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1.
Cell Mol Life Sci ; 77(8): 1551-1569, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31642939

RESUMO

Skeletal muscle plays essential roles in motor function, energy, and glucose metabolism. Skeletal muscle formation occurs through a process called myogenesis, in which a crucial step is the fusion of mononucleated myoblasts to form multinucleated myofibers. The myoblast/myocyte fusion is triggered and coordinated in a muscle-specific way that is essential for muscle development and post-natal muscle regeneration. Many molecules and proteins have been found and demonstrated to have the capacity to regulate the fusion of myoblast/myocytes. Interestingly, two newly discovered muscle-specific membrane proteins, Myomaker and Myomixer (also called Myomerger and Minion), have been identified as fusogenic regulators in vertebrates. Both Myomaker and Myomixer-Myomerger-Minion have the capacity to directly control the myogenic fusion process. Here, we review and discuss the latest studies related to these two proteins, including the discovery, structure, expression pattern, functions, and regulation of Myomaker and Myomixer-Myomerger-Minion. We also emphasize and discuss the interaction between Myomaker and Myomixer-Myomerger-Minion, as well as their cooperative regulatory roles in cell-cell fusion. Moreover, we highlight the areas for exploration of Myomaker and Myomixer-Myomerger-Minion in future studies and consider their potential application to control cell fusion for cell-therapy purposes.


Assuntos
Proteínas de Membrana/metabolismo , Desenvolvimento Muscular , Proteínas Musculares/metabolismo , Músculo Esquelético/fisiologia , Regeneração , Sequência de Aminoácidos , Animais , Fusão Celular , Regulação da Expressão Gênica , Humanos , Proteínas de Membrana/análise , Proteínas de Membrana/genética , Proteínas Musculares/análise , Proteínas Musculares/genética , Mioblastos Esqueléticos/fisiologia , Alinhamento de Sequência
2.
Metabolism ; 102: 153996, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31678069

RESUMO

BACKGROUND: We have recently shown that a novel signalling kinase, inositol hexakisphosphate kinase 1 (IP6K1), is implicated in whole-body insulin resistance via its inhibitory action on Akt. Insulin and insulin like growth factor 1 (IGF-1) share many intracellular processes with both known to play a key role in glucose and protein metabolism in skeletal muscle. AIMS: We aimed to compare IGF/IP6K1/Akt signalling and the plasma proteomic signature in individuals with a range of BMIs after ingestion of lean meat. METHODS: Ten lean [Body mass index (BMI) (in kg/m2): 22.7 ±â€¯0.4; Homeostatic model assessment of insulin resistance (HOMAIR): 1.36 ±â€¯0.17], 10 overweight (BMI: 27.1 ±â€¯0.5; HOMAIR: 1.25 ±â€¯0.11), and 10 obese (BMI: 35.9 ±â€¯1.3; HOMAIR: 5.82 ±â€¯0.81) adults received primed continuous L-[ring-13C6]phenylalanine infusions. Blood and muscle biopsy samples were collected at 0 min (post-absorptive), 120 min and 300 min relative to the ingestion of 170 g pork loin (36 g protein and 5 g fat) to examine skeletal muscle protein signalling, plasma proteomic signatures, and whole-body phenylalanine disappearance rates (Rd). RESULTS: Phenylalanine Rd was not different in obese compared to lean individuals at all time points and was not responsive to a pork ingestion (basal, P = 0.056; 120 & 300 min, P > 0.05). IP6K1 was elevated in obese individuals at 120 min post-prandial vs basal (P < 0.05). There were no acute differences plasma proteomic profiles between groups in the post-prandial state (P > 0.05). CONCLUSIONS: These data demonstrate, for the first time that muscle IP6K1 protein content is elevated after lean meat ingestion in obese adults, suggesting that IP6K1 may be contributing to the dysregulation of nutrient uptake in skeletal muscle. In addition, proteomic analysis showed no differences in proteomic signatures between obese, overweight or lean individuals.


Assuntos
Proteínas Sanguíneas/metabolismo , Ingestão de Alimentos/fisiologia , Carne , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Fosfotransferases (Aceptor do Grupo Fosfato)/metabolismo , Proteoma/metabolismo , Adulto , Fatores Etários , Proteínas Sanguíneas/análise , Índice de Massa Corporal , Gorduras na Dieta/farmacologia , Metabolismo Energético/fisiologia , Feminino , Glucose/metabolismo , Humanos , Resistência à Insulina/fisiologia , Masculino , Pessoa de Meia-Idade , Proteínas Musculares/análise , Proteínas Musculares/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/patologia , Obesidade/sangue , Obesidade/patologia , Fosfotransferases (Aceptor do Grupo Fosfato)/análise , Período Pós-Prandial/fisiologia , Proteoma/análise , Magreza/sangue , Magreza/metabolismo , Magreza/patologia , Adulto Jovem
3.
Proc Natl Acad Sci U S A ; 116(47): 23562-23572, 2019 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-31690665

RESUMO

Primary cilia carry out numerous signaling and sensory functions, and defects in them, "ciliopathies," cause a range of symptoms, including blindness. Understanding of their nanometer-scale ciliary substructures and their disruptions in ciliopathies has been hindered by limitations of conventional microscopic techniques. We have combined cryoelectron tomography, enhanced by subtomogram averaging, with superresolution stochastic optical reconstruction microscopy (STORM) to define subdomains within the light-sensing rod sensory cilium of mouse retinas and reveal previously unknown substructures formed by resident proteins. Domains are demarcated by structural features such as the axoneme and its connections to the ciliary membrane, and are correlated with molecular markers of subcompartments, including the lumen and walls of the axoneme, the membrane glycocalyx, and the intervening cytoplasm. Within this framework, we report spatial distributions of key proteins in wild-type (WT) mice and the effects on them of genetic deficiencies in 3 models of Bardet-Biedl syndrome.


Assuntos
Síndrome de Bardet-Biedl/patologia , Cílios/ultraestrutura , Microscopia Crioeletrônica/métodos , Tomografia com Microscopia Eletrônica/métodos , Microscopia de Fluorescência/métodos , Nanotecnologia/métodos , Cílio Conector dos Fotorreceptores/ultraestrutura , Segmento Externo da Célula Bastonete/ultraestrutura , Imagem Individual de Molécula/métodos , Animais , Axonema/química , Axonema/ultraestrutura , Centríolos/ultraestrutura , Modelos Animais de Doenças , Proteínas do Olho/análise , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Associadas aos Microtúbulos/análise , Microtúbulos/ultraestrutura , Complexos Multiproteicos , Proteínas Musculares/análise , Cílio Conector dos Fotorreceptores/química , Proteínas Qa-SNARE/análise , Proteínas Supressoras de Tumor/análise
4.
Nutrients ; 11(8)2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31394788

RESUMO

Plant-sourced proteins offer environmental and health benefits, and research increasingly includes them in study formulas. However, plant-based proteins have less of an anabolic effect than animal proteins due to their lower digestibility, lower essential amino acid content (especially leucine), and deficiency in other essential amino acids, such as sulfur amino acids or lysine. Thus, plant amino acids are directed toward oxidation rather than used for muscle protein synthesis. In this review, we evaluate the ability of plant- versus animal-based proteins to help maintain skeletal muscle mass in healthy and especially older people and examine different nutritional strategies for improving the anabolic properties of plant-based proteins. Among these strategies, increasing protein intake has led to a positive acute postprandial muscle protein synthesis response and even positive long-term improvement in lean mass. Increasing the quality of protein intake by improving amino acid composition could also compensate for the lower anabolic potential of plant-based proteins. We evaluated and discussed four nutritional strategies for improving the amino acid composition of plant-based proteins: fortifying plant-based proteins with specific essential amino acids, selective breeding, blending several plant protein sources, and blending plant with animal-based protein sources. These nutritional approaches need to be profoundly examined in older individuals in order to optimize protein intake for this population who require a high-quality food protein intake to mitigate age-related muscle loss.


Assuntos
Anabolizantes , Proteínas na Dieta/administração & dosagem , Proteínas Musculares/administração & dosagem , Músculo Esquelético/fisiologia , Proteínas de Plantas/administração & dosagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Aminoácidos/análise , Animais , Proteínas na Dieta/farmacologia , Digestão , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Musculares/análise , Proteínas Musculares/metabolismo , Fenômenos Fisiológicos da Nutrição , Proteínas de Plantas/análise , Proteínas de Plantas/metabolismo , Biossíntese de Proteínas/fisiologia
5.
Genes Genomics ; 41(10): 1147-1163, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31256337

RESUMO

BACKGROUND: As cattle represent one of the most important livestock species for meat production, control of muscle development in regards to quality is an important research focus. OBJECTIVES: In this study, the phenotypic quality traits and its associations with DNA methylation levels of the longissimus muscle in two cattle breeds were studied. METHODS: The pH value, water loss rate, fat and protein and fatty acid content were measured in three beef cattle breeds of longissimus mucle; The longissimus mucle was analyzed by MethylRAD-seq and RNA-seq. The differentially methylated and differentially expressed related genes were subjected to BSP. RESULTS: Methylation status of longissimus mucle was analyzed by MethylRAD-seq. Compared with Simmental, there were 39 differentially methylated and expressed genes in muscle of Yunling cattle, and 123 differentially methylated and expressed genes in Wenshan muscle. A combined analysis of MethylRAD-seq and RNA-seq results revealed differential methylation and expression level of 18 genes between Simmental and Wenshan cattle, and 14 genes between Simmental and Yunling cattle. In addition, 28 genes were differentially methylated between Wenshan and Yunling cattle. Results of promoter methylation analysis of ACAD11, FADS6 and FASN showed that the overall degree of DNA methylation of FADS6 and FASN was negatively correlated with their expression levels. Methylation level of FASN in Simmental was greater than Yunling and Wenshan. The degree of methylation at the FADS6 CpG4 site was significantly higher in Simmental than that in Yunling. The levels of methylation at the CpG7 locus of the Simmental and Yunling breeds were greater than Wenshan cattle. A negative correlation was detected between the methylation levels and the expression of FASN CpG1, CpG2, CpG3, CpG5, CpG7, and CpG10. CONCLUSION: The functional and molecular regulatory mechanism of the genes related to meat quality can be revealed systematically from aspects of the genetic and epigenetic regulation. These studies will help to further explore the molecular mechanisms and phenotypic differences that regulate growth and quality of different breeds of cattle.


Assuntos
Bovinos/genética , Metilação de DNA , Carne/análise , Desenvolvimento Muscular/genética , Músculos , Acil-CoA Desidrogenase/genética , Animais , Cruzamento , China , Epigênese Genética , Ácidos Graxos Dessaturases/genética , Ácido Graxo Sintase Tipo I/genética , Ácidos Graxos/análise , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Concentração de Íons de Hidrogênio , Proteínas Musculares/análise , Músculo Esquelético/metabolismo , Fenótipo , Regiões Promotoras Genéticas , RNA Mensageiro
6.
Food Chem ; 297: 125006, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31253309

RESUMO

Muscle foods, particularly fish products are highly exposed to oxidative stress during processing and storage, resulting in oxidative modification of proteins. Protein carbonyls content has been used as one of the measures of oxidative stress. Generally, the resulting carbonylated proteins (CPs) have so far been labeled with 2,4-dinitrophenyl (DNP) hydrazine and detected with anti-DNP antibody. However, the applicability of this method to food samples is limited by its high price, time-consuming procedure and possibility to perform the measurements just on soluble protein fractions. We developed a simpler, faster and cheaper method to assess CP level in muscle foods, including both soluble and insoluble protein fractions, which is based on a direct reaction of protein carbonyls with 7-(diethylamino)coumarin-3-carbohydrazide (CHH). The paper describes a novel technique to label both soluble and insoluble carbonylated proteins with CHH and determine carbonyl content by fluorescence microscopy assay which correlates (R = 0.911) with conventional ELISA method.


Assuntos
Fluorometria/métodos , Proteínas Musculares/análise , Animais , Anticorpos/imunologia , Ensaio de Imunoadsorção Enzimática , Peixes/metabolismo , Congelamento , Microscopia de Fluorescência , Proteínas Musculares/química , Estresse Oxidativo , Fenil-Hidrazinas/química , Fenil-Hidrazinas/imunologia , Carbonilação Proteica , Reprodutibilidade dos Testes
7.
Dev Genes Evol ; 229(5-6): 147-159, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31214772

RESUMO

Our studies conducted on reptilian limb muscle development revealed, for the first time, early forelimb muscle differentiation at the morphological and molecular level. Sand lizard skeletal muscle differentiation in the early forelimb bud was investigated by light, confocal, and transmission electron microscopy as well as western blot. The early forelimb bud, filled with mesenchymal cells, is surrounded by monolayer epithelium cells. The immunocytochemical analysis revealed the presence of Pax3- and Lbx-positive cells in the vicinity of the ventro-lateral lip (VLL) of the dermomyotome, suggesting that VLL is the source of limb muscle progenitor cells. Furthermore, Pax3- and Lbx-positive cells were observed in the dorsal and ventral myogenic pools of the forelimb bud. Skeletal muscle development in the early limb bud is asynchronous, which is manifested by the presence of myogenic cells in different stages of differentiation: multinucleated myotubes with well-developed contractile apparatus, myoblasts, and mitotically active premyoblasts. The western blot analysis revealed the presence of MyoD and Myf5 proteins in all investigated developmental stages. The MyoD western blot analysis showed two bands corresponding to monomeric (mMyoD) and dimeric (dMyoD) fractions. Two separate bands were also detected in the case of Myf5. The observed bands were related to non-phosphorylated (Myf5) and phosphorylated (pMyf5) fractions of Myf5. Our investigations on sand lizard forelimb myogenesis showed that the pattern of muscle differentiation in the early forelimb bud shares many features with rodents and chicks.


Assuntos
Lagartos/embriologia , Desenvolvimento Muscular , Animais , Feminino , Imunofluorescência , Membro Anterior/embriologia , Botões de Extremidades/citologia , Botões de Extremidades/crescimento & desenvolvimento , Lagartos/genética , Microscopia Confocal , Proteínas Musculares/análise , Proteínas Musculares/genética , Músculo Esquelético/citologia , Músculo Esquelético/embriologia
8.
Ann Biol Clin (Paris) ; 77(4): 415-421, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31225801

RESUMO

Acute mesenteric ischemia (AMI) is one of the most severe diagnostic and therapeutic vital emergencies. This affection is characterized by the insufficient blood supply to the gastrointestinal tract, related to an occlusive or non-occlusive mechanism, resulting in an ischemic and inflammatory injury that may progress to necrosis of the intestinal wall. The clinical picture is nonspecific, dominated by acute abdominal pain. At present, no early biological marker is commonly used in clinical practice for diagnostic purposes. The purpose of this review was to review the markers that have been evaluated in this condition. Among the biological blood markers which have shown a diagnostic interest in the IMA, there are notably the two stereoisomers of lactate (D and L), D-dimers, and alpha glutathione transferase. More specific markers include the intestinal fatty acid binding protein or I-FABP, which is a marker of enterocyte necrosis, citrulline, a marker of enterocyte mass, or Smooth muscle protein 22 (SM22) marker for muscle damage. The early diagnosis of intestinal ischemia remains a challenge. It is likely that in the future IMA's biomarker research will be better customized and adapted to the physiopathological mechanism. More global approaches (proteomics, metabolomics) should also make it possible to identify new biomarkers.


Assuntos
Biomarcadores/sangue , Técnicas de Laboratório Clínico/métodos , Isquemia Mesentérica/diagnóstico , Doença Aguda , Biomarcadores/análise , Citrulina/análise , Citrulina/sangue , Técnicas de Laboratório Clínico/tendências , Diagnóstico Precoce , Proteínas de Ligação a Ácido Graxo/análise , Proteínas de Ligação a Ácido Graxo/sangue , Glutationa Transferase/análise , Glutationa Transferase/sangue , Humanos , Mediadores da Inflamação/análise , Mediadores da Inflamação/sangue , Isoenzimas/análise , Isoenzimas/sangue , Isquemia Mesentérica/sangue , Proteínas dos Microfilamentos/análise , Proteínas dos Microfilamentos/sangue , Proteínas Musculares/análise , Proteínas Musculares/sangue , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
9.
Poult Sci ; 98(7): 3067-3080, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-30938805

RESUMO

This study was performed to determine the slaughter and carcass traits, meat color, pH, nutrient composition, fatty acids, amino acids, and cholesterol content of local Turkish goose varieties. In the present study, a total of 80 geese from white, black, gray, and multicolored varieties were reared in an extensive production system at 28 wk of age. Quality analysis was performed on breast and thigh meat, as well as on the skin and abdominal fat of studied geese. Slaughter and carcass weights were significantly higher in the white variety (P < 0.01). Measured abdominal fat was lower in black and gray geese. Furthermore, the color and pH values were significantly different on breast, thigh, and skin among varieties. Additionally, measured breast meat b*, thigh meat L*, and b* values were higher in male geese. Breast meat protein levels were lower in black and gray geese, and also lower in the thigh meat of gray geese. Total breast meat amino acids were lower in white geese, and the amino acid content of thigh meat was lower in multicolored varieties. Moreover, differences in fatty acid composition were found among the studied varieties. However, cholesterol levels did not differ for either sex or variety.


Assuntos
Gansos , Carne/análise , Tecido Adiposo , Aminoácidos/análise , Criação de Animais Domésticos/métodos , Animais , Composição Corporal , Colesterol/análise , Cor , Ácidos Graxos/análise , Feminino , Masculino , Proteínas Musculares/análise , Valor Nutritivo , Especificidade da Espécie , Turquia
10.
Cardiovasc Pathol ; 41: 29-37, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31029755

RESUMO

BACKGROUND: A bicuspid aortic valve (BAV) is the most common congenital cardiac malformation and is associated with ascending aortic dilation in 60%-80% of patients. In this study, we aimed to address the role of hemodynamic influences on the development of aortopathy in BAV patients. PATIENT AND METHODS: BAV (n=36) and tricuspid aortic valve (TAV) patients (n=17) undergoing aortic valve replacement underwent preoperative flow magnetic resonance imaging (MRI) assessment to detect the area of maximal flow-induced stress in the proximal aorta. Based on these MRI data, paired ascending aortic wall samples [i.e., area of maximal jet impact (jet sample) and the opposite aortic wall (nonjet sample)] were collected during surgery. To study and describe the effects of jet stream on the complete vascular wall, a pathology score was developed based on the recently published aortic consensus paper statement on surgical pathology of the aorta using routine histologic stainings (resorcin fuchsin, hematoxylin-eosin, and Movat) and immunohistochemistry (alpha smooth muscle actin, smooth muscle 22 alpha, platelet endothelial cell adhesion molecule). RESULTS: Comparing the jet and nonjet samples in both BAV and TAV, regions of maximal jet impact did not show any difference in the pathology score in the adventitia and the middle and outer media. In the jet samples, the inner media however showed loss of actin expression in both BAV (P<.0001) and the TAV (P=.0074), and the intimal thickness was significantly enlarged in both patient groups (BAV P=.0005, TAV P=.0041), which was not accompanied by loss of elastic lamellae or vascular smooth muscle cell nuclei. CONCLUSIONS: In our study population, we could not demonstrate a potential distinct role for hemodynamics in the development of aortopathy in BAV patients even if corrected for aortic diameter, raphe position, or whether the valve is stenotic or regurgitant. The intimal layer and inner media however showed alterations in all jet specimens.


Assuntos
Aorta/fisiopatologia , Aneurisma Aórtico/fisiopatologia , Valva Aórtica/anormalidades , Doenças das Valvas Cardíacas/fisiopatologia , Hemodinâmica , Actinas/análise , Idoso , Aorta/química , Aorta/diagnóstico por imagem , Aorta/patologia , Aneurisma Aórtico/diagnóstico por imagem , Aneurisma Aórtico/etiologia , Aneurisma Aórtico/metabolismo , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/fisiopatologia , Insuficiência da Valva Aórtica/etiologia , Insuficiência da Valva Aórtica/fisiopatologia , Estenose da Valva Aórtica/etiologia , Estenose da Valva Aórtica/fisiopatologia , Dilatação Patológica , Feminino , Doenças das Valvas Cardíacas/complicações , Doenças das Valvas Cardíacas/diagnóstico por imagem , Humanos , Imuno-Histoquímica , Imagem por Ressonância Magnética , Masculino , Proteínas dos Microfilamentos/análise , Pessoa de Meia-Idade , Proteínas Musculares/análise , Molécula-1 de Adesão Celular Endotelial a Plaquetas/análise
11.
J Am Soc Mass Spectrom ; 30(12): 2460-2469, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30834509

RESUMO

Sarcomeric proteins, including myofilament and Z-disk proteins, play critical roles in regulating muscle contractile properties. A variety of isoforms and post-translational modifications (PTMs) of sarcomeric proteins have been shown to be associated with modulation of muscle functions and the occurrence of muscle diseases. Non-human primates (NHPs) are excellent research models for sarcopenia, a disease associated with alterations in sarcomeric proteins, due to their marked similarities to humans. However, the sarcomeric proteins in NHP skeletal muscle have not been well characterized. To gain a deeper understanding of sarcomeric proteins in NHP skeletal muscle, we employed top-down mass spectrometry (MS) to conduct a comprehensive analysis on isoforms and PTMs of sarcomeric proteins in rhesus macaque skeletal muscle. We identified 23 protein isoforms with 46 proteoforms of sarcomeric proteins, including 6 isoforms with 18 proteoforms from fast skeletal troponin T. Particularly, for the first time, a novel PDZ/LIM domain protein isoform, PDLIM7, was characterized with a newly identified protein sequence. Moreover, we also identified multiple PTMs on these proteins, including deamidation, methylation, acetylation, tri-methylation, phosphorylation, and S-glutathionylation. Most PTM sites were localized, including Asn13 deamidation on MLC-2S; His73 methylation on αactin; N-terminal acetylation on most identified proteins; N-terminal tri-methylation on MLC-1S, MLC-1F, MLC-2S, and MLC-2F; Ser14 phosphorylation on MLC-2S; and Ser15 and Ser16 phosphorylation on MLC-2F. In summary, a comprehensive characterization of sarcomeric proteins including multiple isoforms and PTMs in NHP skeletal muscle was achieved by analyzing intact proteins in the top-down MS approach.


Assuntos
Proteínas Musculares/análise , Músculo Esquelético/química , Sarcômeros/química , Espectrometria de Massas em Tandem/métodos , Sequência de Aminoácidos , Animais , Macaca mulatta , Domínios PDZ , Isoformas de Proteínas/análise , Processamento de Proteína Pós-Traducional
12.
Meat Sci ; 152: 41-48, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30802817

RESUMO

To investigate the potential of eland for meat production, the yield, physical quality and chemical composition of seven muscles were characterized from both male (n = 6) and female (n = 6) carcasses. Although carcass weights did not differ, cows had greater biceps femoris (BF), semitendinosus (ST) and supraspinatus (SS) muscle yields. The semimembranosus (SM) from cows had the greatest drip loss and were more yellow (b*) than all muscles evaluated from both sexes. Shear force measurements showed no effect of sex but large variations between muscles existed (65.5 to 97.6 N). Moisture content was greater within the BF, SS, ST and infraspinatus (IS) than the longissimus thoracis et lumborum (LTL) and SM. The LTL and IS had the lowest intramuscular fat content, while the LTL and SM muscle had the highest crude protein content. Sex had little effect on eland meat composition, showing potential for the meat of both sexes to be marketed as a lean meat source.


Assuntos
Antílopes , Carne/análise , Músculo Esquelético/anatomia & histologia , Músculo Esquelético/química , Animais , Cor , Feminino , Lipídeos/análise , Masculino , Proteínas Musculares/análise , Fatores Sexuais , Resistência ao Cisalhamento
13.
Scand J Med Sci Sports ; 29(5): 669-677, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30676666

RESUMO

This study investigated the in-season effect of intensified training comparing the efficacy of duration-matched intense intermittent exercise training with sprint interval training in increasing intermittent running performance, sprint ability, and muscle content of proteins related to ion handling and metabolism in football players. After the first two weeks in the season, 22 sub-elite football players completed either 10 weeks of intense intermittent training using the 10-20-30 training concept (10-20-30, n = 12) or sprint interval training (SIT, n = 10; work/rest ratio: 6-s/54-s) three times weekly, with a ~20% reduction in weekly training time. Before and after the intervention, players performed a Yo-Yo intermittent recovery test level 1 (Yo-Yo IR1) and a 30-m sprint test. Furthermore, players had a muscle biopsy taken from the vastus lateralis. Yo-Yo IR1 performance increased by 330 m (95%CI: 178-482, P ≤ 0.01) in 10-20-30, whereas no change was observed in SIT. Sprint time did not change in 10-20-30 but decreased by 0.04 second (95%CI: 0.00-0.09, P ≤ 0.05) in SIT. Muscle content of HADHA (24%, P ≤ 0.01), PDH-E1α (40%, P ≤ 0.01), complex I-V of the electron transport chain (ETC) (51%, P ≤ 0.01) and Na+ , K+ -ATPase subunits α2 (33%, P ≤ 0.05) and ß1 (27%, P ≤ 0.05) increased in 10-20-30, whereas content of DHPR (27%, P ≤ 0.01) and complex I-V of the ETC (31%, P ≤ 0.05) increased in SIT. Intense intermittent training, combining short sprints and a high aerobic load, is superior to regular sprint interval training in increasing intense intermittent running performance during a Yo-Yo IR1 test and muscle content of PDH-E1α and HADHA in sub-elite football players.


Assuntos
Adaptação Fisiológica , Desempenho Atlético/fisiologia , Treinamento Intervalado de Alta Intensidade/métodos , Corrida/fisiologia , Futebol/fisiologia , Adulto , Atletas , Humanos , Proteínas Musculares/análise , Cadeias Pesadas de Miosina/análise , Músculo Quadríceps/fisiologia , Adulto Jovem
14.
Meat Sci ; 148: 127-136, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30388477

RESUMO

Thirty Nellore crossbred male sheep (Ovis aries) were divided into two groups of 15 animals each and subjected to either pre-slaughter electrical stunning followed by slaughter (ST) or traditional halal slaughter without stunning (NST) to investigate the changes in blood biochemical parameters, meat quality and proteomic profile. Higher (P < .05) pH, water holding capacity and Warner-Bratzler shear force were observed in meat from stunned sheep. Quantitative proteomic approach using DIGE was employed to find a panel of protein markers that could differentiate ST and NST muscle proteome. Comparison of muscle proteome of ST and NST samples by 2D-DIGE and MALDI-TOF/TOF MS analysis revealed 46 significant (P < .05) differentially expressed proteins. Our analysis revealed changes in the abundance of proteins involved in catalytic, structural, and stress related process. Current study has demonstrated variation meat quality and identified important proteins that correlate with meat texture and pre-slaughter stress in sheep that are slaughtered without and with electrical stunning.


Assuntos
Matadouros , Anestesia/veterinária , Manipulação de Alimentos/métodos , Carne Vermelha/análise , Bem-Estar do Animal , Animais , Qualidade dos Alimentos , Islamismo , Masculino , Proteínas Musculares/análise , Proteômica , Carneiro Doméstico , Estresse Fisiológico , Eletroforese em Gel Diferencial Bidimensional/métodos
15.
J Proteomics ; 193: 123-130, 2019 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-30312679

RESUMO

Ultrasound can intensify the heating process used to correct texture defects in dry-cured hams. The effect of ultrasound-assisted heating on the proteome of sliced dry-cured ham was evaluated. Dry-cured hams with high proteolysis index (PI > 36) were sliced, vacuum packed and subjected to conventional (CV) and ultrasound-assisted (US) thermal treatments. Comparative proteome profiling between sample groups was assessed by two-dimensional electrophoresis (2-DE) coupled to tandem mass spectrometry. It was found that protein fragmentation increased markedly after US thermal treatment. Specifically, fragments of the major myofibrillar protein, actin, were abundantly over-represented following US heating. In addition, five unfragmented sarcoplasmic proteins (i.e. fatty acid-binding protein, peroxiredoxin-6, superoxide dismutase, carbonyl reductase and aminoacylase) showed increased abundance in the US sample group. These results suggest candidate biomarkers to monitor proteolysis intensity and proteolysis-independent effects linked to cured ham quality by ultrasound application. SIGNIFICANCE: The present proteome profiling study of treated dry-cured ham demonstrates the impact of ultrasound action on proteins. Moreover, negative organoleptic effects can be appearing with ultrasound treatment due to proteolysis increase. Therefore, the proteolysis monitoring could help to control these effects. In this regards, our results suggest that actin can be a candidate biomarker to monitor proteolysis intensity.


Assuntos
Análise de Alimentos , Manipulação de Alimentos , Proteínas Musculares/análise , Carne de Porco/análise , Proteômica , Ondas Ultrassônicas
16.
Poult Sci ; 98(1): 480-490, 2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30165657

RESUMO

The muscle syndrome woody breast (WB) impairs quality of chicken fillets and is a challenge to the poultry meat industry. There is a need for online detection of affected fillets for automatic quality sorting in process. Near-infrared spectroscopy (NIRS) is a promising method, and in this study we elucidate the spectral properties of WB versus normal fillets. On a training set of 50 chicken fillets (20 normal, 30 WB), we measured NIR, nuclear magnetic resonance (NMR) T2 relaxation distributions, and crude chemical composition. NIRS could estimate protein in the fillets with an accuracy of ±0.64 percentage points. T2 distributions showed that there was a larger share of free water in WB fillets. This difference in water binding generated a shift and narrowing of the water absorption peak in NIR around 980 nm, quantified by a bound water index (BWI). The correlation between BWI and T2 distributions was 0.78, indicating that NIRS contains information about degree of water binding. Discriminant analysis showed that NIRS obtained 100% correct classification of normal versus WB on the training set, and 96% correct classification on a test set of 52 fillets. The main reason for why NIRS can successfully discriminate between WB and normal fillets is the methods sensitivity to both protein content and degree of water binding in the muscle, both established markers for WB. The classification model can be based on NIR spectra only, calibration against protein is not needed. The affected muscle tissue associated with the WB syndrome is unevenly distributed in the fillets, and this heterogeneity was characterized by NIRS and NMR. Clear differences in water binding properties were found between the superficial 1 cm layer and the deeper layer at 1 to 2 cm depth. Significant differences in protein estimates by NIRS at different measurement points along the chicken fillets were obtained for WB fillets. The findings suggest how to obtain optimal sampling with NIRS for best possible discrimination between WB and normal breast fillets.


Assuntos
Carne/análise , Doenças Musculares/veterinária , Doenças das Aves Domésticas/diagnóstico , Espectroscopia de Luz Próxima ao Infravermelho/veterinária , Animais , Galinhas , Espectroscopia de Ressonância Magnética/métodos , Proteínas Musculares/análise , Doenças Musculares/diagnóstico , Músculos Peitorais/química , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Água/análise
17.
Sci Rep ; 8(1): 15728, 2018 10 24.
Artigo em Inglês | MEDLINE | ID: mdl-30356055

RESUMO

Nebulin is a very large protein required for assembly of the contractile machinery in muscle. Mutations in the nebulin gene NEB are a common cause of nemaline myopathy. Nebulin mRNA is alternatively-spliced so that each mRNA contains either exon 143 or exon 144. We have produced monoclonal antibodies specific for the regions of nebulin encoded by these two exons, enabling analysis of expression of isoforms at the protein level for the first time. All antibodies recognized a protein of the expected size (600-900 kD) and stained cross-striations of sarcomeres in muscle sections. Expression of exon 143 is developmentally-regulated since newly-formed myotubes in cell culture expressed nebulin with exon 144 only; this was confirmed at the mRNA level by qPCR. In fetal muscle, nebulin with exon 143 was expressed in some myotubes by 12-weeks of gestation and strongly-expressed in most myotubes by 17-weeks. In mature human muscle, the exon 144 antibody stained all fibres, but the exon 143 antibody staining varied from very strong in some fibres to almost-undetectable in other fibres. The results show that nebulin containing exon 144 is the default isoform early in myogenesis, while regulated expression of nebulin containing exon 143 occurs at later stages of muscle development.


Assuntos
Éxons , Proteínas Musculares/química , Isoformas de Proteínas/genética , Processamento Alternativo , Anticorpos Monoclonais , Células Cultivadas , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Desenvolvimento Muscular , Fibras Musculares Esqueléticas/metabolismo , Proteínas Musculares/análise , Proteínas Musculares/metabolismo , Isoformas de Proteínas/análise , Isoformas de Proteínas/metabolismo
18.
Gene ; 675: 119-127, 2018 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-30180961

RESUMO

Type 2 diabetes mellitus (T2DM) is a complex, multifactorial metabolic disease, and the number of patients with T2DM has continued to increase in recent years. Large-scale proteomic studies on animal models of T2DM are of great importance to understand the pathophysiology of T2DM. Therefore, in our study, Isobaric tags for relative and absolute quantification (iTRAQ) and Parallel reaction monitoring (PRM) were used for proteomic analysis of skeletal muscles from T2DM-susceptible and -tolerant Bama mini-pig models induced by a high-fat, high-sugar diet. In our proteomic analysis, a total of 1646 proteins and 13 differentially expressed proteins (DEPs) were identified by iTRAQ-mass spectrometry, and 6 differentially expressed proteins were validated by PRM. Gene Ontology (GO) analysis revealed that most DEPs were extracellular matrix (ECM) proteins and participated in several biological processes, such as negative regulation of JAK-STAT cascade, negative regulation of STAT cascade, roundabout signaling pathway and peptide cross-linking via chondroitin 4-sulfate glycosaminoglycan, and the molecular functions of roundabout binding, glycosaminoglycan binding, heparin binding, sulfur compound binding, collagen binding, and kinase inhibitor activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis results showed that the differentially expressed proteins were involved in 14 pathways, including human disease pathways, metabolic pathways, signal transduction pathways, signaling molecules and interaction pathways, and the cellular process pathways associated with phagosomes and focal adhesion. In conclusion, the proteomics based on iTRAQ and PRM in T2DM-susceptible and -tolerant Bama mini-pig models showed that changes in amino acid metabolism, inflammation-associated pathways and the impaired function and environment of extracellular matrix are risk factors associated with increased pathogenesis of T2DM in Bama mini-pig.


Assuntos
Diabetes Mellitus Tipo 2/metabolismo , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Proteômica/métodos , Animais , Diabetes Mellitus Experimental/etiologia , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 2/etiologia , Dieta Hiperlipídica , Suscetibilidade a Doenças , Proteínas da Matriz Extracelular/metabolismo , Proteínas Musculares/análise , Reprodutibilidade dos Testes , Suínos , Porco Miniatura/metabolismo
19.
Bosn J Basic Med Sci ; 18(4): 297-304, 2018 Nov 05.
Artigo em Inglês | MEDLINE | ID: mdl-30179586

RESUMO

Pancreatic cancer (PC) is a highly aggressive tumor, often difficult to diagnose and treat. Aspartate ß-hydroxylase (ASPH) is a type II transmembrane protein and the member of α-ketoglutarate-dependent dioxygenase family, found to be overexpressed in different cancer types, including PC. ASPH appears to be involved in the regulation of proliferation, invasion and metastasis of PC cells through multiple signaling pathways, suggesting its role as a tumor biomarker and therapeutic target. In this review, we briefly summarize the possible mechanisms of action of ASPH in PC and recent progress in the therapeutic approaches targeting ASPH.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Proteínas de Membrana/metabolismo , Oxigenases de Função Mista/metabolismo , Proteínas Musculares/metabolismo , Neoplasias Pancreáticas/enzimologia , Animais , Antineoplásicos/farmacologia , Proteínas de Ligação ao Cálcio/análise , Proteínas de Ligação ao Cálcio/química , Humanos , Proteínas de Membrana/análise , Proteínas de Membrana/química , Oxigenases de Função Mista/análise , Oxigenases de Função Mista/química , Proteínas Musculares/análise , Proteínas Musculares/química , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/patologia
20.
Scand Cardiovasc J ; 52(5): 281-286, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30043668

RESUMO

OBJECTIVES: A careful selection of reference samples in studies on the pathogenesis of thoracic ascending aorta (TAA) dilation is crucial for reliability, consistency and reproducibility of experimental results. Several studies include control TAA samples from heart donors. Others include samples harvested during coronary artery bypass graft (CABG) procedures or a mix of samples from heart donors and CABG patients. We verified the equivalence/homogeneity of TAA samples from heart donors and CABG patients in terms of basal gene expression and thus their reliability as reference groups in aortopathy studies. DESIGN: We analysed by RT-PCR and Western blot the differential expression of smoothelin, α-smooth muscle actin (α-SMA) and transforming growth factor-ß1 (TGF-ß1), selected as major players in smooth muscle cell and myofibroblast phenotype and remodelling. The mean age and comorbidities of subjects were consistent with data routinely seen in clinical practice. RESULTS: Data revealed the loss of smoothelin in samples from CABG patients, together with a significant increase of α-SMA, while TGF-ß1 dimer showed a marked increase in CABG patients versus heart donors, accompanied by a decrease of the corresponding mRNA. Differences in gene expression were maintained after adjustment for age. However, TGF-ß1 mRNA and CABG patients' age showed a positive correlation (ρ = 0.89, p < .05), while α-SMA mRNA and age showed a negative correlation (ρ = -0.85, p < .05). CONCLUSIONS: We revealed the non-equivalence of samples from heart donors and CABG patients, presumably for the presence of microscopic atherosclerotic lesions in CABG patients, suggesting the necessity of a careful selection of control groups in aortopathy studies.


Assuntos
Aorta Torácica/cirurgia , Doenças da Aorta/patologia , Ponte de Artéria Coronária , Transplante de Coração , Doadores de Tecidos , Coleta de Tecidos e Órgãos/métodos , Actinas/análise , Actinas/genética , Adulto , Idoso , Aorta Torácica/química , Aorta Torácica/patologia , Doenças da Aorta/metabolismo , Biomarcadores/análise , Estudos de Casos e Controles , Proteínas do Citoesqueleto/análise , Proteínas do Citoesqueleto/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Musculares/análise , Proteínas Musculares/genética , Fator de Crescimento Transformador beta1/análise , Fator de Crescimento Transformador beta1/genética
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