RESUMEN

Although the importance of intestinal hydrolases is recognized, there is little information on the intestinal proteome of lepidopterans such as Anticarsia gemmatalis. Thus, we carried out the proteomic analysis of the A. gemmatalis intestine to characterize the proteases by LC/MS. We examined the interactions of proteins identified with protease inhibitors (PI) using molecular docking. We found 54 expressed antigens for intestinal protease, suggesting multiple important isoforms. The hydrolytic arsenal featured allows for a more comprehensive understanding of insect feeding. The docking analysis showed that the soybean PI (SKTI) could bind efficiently with the trypsin sequences and, therefore, insect resistance does not seem to involve changing the sequences of the PI binding site. In addition, a SERPIN was identified and the interaction analysis showed the inhibitor binding site is in contact with the catalytic site of trypsin, possibly acting as a regulator. In addition, this SERPIN and the identified PI sequences can be targets for the control of proteolytic activity in the caterpillar intestine and serve as a support for the rational design of a molecule with greater stability, less prone to cleavage by proteases and viable for the control of insect pests such as A. gemmatalis.

Asunto(s)

Mariposas Nocturnas/enzimología , Péptido Hidrolasas/metabolismo , Secuencia de Aminoácidos , Animales , Intestinos/enzimología , Larva/enzimología , Simulación del Acoplamiento Molecular , Mariposas Nocturnas/genética , Péptido Hidrolasas/química , Péptido Hidrolasas/genética

RESUMEN

A raw starch digesting α-amylase from Nile tilapia (Oreochromis niloticus) intestine was identified. The α-amylase, AMY-T, had an estimated molecular weight of 60 kDa and purified to near homogeneity. AMY-T showed an apparent KM 4.78 mg/mL and Vmax 0.44 mg/mL/min) towards soluble starch. It was highly stable for 24 h in the pH range 3.0-10.0, and to solvents like methanol, isopropanol, butanol, dimethylformamide, DMSO and ethyl-ether. AMY-T was able to digest different carbohydrates, mainly showing endo-activity. Importantly, AMY-T was catalytically efficient and adsorbing towards raw potato starch at temperature documented for other raw starch digesting α-amylases. Thin layer and anion exchange chromatography characterization showed that the end products of raw starch hydrolysis were glucose, maltose and maltodextrins, with degree of polymerisation ranging 1-8. Scanning electron microscopy analysis of the AMY-T treated starch granules documented both granular exo- and endo-attack by AMY-T. These catalytic capabilities suggest high potential for AMY-T for industrial use.

Asunto(s)

Cíclidos/metabolismo , Intestinos/enzimología , Almidón/metabolismo , alfa-Amilasas/metabolismo , Animales , Cromatografía por Intercambio Iónico , Cromatografía en Capa Delgada , Concentración de Iones de Hidrógeno , Hidrólisis , Maltosa/análisis , Polisacáridos/análisis , Almidón/análisis , Temperatura

RESUMEN

The family Mugilidae consists mainly of diadromous species, whose reproduction occurs in offshore waters. Pre-juveniles shift their diet in the surf zone (zooplanktophagous to iliophagous). Later, during their recruitment into estuaries, huge changes take place in their digestive system. However, digestive and metabolic characteristics and some morphological traits at recruitment are unknown for Mugilidae. We performed comparative studies on early and late pre-juveniles of Mugil liza recruited in Mar Chiquita Coastal Lagoon (37°32'-37°45'S, 57°19'-57°26'W, Argentina). We determined digestive enzyme activities (intestine), energy reserves (liver/muscle), total/standard length, total weight, intestinal coefficient, hepatosomatic index and retroperitoneal fat. Pre-juveniles exhibited amylase, maltase, sucrase, lipase, trypsin and aminopeptidase-N (APN) activities, which were maintained over a wide range of pH and temperature, and exhibited Michaelis-Menten kinetics. In late pre-juveniles, amylase (422 ± 131 µmol maltose min-1 mgprot-1 ), sucrase (86 ± 14 mg glucose min-1 mgprot-1 ), trypsin (84 ± 9 µmoles min-1 mgprot-1 ) and APN (0.58 ± 0.08 µmoles min-1 mgprot-1 ) activities were higher (42%, 28%, 35% and 28%, respectively) than in the early stage. Also, the intestinal coefficient was higher in late (3.04) compared to early (2.06) pre-juveniles. Moreover, the liver appeared to be a main site of glycogen and triglyceride storage in late pre-juveniles, muscle being the site of storage in early pre-juveniles, exhibiting higher glycogen, free glucose and protein concentrations (92%, 82%, 32%, respectively). The results suggest that pre-juveniles of M. liza exhibit an adequate digestive battery to perform complete hydrolysis of various dietary substrates, availability of energy reserves and morphological characteristics to support their feeding habit and growth after recruitment. Our results represent an important contribution to knowledge of the ecology and digestive physiology of pre-juveniles of Mugilidae in the wild.

Asunto(s)

Digestión/fisiología , Metabolismo Energético , Metaboloma , Smegmamorpha/crecimiento & desarrollo , Smegmamorpha/metabolismo , Animales , Argentina , Dieta , Enzimas/metabolismo , Estuarios , Glucógeno/metabolismo , Intestinos/enzimología , Hígado/metabolismo , Músculos/metabolismo , Smegmamorpha/anatomía & histología

RESUMEN

Two trials were conducted to evaluate the effects of soyabean meal replacement by maize distillers dried grains with solubles (DDGS) in diets for pacu juveniles. Five diets were formulated with 0, 100, 200, 300 and 400 g of DDGS/kg diet replacing up to total dietary soyabean meal. In trial 1, the experimental diets were fed to five groups of fish to evaluate the apparent digestibility coefficients (ADC). In trial 2, four groups of fish were fed each experimental diet for 100 d to evaluate the effects of these diets on digestive enzyme activity, intestine oxidative stress and intestine morphology. The ADC of DM and energy was reduced with dietary DDGS inclusion, while the ADC of lipids was increased, and no differences were observed for the ADC of protein. Independent of dietary treatment, pH increased from anterior to the distal intestine with dietary DDGS inclusion. Digestive enzyme activities were higher on anterior than the distal intestine. Dietary DDGS decreased lipase, amylase, chymotrypsin and trypsin activities, while no differences were observed for total protease activity. Intestine glucose-6-phosphate dehydrogenase was reduced in fish fed the DDGS diets, while catalase activity increased. Lipid peroxidation was lower in fish fed DDGS diets than the control. Intestine histomorphology improved with dietary DDGS inclusion. Overall, the negative effects of soyabean meal could be decreased by dietary replacement with maize DDGS which may have a prebiotic effect, improving intestine health.

Asunto(s)

Alimentación Animal , Dieta , Digestión , Grano Comestible , Peces/fisiología , Intestinos/anatomía & histología , Intestinos/fisiología , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Grasas de la Dieta/metabolismo , Peces/anatomía & histología , Concentración de Iones de Hidrógeno , Hidrolasas/metabolismo , Mucosa Intestinal/anatomía & histología , Intestinos/enzimología , Metabolismo de los Lípidos , Peroxidación de Lípido , Oxidorreductasas/metabolismo , Péptido Hidrolasas/metabolismo , Glycine max , Zea mays

RESUMEN

In fish of freshwaters environments, the accumulation and toxic effects of arsenite (AsIII) can be attenuated by detoxification proteins such as GST and ABCC transporters. We studied the effects of AsIII on the middle intestine of O. mykiss in ex-vivo and in vivo/ex vivo assays. For the ex vivo assays, we measured the transport rate of the ABCC substrate DNP-SG and GST activity in intestinal strips and everted sacs. AsIII inhibited DNP-SG transport in a concentration-dependent manner, specifically when we applied it on the basolateral side. GST activity increased when we applied a maximum concentration of AsIII. For the in vivo/ex vivo assays, we kept fish in water with or without 7.7 µmol L-1 of AsIII for 48 h. Then, we measured DNP-SG transport rate, GST activity, and PP1 activity in intestine strips during one hour. For PP1 activity, we incubated the strips with or without microcystin-LR (MCLR), a toxin excreted through ABCC2 proteins. We also analyzed Abcc2 and Gst-π mRNA expression in intestine and liver tissue. In the group exposed in vivo to AsIII, DNP-SG transport rate and GST activity were higher and the effect of MCLR over PP1 activity was attenuated. AsIII significantly induced only Abcc2 mRNA expression in both middle intestine and liver. Our results suggest that, in the middle intestine of O. mykiss, AsIII is absorbed mainly at the basolateral side of the enterocytes, excreted to the lumen by ABCC2 transporters, and is capable of modulating Abcc2 mRNA expression by a transcriptional mechanism.

Asunto(s)

Transportadoras de Casetes de Unión a ATP/metabolismo , Arsenitos , Gutatión-S-Transferasa pi/metabolismo , Intestinos/enzimología , Hígado/metabolismo , Oncorhynchus mykiss/metabolismo , Animales , Arsenitos/metabolismo , Arsenitos/farmacocinética , Arsenitos/toxicidad , Proteínas de Peces/metabolismo , Regulación de la Expresión Génica , ARN Mensajero , Xenobióticos/metabolismo , Xenobióticos/farmacocinética , Xenobióticos/toxicidad

RESUMEN

Maize silks have been used in Mexico for centuries as a natural-based treatment for various illnesses, including obesity and diabetes. It has been shown in mice that intake of maize silk extracts reduces the levels of blood glucose. However, it is not clear how or what maize silk compounds are involved in such an effect. A hypothesized mechanism is that some maize silk compounds can inhibit carbohydrate hydrolyzing enzymes like α-glucosidases. This work aimed to assess the capability of both saccharides and phenolic compounds from maize silks to inhibit intestinal α-glucosidases. Results showed that saccharides from maize silks did not produce inhibition on intestinal α-glucosidases, but phenolics did. Maize silk phenolics increased the value of Km significantly and decreased the Vmax slightly, indicating a mixed inhibition of α-glucosidases. According to the molecular docking analysis, the phenolics maysin, methoxymaysin, and apimaysin, which had the highest predicted binding energies, could be responsible for the inhibition of α-glucosidases. PRACTICAL APPLICATIONS: The International Diabetes Federation (IDF) reported in 2017 that diabetes affects over 424 million people worldwide, and caused 4 million deaths. Non-insulin-dependent diabetes or type 2 diabetes mellitus (T2DM) accounts for ∼90% of cases. T2DM is characterized by insulin resistance and pancreatic ß-cell failure. Therapy for T2DM includes the use of sulfonylureas, thiazolidinediones, biguanides, and α-glucosidase inhibitors. Regarding the α-glucosidase inhibitors, only few are commercially available, and these have been associated with severe gastrointestinal side effects. This work aimed to assess the capability of both saccharides and phenolic compounds from maize silks to inhibit intestinal α-glucosidases. Results from this work evidenced that maize silk polyphenols acted as effective inhibitors of intestinal rat α-glucosidases. Computational analysis of maize silk polyphenols indicated that maysin, a particular flavonoid from maize silks, could be responsible for the inhibition of α-glucosidases.

Asunto(s)

Diabetes Mellitus Tipo 2/tratamiento farmacológico , Flores/química , Inhibidores de Glicósido Hidrolasas/farmacología , Hipoglucemiantes/farmacología , Fenoles/farmacología , Zea mays/química , alfa-Glucosidasas/metabolismo , Glucemia/efectos de los fármacos , Flavonoides/química , Flavonoides/farmacología , Glucósidos/química , Glucósidos/farmacología , Inhibidores de Glicósido Hidrolasas/química , Hipoglucemiantes/química , Intestinos/enzimología , Cinética , Simulación del Acoplamiento Molecular , Fenoles/química , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Polifenoles/química , Polifenoles/farmacología

RESUMEN

The objective of this study was to evaluate the effect of oxidized soybean oils on the growth performance, metabolic oxidative status and intestinal barrier function of broiler chickens. A total of 240 one-day-old female broiler chickens were assigned to four dietary treatments with six replicates (cages) of 10 birds each. The dietary treatments comprised of a basal diet supplemented with 4% of: non-oxidized (fresh) soybean oil (control treatment, SNX); lowly-oxidized soybean oil (SLX) (oil heated for 10h at 200°C); moderately-oxidized soybean oil (SMX) (oil heated for 18h at 200°C); or highly-oxidized soybean oil (SHX) (oil heated for 30h at 200°C). Diets and water were offered ad libitum. The experiment was lasted 21d.The growth performance of broilers, determined from 1 to 14 d and from 1 to 21 d of age, was not affected by the dietary treatments (p>0.05). Broilers fed oxidized soybean oils presented higher corticosterone serum levels compared with those fed non-oxidized oil (p<0.05). Higher malondialdehyde (MDA) levels onday14 and 21 (p<0.05), and lower total antioxidant capacity (T-AOC) and totalsuperoxide dismutase (T-SOD) values on day 21were obtained in the liver of broiler fed oxidized oils relative to those fed the non-oxidized oil (p<0.05). Broilers fed the highly-oxidized soybean oil had higher (p<0.05) MDA levels in the jejunum on day 21 compared with those fed non-oxidized soybean oil. Chickens fed moderately- and highly-oxidized soybean oil presented lower (p<0.05) T-SOD activity inileal mucosa compared with those fed non-oxidized soybean oil. Ileal mRNA expression of claudin-1 tended to be down regulated by the dietary addition of oxidized oils (p=0.056). The mRNA expression of interleukin-22 (IL-22) of broilers fed moderately-oxidized and highly-oxidized soybean oil was higher (p<0.05), and the mRNA expression of occludin and catalase was lower (p<0.05) than those fed non-oxidized soybean oil. However, the morphology of the jejunal and ileal...

Asunto(s)

Pollos , Intestinos/enzimología , Aceite de Soja/análisis , Aceite de Soja/efectos adversos , Oxidación

RESUMEN

The objective of this study was to evaluate the effect of oxidized soybean oils on the growth performance, metabolic oxidative status and intestinal barrier function of broiler chickens. A total of 240 one-day-old female broiler chickens were assigned to four dietary treatments with six replicates (cages) of 10 birds each. The dietary treatments comprised of a basal diet supplemented with 4% of: non-oxidized (fresh) soybean oil (control treatment, SNX); lowly-oxidized soybean oil (SLX) (oil heated for 10h at 200°C); moderately-oxidized soybean oil (SMX) (oil heated for 18h at 200°C); or highly-oxidized soybean oil (SHX) (oil heated for 30h at 200°C). Diets and water were offered ad libitum. The experiment was lasted 21d.The growth performance of broilers, determined from 1 to 14 d and from 1 to 21 d of age, was not affected by the dietary treatments (p>0.05). Broilers fed oxidized soybean oils presented higher corticosterone serum levels compared with those fed non-oxidized oil (p<0.05). Higher malondialdehyde (MDA) levels onday14 and 21 (p<0.05), and lower total antioxidant capacity (T-AOC) and totalsuperoxide dismutase (T-SOD) values on day 21were obtained in the liver of broiler fed oxidized oils relative to those fed the non-oxidized oil (p<0.05). Broilers fed the highly-oxidized soybean oil had higher (p<0.05) MDA levels in the jejunum on day 21 compared with those fed non-oxidized soybean oil. Chickens fed moderately- and highly-oxidized soybean oil presented lower (p<0.05) T-SOD activity inileal mucosa compared with those fed non-oxidized soybean oil. Ileal mRNA expression of claudin-1 tended to be down regulated by the dietary addition of oxidized oils (p=0.056). The mRNA expression of interleukin-22 (IL-22) of broilers fed moderately-oxidized and highly-oxidized soybean oil was higher (p<0.05), and the mRNA expression of occludin and catalase was lower (p<0.05) than those fed non-oxidized soybean oil. However, the morphology of the jejunal and ileal...(AU)

Asunto(s)

Aceite de Soja/efectos adversos , Aceite de Soja/análisis , Intestinos/enzimología , Pollos , Oxidación

RESUMEN

Epigeic worms modify microbial communities through their digestive processes, thereby influencing the decomposition of organic matter in vermicomposting systems. Nevertheless, the enzyme dynamics within the gut of tropically adapted earthworms is unknown, and the enzymes involved have not been simultaneously studied. The activities of 19 hydrolytic enzymes within three different sections of the intestine of Eisenia fetida were determined over a fasting period and at 24 h and 30, 60, and 90 days of vermicomposting, and data were evaluated by multivariate analyses. There were found positive correlations between the maximal activity of glycosyl hydrolases and one esterase with the anterior intestine (coincident with the reduction of hemicellulose in the substrate) and the activity of the protease α-chymotrypsin with posterior intestine. The results suggest that activities of enzymes change in a coordinated manner within each gut section, probably influenced by selective microbial enzyme enrichment and by the availability of nutrients throughout vermicomposting.

Asunto(s)

Café , Compostaje , Oligoquetos/enzimología , Clima Tropical , Animales , Biodegradación Ambiental , Café/metabolismo , Intestinos/anatomía & histología , Intestinos/enzimología , Suelo/química , Microbiología del Suelo

RESUMEN

Studies on the 'gut origin of sepsis' have suggested that stressful insults, such as surgery, can affect intestinal permeability, leading to bacterial translocation. Symbiotics have been reported to be able to improve gut permeability and modulate the immunologic system, thereby decreasing postoperative complications. Therefore we aimed to evaluate the postoperative use of symbiotics in head and neck cancer surgical patients for intestinal function and permeability, as well as the postoperative outcomes. Patients were double-blind randomised into the symbiotic (n 18) or the control group (n 18). Samples were administered twice a day by nasoenteric tube, starting on the 1st postoperative day until the 5th to 7th day, and comprised 109 colony-forming units/ml each of Lactobacillus paracasei, L. rhamnosus, L. acidophilus, and Bifidobacterium lactis plus 6 g of fructo-oligosaccharides, or a placebo (6 g of maltodextrin). Intestinal function (day of first evacuation, total stool episodes, stool consistency, gastrointestinal tract symptoms and gut permeability by diamine oxidase (DAO) enzyme) and postoperative complications (infectious and non-infectious) were assessed. Results of comparison of the pre- and postoperative periods showed that the groups were similar for all outcome variables. In all, twelve patients had complications in the symbiotic group v. nine in the control group (P>0·05), and the preoperative-postoperative DAO activity ranged from 28·5 (sd 15·4) to 32·7 (sd 11·0) ng/ml in the symbiotic group and 35·2 (sd 17·7) to 34·1 (sd 12·0) ng/ml in the control group (P>0·05). In conclusion, postoperative symbiotics did not impact on intestinal function and postoperative outcomes of head and neck surgical patients.

Asunto(s)

Neoplasias de Cabeza y Cuello/cirugía , Cuidados Posoperatorios/métodos , Prebióticos/administración & dosificación , Probióticos/administración & dosificación , Anciano , Amina Oxidasa (conteniendo Cobre)/metabolismo , Traslocación Bacteriana/fisiología , Bifidobacterium/crecimiento & desarrollo , Recuento de Colonia Microbiana , Método Doble Ciego , Femenino , Microbioma Gastrointestinal/fisiología , Neoplasias de Cabeza y Cuello/fisiopatología , Humanos , Intestinos/enzimología , Intestinos/fisiología , Lactobacillus/crecimiento & desarrollo , Masculino , Persona de Mediana Edad , Terapia Nutricional , Permeabilidad , Placebos , Complicaciones Posoperatorias/epidemiología , Resultado del Tratamiento

RESUMEN

Previous studies have demonstrated a negative correlation between intestinal alkaline phosphatase (IAP) activity and calcium (Ca) absorption in the gut, as IAP acts as a protective mechanism inhibiting high Ca entry into enterocytes, preventing Ca overload. Here we evaluated Ca absorption and bone properties in knockout mice (KO) completely devoid of duodenal IAP (Akp3 -/- mice). Female C57BL/6 control mice (WT, n = 7) and KO mice (n = 10) were used to determine Ca absorption in vivo and by in situ isolated duodenal loops followed by histomorphometric analysis of duodenal villi and crypts. Bone mineral density, morphometry, histomorphometry and trabecular connectivity and biomechanical properties were measured on bones. We observed mild atrophy of the villi with lower absorption surface and a significantly higher Ca uptake in KO mice. While no changes were seen in cortical bone, we found better trabecular connectivity and biomechanical properties in the femurs of KO mice compared to WT mice. Our data indicate that IAP KO mice display higher intestinal Ca uptake, which over time appears to correlate with a positive effect on the biomechanical properties of trabecular bone.

Asunto(s)

Fosfatasa Alcalina/deficiencia , Calcio/metabolismo , Hueso Esponjoso/metabolismo , Intestinos/enzimología , Fosfatasa Alcalina/metabolismo , Animales , Fenómenos Biomecánicos , Densidad Ósea , Calcio/sangre , Duodeno/metabolismo , Femenino , Fracturas del Cuello Femoral/fisiopatología , Ratones Endogámicos C57BL , Ratones Noqueados , Fosfatos/sangre

RESUMEN

This work presents an inexpensive, simple and fast procedure to purify trypsin based on affinity binding with ferromagnetic particles of azocasein composite (mAzo). Crude extract was obtained from intestines of fish Nile tilapia (Oreochromis niloticus) homogenized in buffer (01g tissue/ml). This extract was exposed to 100mg of mAzo and washed to remove unbound proteins by magnetic field. Trypsin was leached off under high ionic strength (3M NaCl). Preparation was achieved containing specific activity about 60 times higher than that of the crude extract. SDS-PAGE showed that the purified protein had molecular weight (24kDa) in concordance with the literature for the Nile tilapia trypsin. The mAzo composite can be reused and applied to purify trypsin from other sources.

Asunto(s)

Caseínas/química , Cíclidos/metabolismo , Intestinos/enzimología , Tripsina/aislamiento & purificación , Animales , Fraccionamiento Químico , Proteínas de Peces/química , Proteínas de Peces/aislamiento & purificación , Hierro/química , Nanopartículas de Magnetita/química , Peso Molecular , Tripsina/química

RESUMEN

This study evaluated the effect of tin exposure on enzyme activity in the sea cucumber (Holothuria grisea Selenka, 1867). After exposure to 0 (control), 0.04, 0.08, or 0.12 mg L-1 tin, we tested the activities of total cholinesterase in longitudinal muscles, acid phosphatase in gonads and the respiratory tree, as well as alkaline phosphatase in the intestines during a 96-h bioassay. Regression analyses showed that all enzyme activities declined with increasing tin concentrations, except for acid phosphatase in the respiratory tree, which were similarly, inhibited at all tin concentrations. These results indicate that H. grisea is a potential bioindicator for seascape habitat monitoring programs, as its biochemical markers show sensitivity to trace elements that can indicate a rise in pollution levels.

Asunto(s)

Fosfatasa Ácida/antagonistas & inhibidores , Fosfatasa Alcalina/antagonistas & inhibidores , Inhibidores de la Colinesterasa/farmacología , Holothuria/enzimología , Estaño/farmacología , Animales , Relación Dosis-Respuesta a Droga , Gónadas/enzimología , Intestinos/enzimología , Músculos/enzimología , Sistema Respiratorio/enzimología

RESUMEN

An outdoor microcosm was performed with tadpoles (Rhinella arenarum) exposed to 125µgL-1 chlorpyrifos and fed two types of food, i.e., lettuce (Lactuca sativa) and a formulated commercial pellet. Acetylcholinesterase (AChE) and carboxylesterase (CbE) activities were measured in liver and intestine after 10 days of pesticide exposure. Non-exposed tadpoles fed lettuce had an intestinal AChE activity almost two-fold higher than that of pellet-fed tadpoles. No significant differences were observed, however, in liver AChE activity between diets. Likewise, intestinal CbE activity - measured using two substrates, i.e. 1-naphthyl acetate (1-NA) and 4-nitrophenyl valerate (4-NPV) - was higher in tadpoles fed lettuce than in those fed pellets. However, the diet-dependent response of liver CbE activity was opposite to that in the intestine. Chlorpyrifos caused a significant inhibition of both esterase activities, which was tissue- and diet-specific. The highest inhibition degree was found in the intestinal AChE and CbE activities of lettuce-fed tadpoles (42-78% of controls) compared with pellet-fed tadpoles (<60%). Although chlorpyrifos significantly inhibited liver CbE activity of the group fed lettuce, this effect was not observed in the group fed pellets. In general, intestinal CbE activity was more sensitive to chlorpyrifos inhibition than AChE activity. This finding, together with the high levels of basal CbE activity found in the intestine, may be understood as a detoxification system able to reduce intestinal OP uptake. Moreover, the results of this study suggest that diet is a determinant factor in toxicity testing with tadpoles to assess OP toxicity, because it modulates levels of this potential detoxifying enzyme activity.

Asunto(s)

Carboxilesterasa/metabolismo , Cloropirifos/toxicidad , Contaminantes Ambientales/toxicidad , Larva/efectos de los fármacos , Plaguicidas/toxicidad , Acetilcolinesterasa/metabolismo , Animales , Argentina , Bufo arenarum , Dieta , Monitoreo del Ambiente , Intestinos/efectos de los fármacos , Intestinos/enzimología , Larva/enzimología , Nitrobencenos , Valeratos

RESUMEN

Inositol hexakisphosphate (IP6) and inositol both regulate insulin secretion, but their combined use in the management of diabetes deserves investigation. The combined effects of IP6 and inositol supplementation were investigated in streptozotocin-induced type 2 diabetic rats. The following groups of rats were studied for 8 weeks: non-diabetic control, non-diabetic high-fat diet control, diabetic untreated, diabetic rats treated with the combination of IP6 and inositol (650 mg/kg bw) and diabetic rats treated with glibenclamide (10 mg/kg bw). High-fat diet and streptozotocin were used to induce type 2 diabetes mellitus in Sprague-Dawley rats. Body weight, blood glucose, glycated haemoglobin, insulin, serum leptin, HOMA-insulin resistance scores, intestinal amylase activity, serum and faecal lipids and food and fluid consumption were measured. Treatment with the combination significantly reduced blood glucose (306 ± 53 mg/dl) and insulin resistance score (1.93 ± 0.45) compared with diabetic controls (522 ± 24 mg/dl and 5.1 ± 0.69 respectively). Serum leptin (2.8 ± 0.6 ng/dl) and faecal triglycerides (108 ± 8 mg/dl) were significantly increased in rats treated with the combination compared with the diabetic control (1.8 ± 0.06 ng/dl and 86 ± 4 mg/dl). Serum triglyceride (47 ± 5.1 mg/dl), total cholesterol (98 ± 3.2 mg/dl) and food intake (26 ± 0.3 g) were significantly reduced by 45%, 25% and 25%, respectively, in rats treated with the combination compared with the diabetic control. Inositol and IP6 combined supplementation may be effective in the management of type 2 diabetes mellitus and related metabolic disorders by regulating some aspects of lipid and carbohydrate metabolism.

Asunto(s)

Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Hipoglucemiantes/uso terapéutico , Inositol/uso terapéutico , Ácido Fítico/uso terapéutico , Amilasas/metabolismo , Animales , Glucemia/metabolismo , Peso Corporal/efectos de los fármacos , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/fisiopatología , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/fisiopatología , Suplementos Dietéticos , Ingestión de Líquidos/efectos de los fármacos , Ingestión de Líquidos/fisiología , Evaluación Preclínica de Medicamentos/métodos , Quimioterapia Combinada , Ingestión de Alimentos/efectos de los fármacos , Ingestión de Alimentos/fisiología , Heces/química , Hipoglucemiantes/farmacología , Inositol/farmacología , Intestinos/enzimología , Leptina/sangre , Metabolismo de los Lípidos/efectos de los fármacos , Lípidos/sangre , Masculino , Ácido Fítico/farmacología , Ratas Sprague-Dawley

RESUMEN

Intestinal mucositis is an inflammatory process occurring in the intestinal mucosa and is a common side effect of irinotecan hydrochloride (CPT-11) based anticancer regimens. The transient receptor potential cation channel, subfamily A, member 1 (TRPA1) receptor is highly expressed in the intestinal mucosa and has the ability to identify cell damage signaling indicates its possible association with intestinal mucositis. Carvacrol is an agonist of the TRPA1 receptor and has anti-inflammatory properties. Thus, the aim of the present study was to verify the supposed anti-inflammatory and protective action of carvacrol via TRPA1 activation against intestinal mucositis induced by CPT-11 in mice. Briefly, mice were treated with either DMSO 2% or CPT-11 (75 mg/kg, per 4 days, i.p.) or the carvacrol (25, 75 or 150 mg/kg, per 8 days, i.p.) before CPT-11. In other group, the animals were pretreated with HC-030031, a TRPA1 antagonist, 30 min before treatment with carvacrol. On day 7, animal survival and bacteremia were assessed, and following euthanasia, samples of the jejunum were obtained for morphometric analysis and measurement of antioxidant and pro-inflammatory markers. Carvacrol was found to exert an anti-inflammatory action against CPT-11-induced intestinal mucositis through strong interactions with TRPA1 receptors; reduction in the production or release or both of pro-inflammatory cytokines (TNF-α, IL-1ß, and KC); and decrease in other indicators of inflammation (MPO, NF-κB, COX-2) and oxidative stress (GSH, MDA, and NOx levels). It also contributed to the restoration of the tissue architecture of the villi and crypts in the small intestine, and improved clinical parameters such as survival, body mass variation, leukogram, and blood bacterial count. Thus, TRPA1 could be a target for future therapeutic approaches in the treatment of intestinal mucositis.

Asunto(s)

Camptotecina/análogos & derivados , Inflamación/tratamiento farmacológico , Intestinos/patología , Monoterpenos/uso terapéutico , Mucositis/inducido químicamente , Mucositis/tratamiento farmacológico , Estrés Oxidativo , Canales de Potencial de Receptor Transitorio/metabolismo , Animales , Antioxidantes/metabolismo , Peso Corporal/efectos de los fármacos , Camptotecina/efectos adversos , Ciclooxigenasa 2/metabolismo , Cimenos , Femenino , Inmunohistoquímica , Inflamación/sangre , Inflamación/complicaciones , Intestinos/efectos de los fármacos , Intestinos/enzimología , Irinotecán , Recuento de Leucocitos , Ratones , Simulación del Acoplamiento Molecular , Monoterpenos/química , Monoterpenos/farmacología , Mucositis/sangre , Mucositis/enzimología , FN-kappa B/metabolismo , Estrés Oxidativo/efectos de los fármacos , Peroxidasa/metabolismo , Análisis de Supervivencia , Canal Catiónico TRPA1 , Canales de Potencial de Receptor Transitorio/agonistas

RESUMEN

The purpose of this study was to determine whether the administration of the feruloyl esterase (FE)-producing strain Lactobacillus fermentum CRL1446 enhances metabolic and oxidative parameters in caloric-restricted (CR) mice. Balb/c male mice were divided into ad libitum fed Group (ALF Group), CR diet Group (CR Group) and CR diet plus L. fermentum Group (CR-Lf Group). CR diet was administered during 45 days and CRL1446 strain was given in the dose of 108 cells/mL/day/mouse. FE activity was determined in intestinal mucosa and content at Day 1, 20 and 45. Triglyceride, total cholesterol, glucose, thiobarbituric acid reactive substances (TBARS) levels and glutathione reductase activity were determined in plasma. Gut microbiota was evaluated by high-throughput sequencing of 16S rRNA gene amplicons. At Day 45, total intestinal FE activity in CR-Lf Group was higher (p = 0.020) than in CR and ALF groups and an improvement in both metabolic (reductions in triglyceride (p = 0.0025), total cholesterol (p = 0.005) and glucose (p < 0.0001) levels) and oxidative (decrease of TBARS levels and increase of plasmatic glutathione reductase activity (p = 0.006)) parameters was observed, compared to ALF Group. CR diet increased abundance of Bacteroidetes and CRL1446 administration increased abundance of Bifidobacterium and Lactobacillus genus. L. fermentun CRL1446 exerted a bifidogenic effect under CR conditions.

Asunto(s)

Proteínas Bacterianas/metabolismo , Restricción Calórica , Hidrolasas de Éster Carboxílico/metabolismo , Microbioma Gastrointestinal , Intestinos/enzimología , Intestinos/microbiología , Limosilactobacillus fermentum/enzimología , Probióticos , Animales , Biomarcadores/sangre , Glucemia/metabolismo , Peso Corporal , Peroxidación de Lípido , Lípidos/sangre , Masculino , Ratones Endogámicos BALB C , Modelos Animales , Estrés Oxidativo , Factores de Tiempo

RESUMEN

PURPOSE: To evaluate the effects of an intraperitoneal solution of methylene blue (MB), lidocaine and pentoxyphylline (PTX) on intestinal ischemic and reperfusion injury. METHODS: Superior mesenteric artery was isolated and clamped in 36 adult male Sprague Dawley rats. After 60 minutes, clamp was removed and a group received intraperitoneally UNITO solution (PTX 25mg/kg + lidocaine 5mg/kg + MB 2mg/kg), while the other group was treated with warm 0.9% NaCl solution. Rats were euthanized 45 min after drug administration. Lung and bowel were collected for histological evaluation (using Park's score) and determination of myeloperoxidase (MPO) and malondialdehyde (MDA) levels. RESULTS: Control samples showed lymphoplasmocytic infiltrate and crypt necrosis of villi. MPO and MDA measurements shown no differences between treated and control groups. CONCLUSION: The combination of lidocaine, methylene blue and pentoxyphylline administered intraperitoneally at the studied dose, did not decreased histological lesion scores and biochemical markers levels in intestinal ischemia/reperfusion injury.

Asunto(s)

Antiinflamatorios/uso terapéutico , Intestinos/irrigación sanguínea , Lidocaína/uso terapéutico , Azul de Metileno/uso terapéutico , Pentoxifilina/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Animales , Antiinflamatorios/administración & dosificación , Combinación de Medicamentos , Sinergismo Farmacológico , Inflamación/tratamiento farmacológico , Inflamación/prevención & control , Infusiones Parenterales , Intestinos/enzimología , Lidocaína/administración & dosificación , Pulmón/irrigación sanguínea , Pulmón/metabolismo , Masculino , Malondialdehído/metabolismo , Azul de Metileno/administración & dosificación , Modelos Animales , Pentoxifilina/administración & dosificación , Peroxidasa/metabolismo , Distribución Aleatoria , Ratas Sprague-Dawley , Estadísticas no Paramétricas

RESUMEN

ABSTRACT PURPOSE: To evaluate the effects of an intraperitoneal solution of methylene blue (MB), lidocaine and pentoxyphylline (PTX) on intestinal ischemic and reperfusion injury METHODS: Superior mesenteric artery was isolated and clamped in 36 adult male Sprague Dawley rats. After 60 minutes, clamp was removed and a group received intraperitoneally UNITO solution (PTX 25mg/kg + lidocaine 5mg/kg + MB 2mg/kg), while the other group was treated with warm 0.9% NaCl solution. Rats were euthanized 45 min after drug administration. Lung and bowel were collected for histological evaluation (using Park's score) and determination of myeloperoxidase (MPO) and malondialdehyde (MDA) levels. RESULTS: Control samples showed lymphoplasmocytic infiltrate and crypt necrosis of villi. MPO and MDA measurements shown no differences between treated and control groups. CONCLUSION: The combination of lidocaine, methylene blue and pentoxyphylline administered intraperitoneally at the studied dose, did not decreased histological lesion scores and biochemical markers levels in intestinal ischemia/reperfusion injury.

Asunto(s)

Animales , Masculino , Pentoxifilina/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Intestinos/irrigación sanguínea , Lidocaína/uso terapéutico , Azul de Metileno/uso terapéutico , Antiinflamatorios/uso terapéutico , Pentoxifilina/administración & dosificación , Distribución Aleatoria , Peroxidasa/metabolismo , Modelos Animales , Combinación de Medicamentos , Sinergismo Farmacológico , Inflamación/prevención & control , Inflamación/tratamiento farmacológico , Infusiones Parenterales , Intestinos/enzimología , Lidocaína/administración & dosificación , Pulmón/irrigación sanguínea , Pulmón/metabolismo , Malondialdehído/metabolismo , Azul de Metileno/administración & dosificación , Antiinflamatorios/administración & dosificación

RESUMEN

The effect of different farming systems (cage, pond) upon digestive enzyme activities of Nile tilapia was evaluated. Juvenile Nile tilapia (87.61 ± 1.52 g) were simultaneously cultured in pond and cage systems during 90 days. Cages used nutritional biphasic plan (35 and 32 % crude protein-CP feeds) and ponds used nutritional triphasic plan (35, 32 and 28 % CP feeds). Biometric measurements were monthly performed for adjustments in feeding regimes and removal of intestine tissues to evaluate the performance of enzyme activities. Total proteolytic, amylase and lipase activities were not statistically different between the treatments throughout the periods analyzed (31, 63 and 94 days of culture). However, trypsin and chymotrypsin activities were higher with 31 and 63 days of culture in fish from pond system, suggesting that natural food may have influenced these activities. A positive correlation was observed between the recommended concentration of essential amino acids for Nile tilapia and specific aminopeptidases activity in fish cage system. Substrate-SDS-PAGE revealed 12 active proteolytic bands in both systems. However, integrated density (ID) values were higher in the bands of ponds. Specimens of either cage or pond exhibited five bands of amylolytic activity. Fish from cage and pond systems showed the highest values of ID within 31 days of cultivation. In this study, the complexity of digestive functions could be verified for animals maintained under commercial conditions. Some of the assessed enzymes may show adaptations of their activities and/or expression that allow the fish to achieve a more efficient nutrient assimilation.

Asunto(s)

Acuicultura/métodos , Cíclidos , Intestinos/enzimología , Aminopeptidasas/metabolismo , Amilasas/metabolismo , Animales , Quimotripsina/metabolismo , Cíclidos/crecimiento & desarrollo , Cíclidos/metabolismo , Digestión/fisiología , Proteínas de Peces/metabolismo , Lipasa/metabolismo , Fitoplancton , Estanques , Tripsina/metabolismo , Zooplancton