RESUMO
Several motor, sensory, cognitive, and behavioral dysfunctions are associated with neural lesions occurring after a hypoxic injury (HI) in preterm infants. Growth hormone (GH) expression is upregulated in several brain areas when exposed to HI conditions, suggesting actions as a local neurotrophic factor. It is known that GH, either exogenous and/or locally expressed, exerts neuroprotective and regenerative actions in cerebellar neurons in response to HI. However, it is still controversial whether GH can cross the blood-brain barrier (BBB), and if its effects are exerted directly or if they are mediated by other neurotrophic factors. Here, we found that in ovo microinjection of Cy3-labeled chicken GH resulted in a wide distribution of fluorescence within several brain areas in the chicken embryo (choroid plexus, cortex, hypothalamus, periventricular areas, hippocampus, and cerebellum) in both normoxic and hypoxic conditions. In the cerebellum, Cy3-GH and GH receptor (GHR) co-localized in the granular and Purkinje layers and in deep cerebellar nuclei under hypoxic conditions, suggesting direct actions. Histological analysis showed that hypoxia provoked a significant modification in the size and organization of cerebellar layers; however, GH administration restored the width of external granular layer (EGL) and molecular layer (ML) and improved the Purkinje and granular neurons survival. Additionally, GH treatment provoked a significant reduction in apoptosis and lipoperoxidation; decreased the mRNA expression of the inflammatory mediators (TNFα, IL-6, IL-1ß, and iNOS); and upregulated the expression of several neurotrophic factors (IGF-1, VEGF, and BDNF). Interestingly, we also found an upregulation of cerebellar GH and GHR mRNA expression, which suggests the existence of an endogenous protective mechanism in response to hypoxia. Overall, the results demonstrate that, in the chicken embryo exposed to hypoxia, GH crosses the BBB and reaches the cerebellum, where it exerts antiapoptotic, antioxidative, anti-inflammatory, neuroprotective, and neuroregenerative actions.
Assuntos
Proteínas Aviárias/metabolismo , Hormônio do Crescimento/metabolismo , Fármacos Neuroprotetores , Animais , Barreira Hematoencefálica/metabolismo , Fator Neurotrófico Derivado do Encéfalo/genética , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Cerebelo/metabolismo , Embrião de Galinha , Galinhas/metabolismo , Humanos , Hipóxia/metabolismo , Recém-Nascido , Recém-Nascido Prematuro , Mediadores da Inflamação/metabolismo , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Interleucina-6/metabolismo , Fármacos Neuroprotetores/metabolismo , Fármacos Neuroprotetores/farmacologia , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
We analyzed a red paint sample from the surface of a gold mask excavated from a Middle Sicán elite tomb in Peru. The mask covered the face of the principal male and dates from ca. 1000 AD, a period when many painted precious metal objects were produced. The paint's inorganic pigment was identified more than 30 years ago as cinnabar (a mercuric sulfide scarlet-red to brown-red mineral), but the identity of the effective organic binder remained a mystery. Fourier transform infrared (FTIR) analysis of the sample indicated a proteinaceous composition, and no lipids were recovered from an N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) derivatized extract of the sample analyzed by gas chromatography-mass spectrometry (GC-MS). Proteomics analysis by nanoLC-MS/MS identified unique peptides in the sample, which were matched to human blood and bird egg proteins via Uniprot database searches. These included immunoglobulin heavy chain, immunoglobulin G, serum albumin, and ovomucoid. Cinnabar-based paints were typically used in the context of social elites and ritually important items. The presence of human blood would support previous ideas that red cinnabar paint may represent "life force" intended to support "rebirth". As the red paint sample came from the first scientifically excavated Sicán gold mask, the results suggest a method to authenticate similar unprovenanced masks now in private and museum collections. Proteomics data set identifier https://doi.org/10.5287/bodleian:1ajYbBgQP.
Assuntos
Proteínas Aviárias , Animais , Aves , Proteínas do Ovo , Ouro , Humanos , Masculino , Pintura/análise , Peru , Espectrometria de Massas em TandemRESUMO
A strategy to mitigate the negative effects of stress on animals is to enhance their ability to beneficially respond to stressful conditions. This study aimed to assess whether prenatal ambient temperature influences the response of Japanese quail (Coturnix coturnix japonica) chicks to environmental challenges during growth. The experiment was conducted in a 2 × 2 factorial arrangement: two temperature conditions for the mothers (thermoneutral and heat stress by continuous exposure to 32 °C) and two offspring ambient temperature conditions (thermoneutral and heat stress by intermittent exposure to 34 °C for 6 h/day from 15 to 35 days of age). Heat stress in mothers led to lower laying rate, egg mass, expression of methionine sulfoxide reductase A (MSRA) gene, and antioxidant capacity as well as higher chick mortality rate (1-15 days of age). Maternal heat stress led to lower weight gain and total antioxidant capacity and higher feed conversion ratio. Maternal temperature × Offspring temperature interaction effects were observed on carbonylated protein content and HSP70, GSS, and MSRA gene expression. It was observed that, for chicks hatched from heat-stressed mothers, exposure to heat stress led to higher carbonylated protein content and HSP70 expression than exposure to thermoneutral conditions. Maternal heat stress was also responsible for increasing GSS expression in chicks grown under thermoneutral conditions. Chicks hatched from non-stressed mothers and subjected to heat stress had higher MSRA expression compared to chicks maintained in a thermoneutral environment. Our results show that, although maternal heat stress had no negative effects on performance or oxidative metabolism of offspring grown under thermoneutral conditions, it was associated with lower performance and higher protein oxidation in offspring exposed to heat stress during growth. These results could be due in part to alterations in the expression of genes related to antioxidant capacity.
Assuntos
Proteínas Aviárias/biossíntese , Coturnix/crescimento & desenvolvimento , Proteínas do Ovo/biossíntese , Resposta ao Choque Térmico , Temperatura Alta , Óvulo/metabolismo , Estresse Oxidativo , Animais , FemininoRESUMO
A novel assay technique that involves quantification of lysozyme (Lys) through machine learning is put forward here. This article reports the tendency of the well- documented Ellington group anti-Lys aptamer, to produce aggregates when exposed to Lys. This property of apta-aggregation has been exploited here to develop an assay that quantifies the Lys using texture and area parameters from a photograph of the elliptical aggregate mass through machine learning. Two assay sets were made for the experimental procedure: one with high Lys concentration between 25-100 mM and another with low concentration between 1-20 mM. The high concentration set had a sample volume of 10 µl while the low concentration set had a higher sample volume of 100 µl, in order to obtain the statistical texture values reliably from the aggregate mass. The platform exhibited an experimental limit of detection of 1 mM and a response time of less than 10 seconds. Further, two potential operating modes for the aptamer were hypothesized for this aggregation property and the more accurate mode among the two was ascertained through bioinformatics studies.
Assuntos
Aptâmeros de Nucleotídeos/química , Proteínas Aviárias/análise , Aprendizado de Máquina , Muramidase/análise , Agregados Proteicos , Animais , GalinhasRESUMO
This study aimed to provide the performance, localization and expression of the epithelial calcium transporter channels Calbindin-D28k (Calb) and TRPV6, and of the morphology of the digestive and reproductive system of laying quail under heat stress (HS), and with methionine supplementation (MS). This study characterized the positivity (immunohistochemistry) and expression (real-time PCR) of calcium channels in the kidneys, intestine and uterus of 504 laying quails under different MS (100, 110 and 120%) and temperatures (20, 24, 28 and 32°C). The animals under HS (32°C) had lower villus height, villus:crypt ratio, and goblet cell index in the duodenum and jejunum, fewer secondary and tertiary uterine folds, smaller hepatic steatosis, and increased number of distal convoluted renal tubules (CT) positive to Calb, and increased positivity in proximal CTs. Deleterious effects of HS were minimized with MS for: duodenal crypts, number of goblet cells of the jejunum, number of uterine folds, decreased Calb positivity in intestines and kidney, increased positivity of Calb in the uterus and increased TRPV6 gene expression in the kidney (P≤0.05). Epithelial calcium transporters were altered due to less need for calcium absorption and reabsorption due to more calcium available with the MS, increasing egg production in HS and quality in termoneutrality (P≤0.05). MS further increased intestinal villus absorption area and height, increased steatosis, decreased Calb positivity in the intestine and kidney, increased uterine positivity of Calb, and increase Calb and TRPV6 expression in the kidney (P≤0.001) under thermoneutrality. It was concluded that the use of MS (120%) is justifiable in order to partially reverse the deleterious effects of HS on the production, in the epithelial calcium carriers, and in the digestory and reproductive morphology of laying quail.
Assuntos
Proteínas Aviárias/biossíntese , Calbindinas/biossíntese , Duodeno , Regulação da Expressão Gênica/efeitos dos fármacos , Resposta ao Choque Térmico/efeitos dos fármacos , Fígado , Metionina/farmacologia , Codorniz , Canais de Cátion TRPV/biossíntese , Útero , Animais , Duodeno/anatomia & histologia , Duodeno/metabolismo , Feminino , Fígado/anatomia & histologia , Fígado/metabolismo , Codorniz/anatomia & histologia , Codorniz/metabolismo , Útero/anatomia & histologia , Útero/metabolismoRESUMO
BACKGROUND: This study aimed to investigate the molecular mechanisms involved in the onset of the white striping (WS) myopathy with particular attention to the role of oxidative stress and protein oxidation in the loss of meat quality. RESULTS: It was found that WS-M (moderate degree; white stripes <1 mm thickness) and WS-S (severe degree; white stripes >1 mm thickness) breast presented higher pH, hardness, redness, lipid, and collagen content, and lower lightness than normal breast. Compared with the latter, WS-S had a more severe loss of protein thiols (70.7% less thiols than in N), reduced activity of antioxidant enzymes such as catalase (23 versus 40 U g-1 ), glutathione peroxidase (0.21 versus 0.54 U g-1 ), and superoxide dismutase (56 versus 73 U g-1 ), and consequently, had greater accretion of thiobarbituric acid reactive substances (0.64 versus 0.22 mg MDAkg-1 muscle), allysine (3.1 versus 1.9 nmol mg-1 protein) and Schiff base structures (645 versus 258 fluorescent units). The analysis of sarcoplasmic proteins revealed that muscles severely affected by the myopathy suffered a chronic impairment of physiological (upregulation of sarcoplasmic reticulum Ca2+ ATPase, sarcalumenin and calsequestrin-2) and metabolic processes (downregulation of pyruvate kinase, creatine kinase, and l-lactate dehydrogenase). CONCLUSION: The overexpression of ribonuclease / angiogenin inhibitor 1 and Kelch-like proteins in WS chicken breasts indicates altered protein turnover plausibly mediated by oxidative stress and accumulation of oxidized proteins. © 2020 Society of Chemical Industry.
Assuntos
Antioxidantes/metabolismo , Proteínas Aviárias/metabolismo , Doenças Musculares/veterinária , Estresse Oxidativo , Doenças das Aves Domésticas/metabolismo , Animais , Galinhas , Carne/análise , Músculo Esquelético/metabolismo , Doenças Musculares/metabolismo , Oxirredução , ProteostaseRESUMO
Lysosomes and acidic compartments are involved in breaking down of macromolecules, membrane recycling, and regulation of signaling pathways. Here, we analyzed the role of acidic compartments during muscle differentiation and the involvement of the Wnt/beta-catenin pathway in lysosomal function during myogenesis. Acridine orange was used to localize and quantify acidic cellular compartments in primary cultures of embryonic muscle cells from Gallus gallus. Our results show an increase in acidic compartment size and area, as well as changes in their positioning during the initial steps of myogenesis. The inhibition of lysosomal function by either the chloroquine Lys05 or the downregulation of LAMP-2 with siRNA impaired chick myogenesis, by inhibiting myoblast fusion. Two activators of the Wnt/beta-catenin pathway, BIO and Wnt3a, were able to rescue the inhibitory effects of Lys05 in myogenesis. These results suggest a new role for the Wnt/beta-catenin pathway in the regulation of acidic compartment size, positioning, and function in muscle cells.
Assuntos
Proteínas Aviárias/metabolismo , Desenvolvimento Muscular , Mioblastos Esqueléticos/metabolismo , Via de Sinalização Wnt , beta Catenina/metabolismo , Animais , Embrião de GalinhaRESUMO
Carotenoids are pigmented compounds acquired through diet that have important functions as antioxidants and immune modulators. We studied the association between immunity and circulating carotenoids in Andean condors (Vultur gryphus). We evaluated the relationship between α-, ß-, and γ-globulin blood concentrations and different circulating carotenoids in two groups of Andean condors that differ in their mean health status, rehabilitating (suffering different pathologies), and wild individuals (trapped when displaying their physiological behavior). In rehabilitating individuals, α-, ß-, and γ-globulin concentrations were higher than in wild individuals. This shows that rehabilitating individuals were developing an immune response associated with the pathologies that they were suffering at the time of sampling. In addition, circulating carotenoids were lower in rehabilitating than in wild individuals. We found negative correlations between α-, ß-, and γ-globulins and different circulating carotenoids in rehabilitating individuals, but not in wild condors. Xanthophylls were strongly related to α-, ß-, and γ-globulin blood concentrations in rehabilitating, but not in wild condors. Our results suggest that there is a potential relationship between circulating carotenoids and immunity in the Andean condor. Given that this species may display a carotenoid-based pigmentation, our results could suggest that a trade-off between the immune system and external coloration could operate in this species, which may have implications in their access to food resources and mate selection and, thus, in their conservation.
Assuntos
Carotenoides/sangue , Imunidade/fisiologia , Aves Predatórias , Animais , Animais Selvagens/imunologia , Animais Selvagens/metabolismo , Proteínas Aviárias/metabolismo , Aves/fisiologia , Conservação dos Recursos Naturais , Globulinas/metabolismo , Pigmentação/fisiologia , Plasma/metabolismo , Aves Predatórias/imunologia , Aves Predatórias/metabolismoRESUMO
Reproductive success requires that individuals acquire sufficient energy resources. Restricting food availability or increasing energy expenditure (e.g., thermoregulation) inhibits reproductive development in multiple avian species, but the nature of the energy-related signal mediating this effect is unclear. To investigate this question, we examined reproductive and metabolic physiology in male house finches that either underwent moderate food restriction (FR) or were exposed to high temperature (HT), in which birds were held at a high, but not locally atypical, ambient temperature cycle (37.8 °C day, 29.4 °C night) compared to a control group (CT; 29.4 °C day, 21.1 °C night). We hypothesized that FR and HT inhibit reproductive development by lowering available metabolic fuel, in particular plasma glucose (GLU) and free fatty acids (FFA). Following FR for 4 weeks, finches lost body mass, had marginally higher plasma FFA, and experienced a 90% reduction in testis mass compared to CT birds. Four weeks of HT exposure resulted in reduced voluntary food consumption and muscle mass, as well as an 80% reduction in testis mass relative to CT birds. Both FR and HT birds expressed less testicular 17ß-hydroxysteroid dehydrogenase (17ß-HSD) mRNA than controls but the expression of other testicular genes measured was unaffected by either treatment. Neither treatment significantly influenced plasma GLU. This study is among the first to demonstrate a negative effect of HT on reproductive development in a wild bird. Further studies are needed to clarify the role of metabolic mediators and their involvement under various conditions of energy availability and demand.
Assuntos
Ingestão de Alimentos , Metabolismo Energético , Tentilhões/fisiologia , Temperatura Alta , Testículo/metabolismo , 17-Hidroxiesteroide Desidrogenases/genética , Animais , Proteínas Aviárias/genética , Glicemia/análise , Peso Corporal , Exposição Ambiental , Ácidos Graxos não Esterificados/metabolismo , Masculino , Músculo Esquelético/anatomia & histologia , Testosterona/sangueRESUMO
Imbalance in nutrients can affect digestibility of amino acids by altering gene expression of amino acid transporters. We investigated digestibility and molecular transporters of essential amino acids in chickens fed a methionine-deficient diet. A total of 40 chicks (23 D old) were randomly assigned to either a control (0.49% methionine) or a deficient (0.28%) diet until 41 D when they were sampled for Pectoralis (P.) major, kidney, ileum, and hypothalamus for mRNA expression analysis. The ileal content was collected for apparent ileal digestibility (AID) analysis. Birds fed the deficient diet had reduced growth and worse feed efficiency compared to control. The AID of methionine was similar between both groups. The AID of other essential amino acids was higher in the deficient group than control. mRNA expression of b0,+ AT and LAT4 were upregulated in the ileum and kidney but LAT1 was downregulated only in kidney of the deficient group compared to control. In the P. major, SNAT1, SNAT2, and CAT1 were upregulated in the deficient group compared to control. A diet deficiency in methionine affects digestibility of essential amino acids and cysteine, but not the digestibility of methionine. The change in digestibility is reflected in the mRNA expression of amino acid transporters across different tissues.