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1.
J Anim Sci ; 98(10)2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32910163

RESUMO

Developed initially for use in fuel ethanol production, Enogen Feed Corn (EFC; Syngenta Crop Protection) is genetically modified to express high concentrations of α-amylase in the corn kernel. Experiments were conducted to evaluate processing characteristics of EFC, in vitro digestion, and effects on feedlot performance, carcass characteristics, and liver abscess incidence. Experiment 1 used a randomized complete block design (3 × 3 × 5 factorial) to evaluate starch availability, in situ dry matter disappearance (ISDMD), in vitro gas production (IVGP), and volatile fatty acid (VFA) profiles of in vitro cultures. Grains (EFC or mill-run control [CON]) were flaked to a density of 360 g/L, and mixtures with 0%, 25%, 50%, 75%, or 100% EFC were prepared. Grains were tempered with added moisture (0%, 3%, or 6%) prior to steam conditioning for 15, 30, or 45 min. No two- or three-way interactions were observed. Adding moisture improved starch availability (linear; P < 0.01), and tended to improve ISDMD (linear, P = 0.06). Steam conditioning for 30 min improved starch availability, IVGP, and production of acetate, propionate, butyrate, valerate, and total VFA (P < 0.01) compared with conditioning for 15 or 45 min. Starch availability, ISDMD, IVGP, acetate, propionate, valerate, and total VFA production increased with an increasing proportion of EFC (linear, P < 0.01). Experiment 2 used 700 beef heifers (394 ± 8.5 kg initial body weight [BW]) fed finishing diets with steam-flaked corn as CON or EFC for 136 d. Targeting similar starch availabilities, grains were processed to 360 g/L (CON) and 390 g/L for CON and EFC, respectively. Heifers were blocked by BW, stratified, and then randomly assigned to 28 dirt-surfaced pens (25 animals per pen). Dry matter intakes were similar between treatments (P = 0.78), but cattle fed EFC had greater average daily gain (P < 0.01), improving feed efficiency by 5% (P < 0.01). Hot carcass weight was 6 kg greater for EFC cattle (P <0.01) than CON. No differences were observed for longissimus muscle area (P = 0.89), 12th-rib fat thickness (P = 0.21), or USDA yield grade (P = 0.13). Cattle fed CON had greater marbling scores than EFC (P = 0.04), but this did not affect the USDA quality grade (P > 0.33). Cattle fed EFC had 23% fewer abscessed livers than CON (P = 0.03). High-amylase corn may be used to improve microbial digestion, mill-throughput, and cattle performance, and it may mitigate liver abscesses.


Assuntos
Amilases/farmacologia , Ração Animal/análise , Bovinos/fisiologia , Dieta/veterinária , Zea mays/enzimologia , Amilases/administração & dosagem , Amilases/química , Amilases/metabolismo , Fenômenos Fisiológicos da Nutrição Animal , Animais , Composição Corporal , Digestão/fisiologia , Ácidos Graxos Voláteis/farmacologia , Feminino , Amido/farmacologia , Vapor
2.
Microb Pathog ; 137: 103740, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31513898

RESUMO

Mastitis is one of the most important diseases that are threatening modern dairy farms. Biofilms of mastitic teat canal have serious clinical implications because of colonized pathogens having the ability to construct an extracellular polymeric substance (EPS) with increased tolerance to antimicrobials leads to difficulty in eradicating the infection. In this study, we investigated the synergistic biofilm disruptive effect of a combination of carbohydrate hydrolases targeting extracellular polysaccharides of biofilm matrix and we termed it as 'Biofilm Clippers (BC)'. Our findings demonstrate that the BC formulation exhibits intense biofilm-disrupting activity against Staphylococcus aureus biofilms. The results of the study showed that BC enables activity equivalent to physiologically achievable concentrations in disrupting biofilms of S. aureus in vitro. The synergistic anti-biofilm activities of BC on S. aureus biofilms demonstrated that the biofilm matrix is predominant of complex polysaccharides. Further, the confocal microscopic analysis demonstrates that the BC formulation is highly effective compared to the single treatment of either of the enzymes in disrupting the biofilm. To the best of our knowledge, this is the first report on the synergistic anti-biofilm activity of a class of enzyme formulation against mastitic biofilm mass. Even though a small study showed a promising effect on mastitic teat canal, further extensive investigation on a large number of bovines for mastitis therapeutic potential of this BC-derived product is now warranted.


Assuntos
Biofilmes/efeitos dos fármacos , Matriz Extracelular de Substâncias Poliméricas/efeitos dos fármacos , Mastite Bovina/tratamento farmacológico , Amilases/farmacologia , Animais , Antibacterianos/farmacologia , Bovinos , Celulases/farmacologia , Esterases/farmacologia , Feminino , Glândulas Mamárias Animais , Manosidases/farmacologia , Mastite Bovina/microbiologia , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/veterinária , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/isolamento & purificação
3.
Food Chem ; 299: 125038, 2019 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-31284248

RESUMO

Wheat is one of the world's most widely consumed staple food. However, the number of people suffering from wheat-related disorders has increased drastically. Amylase-trypsin inhibitors (ATIs) have recently been identified as one of the main triggers of non-celiac wheat sensitivity (NCWS). In this study, an enzymatic assay for the determination of trypsin inhibition activity in hexaploid wheat was developed. This method was optimized with respect to several parameters, such as extraction and incubation procedures, and was validated according to international standards, concerning accuracy, precision and robustness of the method. Results revealed that linear inhibition and thus accuracy occurred only in a narrow concentration range. However, after optimization of settings the novel method was found to be satisfactory for accurate determination of trypsin inhibition in wheat. Purification of the wheat extract with immobilized trypsin beads led to the identification of CM inhibitors (chloroform/methanol soluble proteins) as main contributors of trypsin inhibition.


Assuntos
Amilases/farmacologia , Ensaios Enzimáticos/métodos , Triticum/enzimologia , Inibidores da Tripsina/farmacologia , Tripsina/metabolismo , Alérgenos/farmacologia , Humanos
4.
Matern Child Nutr ; 15(2): e12742, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30375162

RESUMO

Adding amylase to Super Cereal Plus (SC+A) improves energy and nutrient intake as porridge energy density reaches 1.0 kcal/g, meeting the recommended ≥0.8 kcal/g for prepared foods for young children. Caregiver response to SC+A in terms of adjusting porridge preparation using printed pictogram instructions was not yet investigated. The study assessed (a) porridge preparation by caregivers; (b) porridge energy density; (c) sensory porridge acceptability; and (d) understanding of preparation instructions. An 8-day follow-up intervention study was conducted amongst caregivers of children aged 6-23 months (n = 238) in Rwanda. Caregivers prepared porridge using SC+A whilst referring to printed pictogram instructions at the study site on Days 1 and 8 and received flour for preparation at home on Days 2-7. At the site, data were collected on porridge preparation procedures, energy density, consistency, acceptability, and interviews (n = 12), and focus group discussions (n = 6) were conducted. Mean porridge dry matter (DM) increased from 21.3 ± 4.4% (Day 1) to 25.1 ± 4.8% (Day 8; p < 0.0005). Flour and water were mixed before cooking by 95% of the participants, as per printed instructions. Sensory porridge acceptability was high, and the printed instructions enabled caregivers to prepare an accepted and energy dense porridge. The preferred water/flour volume ratio was 2.5 instead of 3. In conclusion, Rwandan caregivers prepared well-accepted SC+A porridges with a preferred consistency and mean DM content of 25.1% (1.0 kcal/g), after 1 week practicing at home. This supports introducing SC+A with the tested instructions at scale.


Assuntos
Amilases/farmacologia , Culinária/métodos , Grão Comestível , Ingestão de Energia/fisiologia , Alimentos Infantis , Valor Nutritivo/fisiologia , Amilases/administração & dosagem , Cuidadores , Feminino , Farinha , Humanos , Lactente , Masculino , Ruanda
5.
J Dairy Sci ; 101(11): 9815-9826, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30146293

RESUMO

Two experiments were carried out to evaluate a blend of essential oils (EO) combined with amylase as an alternative to ionophores and its potential for reducing the use of antibiotics in the dairy industry. In experiment 1, 8 rumen-cannulated Holstein cows (576 ± 100 kg of body weight, 146 ± 35 d in milk, and 35.1 ± 4.0 kg/d of milk yield at the start of the experiment) were assigned to a 4 × 4 Latin square experiment with 21-d periods to determine the influence of feed additives on total apparent digestibility of nutrients, ruminal fermentation, N utilization, microbial protein synthesis, blood glucose and urea concentrations, and milk yield and composition in dairy cows. Treatment sequences assigned to cows in each block included no feed additives (control; CON); monensin (MON) added at 13 mg/kg of diet dry matter (DM); a blend of EO supplemented at 44 mg/kg of diet DM; and EO treatment combined with α-amylase at 330 kilo novo units/kg of diet DM (EOA). Differences among treatments were studied using orthogonal contrasts as follows: CON versus feed additives (MON, EO, and EOA), MON versus EO and EOA, and EO versus EOA. No differences were detected in nutrient intake and digestibility in cows. In general, feed additives decreased ruminal NH3-N concentration of cows, notably when diet was supplemented with MON. Furthermore, feed additives increased ruminal concentrations of acetate, butyrate, and branched-chain fatty acids. Cows fed treatments containing EO and EOA exhibited lower pH, higher NH3-N, and a trend to greater total volatile fatty acid concentration in the ruminal fluid compared with cows fed MON. Treatments containing EO increased ruminal butyrate concentration compared with MON. No treatment × time interaction effect was observed on ruminal fermentation measurements. Cows fed diets supplemented with feed additives had greater efficiency of N transfer into milk (milk N:N intake), whereas cows fed EOA exhibited greater N transfer into milk than those fed EO. Treatments had no effect on milk yield and composition, but feed additives increased the milk yield efficiency (milk yield divided by dry matter intake), whereas treatments containing EO had similar milk yield efficiency compared with MON. For experiment 2, 30 multiparous Holstein cows (574 ± 68 kg of body weight, 152 ± 54 d in milk, and 30.9 ± 4.1 kg/d of milk yield at the start of the experiment) were enrolled to a randomized complete block design experiment. The MON, EO, and EOA treatments were randomly assigned to cows within blocks (n = 10), and feed additives were provided throughout a 9-wk period. No differences were found in nutrient intake and digestibility, but cows fed EOA tended to exhibit greater dry matter intake than those fed EO. Blood metabolites and milk production were not affected by treatments. However, cows fed MON or EOA had greater milk protein content than those cows fed treatments containing EO. Feeding EO with or without amylase had similar response to feeding MON in terms of feed intake and milk yield, with a small negative effect on milk protein yield when feeding EO alone. Feed additives increased the concentrations of acetate, butyrate, and branched-fatty acids in ruminal fluid, whereas treatments containing EO had greater ruminal butyrate and NH3-N concentrations. Therefore, either EO or EOA can replace MON in diets of dairy cows while maintaining performance.


Assuntos
Amilases/farmacologia , Suplementos Nutricionais , Leite/metabolismo , Óleos Voláteis/farmacologia , Ração Animal/análise , Animais , Peso Corporal , Bovinos , Dieta/veterinária , Digestão/efeitos dos fármacos , Ácidos Graxos Voláteis/análise , Feminino , Fermentação/efeitos dos fármacos , Lactação/efeitos dos fármacos , Leite/química , Proteínas do Leite/análise , Monensin/análise , Nutrientes , Distribuição Aleatória , Rúmen/efeitos dos fármacos , Rúmen/metabolismo
6.
J Dairy Sci ; 101(8): 7199-7207, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29753467

RESUMO

Exogenous amylase supplementation can increase starch and fiber digestibility in lactating dairy cows. We evaluated the effect of exogenous amylase supplementation on diets with high starch concentration (32% of dry matter). Twenty-eight Holstein cows (171 ± 80 d in milk, 4 primiparous) received a standard diet for 14 d and then a treatment for 63 d, in a covariate-adjusted randomized block design with repeated measures over time. Treatments were amylase [0.5 g of Ronozyme RumiStar (DSM Nutritional Products, Basel, Switzerland) per kg of total mixed ration dry matter] or control. The diets contained (% of dry matter): 39.4% corn silage, 11.2% rehydrated and ensiled mature corn grain, and 11.7% finely ground mature corn. Amylase increased milk yield (32.3 vs. 33.0 kg/d) and reduced dry matter intake (20.7 vs. 19.7 kg/d), increasing feed efficiency (1.52 vs. 1.63). Amylase also increased milk lactose synthesis (1.49 vs. 1.56 kg/d) and plasma glucose concentration (59.3 vs. 68.6 mg/dL). Secretions of milk fat and protein did not differ. Although milk urea N did not differ, amylase reduced the concentration of urea N in blood, suggesting an increase in ruminal starch degradation. However, the total-tract apparent digestibility of starch (96.3% of intake) and neutral detergent fiber (44.4% of intake), ruminal fermentation profile, and microbial yield estimated by urinary allantoin excretion did not differ. Cows fed amylase sorted in favor of long feed particles and against short particles, had shorter chewing activity (780 vs. 699 min/d), and had fewer meals per day (11.5 vs. 9.7). Amylase improved the feed efficiency of lactating cows fed a high-starch diet; the enzyme increased milk yield and reduced intake.


Assuntos
Amilases/farmacologia , Fenômenos Fisiológicos da Nutrição Animal , Lactação/efeitos dos fármacos , Amido/metabolismo , Animais , Bovinos , Dieta , Digestão , Feminino , Leite , Rúmen , Amido/administração & dosagem , Zea mays
7.
Front Immunol ; 9: 3174, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30740114

RESUMO

Amylase trypsin inhibitors (ATI) can be found in all gluten containing cereals and are, therefore, ingredient of basic foods like bread or pasta. In the gut ATI can mediate innate immunity via activation of the Toll-like receptor 4 (TLR4) on immune cells residing in the lamina propria, promoting intestinal, as well as extra-intestinal, inflammation. Inflammatory conditions can induce formation of peroxynitrite (ONOO-) and, thereby, endogenous protein nitration in the body. Moreover, air pollutants like ozone (O3) and nitrogen dioxide (NO2) can cause exogenous protein nitration in the environment. Both reaction pathways may lead to the nitration of ATI. To investigate if and how nitration modulates the immunostimulatory properties of ATI, they were chemically modified by three different methods simulating endogenous and exogenous protein nitration and tested in vitro. Here we show that ATI nitration was achieved by all three methods and lead to increased immune reactions. We found that ATI nitrated by tetranitromethane (TNM) or ONOO- lead to a significantly enhanced TLR4 activation. Furthermore, in human primary immune cells, TNM nitrated ATI induced a significantly higher T cell proliferation and release of Th1 and Th2 cytokines compared to unmodified ATI. Our findings implicate a causative chain between nitration, enhanced TLR4 stimulation, and adaptive immune responses, providing major implications for public health, as nitrated ATI may strongly promote inhalative wheat allergies (baker's asthma), non-celiac wheat sensitivity (NCWS), other allergies, and autoimmune diseases. This underlines the importance of future work analyzing the relationship between endo- and exogenous protein nitration, and the rise in incidence of ATI-related and other food hypersensitivities.


Assuntos
Imunidade Adaptativa/efeitos dos fármacos , Amilases/farmacologia , Imunidade Inata/efeitos dos fármacos , Triticum/metabolismo , Inibidores da Tripsina/farmacologia , Amilases/química , Biomarcadores , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Humanos , Imunofenotipagem , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Proteínas de Plantas/metabolismo , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Receptor 4 Toll-Like/metabolismo , Inibidores da Tripsina/química
8.
J Anim Physiol Anim Nutr (Berl) ; 102(2): e641-e652, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29030887

RESUMO

Thirty-six cows were blocked by calving date and randomly assigned to one of three treatments. Cows were on treatments 3 weeks prepartum through 8 weeks post-partum. Treatments were as follows: (i) no direct-fed microbial (DFM) or cellulase and amylase enzymes (C), (ii) 45.4 g/day of DFM (D) or (iii) 45.4 g/day of DFM and 18.2 g/day of enzyme (DE). Total mixed ration fed and refused were measured daily to determine dry matter intake (DMI). Blood samples were taken three times weekly and analysed for ß-hydroxybutyrate, glucose and non-esterified fatty acids. Body weight (BW) was measured weekly. Colostrum was weighed and analysed for IgA and IgG concentration. Calves were fed 4 L of colostrum within 2 hr of birth. Calf blood samples were taken at 0 and 24 hr for analysis of IgA and IgG concentrations and apparent efficiency of absorption. Milk yield was measured daily and samples collected weekly. Initial BW was different among treatments with D being lesser than C or DE treatments. Body weight, weight gain, efficiency of gain, DMI and blood parameters were unaffected. Treatment did not affect colostrum yield. Ash percentage of colostrum tended to increase with D and DE, while IgA and total solids yield decreased with D. Colostrum fat yield was decreased in D and DE. Treatments did not impact BW, serum IgA and IgG concentrations or apparent efficiency of absorption of calves. Post-partum BW, DMI, blood parameters, milk production and composition were unaffected by treatment. However, cows on D gained more BW and tended to have greater efficiency of gain compared to those on DE, but were similar to C. Somatic cell scores were greatest for D. Results indicate that DFM and enzyme supplementation did not improve health and performance of dairy cattle during the pre- and post-partum periods under conditions of this study.


Assuntos
Amilases/farmacologia , Ração Animal/análise , Bovinos/fisiologia , Celulase/farmacologia , Colostro/química , Probióticos/administração & dosagem , Amilases/administração & dosagem , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Recém-Nascidos , Celulase/administração & dosagem , Dieta/veterinária , Feminino , Imunoglobulina G/sangue , Gravidez , Fenômenos Fisiológicos da Nutrição Pré-Natal
9.
Surgery ; 160(1): 26-37, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27117578

RESUMO

BACKGROUND: Amylase was the first enzyme to be characterized, and for the previous 200 years, its clinical role has been restricted to a diagnostic aid. Recent interface research has led to a substantial expansion of its role into novel, viable diagnostic, and therapeutic applications to cancer, infection, and wound healing. This review provides a concise "state-of-the-art" overview of the genetics, structure, distribution, and localization of amylase in humans. METHOD: A first-generation literature search was performed with the MeSH search string "Amylase AND (diagnost∗ OR therapeut$)" on OVIDSP and PUBMED platforms. A second-generation search was then performed by forward and backward referencing on Web of Knowledge™ and manual indexing, limited to the English Language. RESULTS: "State of the Art" in amylase genetics, structure, function distribution, localisation and detection of amylase in humans is provided. To the 4 classic patterns of hyperamylasemia (pancreatic, salivary, macroamylasemia, and combinations) a fifth, the localized targeting of amylase to specific foci of infection, is proposed. CONCLUSIONS: The implications are directed at novel therapeutic and diagnostic clinical applications of amylase such as the novel therapeutic drug classes capable of targeted delivery and "smart release" in areas of clinical need. Future directions of research in areas of high clinical benefit are reported.


Assuntos
Amilases , Amilases/química , Amilases/farmacologia , Amilases/fisiologia , Humanos
10.
Anim Sci J ; 86(1): 51-8, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25228428

RESUMO

The aim of this study was to assess the effects of feeding Atriplex halimus (AH) silage treated with two developed enzyme cocktails to sheep on feed intake, nutrient digestibility and ruminal fermentation. The AH silage was treated without or with 2 L of ZAD1(®) or ZAD2(®) /1000 kg with 5% molasses and ensiled for 30 days. Barley grain (300 g/head/day) was fed as an energy supplement once daily at 10.00 hours and AH silage with or without enzyme treatment was offered ad libitum to animals twice daily at 09.00 and 16.00 hours. Sheep were fed on four experimental forage diets comprised of AH silage and barley (D1), AH silage treated with ZAD1(®) and barley (D2), AH silage treated with ZAD2(®) and barley (D3) and AH silage treated with a combination of ZAD1(®) and ZAD2(®) (1:1) and barley (D4). Ensiling AH with enzymes reduced its contents of neutral detergent fiber and acid detergent fiber. The dry matter intake of AH of D2, D3 and D4 decreased (P < 0.001) as compared to D1. However, enzyme-treated diets had greater total digestible nutrients intake (P < 0.001) as compared to D1. The nutrients digestibility for D2, D3 and D4 were higher than those for D1 (P < 0.001), and were higher for D3 as compared to both D2 and D4. Sheep fed on D3 had highest (P < 0.001) ruminal total volatile fatty acids concentration, ammonia nitrogen concentration and microbial protein yield. It could be concluded that AH silage treated with ZAD1(®) or ZAD2(®) improved digestibility and rumen fermentation in sheep.


Assuntos
Amilases/farmacologia , Ração Animal , Atriplex , Celulases/farmacologia , Digestão/efeitos dos fármacos , Digestão/fisiologia , Ingestão de Alimentos/efeitos dos fármacos , Ingestão de Alimentos/fisiologia , Enzimas/farmacologia , Fermentação/efeitos dos fármacos , Fermentação/fisiologia , Alimentos , Peptídeo Hidrolases/farmacologia , Rúmen/fisiologia , Ovinos/fisiologia , Silagem , Xilosidases/farmacologia , Animais , Ácidos Graxos Voláteis/metabolismo , Hordeum , Nitrogênio/metabolismo , Rúmen/metabolismo , Rúmen/microbiologia
11.
Bioresour Technol ; 154: 122-30, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24384319

RESUMO

Effective removal of aging biofilm deserves to receive more attention. This study aimed to characterized aging biofilm from a full-scale moving bed biofilm reactor treating pharmaceutical wastewater and evaluate the hydrolysis effects of biofilm by different enzymatic treatments. Results from FTIR and biochemical composition analyses showed that it was a predominately organic-based biofilm with the ratio of total protein (PN) to polysaccharide (PS) of 20.17. A reticular structure of extracellular polymeric matrix (EPM) with filamentous bacteria as the skeleton was observed on the basal layer through SEM-EDS test. Among the four commercial proteases and amylases from Genencor®, proteases were shown to have better performances than amylases either on the removal of MLSS and PN/MLSS or on DOC (i.e., dissolved organic carbon)/MLSS raising of biofilm pellets. Difference of dynamic fluorescence characteristics of dissolved organic matters after treated by the two proteases indicated distinguishing mechanisms of the treating process.


Assuntos
Amilases/farmacologia , Biofilmes/efeitos dos fármacos , Reatores Biológicos/microbiologia , Peptídeo Hidrolases/farmacologia , Carbono/análise , Compostos Orgânicos/análise , Esgotos/química , Espectrometria de Fluorescência , Fatores de Tempo
12.
JOP ; 14(3): 228-336, 2013 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-23669470

RESUMO

CONTEXT: Our previous studies showed that porcine pancreatic enzymes in Syrian golden hamsters with peripheral insulin resistance normalizes the plasma insulin level, reduces the size of enlarged islets and inhibits the increased DNA synthesis in the beta-cell of islets. OBJECTIVE: In order to exclude the possibility that these effects was attributed to some contaminants of this crude material, we tested the effect of purified fungal pancreatic enzyme (FPE) that contains primarily amylase and lipase without (FPE) and with addition of chymotrypsin (FPE+chy). MATERIAL AND METHODS: In a pilot study we tested the effect of different doses of FPE given in drinking water on insulin level, islet size and DNA synthesis of islet cells in hamsters with induced peripheral insulin resistance by a high fat diet. The most effective dose of FPE on these parameters was used in a long-term experiment with FPE and FPE+chy in hamsters fed a high-fat diet for 36 or 40 weeks. RESULTS: In the pilot study a dose of 2 g/kg body weight was found to be optimal for controlling the body weight, normalizing plasma insulin level, the size of islets, the DNA synthesis and the number of insulin cells in the islets. These data were produced in the long-term study, where steatorrhea was also inhibited. Addition of chymotrypsin had no effects on these parameters. CONCLUSION: Pancreatic lipase and amylase appear to be responsible for the observed effects and offer a safe and effective natural product for the treatment of pancreatic diseases, including acute pancreatitis, chronic pancreatic, cystic fibrosis and any conditions associated with peripheral insulin resistance, including obesity and type 2 diabetes. The possible mechanism of the action is discussed.


Assuntos
Amilases/farmacologia , Proteínas Fúngicas/farmacologia , Ilhotas Pancreáticas/efeitos dos fármacos , Lipase/farmacologia , Amilases/administração & dosagem , Animais , Contagem de Células , Quimotripsina/administração & dosagem , Quimotripsina/farmacologia , Cricetinae , DNA/biossíntese , Diabetes Mellitus Tipo 2/etiologia , Diabetes Mellitus Tipo 2/fisiopatologia , Dieta Hiperlipídica/efeitos adversos , Relação Dose-Resposta a Droga , Feminino , Proteínas Fúngicas/administração & dosagem , Fungos/enzimologia , Insulina/sangue , Resistência à Insulina/fisiologia , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Lipase/administração & dosagem , Mesocricetus , Obesidade/etiologia , Obesidade/fisiopatologia , Tamanho do Órgão/efeitos dos fármacos , Projetos Piloto , Fatores de Tempo
13.
Cell Oncol (Dordr) ; 36(4): 289-301, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23677639

RESUMO

BACKGROUND: Previous research has suggested a putative utility of pancreatic (pro)enzymes in cancer treatment. The aim of the present study was to investigate the in vitro effects of a mixture of two pancreatic pro-enzymes, i.e., Chymotrypsinogen and Trypsinogen, and the enzyme Amylase on three human cancer cell lines, i.e., OE33 (derived from an oesophageal carcinoma), Panc1 (derived from a pancreatic carcinoma) and Caco-2 (derived from a colon carcinoma). RESULTS: After treatment of the three cancer cell lines with different doses of the (pro)enzymes for up to 7 days, we observed (i) growth inhibition in a dose-dependent manner, (ii) enhanced expression of ß-catenin and E-cadherin and decreased expression of several epithelial-mesenchymal transition (EMT)-associated genes, such as Vimentin, Snail and Slug, (iii) differentiation of Caco-2 cells, including the appearance of cell-specific differentiated structures such as microvilli and tight junctions, the acquisition of a more regular polygonal morphology, and an increased expression of the intestinal differentiation markers alkaline phosphatase and cytokeratin 8, and (iv) differentiation of Panc1 cells, including the formation of cell aggregates, an increment on lamellar bodies and an increased expression of the pancreatic differentiation markers glucagon and insulin. CONCLUSIONS: Our results show that the treatment of three different human cancer cell lines with pancreatic (pro)enzymes results in an enhancement of cell adhesion, an attenuation of several EMT-associated markers, and an increase in the expression of several differentiation-associated markers, suggesting the acquisition of a less malignant phenotype and a decrease in proliferative capacity due to lineage-specific cellular differentiation.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Quimotripsinogênio/farmacologia , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Tripsinogênio/farmacologia , Fosfatase Alcalina/metabolismo , Amilases/farmacologia , Animais , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Células CACO-2 , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Transição Epitelial-Mesenquimal/genética , Citometria de Fluxo , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Queratina-8/metabolismo , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição da Família Snail , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Vimentina/genética , Vimentina/metabolismo , beta Catenina/metabolismo
14.
J Dairy Sci ; 96(5): 3075-84, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23498022

RESUMO

The objective of this experiment was to determine the performance and digestibility response of lactating dairy cows fed a reduced-starch diet containing a commercial amylase product. Treatments consisted of a normal-starch total mixed ration (NS-), a reduced-starch total mixed ration (RS-), and a reduced-starch total mixed ration with exogenous amylase (RS+) added to the concentrate. Treatments were assigned according to a replicated 3 × 3 Latin square design with 28-d periods. Twenty-three cows completed the study. Starch concentrations in NS-, RS-, and RS+ total mixed rations were 27.7, 23.5, and 22.7%, respectively. Effects of treatment on intake, milk production, milk composition, and total-tract apparent nutrient digestibility were evaluated during the last week of each period. Effects of amylase on in vitro starch digestibility of the NS- and RS- grain mixes were also measured. We hypothesized that the reduction in dietary starch in the RS- ration would decrease diet digestibility and limit milk production compared with NS- due to a decrease in available energy, and that RS+ would alleviate some of this decrease by increasing nutrient digestibility. Contrary to this hypothesis, the RS- diet did not affect intake or milk production relative to the NS- diet, except for increased milk urea nitrogen and a tendency for a decrease in milk protein yield. This lack of response is attributed to both low milk fat concentrations across treatments and greater than predicted dietary energy content preventing the energy deficit that was intended to occur with the reduced-starch rations. Cows fed the RS+ ration had the lowest production performance, with reduced milk, fat-corrected milk, protein, and lactose yields relative to cows fed NS-. Cows fed RS+ also had reduced lactose yield and tended to have reduced milk and fat-corrected milk relative to cows fed RS-. Despite the negative effects of the RS+ treatment on performance, exogenous amylase did increase both in vitro and in vivo measurements of digestibility. Although amylase increased nutrient digestibility, this did not translate into improved milk performance, likely due to the relatively high energy content of experimental diets compared with cow requirements.


Assuntos
Amilases/farmacologia , Dieta/veterinária , Digestão/efeitos dos fármacos , Lactação/efeitos dos fármacos , Amilases/metabolismo , Ração Animal , Animais , Bovinos , Carboidratos da Dieta/metabolismo , Carboidratos da Dieta/farmacologia , Digestão/fisiologia , Feminino , Lactação/fisiologia , Leite/química , Proteínas do Leite/análise , Amido/metabolismo
15.
J Anim Sci ; 90 Suppl 4: 311-4, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23365364

RESUMO

Use of nutritional components from the milk and eventually from the solid feed relates to the growth and development of gastrointestinal tract (GIT). We studied the effect of pancreatic-like enzymes [porcine pancreatic enzymes (Creon) or microbial-derived amylase, protease, and lipase] on GIT morphology and lipid absorption in suckling piglets. Both enzyme preparations, in low or high dose, were fed via a stomach tube twice a day for 7 d starting at 8 d of age and controls received vehicle, n = 6. The day after treatments ended, lipid absorption was tested after which pigs were euthanized and GIT was examined. Enzyme cocktails, irrespective of their origin, increased (P < 0.001) triglyceride level in blood. Enzyme preparation affected (P < 0.001) small intestinal mucosal thickness, villi length, and crypt depth and (P < 0.01) mitotic division of enterocytes. In addition, the external administration of pancreatic enzymes stimulated pancreatic growth as observed by increased (P < 0.05) mitotic division of pancreatic cells. The study revealed that pancreatic or pancreatic-like enzymes of microbial origin administrated in the early postperinatal period enhance GIT development and may be used to better prepare the GIT of piglets for milk use and weaning.


Assuntos
Amilases/farmacologia , Trato Gastrointestinal/efeitos dos fármacos , Trato Gastrointestinal/crescimento & desenvolvimento , Pancrelipase/farmacologia , Peptídeo Hidrolases/farmacologia , Suínos/crescimento & desenvolvimento , Amilases/metabolismo , Animais , Trato Gastrointestinal/anatomia & histologia , Metabolismo dos Lipídeos , Peptídeo Hidrolases/metabolismo
16.
J Anim Sci ; 90 Suppl 4: 324-6, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23365368

RESUMO

The exocrine pancreatic insufficient (EPI) pigs grow less due to different disturbances in feed digestion, absorption, and retention. Use of pancreatic-like enzymes of microbial origin in pigs may improve feed use and performance in slow-growing pigs. The aim was to study gut recovery and effectiveness of pancreatic-like enzymes of microbial origin supplementation on pig performance. Six male pigs 10 to 12 kg BW underwent pancreatic duct ligation surgery to induce total exocrine pancreatic insufficiency (EPI). Three cannulas to access the gastrointestinal tract content were installed in stomach, duodenum, and ileum in EPI pigs and in 3 control (healthy) pigs. One month after surgery, enzymes were given before feeding and digesta samples were collected for analyses. The BW of EPI pigs did not increase during 1 mo following surgery (11.7 vs. 11.6 kg BW); however, BW increased after 1 wk of enzyme supplementation (12.1 kg BW). Coefficient of fat and N absorption increased (P < 0.05) in EPI pigs after enzyme supplementation. Activity of amylase, lipase, and protease in chyme samples of EPI pigs was very low compared to controls. In EPI pigs after enzyme supplementation, amylase activity increased from 5.32 to 72.9 units/mL but remained lower than that of healthy pigs (162.7 units/mL). Lipase activity increased from 79.1 to 421.6 units/mL, which was similar to that of controls (507.3 units/mL). Proteolytic activity increased from 7.8 to 69.7 units/mL but still did not reach control pigs (164.3 units/mL). In conclusion, exogenous microbial enzymes mimic endogenous pancreatic enzymes being recovered along the lumen of the gastrointestinal tract. These enzymes might be a useful tool to stimulate growth of slower-growing pigs after the weaning period.


Assuntos
Amilases/farmacologia , Insuficiência Pancreática Exócrina/veterinária , Lipase/farmacologia , Ductos Pancreáticos/cirurgia , Peptídeo Hidrolases/farmacologia , Doenças dos Suínos/patologia , Amilases/administração & dosagem , Amilases/metabolismo , Animais , Peso Corporal , Relação Dose-Resposta a Droga , Insuficiência Pancreática Exócrina/metabolismo , Fezes/química , Lipase/administração & dosagem , Lipase/metabolismo , Masculino , Peptídeo Hidrolases/administração & dosagem , Peptídeo Hidrolases/metabolismo , Suínos , Doenças dos Suínos/metabolismo
17.
J Anim Sci ; 90 Suppl 4: 439-41, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23365403

RESUMO

Behavioral changes during pancreatic enzyme therapy have never been studied. The present study investigated behavioral changes in exocrine pancreatic insufficiency (EPI) pigs when their feed was supplemented with pancreatic-like enzymes of microbial origin. A crossover design study was used to test the effect of enzyme supplementation in 2 × 4 EPI pigs that underwent pancreatic duct ligation (PDL). After 40 d of adaptation, the study commenced, comprising 2 control and 2 enzyme feeding periods of 10 d each in sequence. On days 7 and 10 of each experimental period, behavior was monitored for 24 h and feed consumption and BW were recorded. Behavioral observations focused on the pigs' activity-- lying down or passive, or sitting, or standing or active--and were expressed as percentage activity for 24 h. During the adaptation period, BW gain was completely inhibited after PDL whereas for the entire study period, the body weight increased from 10.5 ± 1.1 to 14.0 ± 1.4 kg (P < 0.01). Exocrine pancreatic insufficiency pigs were more active when fed the enzymes (21 vs. 18% per 24 h; P < 0.01). Microbial enzyme supplementation not only improved the growth of the EPI pigs but it also increased their activity. This behavior change contradicts the generally accepted norm that satiety evokes by digestion and subsequent nutrients absorption reduces human or animal motility.


Assuntos
Amilases/farmacologia , Insuficiência Pancreática Exócrina/veterinária , Lipase/farmacologia , Peptídeo Hidrolases/farmacologia , Doenças dos Suínos/tratamento farmacológico , Amilases/administração & dosagem , Animais , Aspergillus/enzimologia , Burkholderia cepacia/enzimologia , Estudos Cross-Over , Insuficiência Pancreática Exócrina/tratamento farmacológico , Lipase/administração & dosagem , Masculino , Peptídeo Hidrolases/administração & dosagem , Suínos
18.
Br Poult Sci ; 49(3): 340-6, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18568759

RESUMO

1. The aim of the present study was to investigate the effect of an enzyme cocktail composed of xylanase, amylase and protease on the performance and nutrient utilisation of broiler starters fed on a maize-based diet. A maize/soy-based control diet was formulated to be marginal in apparent metabolisable energy (AME) and amino acids and this diet was fed without or with two concentrations (250 and 500 g/tonne) of the exogenous enzyme cocktail. 2. Growth performance was measured over a 21-d period after which ileal contents were collected for measurement of digestible energy, nitrogen and dry matter. Excreta were collected from d 17 to 20 post-hatch and used to determine the AME, and retention of nitrogen and dry matter. 3. Supplementation of the control diet with 250 and 500 g/tonne of the enzyme cocktail resulted in enhanced performance of the chicks (1.4 and 8.3%, respectively, for weight gain and 1.2 and 2.2%, respectively, for feed per gain), but the improvements were significant (P < 0.05) only at 500 g/tonne. 4. Addition of both 250 and 500 g/tonne improved AME, nitrogen-corrected AME and dry matter retention compared with the control though the improvements were greater with the higher dose. These effects were also reflected in the apparent ileal digestibility coefficients of dry matter, nitrogen and energy, with 500 g/tonne of the enzyme cocktail improving ileal digestible energy by 3.4%. 5. These results demonstrate that a cocktail of exogenous enzymes containing xylanase, amylase and protease is effective in improving the performance of broiler chicks fed on a maize/soybean-meal-based diet. However, these benefits may be dose-dependent and so an economic analysis involving ingredient and enzyme pricing and the relative performance and digestibility enhancements could allow the end user to select the most economical dose to maximise return on investment.


Assuntos
Amilases/farmacologia , Ração Animal , Galinhas/crescimento & desenvolvimento , Endo-1,4-beta-Xilanases/farmacologia , Peptídeo Hidrolases/farmacologia , Zea mays , Amilases/metabolismo , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Peso Corporal/efeitos dos fármacos , Endo-1,4-beta-Xilanases/metabolismo , Ingestão de Energia , Metabolismo Energético , Conteúdo Gastrointestinal/química , Conteúdo Gastrointestinal/efeitos dos fármacos , Íleo , Peptídeo Hidrolases/metabolismo , Subtilisinas/metabolismo , Subtilisinas/farmacologia
19.
J Anim Sci ; 85(7): 1702-11, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17371787

RESUMO

The overall objective of the studies reported here was to evaluate the growth and nutrient utilization responses of pigs to dietary supplementation of phytate- or nonstarch polysaccharide-degrading enzymes. In Exp. 1, growth performance and nutrient digestibility responses of forty-eight 10-kg pigs to dietary supplementation of phytase or a cocktail of xylanase, amylase, and protease (XAP) alone or in combination were evaluated. The growth response of one hundred fifty 23-kg pigs to dietary supplementation of phytase or xylanase individually or in combination was studied in Exp. 2 in a 6-wk growth trial, whereas Exp. 3 investigated the nutrient digestibility and nutrient retention responses of thirty 24-kg pigs to dietary supplementation of the same enzymes used in Exp. 2. In Exp. 1, the pigs were used in a 28-d feeding trial. They were blocked by BW and sex and allocated to 6 dietary treatments. The treatments were a positive control (PC) diet; a negative control (NC) diet marginally deficient in P and DE; NC with phytase added at 500 or 1,000 phytase units (FTU)/kg; NC with xylanase at 2,500 units (U)/kg, amylase at 400 U/kg, and protease at 4,000 U/kg; and NC with a combination of phytase added at 500 FTU/kg and XAP as above. In Exp. 2 and 3, the 5 dietary treatments were positive control (PC), negative control (NC), NC plus 500 FTU of phytase/kg, NC plus 4,000 U of xylanase/kg, and NC plus phytase and xylanase. In Exp. 1, low levels of nonphytate P and DE in the NC diet depressed (P < 0.05) ADG of the pigs by 16%, but phytase linearly increased (P < 0.05) ADG by up to 24% compared with NC. The cocktail of XAP alone had no effect on ADG of pigs, but the combination of XAP and phytase increased (P < 0.05) ADG by 17% compared with the NC treatment. There was a linear increase (P < 0.01) in Ca and P digestibility in response to phytase. In Exp. 2, ADG was 7% greater in PC than NC (P < 0.05); there were no effects of enzyme addition on any response. In Exp. 3, addition of phytase alone or in combination with xylanase improved (P < 0.05) P digestibility. Phosphorus excretion was greatest (P < 0.01) in the PC and lowest (P < 0.05) in the diet with the combination of phytase and xylanase. The combination of phytase and xylanase improved P retention (P < 0.01) above the NC diet to a level similar to the PC diet. In conclusion, a combination of phytase and carbohydrases improved ADG in 10-kg but not 23-kg pigs, but was efficient in improving P digestibility in pigs of all ages.


Assuntos
6-Fitase/farmacologia , Digestão/fisiologia , Glicosídeo Hidrolases/farmacologia , Fósforo/metabolismo , Suínos/crescimento & desenvolvimento , Amilases/farmacologia , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Cálcio/metabolismo , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Endo-1,4-beta-Xilanases/farmacologia , Feminino , Masculino , Peptídeo Hidrolases/farmacologia , Ácido Fítico/metabolismo , Distribuição Aleatória , Suínos/metabolismo , Desmame , Ganho de Peso
20.
Poult Sci ; 86(3): 496-502, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17297161

RESUMO

A study was designed to determine the effects of enzyme supplementation on poults fed commercially based diets that included corn, soybean meal, and ground wheat with meat and bone meal (0 to 21 d) or Pro-Pak (22 to 56 d). Day-old turkey poults (n = 3,850) were divided into 35 pens and fed 1 of 5 dietary treatments for 56 d. Treatments were a positive control, a negative control (corn matrix adjustment of 140 kcal), and negative control diets supplemented with Avizyme 1502 at 250, 500, or 750 g/tonne. Feed intake, live weight gain, feed conversion ratio, and mortality were measured for the periods 0 to 21 d, 21 to 42 d, and 42 to 56 d, as well as for the cumulative 0 to 56 d. The 0- to 21-d period was further divided into subperiods (0 to 4 d, 4 to 8 d, 8 to 12 d, 12 to 16 d, 16 to 21 d) to evaluate early nutritional development. Ileal contents along with duodenal, jejunal, and ileal sections (n = 7/treatment) were sampled to determine apparent digestibility and morphology. In most instances, production response differences between the positive and negative controls were not significant, making definitive interpretation of enzyme addition difficult. Energy and protein ileal digestibilities of the negative control diets were lower than those of the positive control diet at 4, 8, 12, 16, and 42 d. Enzyme supplementation significantly improved energy and protein beyond that of the PC diet on d 42. Villus height and crypt depth did not respond to dietary treatment, although there was a significant interaction of age by treatment on jejunal villus height. The similarity between the controls, as well as the high inclusion of CuSO(4), may be responsible for the low response with enzyme inclusion.


Assuntos
Amilases/administração & dosagem , Amilases/farmacologia , Dieta/veterinária , Suplementos Nutricionais , Endo-1,4-beta-Xilanases/administração & dosagem , Endo-1,4-beta-Xilanases/farmacologia , Peptídeo Hidrolases/administração & dosagem , Peptídeo Hidrolases/farmacologia , Perus/crescimento & desenvolvimento , Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Animais , Digestão , Relação Dose-Resposta a Droga , Duodeno/anatomia & histologia , Duodeno/efeitos dos fármacos , Íleo/anatomia & histologia , Íleo/efeitos dos fármacos , Íleo/fisiologia , Jejuno/anatomia & histologia , Jejuno/efeitos dos fármacos , Masculino , Soja , Fatores de Tempo , Triticum , Zea mays
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