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1.
Food Chem ; 399: 134004, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36037691

RESUMO

Intensive systems of raising chickens in barns prevail worldwide for financial reasons. In contrast, free-range chickens are raised in better welfare conditions, and preferred by consumers due to their distinctive taste/flavor, having higher market prices. Thus, free-range chickens have been the target of frauds. In this study, 1H NMR metabolic profiles of breasts of free-range and barn-raised broilers (108 individuals) were compared by two discriminant models, based on t-test ranking and partial least squares (PLS-DA). Both models provided 100 % of correct classification in both training and test sets, being the univariate model based on t-test screening simpler and more robust. Among other differences, barn-raised broilers presented lower carnosine and anserine concentrations, and higher free amino acids contents. Univariate discrimination was based on the ratio of two NMR signals assigned to ß-alanine and carnosine + anserine, respectively. As an additional advantage, this profiling method could be adapted to other measurement platforms.


Assuntos
Anserina , Carnosina , Animais , Anserina/análise , Carnosina/análise , Galinhas/metabolismo , Análise Discriminante , Espectroscopia de Ressonância Magnética/métodos
2.
J Comp Neurol ; 531(1): 149-169, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36222577

RESUMO

The Fragile X mental retardation protein (FMRP) is an mRNA binding protein that is essential for neural circuit assembly and synaptic plasticity. Loss of functional FMRP leads to Fragile X syndrome (FXS), a neurodevelopmental disorder characterized by sensory dysfunction including abnormal auditory processing. While the central mechanisms of FMRP regulation have been studied in the brain, whether FMRP is expressed in the auditory periphery and how it develops and functions remains unknown. In this study, we characterized the spatiotemporal distribution pattern of FMRP immunoreactivity in the inner ear of mice, rats, gerbils, and chickens. Across species, FMRP was expressed in hair cells and supporting cells, with a particularly high level in immature hair cells during the prehearing period. Interestingly, the distribution of cytoplasmic FMRP displayed an age-dependent translocation in hair cells, and this feature was conserved across species. In the auditory ganglion (AG), FMRP immunoreactivity was detected in neuronal cell bodies as well as their peripheral and central processes. Distinct from hair cells, FMRP intensity in AG neurons was high both during development and after maturation. Additionally, FMRP was evident in mature glial cells surrounding AG neurons. Together, these observations demonstrate distinct developmental trajectories across cell types in the auditory periphery. Given the importance of peripheral inputs to the maturation of auditory circuits, these findings implicate involvement of FMRP in inner ear development as well as a potential contribution of periphery FMRP to the generation of auditory dysfunction in FXS.


Assuntos
Orelha Interna , Síndrome do Cromossomo X Frágil , Animais , Camundongos , Ratos , Proteína do X Frágil de Retardo Mental/genética , Proteína do X Frágil de Retardo Mental/metabolismo , Gerbillinae , Galinhas/metabolismo , Síndrome do Cromossomo X Frágil/genética , Orelha Interna/metabolismo
3.
Food Chem ; 402: 134435, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36303381

RESUMO

This work was aimed to study the effects of the selenized yeast added in feed on selenium-containing proteins of egg yolk. Two groups of the same little hens were given the ordinary grain feed either unsupplemented selenized yeast (Group O) or supplemented with 0.15% selenized yeast (Group Y), respectively. The water-soluble Se-containing proteins were isolated and purified from the two group eggs yolk using the same conditions. SeP1-1 and SeP1-I were purified from the yolk of Group Y and Group O, respectively. Sequences identified by HPLC-MS/MS showed that SeP1-1 was a highly homologous Se-containing protein with Se-free YGP-42 with 83% match, in which Se species include methylselenocysteine and selenocysteine. SeP1-I was a highly homologous Se-containing protein with Se-free ovalbumin with 78.2% match, in which Se species include selenomethionine and selenocysteine. It can be concluded that the selenized yeast can change the compositions and structures of Se-containing proteins in egg yolk.


Assuntos
Selênio , Animais , Feminino , Selênio/química , Gema de Ovo/química , Saccharomyces cerevisiae/metabolismo , Selenocisteína/análise , Galinhas/metabolismo , Albuminas/análise , Espectrometria de Massas em Tandem , Suplementos Nutricionais/análise , Ração Animal/análise , Dieta
4.
Food Chem ; 404(Pt A): 134414, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36240553

RESUMO

By combining python script invocation, the batch processing of molecular docking was achieved to screen 20 potential umami peptides out of the 208 peptides identified in chicken soup. Nine peptides were dominated by umami taste according to sensory analysis, among which PPQEAAQF (2.56) has the highest umami intensity, followed by AEEHVEAVN (2.43) and NEFGYSNR (2.19). The threshold of nine peptides ranged from 0.08 mM to 0.58 mM. In 0.35 % MSG, the effective threshold of umami-enhancing effect of LPLQD was 0.24 mM. In addition, the molecular docking results indicated that His71, Ser107, and Asp147 of taste receptor type 1 member 1, and Asn68, Asp216, His387, and Ala302 of taste receptor type 1 member 3 play critical roles in the binding with umami peptides by forming hydrogen bonds and hydrophobic force. Thus, the combination of molecular docking and python script invocation was effective and economical for umami peptides screening.


Assuntos
Galinhas , Peptídeos , Animais , Simulação de Acoplamento Molecular , Galinhas/metabolismo , Peptídeos/química , Paladar , Aromatizantes/química
5.
Synapse ; 77(1): e22252, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36099479

RESUMO

Previous studies demonstrated that in vitro preparations of the isolated vestibular system of diverse animal species still exhibit stable resting electrical activity and mechanically evoked synaptic transmission between hair cells and primary afferent endings. However, there are no reports related to their neurodevelopment. Therefore, this research aimed to examine whether NMDA receptors mediate these electrical signals in an isolated preparation of the chicken vestibular system at three developmental stages, E15, E18, and E21. We found that the spontaneous and mechanically evoked discharges from primary afferents of the posterior semicircular canal were modulated by agonists NMDA and glycine, but not by the agonist d-serine applied near the synapses. Moreover, the individually applied by bath perfusion of three NMDA receptor antagonists (MK-801, ifenprodil, and 2-naphthoic acid) or high Mg2+ decreased the resting discharge rate, the NMDA response, and the discharge rate of mechanically evoked activity from these primary afferents. Furthermore, we found that the vestibular ganglion shows a stage-dependent increase in the expression of NMDA receptor subunits GluN1, GluN2 (A-C), and GluN3 (A-B), being greater at E21, except for GluN2D, which was inversely related to the developmental stage. However, in the crista ampullaris, the expression pattern remained constant throughout development. This could suggest the possible existence of presynaptic NMDA receptors. Our results highlight that although the NMDA receptors are functionally active at the early embryonic stages of the vestibular system, NMDA and glycine reach their mature functionality to increase NMDA responses close to hatching (E21).


Assuntos
Galinhas , Receptores de N-Metil-D-Aspartato , Animais , Receptores de N-Metil-D-Aspartato/metabolismo , Galinhas/metabolismo , N-Metilaspartato , Sistema Vestibular , Glicina
6.
Artigo em Inglês | MEDLINE | ID: mdl-36244467

RESUMO

Embryonic morphine exposure (EME) leads to abnormal brain development and behavior in the offspring, and the functional alteration of γ-aminobutyric acid (GABA) system is considered to be one of the important mechanisms. To mimic the problem of susceptibility of human gestational drug abuse on addictive drugs in offspring, we administered morphine exposure on days 5-8 and 13-16 of chicken embryo development and examined the functions of GABA neurons and their receptors in postnatal chicks by neuroelectrophysiology, immunohistochemistry and behavioral methods. We found that morphine exposure during embryonic stages 5-8 (MorphineE5-8) significantly reduced the incidence of spontaneous inhibitory postsynaptic potentiation (IPSP) and the induction of evoked IPSP and the mean amplitude of GABAA agonist muscimol-induced response in the intermediate medial interstitial (IMM) region, compared to naïve controls or saline-exposed chicks. The results of immunocytochemistry further suggest that MorphineE5-8 decreased the synaptic density of GAD-expressing sites in the IMM, while increased the expression of the GABAA receptor subtype γ2 isoform. Behavioral results found that Morphine5-8 treatment de-inhibited morphine-induced psychomotor responses in postnatal chicks. Morphine exposure at embryonic stages 13-16 (MorphineE13-16) showed no significant changes in the above indicators compared to the saline group. Evidence suggests that early embryonic morphine exposure leads to defects in GABAergic function in the IMM, which in turn alters the responsiveness of postnatal chicks to addictive drugs. These results will help to understand the GABA mechanisms by which embryonic addictive drug exposure contributes to offspring susceptibility to addiction.


Assuntos
Galinhas , Morfina , Humanos , Animais , Embrião de Galinha , Morfina/farmacologia , Galinhas/metabolismo , Ácido gama-Aminobutírico/metabolismo , Encéfalo/metabolismo , Neurônios , Receptores de GABA-A/metabolismo
7.
Artigo em Inglês | MEDLINE | ID: mdl-36257570

RESUMO

Selenium deficiency can lead to multiple tissue and organ damage in the body and could coexist with chronic toxic exposures. Contamination from Bisphenol A (BPA) exposure can induce the occurrence of various injuries including pyroptosis. However, it is not clear whether selenium deficiency and BPA exposure affect tracheal tissue pyroptosis in chickens. To investigate whether selenium deficiency and BPA exposure induce chicken tracheal tissue pyroptosis via the NF-κB/NLRP3/Caspase-1 pathway and the effect of their combined exposure on tissue injury, we developed a model of relevant chicken tracheal injury. Sixty broilers were divided into four groups: the control group (C group), selenium-deficient group (SeD group), BPA-exposed group (BPA group) and combined exposure group (SeD + BPA group). The study examined the expression indicators of markers of pyroptosis (NLRP3&GSDMD), NF-κB pathway-related inflammatory factors (NF-κB, iNOS, TNF-α, COX-2), pyroptosis-related factors (ASC, Caspase-1, IL-1ß, IL-18), and some heat shock proteins and interleukins (HSP60, HSP90, IL-6, IL-17) in the samples. The results showed that the expression of the above indicators was significantly upregulated in the different treatment groups (P < 0.05). In addition, the expression levels of the above related indicators were more significantly up-regulated in the combined selenium-deficient and BPA-exposed group compared to the group in which they were individually exposed. It was concluded that selenium deficiency and BPA exposure induced tracheal tissue pyroptosis in chickens through NF-κB/NLRP3/Caspase-1 pathway, and BPA exposure exacerbated selenium deficiency-induced tracheal pyroptosis. The present study provides new ideas into studies related to the co-exposure of organismal micronutrient deficiency and chronic toxicants.


Assuntos
Piroptose , Selênio , Animais , NF-kappa B/metabolismo , Caspase 1/metabolismo , Caspase 1/farmacologia , Galinhas/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Selênio/farmacologia , Traqueia
8.
Nutrients ; 14(21)2022 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-36364942

RESUMO

Black corn has been attracting attention to investigate its biological properties due to its anthocyanin composition, mainly cyanidin-3-glucoside. Our study evaluated the effects of black corn extract (BCE) on intestinal morphology, gene expression, and the cecal microbiome. The BCE intra-amniotic administration was evaluated by an animal model in Gallus gallus. The eggs (n = 8 per group) were divided into: (1) no injection; (2) 18 MΩ H2O; (3) 5% black corn extract (BCE); and (4) 0.38% cyanidin-3-glucoside (C3G). A total of 1 mL of each component was injected intra-amniotic on day 17 of incubation. On day 21, the animals were euthanized after hatching, and the duodenum and cecum content were collected. The cecal microbiome changes were attributed to BCE administration, increasing the population of Bifidobacterium and Clostridium, and decreasing E. coli. The BCE did not change the gene expression of intestinal inflammation and functionality. The BCE administration maintained the villi height, Paneth cell number, and goblet cell diameter (in the villi and crypt), similar to the H2O injection but smaller than the C3G. Moreover, a positive correlation was observed between Bifidobacterium, Clostridium, E. coli, and villi GC diameter. The BCE promoted positive changes in the cecum microbiome and maintained intestinal morphology and functionality.


Assuntos
Galinhas , Zea mays , Animais , Galinhas/metabolismo , Zea mays/metabolismo , Antocianinas/farmacologia , Antocianinas/metabolismo , Escherichia coli/metabolismo , Ceco/metabolismo , Bifidobacterium/metabolismo , Clostridium , Extratos Vegetais/farmacologia
9.
Poult Sci ; 101(12): 102231, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36334428

RESUMO

Fasting-refeeding perturbation has been extensively used to reveal specific genes and metabolic pathways that control energy metabolism in chickens. In this study, 200 chickens were randomly assigned to 2 groups after hatching: the control group (C, fed ad libitum) and the fasting-refeeding group (T, water ad libitum). The chicks in Group T were fasted for 72 h, and then fed for another 48 h. Liver, hypothalamus, and adipose samples were collected at 0 (F0), 24 (F24), 48 (F48), and 72 h (F72) after fasting and 4 (FR4), 12 (FR12), 24 (FR24), and 48 h (FR48) after refeeding, respectively. Results showed that Group T had a significantly higher number of liver vacuoles (P < 0.05 or P < 0.01) and a significantly lower gray value of Sudan IIIstained sections (P < 0.05 or P < 0.01) than Group C at F48-FR48. In addition, compared with the Group C, fasting and refeeding reduced the expression of stearoyl CoA desaturase (SCD) mRNA (P < 0.05 or P < 0.01) in the liver and adipose tissues, the expression of glucocorticoid receptor (GR) mRNA (P < 0.05 or P < 0.01) in the liver, adipose, and hypothalamus tissues, and the expression of fatty acid synthase (FAS) mRNA (P < 0.05 or P < 0.01) in the liver at F24-FR24. Moreover, relative to those in Group C, fasting and refeeding increased the mRNA expression levels of adenosine monophosphate-activated protein kinase (AMPK) α, AMPKß, and AMPKγ in the hypothalamus (P < 0.05 or P < 0.01) at F24-FR24. In conclusion, fasting and refeeding increased the fat content of the liver, and the expression of lipolytic genes in the hypothalamus (e.g., AMPKα, AMPKß, and AMPKγ) but decreased the expression of fat synthesis genes in the liver (e.g., SCD, GR, and FAS), adipose (SCD and GR), and hypothalamus (GR).


Assuntos
Jejum , Metabolismo dos Lipídeos , Animais , Metabolismo dos Lipídeos/genética , Galinhas/genética , Galinhas/metabolismo , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Fígado/metabolismo , RNA Mensageiro/genética
10.
Poult Sci ; 101(12): 102227, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36334429

RESUMO

Photoperiod is an important environmental factor that influence seasonal reproduction behavior in bird and GnIH can play a function in this process through the reproductive axis, and some studies suggest that GnIH may have a direct role at the gonadal level. To investigate the expression of GnIH and its effects on follicle development and steroidogenesis in quail ovaries under different photoperiods, 72 healthy laying quails of 8-wk-old were randomly divided into long day (LD) group [16 light (L): 8 dark (D)] (n = 36) and short day (SD) group (8L:16D) (n = 36). Samples were collected from each group on d1, d11, d22, and d36 of the experiment. The result showed that short day treatment upregulated the level of GnIH in the gonads (P < 0.05), decreased the expression level of CYP19A1,3ß-HSD, StAR, LHR, and FSHR and increased the expression level of AMH, AMHR2, GDF9, and BMP15 to inhibit follicle development and ovulation, thus affecting the egg production performance of quails. In vitro culture of quail granulosa cells and treatment with different concentrations of GnIH (0, 1, 10, and 100 ng/mL) for 24 h. Result showed that GnIH inhibited the levels of FSHR, LHR, and steroid synthesis pathways in granulosa cells, upregulated the levels of AMHR2, GDF9, and BMP15. The results suggest that the inhibition of follicle development and reduced egg production in quail by short day treatment is due to GnIH acting at the gonadal level, and GnIH affected the steroid synthesis by inhibiting gonadotropin receptors.


Assuntos
Hormônios Hipotalâmicos , Fotoperíodo , Feminino , Animais , Codorniz/metabolismo , Ovário/metabolismo , Hormônios Hipotalâmicos/metabolismo , Galinhas/metabolismo
11.
PLoS One ; 17(11): e0277476, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36374928

RESUMO

Histidine is an essential amino acid for broiler chickens and a precursor for the dipeptides carnosine and anserine, but little information is available about its metabolism in modern, fast-growing broilers. We used untargeted metabolomics to investigate the metabolic changes caused by the use of different standardized ileal digestible His:Lys ratios in broiler diets with and without ß-alanine supplementation. A total of 2204 broilers were randomly divided into 96 pens of 23 birds each. The pens were divided into 16 blocks, each containing one pen for all six feeding groups (total of 16 pens per group). These feeding groups were fed three different His:Lys ratios (0.44, 0.54, and 0.64, respectively) without and with a combination of 0.5% ß-alanine supplementation. Five randomly selected chickens of one single randomly selected pen per feeding group were slaughtered on day 35 or 54, blood was collected from the neck vessel, and plasma was used for untargeted metabolomic analysis. Here we show that up to 56.0% of all metabolites analyzed were altered by age, whereas only 1.8% of metabolites were affected by the His:Lys ratio in the diet, and 1.5% by ß-alanine supplementation. Two-factor analysis and metabolic pathway analysis showed no interaction between the His:Lys ratio and ß-alanine supplementation. The effect of the His:Lys ratio in the diet was limited to histidine metabolism with a greater change in formiminoglutamate concentration. Supplementation of ß-alanine showed changes in metabolites of several metabolic pathways; increased concentrations of 3-aminoisobutyrate showed the only direct relationship to ß-alanine metabolism. The supplementation of ß-alanine indicated few effects on histidine metabolism. These results suggest that the supplements used had limited effects or interactions on both His and ß-alanine metabolism. In contrast, the birds' age has the strongest influence on the metabolome.


Assuntos
Galinhas , Histidina , Animais , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , beta-Alanina/farmacologia , beta-Alanina/metabolismo , Galinhas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Histidina/metabolismo , Metaboloma , Plasma/metabolismo
12.
Molecules ; 27(21)2022 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-36364251

RESUMO

The effect of a high incorporation level of Ulva lactuca, individually and supplemented with a Carbohydrate-Active enZyme (CAZyme) on broilers' plasma parameters and liver composition is assessed here. Twenty one-day-old Ross 308 male broilers were randomly assigned to one of four treatments (n = 10): corn/soybean meal based-diet (Control); based-diet with 15% U. lactuca (UL); UL diet with 0.005% of commercial carbohydrase mixture; and UL diet with 0.01% of recombinant ulvan lyase. Supplementing U. lactuca with the recombinant CAZyme slightly compromised broilers' growth by negatively affecting final body weight and average daily gain. The combination of U. lactuca with ulvan lyase also increased systemic lipemia through an increase in total lipids, triacylglycerols and VLDL-cholesterol (p < 0.001). Moreover, U. lactuca, regardless of the CAZyme supplementation, enhanced hepatic n-3 PUFA (mostly 20:5n-3) with positive decrease in n-6/n-3 ratio. However, broilers fed with U. lactuca with ulvan lyase reduced hepatic α- and γ-tocopherol concentrations relative to the control. Conversely, the high amount of pigments in macroalga diets led to an increase in hepatic ß-carotene, chlorophylls and total carotenoids. Furthermore, U. lactuca, alone or combined with CAZymes, enhanced hepatic total microminerals, including iron and manganese. Overall, plasma metabolites and liver composition changed favorably in broilers that were fed 15% of U. lactuca, regardless of enzyme supplementation.


Assuntos
Ulva , Animais , Masculino , Ração Animal/análise , Galinhas/metabolismo , Dieta , Suplementos Nutricionais , Fígado
13.
Genes (Basel) ; 13(11)2022 10 29.
Artigo em Inglês | MEDLINE | ID: mdl-36360215

RESUMO

Circular RNAs (circRNAs) play a significant regulatory role during skeletal muscle development. To identify circRNAs during postnatal skeletal muscle development in chickens, we constructed 12 cDNA libraries from breast muscle tissues of Chinese Gushi chickens at 6, 14, 22, and 30 weeks and performed RNA sequencing. In total, 2112 circRNAs were identified, and among them 79.92% were derived from exons. CircRNAs are distributed on all chromosomes of chickens, especially chromosomes 1-9 and Z. Bioinformatics analysis showed that each circRNA had an average of 38 miRNA binding sites, 61.32% of which have internal ribosomal entry site (IRES) elements. Furthermore, in total 543 differentially expressed circRNAs (DE-circRNAs) were identified. Functional enrichment analysis revealed that DE-circRNAs source genes are engaged in biological processes and muscle development-related pathways; for example, cell differentiation, sarcomere, and myofibril formation, mTOR signaling pathway, and TGF-ß signaling pathway, etc. We also established a competitive endogenous RNA (ceRNA) regulatory network associated with skeletal muscle development. The results in this report indicate that circRNAs can mediate the development of chicken skeletal muscle by means of a complex ceRNA network among circRNAs, miRNAs, genes, and pathways. The findings of this study might help increase the number of known circRNAs in skeletal muscle tissue and offer a worthwhile resource to further investigate the function of circRNAs in chicken skeletal muscle development.


Assuntos
MicroRNAs , RNA Circular , Animais , RNA Circular/genética , Galinhas/genética , Galinhas/metabolismo , Desenvolvimento Muscular/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Músculo Esquelético/metabolismo
14.
Trop Anim Health Prod ; 54(6): 399, 2022 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-36422722

RESUMO

A 42-day study was conducted to consider the effect of composite enzyme (Natuzyme®) supplementation on production parameters, intestinal segment measurements, and nutrient digestibility of broiler chickens fed low energy and protein (LEP) diets. Two hundred male Ross 308 broiler chicks consisting of 4 groups were divided into five pens (10 chicks/pen). The first group (control) received a standard diet without composite enzyme (CE) supplementation, whereas the LEP0, LEP0.25, and LEP0.50 groups received LEP supplemented with CE at 0, 0.25, and 0.50 g/kg feed, respectively. Compared to birds in the control group, supplementation of 0.50 g/kg CE to LEP diet offset (P < 0.05) the poor feed intake (FI), body weight gain (BWG), feed conversion ratio (FCR), and cost/kg gain observed among LEP0 and LEP0.25 groups on days 0-21, 22-42, and 0-42. Birds in the LEP0, followed by the LEP0.25 group, had decreased (P < 0.05) ileum weight, villi height, villus height, and crypt depth ratio (VH/CD), and deeper crypt depth of duodenum and ileum compared to birds in the control group. However, the aforementioned parameters were improved in the LEP0.50 group. Likewise, supplementation of CE at 0.50 g/kg to broiler diet having LEP improved (P < 0.05) digestibility of energy, fiber, and protein on day 42. In conclusion, CE supplemented at 0.50 g/kg diet ameliorated the negative effect caused by feeding LEP diets on production parameters, ileum weight, villi height, VH/CD, and deeper crypt depth of duodenum and ileum as well as energy, fiber, and protein digestibility of broiler chickens.


Assuntos
Ração Animal , Galinhas , Animais , Masculino , Galinhas/metabolismo , Ração Animal/análise , Suplementos Nutricionais , Dieta com Restrição de Proteínas/veterinária , Nutrientes
15.
Discov Med ; 34(172): 103-113, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36333112

RESUMO

Proliferative vitreoretinopathy (PVR) is an intractable condition after rhegmatogenous retinal detachment (RD), which is the primary cause of failure in retinal reattachment surgery. This study aimed to investigate the effects of chicken ovalbumin upstream promoter transcriptional factor 1 (COUP-TF1) in the development of proliferative vitreoretinopathy (PVR) both in vitro and in vivo. Adult retinal pigment epithelium cell line was used for in-vitro experiments. Immunocytochemistry assay, real-time quantitative polymerase chain reaction, and Western blot were used to measure the expression of COUP-TF1, alpha-smooth muscle actin (α-SMA), and E-cadherin. Epithelial-mesenchymal transition (EMT) was observed through cell counting kit-8 assay, wound healing tests, and the expression changes of related proteins. PVR rabbit models were established and evaluated by the images of fundus and vitreous cavity, pathological sections, and COUP-TF1 expression. As shown by our results, the proliferation and migration of the COUP-TF1 knockdown cells were reduced compared with the control cells with or without transforming growth factor-ß1 (TGF-ß1) treatment. After TGF-ß1 treatment, α-SMA expression was upregulated in ARPE-19 cells but kept the same in COUP-TF1 knockdown cells. E-cadherin expression was down-regulated in all the groups but the extent of the decrease in COUP-TF1 knockdown cells was smaller. EMT was attenuated in ARPE-19 cells after COUP-TF1 was knocked down. In the in-vivo experiment, PVR severity was attenuated and the retinal detachment rate decreased on the 14th and 28th day in COUP-TF1 knockdown group. In conclusion, COUP-TF1 is related to the development of PVR, and COUP-TF1 knockdown attenuates the progression of PVR. This suggests that COUP-TF1 can be a promising candidate for the treatment of PVR.


Assuntos
Descolamento Retiniano , Vitreorretinopatia Proliferativa , Animais , Coelhos , Vitreorretinopatia Proliferativa/genética , Vitreorretinopatia Proliferativa/metabolismo , Vitreorretinopatia Proliferativa/patologia , Transição Epitelial-Mesenquimal/genética , Fator de Crescimento Transformador beta1/metabolismo , Galinhas/metabolismo , Ovalbumina/metabolismo , Ovalbumina/farmacologia , Descolamento Retiniano/metabolismo , Descolamento Retiniano/patologia , Epitélio Pigmentado da Retina/metabolismo , Movimento Celular/genética , Células Cultivadas , Caderinas/genética , Caderinas/metabolismo
16.
Sci Rep ; 12(1): 18791, 2022 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-36335156

RESUMO

A five weeks biological experiment was planned to investigate the impacts of dietary supplementation with zinc oxide nanoparticles (ZnONPs) synthesized by the endophytic fungus Alternaria tenuissima on productive performance, carcass traits, organ relative weights, serum biochemical parameters, histological alteration in some internal organs and concentration of this element in the serum, liver, thigh and breast muscle in broiler chicks. A total of 108 3-day-old commercial broiler chicks (Cobb 500) were individually weighed and equally distributed in a completely randomized design arrangement according to the dose of ZnONPs supplementation into 3 dietary experimental groups. There were 6 replications having 6 birds per replicate (n = 36/ treatment) for each treatment. The three experiential dietary treatments received corn-soybean meal-based diets enhanced with 0 (control), 40 and 60 mg/kg diet of ZnONPs respectively with feed and water were provided ad libitum consumption through 5 weeks life span. Present results indicated that after 5 weeks of feeding trial and as compared to control, the ZnONPs supplementation groups recorded higher body weight, improved feed consumption, feed conversion ratio and performance index. Serum biochemical analyses revealed that serum cholesterol, triglyceride, low density lipoprotein and uric acid decreased significantly, while high density lipoprotein and liver enzyme concentrations were increased significantly. Meanwhile, zinc accumulation in serum, liver and breast and thigh muscle were linearly increased with increasing zinc supplementation. It could be concluded that supplementation of ZnONPs to broiler diet at 40 or 60 mg/kg improved productive performance, birds' physiological status and the lower levels Zn (40 mg/kg diet) revealed promising results and can be used as an effective feed additive in broilers.


Assuntos
Nanopartículas , Óxido de Zinco , Animais , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Galinhas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Zinco/farmacologia , Zinco/metabolismo , Óxido de Zinco/farmacologia , Óxido de Zinco/metabolismo
17.
Molecules ; 27(21)2022 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-36364267

RESUMO

Skin hyperpigmentation is an aesthetic problem that leads to psychosocial issues. Thus, skin whitening agents from agro- and poultry-industrial co-products are considered high economic value ingredients of interest for sustainable application. Therefore, this study aimed to determine the cosmeceutical potential of anserine/carnosine-rich chicken extract (ACCE) from the Thai native chicken Pradu Hang Dam Mor Kor 55 (PD) meat. The chemical composition was identified and quantified using the HPLC-UV method. Then, the antioxidation potential of the extract was compared to that of L-anserine and L-carnosine, using 1,1-diphenyl-2-picrylhydrazyl assay and shikonin-induced production of reactive oxygen species in CCD-986Sk cell models, and the anti-melanogenesis effect in the MNT-1 melanoma cell line model was investigated. Furthermore, related mechanisms were identified using colorimetric tyrosinase assay and the Western blot technique. The ACCE was composed of L-anserine and L-carnosine as two major constituents. In a dose-dependent manner, ACCE, L-anserine, and L-carnosine manifested significant antioxidation potential and significant reduction of melanin production. Activation of the extracellular signal-regulated kinase (ERK) signaling pathway and inhibition of tyrosinase activity of ACCE were demonstrated as the mechanisms of the anti-melanogenesis effect. In conclusion, ACCE has been revealed as a potential cosmeceutical agent due to its antioxidation and anti-melanogenic activity in association with L-anserine and L-carnosine composition and biomolecular regulating ability. Therefore, further studies and development should be considered to support the utilization of anserine/carnosine-rich chicken extract in the cosmetic industry for economic value creation and sustainability.


Assuntos
Carnosina , Cosmecêuticos , Animais , Anserina/química , Carnosina/química , Galinhas/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Tailândia , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Transdução de Sinais
18.
J Phys Chem B ; 126(44): 9000-9007, 2022 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-36318974

RESUMO

Protein crystals composed of protein molecules are expected as a novel porous material. They have high porosity, and the knowledge of the diffusion of intracrystalline water is important. In this study, the diffusion coefficient of intracrystalline water in intrinsic hen egg-white lysozyme (HEWL) crystals was determined by a method that combines confocal Raman spectroscopy and air convection with controlled relative humidity. Similar to common porous materials, the drying process of the protein crystals includes three periods: constant-rate drying, falling-rate drying, and equilibrium state. During the falling-rate drying period, the drying rate depends on the diffusion of intracrystalline water in the protein crystal. The gradient of the water content was measured using confocal Raman spectroscopy. The diffusion coefficient of the intrinsic HEWL crystals was determined as 3.1 × 10-7 cm2/s with a water content of 36.3 vol %. The estimated diffusion coefficients of the intrinsic HEWL crystals without cross-linking were in close agreement with those of the cross-linked protein crystals. This study is timely as the knowledge of the intrinsic diffusion coefficient is useful not only for understanding the mechanism of hydration of proteins but also in practical applications such as porous materials, drug binding, and cryoprotectant soaks.


Assuntos
Muramidase , Água , Animais , Muramidase/química , Água/química , Cristalização , Análise Espectral Raman , Galinhas/metabolismo
19.
J Agric Food Chem ; 70(47): 14959-14973, 2022 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-36383077

RESUMO

Peroxisome proliferator-activated receptor γ (PPARγ) is the master regulator of adipogenesis and is expressed as two isoforms, PPARγ1 and PPARγ2. Our previous lentiviral overexpression study showed that PPARγ1 and PPARγ2 differentially regulated proliferation, differentiation, and apoptosis of the immortalized chicken preadipocyte cell line (ICP2). However, we cannot rule out the possibility that the endogenous expression of PPARγ isoforms may compromise our findings. In this study, using the dual sgRNA-directed CRISPR/Cas9 system, we generated PPARγ (PPARγ-/-) and PPARγ2-specific knockout (PPARγ2-/-) ICP2 cell lines and investigated the differences in proliferation and differentiation among PPARγ-/-, PPARγ2-/-, and wild-type ICP2 cells. EdU proliferation assay showed that both PPARγ2-specific and PPARγ knockouts significantly increased the proliferation rates. Consistently, real-time RT-PCR analysis showed that both PPARγ2-specific and PPARγ knockouts significantly upregulated the expression of proliferation marker genes PCNA and cyclinD1. FACS analysis revealed that PPARγ knockout significantly increased the number of cells accumulating in the S phase and decreased the number of cells accumulating in the G1/G0 phase. Oil Red O staining and gene expression analysis showed both PPARγ2-specific and PPARγ knockouts dramatically reduced capacity for adipogenic differentiation. To corroborate our previous findings, PPARγ1 and PPARγ2 expression were restored in PPARγ-/- cells by using the lentiviruses expressing chicken PPARγ1 (LV-PPARγ1) and PPARγ2 (LV-PPARγ2), respectively. Subsequent assays showed that restoration of expression of either PPARγ1 or PPARγ2 suppressed proliferation and stimulated differentiation of the PPARγ-/- cells. By comparison, PPARγ2 had stronger anti-proliferative and pro-adipogenic effects than PPARγ1. To understand the molecular mechanism underlying their differential effects on differentiation of the PPARγ-/- cells, we performed RNA-seq in the PPARγ-/- cells in which individual PPARγ isoform expression was restored at 72 h of differentiation. Transcriptomic analysis revealed that restoring PPARγ1 expression caused far more differentially expressed genes (DEGs) than restoring PPARγ2 expression. GO and KEGG pathway enrichment analyses indicated that PPARγ1 and PPARγ2 had distinct and overlapping functions in adipogenesis. Taken together, our results clearly indicate that PPARγ1 and PPARγ2 differentially impact chicken adipogenesis.


Assuntos
Adipogenia , PPAR gama , Animais , PPAR gama/genética , Adipogenia/genética , Fatores de Transcrição/genética , Galinhas/genética , Galinhas/metabolismo , Células Cultivadas , Isoformas de Proteínas/genética
20.
Int J Mol Sci ; 23(22)2022 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-36430616

RESUMO

Blood is a meat by-product rich in proteins with properties that can be improved after hydrolysis, making it a sustainable alternative for use in the generation of bioactive peptides. The objective of this study was to identify dipeptidyl peptidase IV (DPP-IV) inhibitory peptides obtained from different chicken blood hydrolysates prepared using combinations of four different enzymes. Best results were observed for AP (2% Alcalase + 5% Protana Prime) and APP (2% Alcalase + 5% Protana Prime + 3% Protana UBoost) hydrolysates obtaining inhibition values of 60.55 and 53.61%, respectively, assayed at a concentration of 10 mg/mL. Free amino acids were determined to establish the impact of exopeptidase activity in the samples. A total of 79 and 12 sequences of peptides were identified by liquid chromatography and mass spectrometry in tandem (LC-MS/MS) in AP and APP samples, respectively. Nine of the identified peptides were established as potential DPP-IV inhibitory using in silico approaches and later synthesized for confirmation. Thus, peptides GPF, IGL, and GGGW showed good DPP-IV inhibitory activity with IC50 values of 0.94, 2.22, and 2.73 mM, respectively. This study confirmed the potential of peptides obtained from chicken blood hydrolysates to be used as DPP-IV inhibitors and, therefore, in the control or modulation of type 2 diabetes.


Assuntos
Diabetes Mellitus Tipo 2 , Dipeptidil Peptidase 4 , Animais , Dipeptidil Peptidase 4/metabolismo , Galinhas/metabolismo , Cromatografia Líquida , Espectrometria de Massas em Tandem , Peptídeos/farmacologia , Peptídeos/química , Subtilisinas
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