear striking structural and functional similarities across species lines. They provide primitive, nonspecific means of combating a variety of bacteria, fungi, enveloped viruses, and protozoa. Some are also cytotoxic against host cells, including neoplastic cells. Cationic antimicrobial peptides may play various roles in inflammation and tissue repair. Antimicrobial peptides are found in epithelial tissues regularly exposed to microbial attack as well as in cells whose primary function is defense against potential pathogens. They constitute an important part of the nonoxidative antimicrobial arsenal of leukocytes. They are preformed and/or readily synthesized when the cells are stimulated by exposure to pathogens. They exert their effects directly by inserting into membranes of target cells and forming ion channels which increase membrane permeability; however, antimicrobial peptides can also act as opsonins to facilitate phagocytosis. Resistance to defensins is a virulence factor for organisms such as Salmonella sp. The study of cationic antimicrobial peptides is increasing our understanding of innate immunity, inflammation, and the pathogenesis of genetic diseases such as specific granule disease in humans. Therapeutic applications of antimicrobial peptides are currently under investigation.  相似文献   

Molecular identification of high-affinity glutamate transporters in sheep and cattle forestomach, intestine, liver, kidney, and pancreas   总被引：5，自引：0，他引：5  

Howell JA  Matthews AD  Swanson KC  Harmon DL  Matthews JC 《Journal of animal science》2001,79(5):1329-1336

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Effect of sodium bicarbonate and sodium bentonite on digestion, solid and liquid flow, and ruminal fermentation characteristics of forage sorghum silage-based diets fed to steers     

K A Jacques  D E Axe  T R Harris  D L Harmon  K K Bolsen  D E Johnson 《Journal of animal science》1986,63(3):923-932

Six ruminally cannulated steers, five Holsteins and one Hereford (250 to 295 kg), were fed 84% forage sorghum silage plus 16% supplement or 50% forage sorghum silage plus concentrate and supplement diets containing either no addition (controls), 1% sodium bicarbonate (NaHCO3) or 2% sodium bentonite in a 2 X 3 factorial arrangement of treatments in a 6 X 6 Latin-square experiment with 3-wk periods. Sodium bicarbonate increased dry matter (DM) intake when concentrate was included, but neither compound affected intake of the 84% silage diet. Bentonite lowered DM, neutral detergent fiber (NDF), and acid detergent fiber (ADF) digestibilities, but NDF disappearance from nylon bags was unchanged. Ruminal pH, osmolality and L(+) and D(-) lactate were not affected by treatment. Both NaHCO3 and bentonite tended to lower ruminal NH3-N concentrations. Bentonite lowered the molar proportion of isobutyrate in ruminal fluid relative to controls, but proportions of other volatile fatty acids (VFA) and total VFA concentrations were unchanged. Neither NaHCO3 nor bentonite affected ruminal liquid or solid volumes, dilution rate constants or ruminal outflow rates. Markers overestimated volumes, but correction with measured volumes did not change interpretation of treatment effects. It was concluded that control diets had sufficiently high baseline values of pH, dilution rate and acetate proportion to preclude changes induced by either compound, especially at 1 or 2% of DM intake. An effect on palatibility through neutralization of silage acids may have been responsible for the intake response to NaHCO3.  相似文献   

Evaluation of low-ash poultry meal as a protein source in canine foods     

Yamka RM  Jamikorn U  True AD  Harmon DL 《Journal of animal science》2003,81(9):2279-2284

Eight mature female dogs (18.0 +/- 0.2 kg) were used in a replicated 4 x 4 Latin square experiment to determine the feeding value of low-ash poultry meal (PM) in a complete food fed to dogs. All foods contained graded concentrations of PM (10.4 to 32.5% DM), resulting in foods that were 10, 15, 20, and 25% CP. Daily DMI averaged 284 +/- 14 g/d. An increase in PM resulted in an increase in fecal moisture from 44.7 to 55.1% (linear; P < 0.01), and fecal DM output increased from 24.8 to 31.6 g/d (linear; P < 0.05). Ileal DM flow increased from 27.1 to 40.7 g/d (linear; P < 0.01). Small intestinal DM digestibility decreased from 90.4 to 86.1% (linear; P < 0.01) and total-tract DM digestibility decreased from 91.2 to 89.4% (linear; P < 0.01) as PM increased. Large intestinal DM digestibility increased from 8.4 to 21.1% with increasing PM (linear; P < 0.05). Fecal excretion of CP increased from 5.6 to 10.0 g/d (linear; P < 0.01) and ileal flow of CP increased from 6.9 to 15.6 g/d (linear; P < 0.01) as PM increased. Small intestinal CP digestibility was unaffected with treatment (P > 0.05). Large intestinal CP digestibility increased from 21.6 to 37.1% (linear; P < 0.05) with increasing PM. Total-tract CP digestibility increased from 81.0 to 86.6% (linear; P < 0.01) as PM increased. Arginine had the highest overall digestibility ranging from 88.5 to 91.3%, whereas cysteine had the lowest digestibility, ranging from 67.1 to 71.4%. These data indicate that PM is a highly digestible protein source for canine foods with inclusions of 10.4 to 32.5% of DM.  相似文献   

Pancreatic exocrine secretion in steers infused postruminally with casein and cornstarch   总被引：1，自引：0，他引：1  

Richards CJ  Swanson KC  Paton SJ  Harmon DL  Huntington GB 《Journal of animal science》2003,81(4):1051-1056

Our objective was to evaluate the effect of postruminal protein infusion on pancreatic exocrine secretions. One Holstein, two crossbred, and five Angus steers (305 +/- 5 kg) with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square. All steers were abomasally infused with 1,050 g/d of raw cornstarch with treatments of 0, 60, 120, or 180 g/d of sodium casein suspended in water to yield 6,000 g/d of infusate daily. Steers were limit-fed (1.5 x NEm; 12 equal portions daily) a 90% corn silage, 10% supplement diet formulated to contain 12.5% CP. Periods consisted of 3 d of adaptation to infusion, 7 d of full infusion, 1 d of collection, and 7 d of rest. Pancreatic juice was collected in 30-min fractions continuously for 6 h. Total juice secreted and the pH of individual fractions were recorded, a 10% subsample was retained to form a composite sample, and remaining fluid was returned to the duodenum. Juice composite samples were stored (-30 degrees C) until analyzed for total protein and activities of alpha-amylase, trypsin, and chymotrypsin. Casein infusion linearly increased alpha-amylase concentration (182 to 271 units/mL; P < 0.02; 17.5 to 24.6 units/mg of protein; P < 0.03) and secretion rate (26,847 to 41,894 units/h; P < 0.01). Total juice secretion (155 g/h), pH of pancreatic juice (8.13), secretion rate of protein (1,536 mg/h), and concentration of protein (10.2 mg/mL) in pancreatic secretions were not affected (P > 0.05) by casein infusion. Similarly, casein infusion did not change 0.05) trypsin and chymotrypsin concentrations (1,379 and 349 units/L or 0.134 and 0.033 units/mg of protein, respectively) or secretion rates (206 and 52 units/h, respectively). Abomasal infusion of protein with starch stimulated a greater pancreatic secretion of alpha-amylase activity into the intestine than infusion of starch alone.  相似文献   

Intestinal starch disappearance increased in steers abomasally infused with starch and protein     

Richards CJ  Branco AF  Bohnert DW  Huntington GB  Macari M  Harmon DL 《Journal of animal science》2002,80(12):3361-3368

Steers (379 +/- 10 kg) with ruminal, duodenal, and ileal cannulas were used in a 5 x 5 Latin square digestion trial to quantify and evaluate the relationship between intestinal protein supply and intestinal starch disappearance. Treatments were infusions of 0, 50, 100, 150, or 200 g/d of casein along with 1,042 g/d of raw cornstarch. Abomasal infusions were accomplished by passing tubing and a pliable retaining washer through the reticular-omasal orifice into the abomasum. Steers were fed a 93% corn silage, 7% supplement diet that contained 12% crude protein at 1.65% body weight in 12 equal portions/d. Periods lasted 17 d (12 d for adaptation, 2 d of collections, and 3 d of rest). The quantity and percentage of organic matter and protein disappearance from the small intestine increased linearly (P < 0.03) with infused casein. Greater quantities of starch disappeared with increased casein infusion (P < 0.01). The infusion of 200 g/d of casein increased small intestinal starch disappearance by 226 g/d over the control. Casein infusion did not affect the quantity or percent of organic matter, starch, or protein disappearance in the large intestine. Treatments did not change ruminal ammonia N, ruminal pH, or plasma glucose concentrations. Starch disappearance from the small intestine was increased with greater protein flow to the duodenum of steers.  相似文献   

Evaluation of nitric oxide as an inhibitory neurotransmitter in the equine ventral colon     

Van Hoogmoed LM  Rakestraw PC  Snyder JR  Harmon FA 《American journal of veterinary research》2000,61(1):64-68

OBJECTIVE: To determine the role of nitric oxide and an apamin-sensitive nonadrenergic noncholingeric inhibitory transmitter on contractility of the ventral colon of horses. SAMPLE POPULATION: Strips of the circular and longitudinal muscle layers and taenia of the ventral colon from 14 horses. PROCEDURE: Muscle strips were suspended in tissue baths and attached to force transducers. Contractile activity of circular, longitudinal, and taenia muscle strips in response to electrical field stimulation was measured after addition of apamin and a nitric oxide inhibitor, N-nitro-L-arginine methyl ester (L-NAME). RESULTS: Electrical field stimulation reduced contractile activity in the circular muscle layer and taenia but not the longitudinal muscle layer. Addition of L-NAME significantly reduced inhibitory contractile activity at all frequencies for the circular muscle layer, whereas a significant effect was evident for the taenia only at the highest frequency. The combination of L-NAME and apamin resulted in a significant reduction in inhibition of the taenia at all frequencies but for circular muscle only at lower frequencies. CONCLUSIONS AND CLINICAL RELEVANCE: Nitric oxide and an apamin-sensitive neurotransmitter appear to mediate a component of inhibitory transmission in the circular muscle and taenia, but not the longitudinal muscle layer, of the equine ventral colon. Nitric oxide has a role in regulating contractile activity of the equine ventral colon, and nitric oxide synthase inhibitors may be useful in horses with ileus of the large colon.  相似文献   

The pathogenesis of Pasteurella multocida serotype A:3,4 infection in turkeys: a comparison of two vaccine strains and a field isolate     

M M Prantner  B G Harmon  J R Glisson  E A Mahaffey 《Avian diseases》1990,34(2):260-266

The pathogenesis of avian pasteurellosis caused by two vaccine strains, M-9 and Clemson University (CU), and a highly virulent field isolate, 86-1913, of Pasteurella multocida (serotype A:3,4) was studied in 7-week-old turkeys inoculated by an oculo-nasal-oral technique. Turkeys inoculated with strain CU and isolate 86-1913 developed severe progressive bacteremia that began at 4 hours postinoculation (PI) and peaked at 16-20 hours PI. Turkeys inoculated with strain CU and isolate 86-1913 had significantly higher concentrations of bacteria in blood and tissues, and greater histologic lesion scores for necrosis, heterophil infiltrates, and intralesional bacteria than turkeys inoculated with strain M-9. Immunohistochemical staining specific for P. multocida demonstrated numerous extracellular bacteria in tissues from turkeys inoculated with strain CU and isolate 86-1913. The mortality for turkeys inoculated with isolate 86-1913 was significantly higher than for turkeys receiving the two vaccine strains.  相似文献   

Small intestinal starch digestion in steers: effect of various levels of abomasal glucose, corn starch and corn dextrin infusion on small intestinal disappearance and net glucose absorption   总被引：4，自引：0，他引：4  

K K Kreikemeier  D L Harmon  R T Brandt  T B Avery  D E Johnson 《Journal of animal science》1991,69(1):328-338

Eight Holstein steers (four at 300 kg, four at 406 kg) fitted with an elevated carotid artery, hepatic portal and mesenteric venous catheters, and abomasal and ileal cannulas were used in several 4 x 4 Latin square experiments to evaluate small intestinal starch digestion. They were fed alfalfa hay at 1.5% of BW and abomasally infused with water or glucose, corn starch or corn dextrin (one carbohydrate per Latin square) at 20, 40 or 60 g/h, with subsequent determination of small intestinal disappearance and net portal glucose absorption. Increasing the amount of all three carbohydrates infused abomasally increased the amount of carbohydrate disappearing in the small intestine. Increased infusion of glucose caused a continual increase (linear, P less than .01) in net glucose absorption, whereas net glucose absorption for starch and dextrin was maximal at the 20 g/h infusion (quadratic, P less than .05). With the 60 g/h infusion, 94% of the glucose but only 38% of starch and 29% of small intestinal dextrin disappearance could be accounted for as net glucose absorption, leaving a large portion of starch and dextrin disappearance unaccounted for. Of the infused starch and dextrin passing the ileum, 5.8 and 7.3%, respectively, was unpolymerized glucose, indicating that, at least in the distal small intestine, complete starch hydrolysis exceeded the capacity for glucose disappearance. It is concluded that only about 35% of the raw corn starch disappearing in the steer's small intestine resulted in net portal glucose absorption.  相似文献