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1.
J Dairy Sci ; 103(1): 439-446, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31733869

RESUMO

In early-weaning programs, dietary effects on calf rumen development have been studied extensively, but very little information is available about the effects of a solid diet on hepatic metabolism in preweaned dairy calves. The objective of this experiment was to determine the effect of physical form of oats in calf starter on the expression of key hepatic gluconeogenic, ß-oxidation, and acute phase protein genes in preweaned dairy calves. Samples were analyzed from 3 experiments that fed either ground or whole oats in calf starters. Briefly, 7 calves were slaughtered at 5 wk of age in experiment 1, 6 were slaughtered at 6 wk in experiment 2, and 7 were slaughtered at 7 wk in experiment 3, and liver tissue was collected for gene expression analysis. Calves from experiments 1 and 2 were cannulated, and their rumen pH and volatile fatty acids were measured during treatment periods. The mRNA expression of gluconeogenic enzymes pyruvate carboxylase (PC), cytosolic and mitochondrial phosphoenolpyruvate carboxykinase (PCK1 and PCK2), fatty acid oxidation enzyme carnitine palmitoyltransferase I (CPT1), and positive acute phase protein haptoglobin (HPT) was measured by real-time quantitative reverse-transcriptase PCR. Expression of HPT was greater in calves fed whole oats than in calves fed ground oats; however, PC, PCK1, PCK2, and CPT1 were not affected by the physical form of oats. All enzymes (PCK1, PCK2, HPT, and CPT1) except PC were affected by experiment; PCK1 and CPT1 had greater expression in experiment 2 than in experiments 1 and 3. Expression of PCK2 was similar in experiments 2 and 3 but greater than experiment 1. Expression of HPT was similar in experiments 1 and 2 but greater than experiment 3. The mRNA expression of enzymes PCK1, PCK2, and CPT1 differed between experiments 1 and 2 and was negatively correlated with rumen propionate and butyrate but had a positive relationship with rumen acetate. Similarly, rumen pH was different in experiments 1 and 2, averaging 5.69 in experiment 1 and 4.81 in experiment 2, and there was a negative correlation between mRNA expression of rate-limiting gluconeogenic PCK1, PCK2, and ß-oxidation CPT1 enzymes and rumen pH of calves in experiments 1 and 2. We concluded that the physical form of oats in calf starter did not affect gene expression of gluconeogenic and ß-oxidation enzymes in preweaned dairy calves. However, lower rumen pH may be related to the upregulation of these enzymes.


Assuntos
Avena , Bovinos/fisiologia , Dieta/veterinária , Ácidos Graxos Voláteis/análise , Inflamação/veterinária , Fígado/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Bovinos/genética , Doenças dos Bovinos/metabolismo , Grão Comestível/química , Metabolismo Energético/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Gluconeogênese/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Inflamação/metabolismo , Fosfoenolpiruvato Carboxiquinase (ATP)/metabolismo , Propionatos/metabolismo , Piruvato Carboxilase/metabolismo , Rúmen/química , Rúmen/metabolismo , Desmame
2.
J Dairy Sci ; 103(1): 676-689, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31733877

RESUMO

Mastitis, inflammation of the udder, is one of the most common diseases hampering milk yield of dairy cows. Methionine (Met) and arginine (Arg) are key nutrients with potential to regulate inflammation and oxidative stress. The aim of this study was to evaluate the effect of increased supply of Met and Arg on mRNA and protein abundance associated with innate immune response and redox balance during lipopolysaccharide (LPS) stimulation in primary bovine mammary epithelial cells (BMEC). Primary BMEC (n = 4 replicates per treatment) were pre-incubated for 12 h in media with the following amino acid combinations: ideal profile of amino acids (control; Con), increased Met supply (incMet), increased Arg supply (incArg), and increased supply of Met and Arg (incMetArg). Subsequently, cells were challenged with or without LPS (1 µg/mL) and incubated for 6 h. Data were analyzed as a 2 × 2 × 2 factorial using the MIXED procedure of SAS 9.4 (SAS Institute Inc., Cary, NC). The downregulation of SLC36A1 and SLC7A1 mRNA abundance induced by LPS was attenuated in the incArg cultures. Although challenge with LPS led to lower abundance of proteins related to the antioxidant response (NFE2L2, NQO1, GPX1), lower levels of ATG7, and lower mRNA abundance of GPX3, we found little effect in cultures with incMet or incArg. Cultures with incMet, incArg, or incMetArg led to attenuation of the upregulation of SOD2 and NOS2 induced by LPS. Abundance of phosphorylated p65 (RELA) was greater after LPS stimulation, but the response was attenuated in cultures with incMet. The greater ratio of pRELA to total RELA in responses to LPS was also attenuated in cultures with incMetArg. The greater mRNA abundance of the proinflammatory cytokine IL1B induced by LPS was attenuated in cultures with incMet, and the same trend induced by LPS on CXCL2 was also alleviated in cultures with incArg. Overall, the data suggest that greater supply of Met and Arg alleviated the proinflammatory responses triggered by LPS through controlling the abundance of proinflammatory cytokines and chemokines and activity of NF-κB. Little benefit on oxidative stress induced by LPS challenge in BMEC was detected with greater supply of Met and Arg.


Assuntos
Arginina/uso terapêutico , Células Epiteliais/efeitos dos fármacos , Glândulas Mamárias Animais/efeitos dos fármacos , Mastite Bovina/prevenção & controle , Metionina/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Bovinos , Suplementos Nutricionais , Feminino , Imunidade Inata/efeitos dos fármacos , Inflamação/metabolismo , Inflamação/veterinária , Lipopolissacarídeos , Glândulas Mamárias Animais/citologia , Metionina/metabolismo , Leite , Fosforilação , RNA Mensageiro/metabolismo
3.
J Dairy Sci ; 103(1): 972-991, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31704022

RESUMO

The objective of this study was to test the effects of essential fatty acids (EFA), particularly α-linolenic acid, and conjugated linoleic acid (CLA) supplementation on fatty acid (FA) composition, performance, and systemic and hepatic antioxidative and inflammatory responses in dairy cows. Four cows (126 ± 4 d in milk) were investigated in a 4 × 4 Latin square and were abomasally infused with 1 of the following for 6 wk: (1) coconut oil (control treatment, CTRL; 38.3 g/d; providing saturated FA), (2) linseed and safflower oil (EFA treatment; 39.1 and 1.6 g/d, respectively; providing mainly α-linolenic acid), (3) Lutalin (BASF, Ludwigshafen, Germany; CLA treatment; cis-9,trans-11 and trans-10,cis-12 CLA, 4.6 g/d each), (4) or EFA+CLA. The initial dosage was doubled every 2 wk, resulting in 3 dosages (dosage 1, 2, and 3). Cows were fed a corn silage-based total mixed ration with a high n-6/n-3 FA ratio. Dry matter intake and milk yield were recorded daily, and milk composition was measured weekly. The FA compositions of milk fat and blood plasma were analyzed at wk 0, 2, 4, and 6. The plasma concentration and hepatic mRNA abundance of parameters linked to the antioxidative and inflammatory response were analyzed at wk 0 and 6 of each treatment period. Infused FA increased in blood plasma and milk of the respective treatment groups in a dose-dependent manner. The n-6/n-3 FA ratio in milk fat was higher in CTRL and CLA than in EFA and EFA+CLA. The sum of FA

Assuntos
Antioxidantes/metabolismo , Bovinos , Ácidos Graxos/administração & dosagem , Inflamação/veterinária , Ácidos Linoleicos Conjugados/administração & dosagem , Abomaso/metabolismo , Animais , Dieta/veterinária , Suplementos Nutricionais , Vias de Administração de Medicamentos , Metabolismo Energético , Ácidos Graxos/metabolismo , Feminino , Inflamação/prevenção & controle , Injeções , Lactação/fisiologia , Ácidos Linoleicos Conjugados/farmacologia , Leite/metabolismo
4.
J Vet Emerg Crit Care (San Antonio) ; 29(6): 593-603, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31637812

RESUMO

OBJECTIVE: To quantify plasma cytokine concentrations in dogs with sepsis and noninfectious systemic inflammation and to evaluate the association between plasma cytokines and outcome in dogs with sepsis. DESIGN: Prospective, observational cohort study. SETTING: University teaching hospital. ANIMALS: Forty-five dogs with sepsis, 10 dogs with noninfectious systemic inflammation (nSIRS), and 15 healthy controls were consecutively enrolled from June 2015 to February 2016 and followed to hospital discharge. Dogs with sepsis satisfied ≥2 SIRS criteria and had a documented or highly suspected bacterial infection. Dogs with nSIRS satisfied ≥2 SIRS criteria but had no evidence of infection. Dogs <3 kg and those with documented coagulopathy were excluded. INTERVENTIONS: Measurement of inflammatory cytokines and high-mobility group box-1 (HMGB-1) was performed on each group. MEASUREMENTS AND MAIN RESULTS: High-mobility group box-1 concentrations were analyzed by ELISA. Plasma concentrations of 13 cytokines were measured in singlet using multiplex magnetic bead assays. Kruskal-Wallis with Dunn's multiple comparison tests were used to compare biomarker concentrations between groups. Mann-Whitney U-tests were used to compare biomarker concentrations between survivors and nonsurvivors. Associations between biomarkers were evaluated using Spearman's correlation coefficients. Independent outcome predictors were identified using multivariable logistic regression. Alpha was set at 0.05. Concentrations of interleukin (IL)-6, C-X-C motif chemokine (CXCL)-8, keratinocyte-derived chemokine (KC)-like, C-C motif chemokine ligand 2 (CCL2), and HMGB-1 were significantly greater in dogs with sepsis versus healthy controls (all P ≤ 0.034). In dogs with sepsis, only CCL2 was independently associated with survival (odds ratio [OR] 0.996, 95% CI 0.993-0.999, P = 0.004). A cut-off of 385 pg/mL for CCL2 was 80% sensitive and 91.4% specific for nonsurvival (area under the ROC curve [AUROC] 0.866). CONCLUSIONS: Dogs with sepsis have significantly increased concentrations of HMGB-1 and inflammatory cytokines, including IL-6, CXCL8, and KC-like. Increased CCL2 concentration is a negative prognostic indicator in dogs with sepsis. These findings should be confirmed using duplicate analyses in larger, distinct populations of dogs with sepsis before applying them to clinical patients.


Assuntos
Citocinas/metabolismo , Doenças do Cão/sangue , Sepse/veterinária , Síndrome de Resposta Inflamatória Sistêmica/veterinária , Animais , Biomarcadores/sangue , Estudos de Coortes , Citocinas/genética , Cães , Feminino , Regulação da Expressão Gênica , Humanos , Inflamação/metabolismo , Inflamação/veterinária , Masculino , Prognóstico , Estudos Prospectivos , Sepse/sangue , Síndrome de Resposta Inflamatória Sistêmica/sangue
5.
Fish Shellfish Immunol ; 94: 730-738, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31580934

RESUMO

Atrazine (ATR) causes environmental problems and damages the health of fish and aquatic animals. MicroRNAs (miRNAs) play important roles in immune regulation. However, the immunotoxicity mechanism of ATR in fish lymphocytes and the role of miRNA in this process remain unclear. To further study these mechanisms, spleen lymphocytes were exposed to 20, 40 and 60 µg/ml ATR for 18 h. Fluorescence staining and flow cytometry showed that the number of necrotic lymphocytes increased after ATR exposure. Compared with the control group, the mRNA expression of miR-181-5p was inhibited and the mRNA levels of TNF-α and HK2 were increased after ATR exposure. Additionally, the NF-κB inflammatory pathway and the levels of glycometabolism-related genes were upregulated. These results suggest that ATR induces inflammation and elevates glycometabolism in lymphocytes. We further found that the mRNA levels of receptor-interacting serine-threonine kinase 1 (RIP1), receptor-interacting serine-threonine kinase 3 (RIP3), mixed lineage kinase domain-like pseudokinase (MLKL), cylindromatosis (CYLD) and Fas-Associated protein with Death Domain (FADD) and the protein levels of RIP3 and MLKL in the treatment groups were significantly increased compared to those in control group, suggesting that ATR causes lymphocyte necroptosis. We conclude that miR-181-5p plays a key role in necroptosis in carp lymphocytes exposed to ATR by downregulating the expression of HK and TNF-α, which increases the level of glycometabolism and induces the inflammatory response, respectively.


Assuntos
Atrazina/toxicidade , Carpas/imunologia , Doenças dos Peixes/imunologia , Glicólise/imunologia , Inflamação/veterinária , MicroRNAs/metabolismo , /imunologia , Animais , Relação Dose-Resposta a Droga , Doenças dos Peixes/induzido quimicamente , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Glicólise/efeitos dos fármacos , Herbicidas/toxicidade , Inflamação/induzido quimicamente , Inflamação/imunologia , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Baço/efeitos dos fármacos , Baço/imunologia
6.
J Anim Sci ; 97(10): 4349-4361, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31581301

RESUMO

Weaning is one of the most stressful periods in the life of a ruminant. Several factors entrenched within typical management practices pose challenges to the calf gastrointestinal health. Weaning is associated with losses in BW and feed intake. In addition, increasing highly fermentable carbohydrates in the diet at the expense of physically effective fiber after weaning predisposes the development of rumen acidosis and increases the concentration of endotoxin in rumen fluid and the permeability of the lower gut to luminal contents. Endotoxin translocation can elicit immune activation, shifting the metabolic priorities toward the immune system, which if sustained over time can hinder animal health and performance. Strategic supplementation of additives with anti-inflammatory capacity could represent a suitable approach to decrease systemic inflammation, restoring barrier function to luminal contents. Bioactive extracts from Olea europaea have anti-inflammatory activity and have been shown to reduce systemic inflammation in other animal models. A generalized randomized block design was used to evaluate the impact of feeding an olive oil bioactive extract (OBE) to newly weaned heifers injected intravenously with sequentially increasing doses of lipopolysaccharide (LPS). A total of 36 heifers, distributed across 3 experimental periods, were randomly assigned to 1 of 4 treatments that consisted of intravenous injection of either saline (CTL-) or with 6 sequentially increasing doses of LPS (0.10, 0.25, 0.50, 0.75, 1.00, and 1.25 µg/kg of BW) over a 10-d period (CTL+), and CTL+ plus dietary supplementation with a low (OBE-L; 0.04% of diet DM) or a high (OBE-H; 0.16% of diet DM) dose of OBE. Feeding OBE reduced some of the negative effects of prolonged immune activation with LPS, such as improved DMI and decreased intravaginal temperature in some, but not all of the days of LPS challenge (P < 0.05). In addition, feeding OBE reduced circulating concentration of inflammatory markers such as IL-6 and haptoglobin (P < 0.05). Heifers supplemented with OBE had reduced cell surface expression of the cluster of differentiation 14 (CD14) in monocyte cells (P < 0.01), a key receptor for LPS recognition, which was correlated with a faster recovery of immune cell counts in plasma. In conclusion, dietary supplementation with OBE was successful in mitigating the negative effects of sustained immune activation in newly weaned heifers.


Assuntos
Bovinos/fisiologia , Suplementos Nutricionais/análise , Inflamação/veterinária , Olea/química , Extratos Vegetais/química , Ração Animal/análise , Animais , Metabolismo dos Carboidratos , Bovinos/crescimento & desenvolvimento , Bovinos/imunologia , Dieta/veterinária , Feminino , Fermentação , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Lipopolissacarídeos/efeitos adversos , Distribuição Aleatória , Rúmen/efeitos dos fármacos , Rúmen/metabolismo , Desmame
7.
J Vet Diagn Invest ; 31(6): 879-882, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585512

RESUMO

A large, ill-defined, firm, multinodular mass involving the pancreas was confirmed on postmortem examination of a 5-y-old, male Rottweiler that died following acute respiratory distress syndrome, after a period of anorexia and lethargy. Histologically, the mass consisted of plump spindle cells admixed with a variable number of macrophages, lymphocytes, plasma cells, and neutrophils. Foci of coagulative necrosis and hemorrhage were also observed. Spindle cells strongly reacted to antibodies against vimentin, α-smooth muscle actin, and calponin, whereas desmin was expressed only mildly and focally. Pan-cytokeratin, KIT, glial fibrillary acidic protein, and S100 protein were nonreactive. Variable numbers of MAC 387-positive cells, CD3+ lymphocytes, and numerous blood vessels were also detected throughout the mass. Histologic and IHC findings were consistent with a diagnosis of inflammatory myofibroblastic tumor of the pancreas.


Assuntos
Doenças do Cão/diagnóstico , Inflamação/veterinária , Neoplasias de Tecido Muscular/veterinária , Neoplasias Pancreáticas/veterinária , Animais , Doenças do Cão/patologia , Cães , Evolução Fatal , Inflamação/diagnóstico , Inflamação/patologia , Masculino , Neoplasias de Tecido Muscular/diagnóstico , Neoplasias de Tecido Muscular/patologia , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia
8.
J Anim Sci ; 97(11): 4548-4556, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31603198

RESUMO

The objectives of the current study were to explore the effects of mannan oligosaccharide (MOS) supplementation in the diets of sow and (or) their offspring on intestinal bacteria, intestinal and systemic inflammation in the piglet. A total of 60 multiparous sows (4 ± 1 parity; Landrace × Yorkshire) were fed either control diet (sCON, n = 30) or a diet containing 400 mg kg-1 MOS (sMOS, n = 30) from day 86 of gestation until weaning (day 20 of postpartum). On day 7 of age, offspring (Duroc × Landrace Yorkshire) were assigned within sow treatments and fed control diet (pCON) or diet containing 800 mg kg-1 MOS (pMOS) for 28 d (end at 35 d of age), resulting in four piglet diet groups (n = 15 litters per diet group): sCON-pCON, sCON-pMOS, sMOS-pCON, and sMOS-pMOS. Results found that piglet diet MOS increased or tend to increase Lactobacillus amount in the ileum digesta (P < 0.01) and jejunum digesta (P = 0.07), respectively; while tend to decrease Escherichia coli amount in jejunum digesta (P =0.06) and cecum digesta (P = 0.08). Both sow and piglet diets add MOS (sMOS-pMOS) increased Lactobacillus amount but decreased E. coli amount in jejunum digesta (P < 0.05) compared with the sCON-pCON diet group. In addition, sow diet MOS (rather than piglet diet MOS) increased sIgA content in piglet jejunum mucosa compared with control (P = 0.04). Sow diet MOS decreased toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4), and interleukin 8 (IL-8) mRNA levels (P < 0.05) and tended to decrease nuclear factor-κB p65 (NF-κB p65) mRNA level (P = 0.07) in piglet intestinal lymphatic. The interaction effects between sow and piglet diets were found on the mRNA levels of NF- κB p65 (P = 0.03) and IL-8 (P = 0.02) in piglet jejunum. Finally, the sow diet MOS decreased proinflammatory cytokines IL-2 (P < 0.01) and IL-4 (P < 0.01) concentrations in piglet serum. Piglets diet MOS decreased the contents of IL-2 (P = 0.03), IL-4 (P = 0.01) and interferon (IFN)-γ (P < 0.01) while increased anti-inflammatory cytokine IL-10 (P < 0.01) content in serum. The interaction effects between sows and piglet diets on IL-4 (P = 0.02), IL-10 (P < 0.01), and IFN-γ (P = 0.08) were observed. In conclusion, sow and/or piglet diet MOS could improve intestinal microbiota, enhance intestinal mucosal immune competence, and suppress intestinal and systemic inflammation in the piglet.


Assuntos
Suplementos Nutricionais/análise , Microbioma Gastrointestinal/efeitos dos fármacos , Mananas/administração & dosagem , Oligossacarídeos/administração & dosagem , Suínos/fisiologia , Ração Animal/análise , Animais , Animais Recém-Nascidos , Dieta/veterinária , Feminino , Inflamação/veterinária , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Masculino , Paridade , Gravidez , Suínos/imunologia
9.
Fish Shellfish Immunol ; 94: 761-768, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31585240

RESUMO

This study was designed to evaluate the effects of zinc on inflammation and tight junction (TJ) in different intestinal regions of common carp under sub-chronic arsenic insult. Fish were exposed to zinc (0, 1 mg/L) and arsenic trioxide (0, 2.83 mg/L) in individual or combination for a month. Inflammatory infiltration and TJ structure changes were displayed by H&E staining and transmission electron microscope. To further explore these changes, biochemical indicator (SOD), gene or protein expressions of inflammatory responses (NF-κB, IL-1ß, IL-6 and IL-8) and TJ proteins (Occludin, Claudins and ZOs) were determined. In the anterior intestine, arsenic decreased activity of SOD, mRNA levels of Occludin, Claudins and ZOs, increased mRNA levels of ILs. However, unlike the anterior intestine, arsenic has an upregulation effects of Occludin and Claudin-4 in the mid intestine. These anomalies induced by arsenic, except IL-8, were completely or partially recovered by zinc co-administration. Furthermore, transcription factor (NF-κB) nuclear translocation paralleled with its downstream genes in both intestinal regions. In conclusion, our results unambiguously suggested that under arsenic stress, zinc can partly relieve intestinal inflammation and disruption of tight junction segment-dependently.


Assuntos
Arsênico/efeitos adversos , Carpas , Enterotoxinas/efeitos adversos , Doenças dos Peixes/prevenção & controle , Intestinos/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Zinco/farmacologia , Ração Animal/análise , Animais , Dieta/veterinária , Suplementos Nutricionais/análise , Doenças dos Peixes/induzido quimicamente , Inflamação/induzido quimicamente , Inflamação/prevenção & controle , Inflamação/veterinária , Intestinos/fisiologia , Junções Íntimas/efeitos dos fármacos , Junções Íntimas/fisiologia
10.
Vet Res ; 50(1): 65, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533824

RESUMO

Distinct enzymes, including cyclooxygenase 1 and 2 (COX-1 and COX-2), lipoxygenase (LOXs), and cytochrome P450 monooxygenase (CYP450), produce different stress mediators and mediate inflammation in birds. Bioactive agents such as acetylsalicylic acid (ASA) and vitamin E (vE) may affect enzyme activities and could be used in poultry production to control the magnitude of acute phase inflammation. Here, we characterized COX, LOX, and CYP450 mRNA expression levels in chicken immune tissues in response to Escherichia coli lipopolysaccharide (LPS) challenge and investigated whether ASA and vE could alter gene expression. Additionally, for the first time in chickens, we evaluated oxygen consumption by platelet mitochondria as a biomarker of mitochondria function in response to ASA- and vE. LPS challenge compromised bird growth rates, but neither dietary ASA nor vE significantly ameliorated this effect; however, gradually increasing dietary vE levels were more effective than basal levels. ASA regulated arachidonic acid metabolism, providing an eicosanoid synthesis substrate, whereas gradually increasing vE levels evoked aspirin resistance during challenge. Gene expression in immune tissues was highly variable, indicating a complex regulatory network controlling inflammatory pathways. However, unlike COX-1, COX-2 and CYP450 exhibited increased mRNA expression in some cases, suggesting an initiation of novel anti-inflammatory and pro-resolving signals during challenge. Measuring oxygen consumption rate, we revealed that neither the ASA nor vE levels applied here exerted toxic effects on platelet mitochondria.


Assuntos
Aspirina/metabolismo , Galinhas/imunologia , Inflamação/veterinária , Doenças das Aves Domésticas/imunologia , Vitamina E/metabolismo , Ração Animal/análise , Animais , Aspirina/administração & dosagem , Proteínas Aviárias/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Feminino , Inflamação/induzido quimicamente , Inflamação/imunologia , Lipopolissacarídeos/farmacologia , Lipoxigenases/metabolismo , Doenças das Aves Domésticas/induzido quimicamente , Prostaglandina-Endoperóxido Sintases/metabolismo , Vitamina E/administração & dosagem
11.
Fish Shellfish Immunol ; 94: 258-263, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31513913

RESUMO

Grass carp septicemia is a systemic inflammatory response that develops following a bacterial infection. The hyperinflammatory state develops could lead to septic shock and lethality. There is increasing evidence that microRNAs are involved in the regulation of the inflammatory response. In the present study, miR-21 was confirmed to be involved in the inflammatory response following infection with Aeromonas hydrophila and LPS stimulation. Both jnk and ccr7 were identified as target gene of miR-21 by overexpression, inhibition, and dual luciferase reporter assays experiments. Meanwhile, miR-21 targets the jnk and ccr7 to modulate downstream pro-inflammatory factors tnf-α, il-1ß, il-6, and il-12. Our results provide a theoretical basis for exploring the molecular mechanism of grass carp miR-21 regulating inflammation.


Assuntos
Carpas/genética , Carpas/imunologia , Doenças dos Peixes/microbiologia , Inflamação/veterinária , Proteínas Quinases JNK Ativadas por Mitógeno/genética , MicroRNAs/genética , Receptores CCR7/genética , Aeromonas hydrophila/fisiologia , Animais , Sequência de Bases , Doenças dos Peixes/genética , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Infecções por Bactérias Gram-Negativas/genética , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Inflamação/genética , Inflamação/imunologia , Inflamação/microbiologia , Proteínas Quinases JNK Ativadas por Mitógeno/imunologia , Lipopolissacarídeos/farmacologia , MicroRNAs/imunologia , Receptores CCR7/imunologia
13.
Res Vet Sci ; 126: 164-169, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31499425

RESUMO

Endometritis is one of the most common reproductive diseases caused by bacterial infection in the cow. Ferulic acid is a major effective component extracted from Ligusticum wallichii. Ferulic acid displays pharmacological effects such as anti-inflammation and antioxidation. The aim of the present study was to investigate the protective effects of ferulic acid on inflammation induced by lipopolysaccharide (LPS) in bovine endometrial epithelial cells (BEECs). BEECs were pretreated with ferulic acid followed by LPS treatment. QRT-PCR analysis showed the mRNA expression of LPS-induced proinflammatory cytokines (IL1B, IL6, TNFA, and IL8) was decreased with ferulic acid pretreatment. Western blot analysis showed that ferulic acid inhibited the degradation of IκB and phosphorylation of NF-κB p65. Ferulic acid suppressed the phosphorylation of MAPKs, including p38 and JNK. All of these results indicated that ferulic acid may be considered as a potential anti-inflammatory drug for curing endometritis.


Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Doenças dos Bovinos/tratamento farmacológico , Ácidos Cumáricos/administração & dosagem , Endometrite/veterinária , Inflamação/veterinária , Animais , Bovinos , Doenças dos Bovinos/induzido quimicamente , Endometrite/induzido quimicamente , Endometrite/tratamento farmacológico , Endométrio/efeitos dos fármacos , Endométrio/fisiopatologia , Células Epiteliais/efeitos dos fármacos , Feminino , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Lipopolissacarídeos/farmacologia , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos
14.
Fish Shellfish Immunol ; 94: 479-484, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31472264

RESUMO

The present study was conducted to evaluate the anti-inflammatory activity of florfenicol (FFC) against lipopolysaccharide (LPS)-induced inflammatory responses in Ctenopharyngodon idella in vivo and in vitro. Head-kidney (HK) macrophages were pre-treated with 10 µg/mL LPS and then exposed to different concentrations of FFC to determine its in vitro anti-inflammatory activity. Inhibitory effect of FFC on inflammatory mediators TNF-α, IL-6 and IL-1ß, as well as LPS-induced nitric oxide (NO) and prostaglandin E 2 (PGE 2) production were assayed by ELISA. The expression level of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were investigated by RT-PCR. Expression level of TLR-related genes (TLR1, TLR2, TLR4, TLR7, TLR8) expression, tumor necrosis factor receptor-associated factor 6 (TRAF6), transforming growth factor-b-activated kinase 1 (TAK1), Myeloid differentiation factor 88 (MyD88), nucleus p65, NF-κBα (IκBα) were measured by RT-PCR after grass carp were treated with 50, 100 and 200 mg FFC/kg body weight for 3 days. Results from in vitro tests demonstrated that FFC dose-dependently inhibited LPS-induced inflammatory cytokines TNF-α, IL-6 and IL-1ß, inflammatory factors NO and PGE 2 production in macrophages. In addition, iNOS and COX-2 expression levels decreased significantly as compared with LPS treated group. In vivo test demonstrated that treatment with FFC prevented the LPS-induced upregulation of TNF-α, IL-6, IL-1ß, NO and PGE 2. The expression level of iNOS, and COX-2 in FFC-treated grass carp were also downregulated as compared with LPS treated fish. Besides, FFC blocked the expression of Toll-like receptor 2 (TLR2) and then suppressed the phosphorylation of nuclear transcription factor-kappa B (NF-κB) p65 and degradation inhibitor of IκBα. Furthermore, administration of FFC inhibited the up-regulation of IRAK4, TRAF6 and TAK1 induced by LPS. These results suggest that the anti-inflammatory properties of FFC might be the results from the inhibition of iNOS, COX-2, IL-6, IL-1ß, and TNF-α expressions through the down-regulation of Toll/NF-κB signaling pathways.


Assuntos
Anti-Inflamatórios/farmacologia , Carpas/imunologia , Doenças dos Peixes/tratamento farmacológico , Inflamação/veterinária , Tianfenicol/análogos & derivados , Animais , Carpas/genética , Relação Dose-Resposta a Droga , Doenças dos Peixes/induzido quimicamente , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Lipopolissacarídeos/farmacologia , NF-kappa B/fisiologia , Distribuição Aleatória , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Tianfenicol/farmacologia , Receptores Toll-Like/fisiologia
15.
J Anim Sci ; 97(11): 4475-4481, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31560759

RESUMO

Newly weaned, commercial Angus steers [body weight (BW) = 204 ± 19 kg; n = 24; 12 steers from dams administered an injectable trace mineral (MM; Mulimin90) and 12 steers from control (CON) dams] were utilized to determine the effects of maternal supplementation with an injectable trace mineral on the inflammatory response of subsequent steers subjected to a lipopolysaccharide (LPS) challenge at the initiation of a 42-d receiving period. On day -2 steers were weaned, and the following day, shipped 354 km to the Beef Cattle and Sheep Field Laboratory in Urbana, IL. On day 0, steers were administered an intravenous LPS challenge. Body temperature and blood samples were collected from steers prior to LPS administration (0 h) and again at 0.5, 1, 2, 3, 4, 5, and 6 h. Blood samples were analyzed for trace mineral and cortisol at 0 and 2 h and glucose, insulin, LPS-binding protein (LBP), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), haptoglobin, ceruloplasmin, and fibrinogen at 0, 0.5, 1, 2, 3, 4, 5, and 6 h. Calf BW was collected at trial initiation and subsequently every 14 d. Dry matter intake was collected daily and average daily gain (ADG) and feed efficiency were assessed. Initial plasma Zn tended (P = 0.06) to be greater for MM steers. However, there was no difference (P ≥ 0.31) in trace mineral status or serum cortisol at any other time. Total area under the curve (TAUC) for body temperature was lesser (P > 0.01) for MM steers. Basal LBP concentrations and TAUC for LBP tended (P ≤ 0.10) to be greater for MM steers. Peak concentration of IL-6 tended (P = 0.09) to be reached earlier for CON steers. However, there was no difference (P ≥ 0.11) in glucose, insulin, IL-6, ceruloplasmin, haptoglobin, and fibrinogen concentrations between treatments. Calf performance and feed efficiency did not differ (P ≥ 0.17) between treatments except ADG from day 28 to 42, which was greater (P = 0.03) for CON steers. Maternal supplementation with an injectable trace mineral tended to improve steer plasma Zn status at 0 h and tended to increase basal concentrations of LBP and overall LBP production when steers were administered an LPS challenge. Additionally, MM steers exhibited a more favorable change in body temperature following LPS administration. However, injectable trace mineral supplementation of dams during gestation had minimal to no effect on cytokine and acute-phase protein concentrations, as well as overall calf performance and efficiency during a 42-d receiving period.


Assuntos
Bovinos/fisiologia , Suplementos Nutricionais/análise , Inflamação/veterinária , Oligoelementos/administração & dosagem , Ração Animal/análise , Animais , Peso Corporal/efeitos dos fármacos , Dieta/veterinária , Feminino , Hidrocortisona/sangue , Inflamação/prevenção & controle , Lipopolissacarídeos/administração & dosagem , Masculino , Gravidez , Ovinos , Oligoelementos/sangue
16.
Anim Reprod Sci ; 208: 106128, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31405471

RESUMO

The aim was to evaluate effects of addition of pentoxifylline to skimmed milk semen extender on uterine inflammatory response. Thirty-six estrous cycles of 15 mares were randomly divided into five groups for artificial insemination (AI): Control: mimicking the AI procedure (n = 7); Extender: deposition of skimmed milk based extender (n = 7); Extender + PTX: skimmed milk based extender plus pentoxifylline (7.18 mM; n = 8); Semen: semen diluted with extender without pentoxifylline (n = 7), and Semen + PTX: semen diluted with extender containing pentoxifylline (n = 7). Mares in estrus were examined by trans-rectal palpation and using ultrasonography, and ovulation was induced. Uterine hemodynamics were assessed immediately before ovulation induction (T-30), immediately before AI (T0), 2 (T2), 6 (T6), 12 (T12), 24 (T24) and 48 (T48) h after AI. Endometrial samples were collected 6 h after AI, and slides were stained and examined to determine percentage of PMN. Pentoxifylline had no additional effect on vascular perfusion. There was a major inflammatory response with pentoxifylline treatment that was greater than that of the control group. In the group treated with Extender + PTX, there were more PMN (57.98 ±â€¯9.42%) than in the group treated with Extender (20.20 ±â€¯6.63%) and in the Semen + PTX group more PMN (82.84 ±â€¯5.71%) than in the Semen-treated group (47.83 ±â€¯10.61%). These findings indicate the addition of pentoxifylline does not stimulate blood flow; however, it induces a greater immune defense response because more neutrophils migrate to the uterine lumen.


Assuntos
Doenças dos Cavalos/prevenção & controle , Inflamação/veterinária , Pentoxifilina/farmacologia , Sêmen/efeitos dos fármacos , Doenças Uterinas/veterinária , Animais , Estudos Cross-Over , Endométrio/irrigação sanguínea , Endométrio/efeitos dos fármacos , Feminino , Cavalos , Inflamação/prevenção & controle , Inseminação Artificial/veterinária , Masculino , Leite , Ultrassonografia/veterinária , Doenças Uterinas/prevenção & controle , Vasodilatadores/farmacologia
17.
J Dairy Sci ; 102(11): 10277-10290, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31447141

RESUMO

Nonsteroidal anti-inflammatory drugs are used as supportive therapy with antimicrobial treatments for mastitis in cows to alleviate pain of the inflamed mammary gland. They act mainly by inhibition of cyclooxygenases. Meloxicam (MEL) is a drug designed for cyclooxygenase-2 selectivity, which is upregulated upon inflammation, acting as a key enzyme for the conversion of arachidonic acid to prostaglandins. Although some studies in dairy cows showed positive results in recovery from mastitis when MEL was added to the treatments, direct effects of MEL on the immune system of mastitic cows are unknown. The aim of this study was to investigate effects of MEL on the immune response of bovine mammary epithelial cells (MEC) with or without simultaneous immune stimulation by pathogen-associated molecular patterns of common mastitis pathogens. Mammary epithelial cells from 4 cows were isolated and cultured. To evaluate dose effects of MEL, MEC were challenged with or without 0.2 µg/mL lipopolysaccharide (LPS; serotype O26:B6 from Escherichia coli) with addition of increasing concentrations of MEL (0, 0.25, 0.5, 1.0, 1.5, or 2.0 mg/mL). The addition of MEL prevented the increase of mRNA expression of key inflammatory factors in LPS-challenged MEC in a dose-dependent manner. To investigate the effects of MEL on pathogen-specific immune responses of MEC, treatments included challenges with LPS from E. coli and lipoteichoic acid from Staphylococcus aureus with or without 1.5 mg/mL MEL for 3, 6, and 24 h. Meloxicam prevented the increase of mRNA abundance of key inflammatory mediators in response to LPS and lipoteichoic acid, such as tumor necrosis factor, serum amyloid A, inducible nitric oxide synthase, and the chemokines IL-8 and CXC chemokine ligands 3 and 5. The prostaglandin E2 synthesis in challenged and nonchallenged cells was reduced by MEL within 24 h. Furthermore, MEL reduced the viability and consequently the total RNA yield of the cells. However, mRNA abundance of apoptosis-related enzymes was not affected by any treatment. Meloxicam had clear dose-dependent effects on the immune response of MEC to pathogen-associated molecular patterns of common mastitis pathogens by preventing increased expression of important factors involved in inflammation. This nonsteroidal anti-inflammatory drug also has detrimental effects on cell viability. How these effects would influence the elimination of pathogens from an infected mammary gland during mastitis therapy with meloxicam needs to be further investigated.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Inibidores de Ciclo-Oxigenase 2/uso terapêutico , Inflamação/veterinária , Mastite Bovina/tratamento farmacológico , Meloxicam/uso terapêutico , Animais , Biomarcadores/análise , Bovinos , Células Epiteliais/patologia , Escherichia coli/química , Feminino , Lipopolissacarídeos/efeitos adversos , Glândulas Mamárias Animais/patologia , Mastite Bovina/patologia , Staphylococcus aureus/química , Ácidos Teicoicos/efeitos adversos
18.
J Dairy Sci ; 102(11): 10395-10410, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31447151

RESUMO

Neutrophils are the most important polymorphonuclear leukocytes (PMNL), representing the front-line defense involved in pathogen clearance upon invasion. As such, they play a pivotal role in immune and inflammatory responses. Isolated PMNL from 5 mid-lactating Holstein dairy cows were used to evaluate the in vitro effect of methionine (Met) and choline (Chol) supplementation on mRNA expression of genes related to the Met cycle and innate immunity. The target genes are associated with the Met cycle, cell signaling, inflammation, antimicrobial and killing mechanisms, and pathogen recognition. Treatments were allocated in a 3 × 3 factorial arrangement, including 3 Lys-to-Met ratios (L:M, 3.6:1, 2.9:1, or 2.4:1) and 3 levels of supplemental Chol (0, 400, or 800 µg/mL). Three replicates per treatment group were incubated for 2 h at 37°C and 5% atmospheric CO2. Both betaine-homocysteine S-methyltransferase and choline dehydrogenase were undetectable, indicating that PMNL (at least in vitro) cannot generate Met from Chol through the betaine pathway. The PMNL incubated without Chol experienced a specific state of inflammatory mediation [greater interleukin-1ß (IL1B), myeloperoxidase (MPO), IL10, and IL6] and oxidative stress [greater cysteine sulfinic acid decarboxylase (CSAD), cystathionine gamma-lyase (CTH), glutathione reductase (GSR), and glutathione synthase (GSS)]. However, data from the interaction L:M × Chol indicated that this negative state could be overcome by supplementing additional Met. This was reflected in the upregulation of methionine synthase (MTR) and toll-like receptor 2 (TLR2); that is, pathogen detection ability. At the lowest level of supplemental Chol, Met downregulated GSS, GSR, IL1B, and IL6, suggesting it could reduce cellular inflammation and enhance antioxidant status. At 400 µg/mL Chol, supplemental Met upregulated PMNL recognition capacity [higher TLR4 and L-selectin (SELL)]. Overall, enhancing the supply of methyl donors to isolated unstimulated PMNL from mid-lactating dairy cows leads to a low level of PMNL activation and upregulates a cytoprotective mechanism against oxidative stress. Enhancing the supply of Met coupled with adequate Chol levels enhances the gene expression of PMNL pathogen-recognition mechanism. These data suggest that Chol supply to PMNL exposed to low levels of Met effectively downregulated the entire repertoire of innate inflammatory-responsive genes. Thus, Met availability in PMNL during an inflammatory challenge may be sufficient for mounting an appropriate biologic response.


Assuntos
Bovinos/sangue , Colina/administração & dosagem , Metionina/administração & dosagem , Neutrófilos/metabolismo , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase , Animais , Antioxidantes/metabolismo , Bovinos/imunologia , Bovinos/fisiologia , Colina/genética , Colina/metabolismo , Dieta/veterinária , Regulação para Baixo , Feminino , Expressão Gênica , Imunidade Inata/genética , Inflamação/genética , Inflamação/veterinária , Lactação/efeitos dos fármacos , Metionina/genética , Metionina/metabolismo , Neutrófilos/imunologia , Estresse Oxidativo/genética , RNA Mensageiro/metabolismo
19.
J Dairy Sci ; 102(11): 10599-10605, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31447163

RESUMO

The supply of methionine (Met) in late pregnancy can alter mRNA abundance of genes associated with metabolism and immune response in liver and polymorphonuclear leukocytes (PMN) of the neonatal calf. Whether prenatal supply of Met elicits postnatal effects on systemic inflammation and innate immune response of the calf is not well known. We investigated whether enhancing the maternal supply of Met via feeding ethyl-cellulose rumen-protected Met (RPM) was associated with differences in calf innate immune response mRNA abundance in PMN and systemic indicators of inflammation during the first 50 d of life. Calves (n = 14 per maternal diet) born to cows fed RPM at 0.09% of diet dry matter per day (MET) for the last 28 ± 2 d before calving or fed a control diet with no added Met (CON) were used. Blood for biomarker analysis and isolation of PMN for innate immune function assays and mRNA abundance was harvested at birth (before colostrum feeding) and at 7, 21 and 50 d of age. Whole blood was challenged with enteropathogenic bacteria (Escherichia coli 0118:H8) and phagocytosis and oxidative burst of neutrophils and monocytes were quantified via flow cytometry. Although concentration of haptoglobin and activity of myeloperoxidase among calves from both maternal groups increased markedly between 0 and 7 d of age followed by a decrease to baseline at d 21 the responses were lower in MET compared with CON calves. Nitric oxide concentration decreased markedly between 0 and 7 d regardless of maternal group but MET calves tended to have lower overall concentrations during the study. In vitro phagocytosis in stimulated neutrophils increased markedly over time in both CON and MET calves but responses were overall greater in MET calves. Oxidative burst in both neutrophils and monocytes increased over time regardless of maternal treatment. The mRNA abundance of lactate dehydrogenase (LDHA) signal transducer and activator of transcription 3 (STAT3) and S100 calcium binding protein A8 (S100A8) in PMN was overall greater in MET calves. Overall data suggest that increasing the maternal supply of Met during late pregnancy could affect the neonatal calf inflammatory status and innate immune response. Although changes in mRNA abundance could play a role in coordinating the immune response the exact mechanisms merit further study.


Assuntos
Bovinos , Dieta/veterinária , Imunidade Inata/efeitos dos fármacos , Metionina/farmacologia , Neutrófilos/imunologia , RNA Mensageiro/metabolismo , Animais , Bovinos/imunologia , Suplementos Nutricionais , Feminino , Inflamação/prevenção & controle , Inflamação/veterinária , Contagem de Leucócitos , Fígado/metabolismo , Metionina/metabolismo , Fagocitose , Gravidez , Complicações na Gravidez/prevenção & controle , Complicações na Gravidez/veterinária , Rúmen/metabolismo
20.
J Vet Intern Med ; 33(5): 2183-2192, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31454107

RESUMO

BACKGROUND: Brachycephalic dogs have abnormal breathing patterns similar to those in humans with obstructive sleep apnea syndrome. Obstructive sleep apnea syndrome is associated with dyslipidemia, hyperglycemia, and insulin resistance. Despite the fact that anatomic and functional alterations are well described in brachycephalic dogs, little is known about the consequences of upper airway obstruction on systemic inflammatory response and metabolic profile. OBJECTIVES: To describe history, clinical presentation, and anatomic abnormalities; to evaluate systemic inflammatory response and metabolic profile; and to identify possible associations among clinical signs, anatomic abnormalities, inflammatory response, and metabolic profile in brachycephalic dogs with airway obstruction. ANIMALS: Thirty purebred brachycephalic dogs with brachycephalic airway obstructive syndrome (BAOS). METHODS: Prospective study. The following information was recorded and studied: respiratory and digestive signs, airway and digestive endoscopic anomalies, presence or absence of tracheal hypoplasia, histologic evaluation of gastrointestinal tract biopsy specimens, serum concentrations of C-reactive protein (CRP), fructosamine, insulin, glucose, triglyceride, cholesterol, and plasma concentrations of lipoprotein classes. RESULTS: A high proportion of dogs (76.7%) had gastrointestinal signs. Esophageal deviation, atony of the cardia of the stomach, and distal esophagitis were the most common endoscopic anomalies detected. Twenty-six (86.6%) dogs had different degrees of laryngeal collapse. Gastrointestinal histologic evaluation identified mostly chronic inflammation. Glucose, fructosamine, triglycerides, cholesterol, CRP, pre-beta, beta lipoproteins, and chylomicrons were increased to a variable extent. Significant associations among clinical signs, anatomic abnormalities, CRP, and metabolic profile were not found. CONCLUSION AND CLINICAL IMPORTANCE: Despite the presence of inflammation and some mild metabolic derangements, the clinicopathological variables evaluated did not offer valuable information in dogs with BAOS.


Assuntos
Obstrução das Vias Respiratórias/veterinária , Proteína C-Reativa/metabolismo , Doenças do Cão/metabolismo , Metaboloma , Obstrução das Vias Respiratórias/genética , Obstrução das Vias Respiratórias/metabolismo , Animais , Craniossinostoses/genética , Craniossinostoses/veterinária , Doenças do Cão/sangue , Doenças do Cão/genética , Cães , Feminino , Gastroenteropatias/veterinária , Inflamação/veterinária , Masculino , Estudos Prospectivos
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