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1.
Int. j. morphol ; 37(4): 1493-1497, Dec. 2019. tab
Artigo em Espanhol | LILACS | ID: biblio-1040159

RESUMO

Uno de los principales factores genéticos que influenciarían el rendimiento muscular humano es el gen ACTN3, que codifica la proteína estructural del sarcómero α-actinina-3. El polimorfismo R577X (rs1815739) del gen ACTN3 ha sido asociado con varios indicadores de rendimiento muscular y físico en deportistas y población general, pero este fenómeno ha sido escasamente descrito en poblaciones de Latinoamérica y Chile. Por lo tanto, el objetivo del presente estudio fue describir la frecuencia genotípica y distribución alélica de los genotipos de ACTN3 R577X en deportistas universitarios chilenos. 129 deportistas universitarios chilenos representantes de diferentes selecciones deportivas (halterofilia, balonmano, voleibol, rugby, basquetbol, futbol y futsal) participaron como voluntarios. Los análisis moleculares del polimorfismo R577X del gen ACTN3 fueron realizados mediante reacción en cadena de la polimerasa (PCR) y restricción enzimática (RFLP). La distribución de genotipos del polimorfismo ACTN3 R577X fue RR: 34,8 % (n=45), RX: 50,4 % (n=65), XX: 14,7 % (n=19), y la frecuencia relativa de alelos fue R: 0,601 y X: 0,399. Además, se encontró asociación entre distribución de genotipos (c2= 12,26; 2 gl; p=0,002) y frecuencia relativa de alelos (c2= 11.02; 1 gl; p=0.0009) con el sexo de los participantes. Sin embargo, no hubo asociación al realizar análisis por tipo de deporte practicado. Los hallazgos de la presente investigación sugieren que el polimorfismo R577X del gen ACTN3 está asociado con el sexo en deportistas universitarios chilenos. Además, estos resultados describen de forma inédita la distribución genotípica y frecuencia alélica de esta variante genética en población chilena, mostrando una distribución similar a otros estudios realizados en poblaciones de deportistas en Brasil, Rusia, Estados Unidos y Turquía. No obstante, también muestra diferencias con otras poblaciones generales y de deportistas.


One of the main genetic factors that influence the muscular performance is the gene that encodes the structural protein α-actinin-3 (ACTN3). The R577X polymorphism (rs1815739) of ACTN3 has been associated with indicators of muscle and physical performance in athletes and general population, but this has been scarcely described in the Latin American and Chilean population. Thus, the aim of the present study was to describe the genotypic frequency and allelic distribution of ACTN3 R577X genotypes in college athletes. A total of 129 unrelated Chilean college athletes representing various sport disciplines (weightlifting, handball, volleyball, rugby, basketball, soccer and futsal) were volunteered for the study. ACTN3 R577X gene polymorphism was analysed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For the total sample the genotypes distribution for R577X polymorphism was RR: 34.8 % (n=45), RX: 50.4 % (n=65), XX: 14.7 % (n=19), and the relative frequency of alleles was R: 0,601 and X: 0,399. Moreover, an association was found between genotype distribution (c2= 12.26; 2 df; p=0.002) and allele frequencies (c2= 11.02; 1 df; p=0.0009) with the sex of the participants. However, there were no associations when performing analysis by type of sports. These findings suggest that the R577X polymorphism of the ACTN3 gene is associated with sex in Chilean college athletes. Furthermore, these results describe in an unprecedented manner, the genotypic distribution and allelic frequency of this genetic variant in Chilean population, showing a similar distribution to other studies conducted in populations of athletes in Brazil, Russia, the United States and Turkey. However, it also shows differences with other general and athletes populations.


Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Adulto Jovem , Polimorfismo Genético , Estudantes , Actinas/genética , Atletas , Universidades , Chile , Desempenho Atlético/fisiologia
2.
Int. j. morphol ; 36(3): 835-840, Sept. 2018. graf
Artigo em Inglês | LILACS | ID: biblio-954194

RESUMO

Tyrosine phosphorylated proteins have been localized and identified in male reproductive tissues such as testis and capacitated/ acrosome reacted sperm except epididymis. The changes of such proteins are associated with decreased sperm quality of valproic acid treatment. This study aimed to investigate the presence and alterations of protein phosphorylation in epididymal epithelium and fluid of rats treated VPA. Sixteen adult male rats were divided into control and VPA-treated groups (n=8/ each). Treated rats were injected with VPA (500 mg/ kgBW, intraperitoneally) for 10 consecutive days. At the end of experiment, the monoclonal antiphosphotyrosine (clone 4G10) was used for immunohistochemistry to probe tyrosine phosphorylated proteins and also to examine the expression of such proteins using immuno-Western blotting in epididymal tissue and fluid. The result showed that positive reactivity of phosphorylated proteins was clearly observed in cytoplasmic principle cells, nuclei of apical & basal cells and sperm mass surrounded with epididymal fluids. The profiles of phosphorylated proteins in epididymal fluid were 182, 127, 80, 70, 57, 45, 34, and 31 kDas, respectively. Interestingly, VPA affected the changes of phosphorylated proteins and β actin in head, body, and tail epididymal fluids. We conclude that tyrosine phosphorylated proteins were detected in epididymal epithelium and fluid. The expressions of those proteins and actin were altered under VPA treating.


Las proteínas tirosina fosforiladas han sido localizadas e identificadas en tejidos reproductores masculinos tales como testículos y espermatozoides, capacitados a nivel acrosómico, excepto en el epidídimo. Los cambios de estas proteínas están asociadas con una disminución de la calidad del esperma en el tratamiento con ácido valproico (AVP). Este estudio tuvo como objetivo investigar la presencia y las alteraciones de la fosforilación de proteínas en el epitelio epididimal y en el fluido espermático de ratas tratadas con AVP. Dieciséis ratas macho adultas se dividieron en dos grupos: control y tratadas con AVP (n = 8 / cada uno). A las ratas tratadas se les inyectó AVP por vía intraperitoneal (500 mg / kg de peso corporal) durante 10 días consecutivos. Al final del experimento, se realizó inmunohistoquímica con la anti-fosfotirosina monoclonal (clon 4G10) para sondear las proteínas tirosina fosforiladas y también para examinar la expresión de tales proteínas usando inmunotransferencia Western, en tejido y fluido epididimarios. El resultado mostró reactividad positiva de proteínas fosforiladas en células citoplásmicas principales, en los núcleos de las células apicales y basales y en la masa de esperma rodeada por fluidos epididimarios. Los perfiles de proteínas fosforiladas en el fluido epididimal fueron 182, 127, 80, 70, 57, 45, 34 y 31 kDas, respectivamente. El AVP provocó cambios en las proteínas fosforiladas y en la β actina de los fluidos epididimarios de cabeza, cuerpo y cola del epidídimo. Concluimos que las proteínas tirosina fosforiladas se detectaron en el epitelio y el fluido epididimarios. Las expresiones de esas proteínas y de la β actina se alteraron bajo tratamiento con AVP.


Assuntos
Animais , Masculino , Ratos , Fosfoproteínas/efeitos dos fármacos , Tirosina/efeitos dos fármacos , Ácido Valproico/administração & dosagem , Actinas/efeitos dos fármacos , Anticonvulsivantes/administração & dosagem , Fosfoproteínas/metabolismo , Fosforilação , Tirosina/metabolismo , Imuno-Histoquímica , Western Blotting , Actinas/metabolismo , Ratos Sprague-Dawley , Fosfotirosina , Epididimo
3.
Acta cir. bras ; 33(7): 629-640, July 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-949364

RESUMO

Abstract Purpose: To investigate the effect of melatonin on uterine tissue in the ovariectomized rat model. Methods: Fourty Wistar albino rats were divided into four groups for histologic and immunohistochemical examination. The rats were first numbered randomly and then randomly divided into 4 equal groups: control (group 1), torsion (group 2), torsion+detorsion (group 3) and torsion+detorsion+melatonin (group 4) groups. In addition, four Wistar albino rats were used for western blot analysis in each group. And also, malondialdehyde (MDA) levels were measured biochemically in all rats. Results: The histopathological examination of the uterine tissue in rats ovarectomized showed a degeneration in uterine glands, dilation of blood vessels in the internal layer with a thrombosis and bleeding, abnormal nucleuses and vacuolated cytoplasm above and below the nucleus. In torsion group, the apoptotic cells increased in luminal epithelium and gland cells. In the melatonin group showed that the Bcl2 negative effect on the uterine epithelium and did not lead to apoptotic cells. Conclusion: The increase in vascular endothelial growth factor expression resulted in the rearrangement of endothelial cell growth and the induction of angiogenesis.


Assuntos
Animais , Feminino , Útero/efeitos dos fármacos , Útero/patologia , Estro/efeitos dos fármacos , Genes bcl-2/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/análise , Melatonina/farmacologia , Antioxidantes/farmacologia , Imuno-Histoquímica , Ovariectomia , Distribuição Aleatória , Western Blotting , Actinas/análise , Fator A de Crescimento do Endotélio Vascular/efeitos dos fármacos , Malondialdeído/análise
4.
An. bras. dermatol ; 93(2): 262-264, Mar.-Apr. 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-887177

RESUMO

Abstract: Leiomyosarcoma is a rare skin tumor, most common in white men in the fifth to eighth decades of life. Primary tumors are classified in dermal or subcutaneous, that differ by clinical and prognostic features. They may appear on any site of the body, but are rare on the face. A 54-year-old female was admitted with a 5cm exophytic nodular lesion of 8 months duration on the right cheek, site of previous chronic radiodermatitis. Histopathology revealed spindle-shaped cell neoplasia, positive for smooth muscle actin on immunohistochemistry. Cutaneous leiomyosarcomas on the face are rare and may occur in previously irradiated areas. Immunohistochemistry is mandatory for an accurate diagnosis. Its similarity with other tumors may complicate the diagnosis, with delay expansion of the tumor.


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Neoplasias Cutâneas/patologia , Neoplasias Faciais/patologia , Leiomiossarcoma/patologia , Neoplasias Cutâneas/diagnóstico , Neoplasias Faciais/diagnóstico , Imuno-Histoquímica , Actinas/análise , Doenças Raras/patologia , Diagnóstico Diferencial , Leiomiossarcoma/diagnóstico , Músculo Liso/patologia
5.
Braz. oral res. (Online) ; 32: e004, 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889501

RESUMO

Abstract Radiation combined injury, a life-threatening condition, has higher mortality than simple radiation injury. The aim of the present study was to analyze the efficiency of Aloe vera and silver nanoparticles in improving the healing of ulcerated oral mucosa after irradiation. Thirty male Albino mice were divided into five groups: control, radiation, Aloe vera (AV), silver nanoparticles (NS), and AV+NS. The mice were exposed to whole body 6Gy gamma-radiation. After one hour, 20% acetic acid was injected into the submucosal layer of the lower lip for ulcer induction. The animals received topical treatment with the assigned substances for 5 days. Lip specimens were subjected to hematoxylin and eosin and anti alpha-smooth muscle actin immunohistochemical staining. Results demonstrated occurance of ulcer three days post irradiation in all groups except in the AV+NS group where only epithelial detachment was developed. After seven days, data revealed persistent ulcer in radiation group, and almost normal epithelium in the AV+NS group. A significant reduction of epithelial thickness was detected in all groups at the third day as compared to control. At the seventh day, only the AV+NS group restored the epithelial thickness. Area percent of alpha-smooth muscle actin expression was significantly decreased in radiation group at the third day followed by significant increase at the seventh day. However, all treatment groups showed significant increase in alpha-smooth muscle actin at the third day, which decreased to normal level at the seventh day. Our study demonstrated the efficiency of Aloe vera and silver nanoparticles in enhancing ulcer healing after irradiation.


Assuntos
Animais , Masculino , Camundongos , Aloe/química , Raios gama/efeitos adversos , Nanopartículas Metálicas/uso terapêutico , Úlceras Orais/tratamento farmacológico , Úlceras Orais/etiologia , Lesões Experimentais por Radiação/tratamento farmacológico , Prata/uso terapêutico , Ácido Acético , Actinas/análise , Administração Tópica , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/efeitos da radiação , Fibroblastos/efeitos dos fármacos , Imuno-Histoquímica , Microscopia Eletrônica de Transmissão , Úlceras Orais/patologia , Lesões Experimentais por Radiação/patologia , Valores de Referência , Reprodutibilidade dos Testes , Fatores de Tempo , Resultado do Tratamento , Cicatrização/efeitos dos fármacos
6.
Arq. bras. med. vet. zootec. (Online) ; 69(6): 1398-1406, nov.-dez. 2017. ilus, tab
Artigo em Português | LILACS, VETINDEX | ID: biblio-909825

RESUMO

A endometrose é uma alteração degenerativa das glândulas uterinas e do estroma circundante, caracterizada pelo arranjo periglandular de miofibroblastos e pela deposição de matriz extracelular (ECM). O presente trabalho objetivou avaliar a expressão de colágenos tipos I, III e IV e α-actina de músculo liso (α-SMA) nas endometroses equinas, procurando esclarecer a participação dos miofibroblastos na progressão desses processos. Foram utilizadas 24 biópsias uterinas com diagnóstico de endometrose, recebidas pelo Serviço de Patologia Veterinária e de Reprodução Animal da FMVZ, Unesp, Botucatu, SP. Cortes histológicos foram submetidos às técnicas histoquímicas de tricrômico de Masson, picrosirius red sob luz polarizada e ácido periódico de Schiff (PAS) e imuno-histoquímicas para os três tipos de colágeno citados e α-SMA. Ainda, traçou-se um paralelo entre a técnica de picrosirius red e a imunomarcação dos colágenos tipos I e III. A análise histológica revelou que as fibras de colágeno denso correspondem ao colágeno tipo I, predominantes nas endometroses inativa e inativa destrutiva. As fibras de colágeno frouxo correspondem ao colágeno tipo III, predominantes nas endometroses ativas e ativas destrutivas. Nesses mesmos processos, a membrana basal revelou espessamento, aparentemente não relacionado ao colágeno tipo IV, e uma maior imunomarcação de miofibroblastos periglandulares em relação às endometroses inativa e inativa destrutiva. Dessa forma, nota-se que os miofibroblastos estão relacionados ao aumento na deposição de colágeno tipo III nos ninhos fibróticos ativos.(AU)


Endometriosis is a degenerative change of the uterine glands and surrounding stroma, characterized by periglandular arrangement of myofibroblasts and deposition of extracellular matrix (ECM). The aim of this study was to evaluate the expression of collagen type I, III and IV and α-smooth muscle actin (α-SMA) in equine endometriosis, and investigate the role of myofibroblasts in the progression of these processes. A parallel was made with histochemical techniques of Masson's trichrome, Picrosirius Red under polarized light and Periodic Acid-Schiff (PAS). Twenty four uterine biopsies received by the Veterinary Pathology Service and Animal Reproduction of FMVZ, UNESP, Botucatu, SP, were diagnosed with endometriosis. Histological analysis revealed that the orange dense collagen fibers correspond to type I collagen, being prevalent in inactive and inactive destructive endometriosis. The green loose collagen fibers correspond to type III collagen, and are predominant in active and active destructive endometriosis. In the same processes, a greater amount of periglandular myofibroblasts were observed in comparison to inactive and inactive destructive endometriosis. The presence of these cells in active processes are strongly related to an increased deposition of collagen type III in fibrotic nests. Regarding the basement membrane, the active destructive and active endometriosis shows thickening, apparently not related to an increase in expression of type IV collagen. The active destructive and inactive destructive endometriosis exhibited disruption areas in type IV collagen fibers. Thus, it is noted that the myofibroblasts are related to increased deposition of type III collagen in active fibrotic nests.(AU)


Assuntos
Animais , Feminino , Actinas/análise , Colágeno Tipo III/análise , Colágeno Tipo IV/análise , Colágeno Tipo I/análise , Endometriose/fisiopatologia , Cavalos , Miofibroblastos , Imuno-Histoquímica/veterinária
7.
Rev. Col. Bras. Cir ; 44(4): 367-373, jul.-ago. 2017. tab, graf
Artigo em Português | LILACS | ID: biblio-896591

RESUMO

RESUMO Objetivo: avaliar a eficácia de três marcadores imunoistoquímicos envolvidos no processo de cicatrização de ferida cirúrgica. Métodos: estudo experimental em 40 ratos da raça Wistar, dos marcadores metaloproteinases e metaloproteinase da matriz 9 (MMP-9), fator de transformação do crescimento beta (TGF-β) e miofibroblasto e alfa actina de músculo liso (α-AML), estudados a partir de fragmentos de cicatriz cirúrgica de incisão abdominal envolvendo pele, aponeurose e peritônio. Os animais foram distribuídos em quatro subgrupos de dez de acordo com o dia da morte, programada em três, sete, 14 e 21 dias. Resultados: na expressão da MMP-9 ocorreu aumento progressivo de sua concentração, mais evidente do 7º ao 14º dias variando a imuno-expressão tecidual entre 2,65% e 11,50%.TGF- β mostrou expressão em nível alto no 3º dia, caiu no 7º, voltando a subir no 14º, com pequena queda no 21º dia variando a imuno-expressão tecidual entre 0,03% e 2,92%. A α-AML apresentou níveis com pouca variação e discreto aumento variando a imuno-expressão tecidual entre 0,88% e 3,23%. Conclusão: a MMP-9 se apresentou como melhor marcador, seguido pela TGF-β. Já o α-AML não se mostrou um bom sinalizador da evolução da reparação tissular.


ABSTRACT Objective: to evaluate the efficacy of three immunohistochemical markers involved in the wound healing process. Methods: experimental study of 40 Wistar rats of the markers metalloproteinases and matrix metalloproteinase 9 (MMP-9), beta transforming growth factor (TGF-β) and myofibroblasts and smooth muscle actin alpha (α-MLA) markers, studied from fragments of surgical scar of abdominal incision involving skin, aponeurosis and peritoneum. The animals were divided into four subgroups of ten according to the day of death, scheduled in three, seven, 14 and 21 days. Results: MMP-9 expression showed a progressive increase of its concentration, more evident from 7th to 14th days, varying the tissue immunoexpression between 2.65% and 11.50% . TGF- β showed expression at high level on the 3rd day, fell in the 7th, rising again in the 14th, with a small decrease in the 21st day, varying the tissue immunoexpression between 0.03% and 2.92%. The α-AML presented levels with little variation and a slight increase, varying the tissue immunoexpression between 0.88% and 3.23%. Conclusion: MMP-9 presented as the best marker, followed by TGF-β. However, α-AML was not a good indicator of the evolution of tissue repair.


Assuntos
Animais , Masculino , Ratos , Cicatrização , Fator de Crescimento Transformador beta/análise , Actinas/análise , Metaloproteinase 9 da Matriz/biossíntese , Ferida Cirúrgica/imunologia , Ferida Cirúrgica/patologia , Imuno-Histoquímica , Biomarcadores/análise , Fator de Crescimento Transformador beta/fisiologia , Actinas/fisiologia , Ratos Wistar , Metaloproteinase 9 da Matriz/fisiologia
8.
Acta cir. bras ; 31(1): 8-14, Jan. 2016. graf
Artigo em Inglês | LILACS | ID: lil-771854

RESUMO

PURPOSE: To investigate the protective effects of dexmedetomidine (Dex) against renal ischemia/reperfusion injury (IRI). METHODS: Sprague-Dawley rats were randomly divided to sham group, IRI group and Dex group. The SD rats were subjected to 45 min of ischemia followed by eight weeks of reperfusion. Prior to ischemia, rats were either treated with Dex or not. Blood samples were collected for the detection of blood urea nitrogen (BUN) and creatinine (Cr) levels. Immunohistochemistry was performed for CD3 T-cell infiltrates. Real-time PCR and western blot were detected for the expression of TNF-α, IL-1β, ICAM-1, HMGB1 and TLR4. RESULTS: Compared with sham group, renal IRI significantly increased the serum levels of BUN and Cr. The H&E staining indicated that renal IRI resulted in obvious renal injury and immunohistochemistry found that there were more CD3 T-cell infiltrates in IRI group. Also, renal IRI upregulated the expression of TNF-α, IL-1β, ICAM-1, HMGB1 and TLR4. However, all these changes were alleviated by the treatment with Dex. CONCLUSIONS: Dexmedetomidine has beneficial effects on long term inflammation induced by renal ischemia/reperfusion injury. Its mechanisms may be achieved through inhibiting the HMGB1/TLR4 pathway to exert protective effects.


Assuntos
Animais , Masculino , Lesão Renal Aguda/patologia , /farmacologia , Dexmedetomidina/farmacologia , Rim/irrigação sanguínea , Traumatismo por Reperfusão/complicações , Actinas/análise , Lesão Renal Aguda/etiologia , Lesão Renal Aguda/metabolismo , Nitrogênio da Ureia Sanguínea , Western Blotting , Creatinina/sangue , Proteína HMGB1/análise , Imuno-Histoquímica , Inflamação/etiologia , Inflamação/metabolismo , Molécula 1 de Adesão Intercelular/análise , Interleucina-1beta/análise , Rim/química , Distribuição Aleatória , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , RNA , Traumatismo por Reperfusão/patologia , /análise , Fator de Necrose Tumoral alfa/análise
9.
Braz. j. med. biol. res ; 49(10): e5526, 2016. graf
Artigo em Inglês | LILACS | ID: lil-792523

RESUMO

Pseudomonas aeruginosa is one of the common colonizing bacteria of the human body and is an opportunistic pathogen frequently associated with respiratory infections. Inactivated P. aeruginosa (IPA) have a variety of biological effects against inflammation and allergy. Transforming growth factor-β (TGF-β) signaling plays a critical role in the regulation of cell growth, differentiation, and development in a wide range of biological systems. The present study was designed to investigate the effects of IPA on TGF-β/Smad signaling in vivo, using a hypoxia-induced pulmonary hypertension (PH) rat model. Sprague Dawley rats (n=40) were exposed to 10% oxygen for 21 days to induce PH. At the same time, IPA was administered intravenously from day 1 to day 14. Mean pulmonary artery pressure (mPAP) and the right ventricle (RV) to left ventricle plus the interventricular septum (LV+S) mass ratio were used to evaluate the development of PH. Vessel thickness and density were measured using immunohistochemistry. Primary arterial smooth muscle cells (PASMCs) were isolated and the proliferation of PASMCs was assayed by flow cytometry. The production of TGF-β1 in cultured supernatant of PASMCs was assayed by ELISA. The expression levels of α-smooth muscle actin (α-SMA), TGF-β1 and phospho-Smad 2/3 in PASMCs were assayed by western blot. Our data indicated that IPA attenuated PH, RV hypertrophy and pulmonary vascular remodeling in rats, which was probably mediated by restraining the hypoxia-induced overactive TGF-β1/Smad signaling. In conclusion, IPA is a promising protective treatment in PH due to the inhibiting effects on TGF-β1/Smad 2/3 signaling.


Assuntos
Animais , Masculino , Hipertensão Pulmonar/metabolismo , Hipertensão Pulmonar/prevenção & controle , Hipóxia/metabolismo , Miócitos de Músculo Liso/fisiologia , Pseudomonas aeruginosa/fisiologia , Proteínas Smad/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Actinas/análise , Actinas/metabolismo , Western Blotting , Proliferação de Células/fisiologia , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Hipertensão Pulmonar/etiologia , Hipóxia/complicações , Imuno-Histoquímica , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Transdução de Sinais/fisiologia , Proteínas Smad/análise , Fator de Crescimento Transformador beta1/análise
10.
Biol. Res ; 49: 1-11, 2016. ilus, graf, tab
Artigo em Inglês | LILACS | ID: lil-774428

RESUMO

BACKGROUND: Mammary cell cultures are convenient tools for in vitro studies of mammary gland biology. However, the heterogeneity of mammary cell types, e.g., glandular milk secretory epithelial or myoepithelial cells, often complicates the interpretation of cell-based data. The present study was undertaken to determine the relevance of bovine primary mammary epithelial cells isolated from American Holstein (bMEC US) or Swiss Holstein-Friesian (bMEC CH) cows, and of primary bovine mammary alveolar epithelial cells stably transfected with simian virus-40 (SV-40) large T-antigen (MAC-T) for in vitro analyses. This was evaluated by testing their expression pattern of cytokeratin (CK) 7, 18, 19, vimentin, and α-smooth muscle actin (α-SMA. RESULTS: The expression of the listed markers was assessed using real-time quantitative PCR, flow cytometry and immunofluorescence microscopy. Characteristic markers of the mesenchymal (vimentin), myoepithelial (α-SMA) and glandular secretory cells (CKs) showed differential expression among the studied cell cultures, partly depending on the analytical method used. The relative mRNA expression of vimentin, CK7 and CK19, respectively, was lower (P < 0.05) in immortalized than in primary mammary cell cultures. The stain index (based on flow cytometry) of CK7 and CK19 protein was lower (P < 0.05) in MAC-T than in bMECs, while the expression of α-SMA and CK18 showed an inverse pattern. Immunofluorescence microscopy analysis mostly confirmed the mRNA data, while partly disagreed with flow cytometry data (e.g., vimentin level in MAC-T). The differential expression of CK7 and CK19 allowed discriminating between immortal and primary mammary cultures. CONCLUSIONS: The expression of the selected widely used cell type markers in primary and immortalized MEC cells did not allow a clear preference between these two cell models for in vitro analyses studying aspects of milk composition. All tested cell models exhibited to a variable degree epithelial and mesenchymal features. Thus, based on their characterization with widely used cell markers, none of these cultures represent an unequivocal alveolar mammary epithelial cell model. For choosing the appropriate in vitro model additional properties such as the expression profile of specific proteins of interest (e.g., transporter proteins) should equally be taken into account.


Assuntos
Animais , Bovinos , Feminino , Actinas/análise , Células Epiteliais/citologia , Queratinas/análise , Glândulas Mamárias Animais/citologia , Vimentina/análise , Análise de Variância , Antígenos Virais de Tumores , Linhagem Celular , Células Cultivadas , Células Epiteliais/química , Citometria de Fluxo/métodos , Glândulas Mamárias Animais/química , Microscopia de Fluorescência/métodos , Cultura Primária de Células , Reação em Cadeia da Polimerase em Tempo Real
11.
Rev. bras. cir. cardiovasc ; 30(6): 644-649, Nov.-Dec. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-774551

RESUMO

ABSTRACT α-smooth muscle actin, encoded by ACTA2 gene, is an isoform of the vascular smooth muscle actins, typically expressed in the vascular smooth muscle cells contributing to vascular motility and contraction. ACTA2 gene mutations cause a diversity of diffuse vasculopathies such as thoracic aortic aneurysms and dissections as well as occlusive vascular diseases, including premature coronary artery disease and ischemic stroke. Dynamics of differentiation-specific α-smooth muscle actin in arterial smooth muscle cells and proliferation of the proteins have been well described. Although a variety of research works have been undertaken in terms of modifications of α-smooth muscle actin and mutations of ACTA2 gene and myosin, the underlying mechanisms towards the pathological processes by way of gene mutations are yet to be clarified. The purpose of the present article is to describe the phenotypes of α-smooth muscle actin and implications of ACTA2 mutations in vasculopathies in order to enhance the understanding of potential mechanisms of aortic and coronary disorders.


Assuntos
Humanos , Actinas/genética , Doenças da Aorta/metabolismo , Doença das Coronárias/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Actinas/metabolismo , Doenças da Aorta/genética , Doença das Coronárias/genética , Expressão Gênica , Contração Muscular/fisiologia , Mutação/genética , Fenótipo
12.
Int. braz. j. urol ; 41(5): 849-858, Sept.-Oct. 2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-767051

RESUMO

ABSTRACT Introduction and Objectives: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. Materials and Methods: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: α-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Erα and ERβ. Results: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERα. Conclusions: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.


Assuntos
Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Adenocarcinoma/patologia , Células Epiteliais/patologia , Células-Tronco Neoplásicas/patologia , Neoplasias da Próstata/patologia , Células Estromais/patologia , Actinas/análise , Adenocarcinoma/química , Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Progressão da Doença , Proteínas de Ligação a DNA/análise , Células Epiteliais/química , Receptor alfa de Estrogênio/análise , /análise , Proteínas Ligadas por GPI/análise , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/análise , /análise , Gradação de Tumores , Proteínas de Neoplasias/análise , Células-Tronco Neoplásicas/química , Neoplasias da Próstata/química , Células Estromais/química , Microambiente Tumoral , Fatores de Transcrição/análise , Vimentina/análise
13.
Rev. bras. enferm ; 68(2): 253-260, Mar-Apr/2015. tab
Artigo em Português | LILACS, BDENF - Enfermagem | ID: lil-752516

RESUMO

RESUMO Objetivo: construir e validar um instrumento para monitorar a qualidade dos registros de enfermagem no Programa de Assistência Domiciliar (PAD) em um hospital universitário. Método: estudo metodológico envolvendo a elaboração de um manual e submetido à validação de conteúdo por seis juízes sob consenso ≥ 80%. A coleta ocorreu em 2012 por meio de questionário contendo: evolução de enfermagem, diagnóstico e prescrição de enfermagem e normas para os registros da equipe de enfermagem preconizadas pelo Conselho Regional de Enfermagem-SP e pela instituição. Os itens do manual foram julgados de acordo com as variáveis - relevância, pertinência, clareza e simplicidade. Resultados: das 39 proposições 100% atingiram consenso ≥ 80% em relevância, pertinência e clareza; 92,3% em simplicidade. Os itens sono/repouso, mobilidade e checagem nas atividades prescritas não atingiram consenso mínimo favorável, sendo aprimorados pelas sugestões dos juízes. Conclusão: acreditamos que o instrumento possibilitará a melhoria dos processos de trabalho no PAD. .


RESUMEN Objetivo: construir y validar un instrumento para monitorear la calidad del registros de enfermería en Programa de Atención Domiciliaria (PAD) de un Hospital Universitario. Metodo: estudio metodológico. Fue construido un manual y sometió a validación de contenido por seis jueces bajo el consenso ≥80%. La recogida currió en 2012, con un cuestionario que contiene: evolución de enfermería, diagnóstico y prescripción de enfermería y normas para los registros del personal de enfermaria estabelecidas por Consejo Regional de Enfermería-SP y por la institución. Los artículos del manual fueran juzgadso conforme las variables relevancia, pertinencia, claridad y sencillez. Resultados: de las 39 proposiciones 100% alcanzó consenso ≥ 80% en la relevancia, pertinencia y claridad; 92,3% en la simplicidad. Los itens sueño/resto, movilidad y verificar las actividades prescritas no alcanzó consenso favorable, siendo mejoradas por las sugerencias de los jueces. Conclusión: creemos que el instrumento permitirá la mejora de los procesos de trabajo en PAD. .


ABSTRACT Objective: to build and validate an instrument aimed at monitoring the quality of nursing records in the Home Care Program (HCP) of a university hospital. Method: methodological study involving the elaboration of a manual, whose content was later submitted to six experts for validation, reaching a ≥ 80% consensus. The data collection process was carried out in 2012 by means of a questionnaire comprised of the following issues: nursing evolution, nursing diagnosis, and nursing prescription, and standards for the nursing team recommended by the Regional Nursing Council of São Paulo and by the assessed institution. Manual items were judged according to the following variables: relevance, pertinence, clarity and simplicity. Results: of the 39 propositions, 100% achieved ≥ 80% agreement in the relevance, pertinence and clarity variables; 92.3% in the simplicity variable. Sleep/rest, Mobility and Check-out variables did not reach a favorable minimum consensus in the prescribed activities and were improved following suggestions from the experts. Conclusion: we believe that the instrument will enable the improvement of the HCP’s work process. .


Assuntos
Humanos , Actinas/metabolismo , Cofilina 1/metabolismo , Disenteria Bacilar/microbiologia , Proteína Adaptadora de Sinalização NOD1/metabolismo , Fosfoproteínas Fosfatases/metabolismo , Shigella flexneri/fisiologia , Actinas/química , Western Blotting , Células Cultivadas , Cofilina 1/genética , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Regulação da Expressão Gênica , Células HeLa , Ensaios de Triagem em Larga Escala , Técnicas Imunoenzimáticas , Imunoprecipitação , Inflamação , Mediadores da Inflamação/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Proteína Adaptadora de Sinalização NOD1/antagonistas & inibidores , Proteína Adaptadora de Sinalização NOD1/genética , Fosforilação , Fosfoproteínas Fosfatases/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Transdução de Sinais
14.
Cad. saúde pública ; 31(3): 597-606, 03/2015. tab
Artigo em Português | LILACS | ID: lil-744836

RESUMO

Foi avaliada a associação entre menopausa e insônia e a influência de variáveis socioeconômicas e psicossociais, em estudo transversal com 2.190 funcionárias de uma universidade (Estudo Pró-Saúde), a partir de um questionário autopreenchível com variáveis sobre menopausa, insônia, transtorno mental comum, eventos de vida estressantes, apoio social e variáveis socioeconômicas. Odds ratios foram calculados por meio de regressão logística multivariada, com desfecho politômico. Após ajuste para potenciais confundidoras sociodemográficas, mulheres na menopausa há mais de 60 meses apresentaram maior chance de reportar queixas de sono frequentes (OR entre 1,53 e 1,86) do que as que estavam na menopausa há menos de 60 meses. Após os ajustes, no primeiro grupo, para as variáveis psicossociais, a magnitude dos ORs reduziu para 1,53 (IC95%: 0,92-2,52) para dificuldade em iniciar o sono, 1,81 (IC95%: 1,09-2,98) para dificuldade em manter o sono e 1,71 (IC95%: 1,08-2,73) para queixa geral de insônia. Fatores psicossociais podem mediar a manifestação da insônia em mulheres na menopausa.


This study evaluated the association between insomnia and menopausal status and the influence of socioeconomic and psychosocial variables on this association in a cross-sectional analysis of 2,190 university employees (the Pró-Saúde Study). A self-administered questionnaire was used, covering menopausal status, complaints of insomnia, common mental disorders, stressful life events, social support, and socioeconomic variables. Odds ratios were calculated using logistic regression with a polytomous outcome. After adjusting for potential socio-demographic confounders, women who had entered menopause more than 60 months previously were more likely to report complaints with sleep (OR 1.53-1.86) as compared to women in menopause for less than 60 months. After adjusting for psychosocial variables, in the first group the ORs decreased to 1.53 (95%CI: 0.92-2.52) for difficulty initiating sleep, 1.81 (95%CI: 1.09-2.98) for difficulty maintaining sleep, and 1.71 (95%CI: 1.08-2.73) for general complaints of insomnia. Psychosocial factors can mediate the manifestation of insomnia among menopausal women.


En este estudio se evaluó la asociación entre insomnio y menopausia y la influencia de las variables socioeconómicas y psicosociales, en un estudio transversal con 2.190 mujeres de una universidad (Estudio Pro-Salud), a partir de un cuestionario autoadministrado con variables de la menopausia, insomnio, trastornos mentales, situaciones de estrés vital, apoyo social y variables socioeconómicas. Se calcularon los odds ratio mediante regresión logística multivariante con desenlace politómico. Después de ajustar por factores de confusión sociodemográficos potenciales, las mujeres menopáusicas desde hace más de 60 meses fueron más propensas a reportar quejas frecuentes de sueño (OR entre 1,53 y 1,86) que las menopáusicas hace menos de 60 meses. Después de los ajustes, en el primer grupo, para las variables psicosociales la magnitud de los OR se redujo a 1,53 (IC95%: 0,92-2,52) para la dificultad para iniciar el sueño, un 1,81 (IC95%: 1,09-2,98) para mantener el sueño y un 1,71 (IC95%: 1,08-2,73) para las quejas de insomnio en general. Los factores psicosociales pueden mediar en la manifestación del insomnio en las mujeres menopáusicas.


Assuntos
Animais , Camundongos , Córtex Cerebral/metabolismo , Proteínas de Drosophila/metabolismo , Proteínas dos Microfilamentos/metabolismo , Microtúbulos/metabolismo , Neurogênese , Neurônios/metabolismo , Pseudópodes/metabolismo , Actinas/metabolismo , Linhagem Celular Tumoral , Células Cultivadas , Córtex Cerebral/embriologia , Drosophila , Proteínas de Drosophila/genética , /metabolismo , Cones de Crescimento/metabolismo , Mutação , Proteínas dos Microfilamentos/genética , Interferência de RNA
15.
Einstein (Säo Paulo) ; 13(1): 89-95, Jan-Mar/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-745883

RESUMO

Objective Evaluate the effects of VEGF165 gene transfer in the process of remodeling of the extracellular matrix after an acute myocardial infarct. Methods Wistar rats were submitted to myocardial infarction, after the ligation of the left descending artery, and the left ventricle ejection fraction was used to classify the infarcts into large and small. The animals were divided into groups of ten, according to the size of infarcted area (large or small), and received or not VEGF165 treatment. Evaluation of different markers was performed using immunohistochemistry and digital quantification. The primary antibodies used in the analysis were anti-fibronectin, anti-vimentin, anti-CD44, anti-E-cadherin, anti-CD24, anti-alpha-1-actin, and anti-PCNA. The results were expressed as mean and standard error, and analyzed by ANOVA, considering statistically significant if p≤0.05. Results There was a significant increase in the expression of undifferentiated cell markers, such as fibronectin (protein present in the extracellular matrix) and CD44 (glycoprotein present in the endothelial cells). However, there was decreased expression of vimentin and PCNA, indicating a possible decrease in the process of cell proliferation after treatment with VEGF165. Markers of differentiated cells, E-cadherin (adhesion protein between myocardial cells), CD24 (protein present in the blood vessels), and alpha-1-actin (specific myocyte marker), showed higher expression in the groups submitted to gene therapy, compared to non-treated group. The value obtained by the relation between alpha-1-actin and vimentin was approximately three times higher in the groups treated with VEGF165, suggesting greater tissue differentiation. Conclusion The results demonstrated the important role of myocytes in the process of tissue remodeling, confirming that VEGF165 seems to provide a protective effect in the treatment of acute myocardial infarct. .


Objetivo Avaliar os efeitos da transferência gênica do VEGF165 no processo de remodelamento da matriz extracelular após infarto agudo do miocárdio. Métodos Ratos Wistar foram submetidos ao infarto do miocárdio por ligação da artéria coronária descendente esquerda, e a fração de ejeção de ventrículo esquerdo foi utilizada para classificar os infartos em grandes e pequenos. Os animais foram divididos em grupos de dez animais, de acordo com o tamanho do infarto (grande ou pequeno), e receberam ou não tratamento com o VEGF165. A avaliação dos diferentes marcadores foi realizada por imuno-histoquímica e quantificação digital. Os anticorpos primários utilizados foram antifibronectina, antivimentina, anti- CD44, anti-E-caderina, anti-CD24, anti-alfa-1-actina e anti-PCNA. Os resultados foram representados como média e erro padrão, e analisados por ANOVA, sendo considerado estatisticamente significativo se p≤0,05. Resultados Houve aumento significativo da expressão de marcadores de células indiferenciadas, como fibronectina (proteína presente na matriz extracelular) e CD44 (glicoproteína presente nas células endoteliais). Entretanto, houve diminuição da expressão de vimentina e PCNA, indicando possível diminuição do processo de proliferação celular após o tratamento com VEGF165. Os marcadores de células diferenciadas, E-caderina (proteína de adesão entre as células do miocárdio), CD24 (proteína presente nos vasos sanguíneos) e alfa-1-actina (marcador especifico de miócitos) também apresentaram maior expressão nos grupos submetidos à terapia gênica, comparativamente com o grupo não tratado. O valor obtido pela relação entre alfa-1-actina e vimentina foi aproximadamente três vezes maior nos grupos tratados com VEGF165, indicando maior diferenciação tecidual. Conclusão O papel dos miócitos se mostrou importante no processo de remodelamento tecidual, confirmando que o VEGF165 parece conferir um efeito protetor no tratamento do infarto agudo do miocárdio. .


Assuntos
Animais , Feminino , Matriz Extracelular/fisiologia , Técnicas de Transferência de Genes , Infarto do Miocárdio/terapia , Fator A de Crescimento do Endotélio Vascular/uso terapêutico , Actinas/análise , /análise , /análise , Caderinas/análise , Proliferação de Células/fisiologia , Modelos Animais de Doenças , Fibronectinas/análise , Terapia Genética/métodos , Imuno-Histoquímica , Miócitos Cardíacos/fisiologia , Ratos Wistar , Reprodutibilidade dos Testes , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/genética , Vimentina/análise
16.
Mem. Inst. Oswaldo Cruz ; 110(1): 56-64, 03/02/2015. tab, graf
Artigo em Inglês | LILACS | ID: lil-741619

RESUMO

Histology is the gold standard for diagnosing acute rejection and hepatitis C recurrence after liver transplantation. However, differential diagnosis between the two can be difficult. We evaluated the role of C4d staining and quantification of hepatitis C virus (HCV) RNA levels in liver tissue. This was a retrospective study of 98 liver biopsy samples divided into four groups by histological diagnosis: acute rejection in patients undergoing liver transplant for hepatitis C (RejHCV+), HCV recurrence in patients undergoing liver transplant for hepatitis C (HCVTx+), acute rejection in patients undergoing liver transplant for reasons other than hepatitis C and chronic hepatitis C not transplanted (HCVTx-). All samples were submitted for immunohistochemical staining for C4d and HCV RNA quantification. Immunoexpression of C4d was observed in the portal vessels and was highest in the HCVTx- group. There was no difference in C4d expression between the RejHCV+ and HCVTx+ groups. However, tissue HCV RNA levels were higher in the HCVTx+ group samples than in the RejHCV+ group samples. Additionally, there was a significant correlation between tissue and serum levels of HCV RNA. The quantification of HCV RNA in liver tissue might prove to be an efficient diagnostic test for the recurrence of HCV infection.


Assuntos
Animais , Humanos , Camundongos , Anexina A1/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Neutrófilos/citologia , Neutrófilos/imunologia , Apoptose , Actinas/metabolismo , Anexina A1/deficiência , Anexina A1/genética , Anexina A1/imunologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Dexametasona/farmacologia , Técnicas In Vitro , /biossíntese , Camundongos Knockout , Macrófagos/metabolismo , Peptídeos , Fagocitose/efeitos dos fármacos , Fator de Crescimento Transformador beta/biossíntese
19.
Braz. j. med. biol. res ; 47(10): 850-857, 10/2014. tab, graf
Artigo em Inglês | LILACS | ID: lil-722172

RESUMO

We previously described a selective bile duct ligation model to elucidate the process of hepatic fibrogenesis in children with biliary atresia or intrahepatic biliary stenosis. Using this model, we identified changes in the expression of alpha smooth muscle actin (α-SMA) both in the obstructed parenchyma and in the hepatic parenchyma adjacent to the obstruction. However, the expression profiles of desmin and TGF-β1, molecules known to be involved in hepatic fibrogenesis, were unchanged when analyzed by semiquantitative polymerase chain reaction (RT-PCR). Thus, the molecular mechanisms involved in the modulation of liver fibrosis in this experimental model are not fully understood. This study aimed to evaluate the molecular changes in an experimental model of selective bile duct ligation and to compare the gene expression changes observed in RT-PCR and in real-time quantitative PCR (qRT‐PCR). Twenty-eight Wistar rats of both sexes and weaning age (21-23 days old) were used. The rats were separated into groups that were assessed 7 or 60 days after selective biliary duct ligation. The expression of desmin, α-SMA and TGF-β1 was examined in tissue from hepatic parenchyma with biliary obstruction (BO) and in hepatic parenchyma without biliary obstruction (WBO), using RT-PCR and qRT‐PCR. The results obtained in this study using these two methods were significantly different. The BO parenchyma had a more severe fibrogenic reaction, with increased α-SMA and TGF-β1 expression after 7 days. The WBO parenchyma presented a later, fibrotic response, with increased desmin expression 7 days after surgery and increased α-SMA 60 days after surgery. The qRT‐PCR technique was more sensitive to expression changes than the semiquantitative method.


Assuntos
Animais , Feminino , Masculino , Actinas/metabolismo , Colestase/complicações , Desmina/metabolismo , Cirrose Hepática/etiologia , Fígado/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos , Fator de Crescimento Transformador beta1/metabolismo , Análise de Variância , Actinas/genética , Atresia Biliar , Ductos Biliares/cirurgia , Colágeno Tipo I/biossíntese , Modelos Animais de Doenças , Desmina/genética , Expressão Gênica , Ligadura , Cirrose Hepática/metabolismo , Fígado/cirurgia , Ratos Wistar , Fator de Crescimento Transformador beta1/genética
20.
Acta cir. bras ; 29(9): 596-602, 09/2014. tab, graf
Artigo em Inglês | LILACS | ID: lil-722126

RESUMO

PURPOSE: To assess the evolution profile of the immunohistochemical expression of stromal constituents over the time-course of wound healing in a murine model. METHODS: Surgical wounds were performed in the back of 24 Wistar rats. After three, seven, 14 and 21 days, six rats were euthanized and the wounded histologically processed to assess the immunohistochemical expression of CD3, CD20, CD31, α-SMA and type-I collagen. Non-injured skin samples (NSS) were used as control. Data were subjected to statistical analysis using ANOVA and Tukey test. RESULTS: The mean of CD3 and CD20 positive cells in the wounds was significantly higher than in NSS at seven and 14 days (p<0.001). The blood vessels content was significantly lower than in NSS (p<0.05) at three days, but increased at seven and 14 days (p<0.01). The mean of α-SMA positive cells at seven, 14 and 21 days was higher than in NSS (p<0.05). The relative content of type I collagen increased from three to 21 days, but remained lower than in NSS (p<0.05). CONCLUSIONS: Lymphoid cells, myofibroblasts and microvessels contents varied over the time-course of wound healing, with peak at seven days and progressive reduction until 21 days. The type I collagen content increased over time. .


Assuntos
Animais , Masculino , Actinas/metabolismo , Antígenos CD/metabolismo , Colágeno Tipo I/metabolismo , Modelos Animais de Doenças , Linfócitos/patologia , Pele/lesões , Cicatrização/fisiologia , /metabolismo , /metabolismo , /metabolismo , Imuno-Histoquímica , Miofibroblastos/patologia , Ratos Wistar , Pele/metabolismo , Células Estromais/metabolismo , Células Estromais/patologia , Fatores de Tempo
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