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1.
Life Sci ; 241: 117119, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31794771

RESUMO

AIM: Mechanoelectric feedback (MEF) was related to malignant arrhythmias in heart failure (HF). Desmin is a cytoskeleton protein and could be involved in MEF as a mechanoelectrical transducer. In this study, we will discuss the role of desmin alterations in mechanical electrical feedback in heart failure and its mechanisms. METHODS: We used both an in vivo rat model and an in vitro cardiomyocyte model to address this issue. For the in vivo experiments, we establish a sham group, an HF group, streptomycin (SM) group, and an MDL-28170 group. The occurrence of ventricular arrhythmias (VA) was recorded in each group. For the in vitro cardiomyocyte model, we established an NC group, a si-desmin group, and a si-desmin + NBD IKK group. The expression of desmin, IKKß, p-IKKß, IKBα, p-NF-κB, and SERCA2 were detected in both in vivo and in vitro experiments. The content of Ca2+ in cytoplasm and sarcoplasmic were detected by confocal imaging in vitro experiments. RESULTS: An increased number of VAs were found in the HF group. SM and MDL-28170 can reduce desmin breakdown and the number of VAs in heart failure. The knockdown of desmin in the cardiomyocyte can activate the NF-κB pathway, decrease the level of SERCA2, and result in abnormal distribution of Ca2+. While treatment with NF-κB inhibitor can elevate the level of SERCA2 and alleviate the abnormal distribution of Ca2+. SIGNIFICANCE: Overall, desmin may participate in MEF through the NF-κB pathway. This study provides a potential therapeutic target for VA in HF.


Assuntos
Cálcio/metabolismo , Desmina/metabolismo , Insuficiência Cardíaca/etiologia , NF-kappa B/metabolismo , Animais , Calpaína/metabolismo , Células Cultivadas , Desmina/genética , Dipeptídeos/farmacologia , Modelos Animais de Doenças , Eletrocardiografia , Retroalimentação Fisiológica , Técnicas de Silenciamento de Genes , Masculino , Miócitos Cardíacos , NF-kappa B/genética , Ratos Sprague-Dawley , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/genética , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático/metabolismo , Taquicardia Ventricular/complicações , Taquicardia Ventricular/fisiopatologia
2.
Yonsei Med J ; 61(1): 85-93, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31887804

RESUMO

PURPOSE: The aim of this study was to investigate the effect of FST gene on the inhibition of fibrosis in fibroblastic cells from scar tissue around repaired zone II flexor tendons. MATERIALS AND METHODS: Immunohistochemistry was conducted on fibroblast cells transfected with adenovirus-LacZ (Ad-LacZ) as a marker gene (control), or with adenovirus-FST (Ad-FST) as a therapeutic gene. Fibroblast cultures without adenoviral exposure served as controls. RESULTS: Fibroblastic cells transfected with Ad-FST demonstrated significant decrease in collagen type I, MMP-1, MMP2, and α-SMA mRNA expressions compared to those transfected with Ad-LacZ. In addition, fibroblastic cells transfected with Ad-FST exhibited significant decrease in MMP-1, TIMP-1, fibronectin, PAI-1, TRPV4, α-SMA, desmin, and PAX7 protein expressions. CONCLUSION: Based on these findings, we conclude that FST may be a novel therapeutic strategy for preventing scar adhesions around repaired tendons by inhibiting fibroblasts from differentiating into myofibroblasts, in addition to producing type I collagen and regulating extracellular matrix turnover via the downregulation of MMP-1 and TIMP-1. FST may also decrease contracture of the scar by inhibiting Ca2+-dependent cell contraction.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Cicatriz/metabolismo , Cicatriz/patologia , Colágeno Tipo I/biossíntese , Fibroblastos/metabolismo , Folistatina/metabolismo , Miofibroblastos/patologia , Traumatismos dos Tendões/patologia , Actinas/metabolismo , Animais , Células Cultivadas , Desmina/metabolismo , Feminino , Fibronectinas/genética , Fibronectinas/metabolismo , Fibrose , Regulação da Expressão Gênica , Humanos , Metaloproteinase 1 da Matriz/genética , Metaloproteinase 1 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Fator de Transcrição PAX7/genética , Fator de Transcrição PAX7/metabolismo , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Tendões/patologia
3.
Food Chem ; 306: 125616, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31622832

RESUMO

This research aimed to explore the role of protein S-nitrosylation in regulating the tenderness of postmortem beef, from the perspective of µ-calpain autolysis and protein proteolysis. Five bovine semimembranosus muscles were incubated with three treatments including S-nitrosoglutathione (GSNO, nitric oxide donor), normal saline and Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME, nitric oxide synthase inhibitor). The results showed that the level of protein S-nitrosylation was improved by GSNO treatment and reduced by L-NAME treatment (p < 0.05). Compared to the control, GSNO treatment had higher shear force while L-NAME treatment presented lower shear force at 7 d postmortem (p < 0.05). In addition, µ-calpain autolysis, myofibrillar protein and desmin degradation were reduced by GSNO treatment and accelerated by L-NAME treatment (p < 0.05). Therefore, it can be speculated that protein S-nitrosylation could affect beef tenderization by regulating the autolysis of µ-calpain and the degradation of myofibrillar proteins.


Assuntos
Proteína S/metabolismo , Carne Vermelha , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Calpaína/metabolismo , Bovinos , Desmina/metabolismo , Proteína S/química , Proteólise
4.
Food Chem ; 301: 125278, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31387033

RESUMO

Softening is always a problem in fish preservation. This study was aimed to investigate the role of myofibrillar structural proteins degradation in fish softening. The changes of myofibrillar structural proteins, muscle ultrastructure, myofibril fragmentation, and shear force were studied. The results indicated that during the superchilled preservation of grass carp (Ctenopharyngodon idella), small (low-molecular-weight) myofibrillar structural proteins like desmin and troponin-T initiated textural deterioration, leading to Z-disk weakening and actin loosening. In contrast, giant (high-molecular-weight) myofibrillar structural proteins like titin and nebulin were degraded in more amount in the later storage, contributing to Z-disk and M-band disassembly and vague of light and dark regions (I and A bands). Compared to each other, desmin and titin played more important part in softening. All these changes were involved in the increase of muscle fibril segments and the sharp decrease of shear force.


Assuntos
Carpas/metabolismo , Produtos Pesqueiros , Proteínas de Peixes/química , Miofibrilas/química , Animais , Conectina/química , Conectina/metabolismo , Desmina/química , Desmina/metabolismo , Proteínas de Peixes/metabolismo , Proteínas Musculares/química , Proteínas Musculares/metabolismo , Miofibrilas/metabolismo , Proteólise , Troponina T/química , Troponina T/metabolismo
5.
Anim Sci J ; 90(8): 1050-1059, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31199034

RESUMO

The objective of this study was to create various pH/temp decline rates in hot-boned bull beef M. longissimus lumborum (LL) through a combination of electrical stimulation (ES) and pre-rigor holding temperature. The relationship between the pre-rigor interventions, the activities of µ-calpain and small heat shock proteins (sHSP), and the impacts on meat product quality were determined. Paired LL loins from 13 bulls were hot-boned within 40 min of slaughter, immediately ES and subjected to various holding temperatures (5, 15, 25, and 35°C) for 3 hr. The rate of muscle pH decline, sarcomere length, shear force, and proteolysis of muscle proteins were measured. ES-25°C had a longer sarcomere length compared to non-electrical stimulation samples. ES-25°C and ES-35°C samples had lower shear force values, higher µ-calpain activity and higher desmin, troponin-T, and sHSP degradation. The above findings suggest that pH/temp decline rates created in hot-boned muscle impacted muscle protein proteolysis by increasing the activity of proteases and degradation of sHSP.


Assuntos
Calpaína/análise , Estimulação Elétrica , Qualidade dos Alimentos , Proteínas de Choque Térmico/análise , Músculo Esquelético/metabolismo , Carne Vermelha/análise , Temperatura Ambiente , Animais , Bovinos , Desmina/metabolismo , Concentração de Íons de Hidrogênio , Masculino , Peptídeo Hidrolases/metabolismo , Proteólise , Sarcômeros/patologia , Resistência ao Cisalhamento , Fatores de Tempo , Troponina T/metabolismo
6.
Leg Med (Tokyo) ; 38: 36-44, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30959395

RESUMO

In order to investigate the sequence and time course of fibronectin (Fn), fibrinogen (Fg), complement (C5), myoglobin (Mb), actin (HHF35), and desmin (Dm) for the diagnosis of early myocardial ischemia, the myocardial ischemia model was established in rats, the positive reaction areas of Fn, Fg and C5 and the depletion areas of Mb, HHF35 and Dm in the ischemic cardiomyocytes were studied with immunohistochemistry, image analysis technique and statistical system. The results showed that the depletion of Dm, HHF35 and Mb, and the positive staining of Fg and C5 in ischemic cardiomyocytes were found as early as 15 min after the myocardial ischemia, but the positive staining of Fn occurred till 3 h after myocardial ischemia. With the prolongation of ischemia, the areas of the depletion of Dm, HHF35, Mb and the positive staining of Fg, C5 and Fn gradually enlarged. It is suggested that all the six immunohistochemical markers are more sensitive than routine H&E staining, and that Dm, HHF35, Mb, Fg, C5 are more sensitive markers than Fn for detection of early myocardial ischemia.


Assuntos
Actinas/metabolismo , Complemento C5/metabolismo , Desmina/metabolismo , Fibrinogênio/metabolismo , Fibronectinas/metabolismo , Isquemia Miocárdica/diagnóstico , Isquemia Miocárdica/metabolismo , Miocárdio/metabolismo , Mioglobina/metabolismo , Animais , Biomarcadores/metabolismo , Proteínas Sanguíneas , Modelos Animais de Doenças , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Ratos Sprague-Dawley , Fatores de Tempo
7.
Meat Sci ; 153: 144-151, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30946977

RESUMO

A study was conducted to elucidate the impact of pulsed electric field (PEF) on the activity of calpains, proteolytic activity of desmin and troponin-T and physicochemical properties of beef Biceps femoris during ageing. The meat samples (meat blocks) were subjected to two PEF treatments; T1 (5 kV, 90 Hz, 0.38 kV/cm) and T2 (10 kV, 20 Hz, 0.61 kV/cm) and a non-treated control was run in parallel. The samples were vacuum packaged and aged for 1, 7 and 14 days at 4 ±â€¯1 °C. This study reports for the first time the impact of PEF-processing on the calpain activity in beef. Early post-mortem activation of calpain 2 was observed in PEF-treated samples. An increase in the calpain activity and proteolysis of desmin and troponin-T was observed. No significant effect of PEF was observed on the shear force of tough muscles from culled dairy animals during the entire ageing period.


Assuntos
Calpaína/metabolismo , Eletricidade , Manipulação de Alimentos/métodos , Carne Vermelha/análise , Animais , Bovinos , Desmina/metabolismo , Músculo Esquelético/metabolismo , Proteólise , Resistência ao Cisalhamento , Troponina T/metabolismo
8.
Genes Chromosomes Cancer ; 58(9): 643-649, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30920708

RESUMO

Ossifying fibromyxoid tumor (OFMT) is an uncommon mesenchymal neoplasm of uncertain differentiation and intermediate malignant potential. Recurrent PHF1 gene rearrangements are detected in up to 80% of OFMTs. We describe the clinicopathologic features of five OFMTs harboring a novel PHF1-TFE3 fusion. In two cases, RNA sequencing identified a fusion transcript composed of PHF1 exon 11 fused to TFE3 exon 3, whereas in a third case PHF1 exon 12 was fused to TFE3 exon 7. A FISH break-apart assay revealed rearrangements in both PHF1 and TFE3 genes in all cases. The cohort included three males and two females with a median age of 64 years. One OFMT originated in the scapula, while four occurred in the deep soft tissues. Two OFMTs had typical features, whereas three were classified as malignant. Despite uniform cytologic features and fibromyxoid stroma compatible with an OFMT diagnosis, none showed a peripheral shell of lamellar bone. S100 expression was focally present in only one case, while desmin was positive in three cases. All tumors showed strong nuclear immunopositivity for TFE3. All three malignant OFMTs developed metastases, either regionally or to the lung. One patient died of disease 1 year after diagnosis, while the remaining two are alive with disease. In summary, we report novel recurrent PHF1-TFE3 fusions in a subset of OFMTs with aggressive clinical behavior. The PHF1-TFE3 fusions resulted in consistent protein TFE3 overexpression which can be used as a reliable screening tool, adding OFMT as another tumor driven by TFE3 oncogenic activation pathway.


Assuntos
Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Biomarcadores Tumorais/genética , Proteínas de Ligação a DNA/genética , Fibroma Ossificante/genética , Fusão Oncogênica , Proteínas do Grupo Polycomb/genética , Neoplasias de Tecidos Moles/genética , Idoso , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Biomarcadores Tumorais/metabolismo , Proteínas de Ligação a DNA/metabolismo , Desmina/genética , Desmina/metabolismo , Feminino , Fibroma Ossificante/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Proteínas do Grupo Polycomb/metabolismo , Proteínas S100/genética , Proteínas S100/metabolismo , Neoplasias de Tecidos Moles/patologia
9.
Reprod Domest Anim ; 54(6): 835-845, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30907027

RESUMO

Equine endometrosis, a frequent cause of subfertility, is characterized by periglandular fibrosis, and no treatment exists. Endometrial biopsies not only contain diseased glands, but also contain healthy glands and stroma. Myoepithelial (ME) and myofibroblastic (MF) markers are calponin, smooth muscle actin (SMA), desmin and glial fibrillary acidic protein (GFAP). Epithelial vimentin expression indicates epithelial to mesenchymal transition (EMT). The aim of this immunohistochemical study was to investigate whether biopsies with endometrosis express MF and ME markers and vimentin. Compared to healthy areas, significantly higher percentages of endometrotic glands were lined by calponin- and vimentin-positive epithelial cells, whereas periglandular fibrosis contained significantly higher percentages of stromal cells positive for vimentin, desmin and SMA and significantly less calponin-positive stromal cells. The rare GFAP expression was restricted to endometrotic glands. Of these, the most frequent features of endometrotic glands were higher percentages of SMA- and vimentin-positive stromal cells and the prominent epithelial calponin staining that occurred in 100%, 93% and 95% of examined biopsies. Results indicate plasticity of equine endometrial epithelial and stromal cells. Particularly, endometrotic glands show evidence for ME differentiation and EMT. The different expression of MF markers between stromal cells from healthy and endometrotic areas suggests functional differences. The characteristic changes in the expression of SMA, vimentin and calponin between endometrotic glands and healthy areas can be helpful to confirm early stages of endometrosis. The characterization of cellular differentiation may help to decipher the pathogenesis of endometrosis and could lead to therapeutic strategies.


Assuntos
Endometriose/veterinária , Transição Epitelial-Mesenquimal , Doenças dos Cavalos/patologia , Actinas/metabolismo , Animais , Biomarcadores , Biópsia/veterinária , Proteínas de Ligação ao Cálcio/metabolismo , Desmina/metabolismo , Endometriose/metabolismo , Endometriose/patologia , Feminino , Doenças dos Cavalos/metabolismo , Cavalos , Imuno-Histoquímica/veterinária , Proteínas dos Microfilamentos/metabolismo , Células Estromais , Vimentina/metabolismo
10.
Saudi Med J ; 40(2): 126-130, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30723856

RESUMO

OBJECTIVES: To evaluate the clinical presentations and immunohistochemical (IHC) properties of gastrointestinal stromal tumors (GISTs) and to compare them to internationally published data. METHODS: Thirty-six patients diagnosed with GISTs between January 1997 and December 2015 were retrospectively studied in 2 tertiary hospitals. Immunohistochemical staining was carried out prospectively when it has not been completed fully at the beginning. Results: The median age of patients was 54 years  (range; 17-81 years). Predominantly, we found more females were affected. The male to female ratio was 1:1.7. The most frequently affected organs were the stomach (63.8%) followed by small bowel (25%) and colorectal region (8.4%). Abdominal pain was the most frequent presentation in 33.3% of the patients then gastrointestinal (GI) bleeding in 30.5%. Most of the gastric GISTs were at early stages at presentation: stage 1 and II (60.8%), while in non-gastric GISTs, the tumor stage was advanced: stage III and IV (69.3%). The IHC characteristic of GIST in descending order showed positivity for vimentin (88.9%), CD117 (83.3%), CD34 (77.8%), Ki67 (63.9%), SMA (38.9%), desmin (27.8%), and S100 (19.4%). CONCLUSION: Gastrointestinal stromal tumors in our study demonstrates a major similar feature as the published international data. However, minor differences do exist in terms of clinical features and immunohistochemistry.


Assuntos
Dor Abdominal/etiologia , Hemorragia Gastrointestinal/etiologia , Neoplasias Gastrointestinais/metabolismo , Tumores do Estroma Gastrointestinal/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos CD34/metabolismo , Colo , Desmina/metabolismo , Feminino , Neoplasias Gastrointestinais/complicações , Neoplasias Gastrointestinais/patologia , Tumores do Estroma Gastrointestinal/complicações , Tumores do Estroma Gastrointestinal/secundário , Humanos , Imuno-Histoquímica , Intestino Delgado , Antígeno Ki-67/metabolismo , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Proteínas Proto-Oncogênicas c-kit/metabolismo , Reto , Estudos Retrospectivos , Proteínas S100/metabolismo , Estômago , Vimentina/metabolismo , Adulto Jovem
11.
Food Res Int ; 116: 354-361, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30716956

RESUMO

The objective of this study was to unravelling the possible mechanism underlying drip loss and identify the protein markers associated with water holding capacity (WHC) of meat. Pectoralis major muscles from geese were assigned to high and low drip loss groups. The physio-chemical properties and proteome profiles were compared between these two groups. Label-free quantitative mass spectrometry was applied to investigate the differentially expressed proteins, and the results were confirmed with parallel reaction monitoring (PRM). 21 differential proteins were identified in abundance between high and low drip loss groups, and they fall generally into the structural proteins, metabolic enzymes, antioxidant enzymes and stress response proteins. The results may provide a perspective to the discovery of new biomarkers for drip loss.


Assuntos
Gansos , Proteínas Musculares/metabolismo , Músculos Peitorais/metabolismo , Proteômica , Água/análise , Animais , Culinária , Desmina/metabolismo , Glutationa Redutase/metabolismo , Gliceraldeído-3-Fosfato Desidrogenase (Fosforiladora)/metabolismo , Proteínas de Choque Térmico/metabolismo , Concentração de Íons de Hidrogênio , Queratinas/metabolismo , Cadeias Leves de Miosina/metabolismo , Superóxido Dismutase/metabolismo , Espectrometria de Massas em Tandem
12.
Respir Res ; 20(1): 31, 2019 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-30764835

RESUMO

BACKGROUND: The pathophysiology of obstruction and swallowing dysfunction in snores and sleep apnea patients remains unclear. Neuropathy and to some extent myopathy have been suggested as contributing causes. Recently we reported an absence and an abnormal isoform of two cytoskeletal proteins, desmin, and dystrophin, in upper airway muscles of healthy humans. These cytoskeletal proteins are considered vital for muscle function. We aimed to investigate for muscle cytoskeletal abnormalities in upper airways and its association with swallowing dysfunction and severity of sleep apnea. METHODS: Cytoskeletal proteins desmin and dystrophin were morphologically evaluated in the uvula muscle of 22 patients undergoing soft palate surgery due to snoring and sleep apnea and in 10 healthy controls. The muscles were analysed with immunohistochemical methods, and swallowing function was assessed using videoradiography. RESULTS: Desmin displayed a disorganized pattern in 21 ± 13% of the muscle fibres in patients, while these fibers were not present in controls. Muscle fibres lacking desmin were present in both patients and controls, but the proportion was higher in patients (25 ± 12% vs. 14 ± 7%, p = 0.009). The overall desmin abnormalities were significantly more frequent in patients than in controls (46 ± 18% vs. 14 ± 7%, p < 0.001). In patients, the C-terminus of the dystrophin molecule was absent in 19 ± 18% of the desmin-abnormal muscle fibres. Patients with swallowing dysfunction had 55 ± 10% desmin-abnormal muscle fibres vs. 22 ± 6% in patients without swallowing dysfunction, p = 0.002. CONCLUSION: Cytoskeletal abnormalities in soft palate muscles most likely contribute to pharyngeal dysfunction in snorers and sleep apnea patients. Plausible causes for the presence of these abnormalities is traumatic snoring vibrations, tissue stretch or muscle overload.


Assuntos
Desmina/metabolismo , Distrofina/metabolismo , Músculos Respiratórios/metabolismo , Síndromes da Apneia do Sono/metabolismo , Ronco/metabolismo , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Citoesqueleto/patologia , Transtornos de Deglutição/metabolismo , Transtornos de Deglutição/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Fibras Musculares de Contração Rápida/metabolismo , Fibras Musculares de Contração Rápida/patologia , Fibras Musculares de Contração Lenta/metabolismo , Fibras Musculares de Contração Lenta/patologia , Palato Mole/metabolismo , Palato Mole/patologia , Músculos Respiratórios/patologia , Síndromes da Apneia do Sono/patologia , Ronco/patologia , Úvula/metabolismo , Úvula/patologia , Adulto Jovem
13.
Dev Biol ; 448(1): 36-47, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30695685

RESUMO

Joubert syndrome (JBTS) is a predominantly autosomal recessive neurodevelopmental disorder that presents with characteristic malformations of the cerebellar vermis, superior cerebellar peduncles and midbrain in humans. Accompanying these malformations are a heterogeneous set of clinical symptoms, which frequently include deficits in motor and muscle function, such as hypotonia (low muscle tone) and ataxia (clumsiness). These symptoms are attributed to improper development of the hindbrain, but no direct evidence has been reported linking these in JBTS. Here, we describe muscle developmental defects in a mouse with a targeted deletion of the Abelson helper integration site 1 gene, Ahi1, one of the genes known to cause JBTS in humans. While FVB/NJ Ahi1-/- mice display no gross malformations of the cerebellum, deficits are observed in several measures of motor function, strength, and body development. Specifically, Ahi1-/- mice show delayed physical development, delays in surface reflex righting as neonates, and reductions in grip strength and spontaneous locomotor activity as adults. Additionally, Ahi1-/- mice showed evidence of muscle-specific contributions to this phenotype, such as reductions in 1) myoblast differentiation potential in vitro, 2) muscle desmin expression, and 3) overall muscle mass, myonuclear domain, and muscle fiber cross-sectional area. Together, these data suggest that loss of Ahi1 may cause abnormalities in the differentiation of myoblasts to mature muscle cells. Moreover, Ahi1 loss impacts muscle development directly, outside of any indirect impact of cerebellar malformations, revealing a novel myogenic cause for hypotonia in JBTS.


Assuntos
Anormalidades Múltiplas/embriologia , Diferenciação Celular , Cerebelo/anormalidades , Anormalidades do Olho/embriologia , Doenças Renais Císticas/embriologia , Desenvolvimento Muscular , Transtornos do Neurodesenvolvimento/metabolismo , Proteínas Proto-Oncogênicas/deficiência , Retina/anormalidades , Anormalidades Múltiplas/genética , Anormalidades Múltiplas/patologia , Proteínas Adaptadoras de Transporte Vesicular , Animais , Cerebelo/embriologia , Cerebelo/patologia , Desmina/genética , Desmina/metabolismo , Anormalidades do Olho/genética , Anormalidades do Olho/patologia , Doenças Renais Císticas/genética , Doenças Renais Císticas/patologia , Locomoção/genética , Camundongos , Camundongos Knockout , Força Muscular/genética , Mioblastos/metabolismo , Mioblastos/patologia , Transtornos do Neurodesenvolvimento/genética , Transtornos do Neurodesenvolvimento/patologia , Proteínas Proto-Oncogênicas/metabolismo , Reflexo de Endireitamento/genética , Retina/embriologia , Retina/patologia
15.
Mol Biol Cell ; 30(5): 579-590, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30601711

RESUMO

Clathrin plaques are stable features of the plasma membrane observed in several cell types. They are abundant in muscle, where they localize at costameres that link the contractile apparatus to the sarcolemma and connect the sarcolemma to the basal lamina. Here, we show that clathrin plaques and surrounding branched actin filaments form microdomains that anchor a three-dimensional desmin intermediate filament (IF) web. Depletion of clathrin plaque and branched actin components causes accumulation of desmin tangles in the cytoplasm. We show that dynamin 2, whose mutations cause centronuclear myopathy (CNM), regulates both clathrin plaques and surrounding branched actin filaments, while CNM-causing mutations lead to desmin disorganization in a CNM mouse model and patient biopsies. Our results suggest a novel paradigm in cell biology, wherein clathrin plaques act as platforms capable of recruiting branched cortical actin, which in turn anchors IFs, both essential for striated muscle formation and function.


Assuntos
Actinas/metabolismo , Clatrina/metabolismo , Músculo Esquelético/metabolismo , Animais , Desmina/metabolismo , Dinamina II/metabolismo , Humanos , Filamentos Intermediários/metabolismo , Filamentos Intermediários/ultraestrutura , Camundongos Knockout , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/ultraestrutura , Mutação/genética , Miopatias Congênitas Estruturais/genética , Proteína da Síndrome de Wiskott-Aldrich/metabolismo
16.
Int Urogynecol J ; 30(3): 465-476, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29564513

RESUMO

INTRODUCTION AND HYPOTHESIS: An animal model of vaginal distention (VD) was developed to reproduce the acute urethral injury and deficiency underlying stress urinary incontinence (SUI). Data on the chronic effects of urethral trauma and the recovery process are still scarce. We investigated acute, short- and long-term histomorphological and molecular changes in the urethra of rats post 12-h intermittent VD. METHODS: We evaluated the urethra of four groups of female rats (n = 72): control without trauma, 1 h, 7 days and 30 days post VD. We compared the gene and protein expression of the VEGF and NGF growth factors, collagens (COL1a1 and COL3a1), desmin, smooth muscle myosin (MYH11), skeletal muscle myosins (MYH1, MYH2 and MYH3) and cell proliferation marker MKi67. We used real-time RT-qPCR, and immunohistochemistry. RESULTS: Histology showed urethral damage after VD mainly involving the muscular layers. VEGF, NGF, desmin and MKi67 mRNA were significantly upregulated in the urethras of rats 1-h post VD compared with controls (P < 0.05 for all). By 7 days post trauma, COL1a1, MYH11 and MYH3 genes were overexpressed compared with controls (p < 0.05 for all). The COL3a1 protein level was increased by 2.6 times by day 7, while MYH2 protein was significantly decreased (around two times) from 7 to 30 days post VD compared with controls (p < 0.05 for both). CONCLUSIONS: The 12-h intermittent VD causes chronic alterations in the urethra represented by increased COL3a1 and decreased MYH2 protein levels in the long term. The model can potentially be used to study the mechanisms of urethral injury and recovery as well as the physiopathology of SUI.


Assuntos
Uretra/metabolismo , Uretra/patologia , Incontinência Urinária por Estresse/metabolismo , Incontinência Urinária por Estresse/patologia , Animais , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo III/genética , Colágeno Tipo III/metabolismo , Desmina/genética , Desmina/metabolismo , Modelos Animais de Doenças , Feminino , Antígeno Ki-67/genética , Antígeno Ki-67/metabolismo , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Fator de Crescimento Neural/genética , Fator de Crescimento Neural/metabolismo , RNA Mensageiro/metabolismo , Ratos , Fatores de Tempo , Uretra/lesões , Incontinência Urinária por Estresse/genética , Vagina , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
17.
Am J Med Sci ; 357(1): 16-22, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30466735

RESUMO

BACKGROUND: This study aimed to investigate the correlation between pancreatic stellate cell activation, matrix metallopeptidase 2 (MMP2) expression and lymph node metastasis in pancreatic carcinoma. METHODS: Alpha-smooth muscle actin (ACTA2), Desmin (DES) and MMP2 were detected in 40 pancreatic carcinoma patients and 10 cases of normal pancreas tissues using immunohistochemistry. Then MMP2 and ACTA2 expression profiles in pancreatic cancer were obtained from UCSC (University of California, Santa Cruz) and SurvExpress. RESULTS: A total of 67.5% and 55.0% of cases positively expressed ACTA2 and DES in pancreatic carcinoma, respectively. MMP2 in pancreatic carcinoma was expressed in 55.0% of cases, and there were significant differences between the lymph node metastasis group and the lymph node nonmetastasis group, as well as invasion and noninvasion to the peripheral tissue group (P < 0.01). High throughput sequencing databases verified that ACTA2 and MMP2 gene expression were both upregulated in pancreatic carcinoma tissues. CONCLUSIONS: The coexpression of ACTA2 and DES was related to the expression of MMP2, and positively correlated with lymph node metastasis. Activation of pancreatic stellate cells may promote the expression of MMP2 and enhance the invasion and metastasis of pancreatic carcinoma.


Assuntos
Actinas/genética , Desmina/genética , Metástase Linfática/patologia , Metaloproteinase 2 da Matriz/genética , Pâncreas/patologia , Neoplasias Pancreáticas/patologia , Células Estreladas do Pâncreas/fisiologia , Actinas/metabolismo , Adulto , Idoso , Desmina/metabolismo , Feminino , Humanos , Metástase Linfática/genética , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Pessoa de Meia-Idade , Neoplasias Pancreáticas/genética
18.
J Investig Med ; 67(1): 11-19, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30097466

RESUMO

Desmin expression depends on desmin messenger RNA (mRNA) and ubiquitin proteasome system. This process is poorly understood in dilated cardiomyopathy. The aim of the study was to investigate whether changes of desmin mRNA and ubiquitin expression correlate with types of desmin expression in cardiomyocytes. Endomyocardial biopsy was performed in 60 patients (85% men, mean age 46±14 years) with heart failure (HF; left ventricular ejection fraction <45%). Desmin and ubiquitin expression were analysed in histological sections by immunohistochemistry and in Western blot. Desmin mRNA expression was determined by fluorescent in situ hybridization methods. In patients with weak/even desmin expression, weak/even expression of ubiquitin in the cytosol and low desmin mRNA expression in the cytosol and nuclei of cardiomyocytes were observed. Expression of ubiquitin and desmin mRNA increased along with the progression of desmin cytoskeleton remodeling. Desmin mRNA and ubiquitin were weakly expressed/absent in cardiomyocytes with low/lack of desmin expression. Variations in desmin mRNA, desmin and ubiquitin expression were associated with gradual changes in myocardial structure and clinical parameters. To conclude, changes in ubiquitin and desmin mRNA expression are related to patterns of desmin expression. An increase in the expression of ubiquitin and desmin mRNA may be a protective feature against unfavorable cell remodeling. This may reduce the adverse effects of cytoskeleton damage in the early stages of HF. Low/lack ubiquitin and/or desmin mRNA expression may be markers of end-stage HF.


Assuntos
Cardiomiopatia Dilatada/genética , Desmina/genética , Regulação da Expressão Gênica , Ubiquitina/genética , Cardiomiopatia Dilatada/diagnóstico por imagem , Cardiomiopatia Dilatada/patologia , Desmina/metabolismo , Feminino , Fibrose , Humanos , Masculino , Pessoa de Meia-Idade , Miócitos Cardíacos/metabolismo , Miócitos Cardíacos/patologia , Peptídeo Natriurético Encefálico/metabolismo , Fragmentos de Peptídeos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ubiquitina/metabolismo
19.
Gen Comp Endocrinol ; 273: 236-248, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30292702

RESUMO

The broad-snouted caiman (Caiman latirostris) is a crocodilian species that inhabits South American wetlands. As in all other crocodilians, the egg incubation temperature during a critical thermo-sensitive window (TSW) determines the sex of the hatchlings, a phenomenon known as temperature-dependent sex determination (TSD). In C. latirostris, we have shown that administration of 17-ß-estradiol (E2) during the TSW overrides the effect of the male-producing temperature, producing phenotypic females (E2SD-females). Moreover, the administration of E2 during TSW has been proposed as an alternative way to improve the recovery of endangered reptile species, by skewing the population sex ratio to one that favors females. However, the ovaries of E2SD-female caimans differ from those of TSD-females. In crocodilians, the external genitalia (i.e. clitero-penis structure or phallus) are sexually dimorphic and hormone-sensitive. Despite some morphological descriptions aimed to facilitate sexing, we found no available data on the C. latirostris phallus histoarchitecture or hormone dependence. Thus, the aims of this study were: (1) to establish the temporal growth pattern of the phallus in male and female caimans; (2) to evaluate histo-morphological features and the expression of estrogen receptor alpha (ERα) and androgen receptor (AR) in the phallus of male and female pre-pubertal juvenile caimans; and (3) to determine whether the phallus of TSD-females differs from the phallus of E2SD-females. Our results demonstrated sexually dimorphic differences in the size and growth dynamics of the caiman external genitalia, similarities in the shape and spatial distribution of general histo-morphological compartments, and sexually dimorphic differences in innervation, smooth muscle fiber distribution, collagen organization, and ERα and AR expressions. The external genitalia of E2SD-females differed from that of TSD-females in many histological features and in the expression of ERα and AR, resembling patterns described in males. Our results alert on the effects of estrogen agonist exposure during TSW and suggest that caution must be taken regarding the use of E2SD as a procedure for wildlife population management.


Assuntos
Jacarés e Crocodilos/fisiologia , Genitália Feminina/fisiologia , Hormônios Esteroides Gonadais/metabolismo , Processos de Determinação Sexual , Temperatura Ambiente , Jacarés e Crocodilos/anatomia & histologia , Animais , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Desmina/metabolismo , Feminino , Genitália Feminina/citologia , Genitália Feminina/crescimento & desenvolvimento , Masculino , Receptores de Superfície Celular/metabolismo
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