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1.
Cartilage ; 11(3): 385-394, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-30146893

RESUMO

OBJECTIVE: Synovial fluid (SF) plays an important role in the maintenance of articular cartilage. SF is a dynamic reservoir of proteins derived from cartilage and synovial tissue; thus, its composition may serve as a biomarker that reflects the health and pathophysiological condition of the joint. The purpose of the current study was to evaluate the osteoarthritic synovial fluid (OASF) and transforming growth factor-ß1 (TGF-ß1) activity in articular chondrocytes catabolic and inflammatory responses. DESIGN: Chondrocytes were seeded at passage 2 and cultured for 72 hours under different conditions. Human chondrocytes were subjected to OASF while rat chondrocytes were subjected to either healthy synovial fluid (rSF) or TGF-ß1 and then assigned for cell viability analysis. In addition, the effects of OASF and TGF-ß1 on chondrocytes metalloprotease (MMP)-3 and MMP-13 and interleukin-18 (IL-18) expression were evaluated by immunocytochemistry, ELISA, and reverse transcriptase-polymerase chain reaction. RESULTS: SF from osteoarthritic patients significantly induced MMP-3, MMP-13, and IL-18 receptor expression in chondrocytes. To put in evidence the inflammatory activity of OASF, healthy chondrocytes from rat were cultured with TGF-ß1. In the presence of TGF-ß1 these cells started to express MMP-3, MMP-13, and IL-18 genes and attached to each other forming a chondrocyte aggregated structure. Healthy SF was able to maintain a typical monolayer of rounded chondrocytes with no inflammatory response. CONCLUSION: In summary, these observations demonstrated that TGF-ß1, one of the components of OASF, has a dual effect, acting in chondrocyte maintenance and also inducing inflammatory and catabolic properties of these cells.


Assuntos
Condrócitos/metabolismo , Interleucina-18/metabolismo , Osteoartrite/metabolismo , Líquido Sinovial/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Animais , Cartilagem Articular/citologia , Células Cultivadas , Humanos , Inflamação , Ratos , Membrana Sinovial/metabolismo
2.
Sci Rep ; 6: 23795, 2016 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-27029648

RESUMO

We characterized cerebral Oil Red O-positive lipid-laden cells (LLC) of aging mice evaluating their distribution, morphology, density, functional activities and inflammatory phenotype. We identified LLC in meningeal, cortical and neurogenic brain regions. The density of cerebral LLC increased with age. LLC presenting small lipid droplets were visualized adjacent to blood vessels or deeper in the brain cortical and striatal parenchyma of aging mice. LLC with larger droplets were asymmetrically distributed in the cerebral ventricle walls, mainly located in the lateral wall. We also found that LLC in the subventricular region co-expressed beclin-1 or LC3, markers for autophagosome or autophagolysosome formation, and perilipin (PLIN), a lipid droplet-associated protein, suggesting lipophagic activity. Some cerebral LLC exhibited ß galactosidase activity indicating a senescence phenotype. Moreover, we detected production of the pro-inflammatory cytokine TNF-α in cortical PLIN(+) LLC. Some cortical NeuN(+) neurons, GFAP(+) glia limitans astrocytes, Iba-1(+) microglia and S100ß(+) ependymal cells expressed PLIN in the aging brain. Our findings suggest that cerebral LLC exhibit distinct cellular phenotypes and may participate in the age-associated neuroinflammatory processes.


Assuntos
Envelhecimento/metabolismo , Astrócitos/ultraestrutura , Gotículas Lipídicas/ultraestrutura , Microglia/ultraestrutura , Neurônios/ultraestrutura , Animais , Astrócitos/metabolismo , Autofagia , Proteína Beclina-1/genética , Proteína Beclina-1/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Contagem de Células , Córtex Cerebral/metabolismo , Córtex Cerebral/ultraestrutura , Feminino , Expressão Gênica , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/metabolismo , Histocitoquímica , Gotículas Lipídicas/metabolismo , Meninges/metabolismo , Meninges/ultraestrutura , Camundongos , Camundongos Endogâmicos BALB C , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Microglia/metabolismo , Proteínas Associadas aos Microtúbulos/genética , Proteínas Associadas aos Microtúbulos/metabolismo , Neurogênese , Neurônios/metabolismo , Perilipina-1/genética , Perilipina-1/metabolismo , Fenótipo , Subunidade beta da Proteína Ligante de Cálcio S100/genética , Subunidade beta da Proteína Ligante de Cálcio S100/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , beta-Galactosidase/genética , beta-Galactosidase/metabolismo
3.
Cell Tissue Res ; 363(2): 411-26, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26003178

RESUMO

Galectin-3 is a ß-galactoside-binding protein with an inhibitory role in B cell differentiation into plasma cells in distinct lymphoid tissues. We use a model of chronic schistosomiasis, a well-characterized experimental disease hallmarked by polyclonal B cell activation, in order to investigate the role of galectin-3 in controlling IgA production through peritoneal B1 cells. Chronically infected, galectin-3-deficient mice (Lgals3(-/-)) display peritoneal fluid hypercellularity, increased numbers of atypical peritoneal IgM(+)/IgA(+) B1a and B1b lymphocytes and histological disturbances in plasma cell niches when compared with Lgals3(+/+) mice. Similar to our infection model, peritoneal B1 cells from uninfected Lgals3(-/-) mice show enhanced switching to IgA after in vitro treatment with interleukin-5 plus transforming growth factor-ß (IL-5 + TGF-ß1). A higher number of IgA(+) B1a lymphocytes was found in the peritoneal cavity of Lgals3(-/-)-uninfected mice at 1 week after i.p. injection of IL-5 + TGF-ß1; this correlates with the increased levels of secreted IgA detected in the peritoneal fluid of these mice after cytokine treatment. Interestingly, a higher number of degranulated mast cells is present in the peritoneal cavity of uninfected and Schistosoma mansoni-infected Lgals3(-/-) mice, indicating that, at least in part, mast cells account for the enhanced differentiation of B1 into IgA-producing B cells found in the absence of galectin-3. Thus, a novel role is revealed for galectin-3 in controlling the expression of surface IgA by peritoneal B1 lymphocytes; this might have important implications for manipulating the mucosal immune response.


Assuntos
Linfócitos B/citologia , Linfócitos B/metabolismo , Diferenciação Celular , Galectina 3/deficiência , Imunoglobulina A/metabolismo , Peritônio/citologia , Regulação para Cima , Animais , Contagem de Células , Degranulação Celular , Proliferação de Células , Forma Celular , Doença Crônica , Galectina 3/metabolismo , Imunoglobulina A/sangue , Switching de Imunoglobulina , Imunoglobulina M/sangue , Interleucina-5 , Mastócitos/fisiologia , Mesentério/metabolismo , Camundongos Endogâmicos C57BL , Omento/metabolismo , Fenótipo , Plasmócitos/metabolismo , Esquistossomose/sangue , Esquistossomose/imunologia , Esquistossomose/parasitologia , Esquistossomose/patologia , Fator de Crescimento Transformador beta1/metabolismo
4.
Biol Open ; 4(9): 1180-93, 2015 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-26319582

RESUMO

Human adipose-derived stromal cells (hADSC) are a heterogeneous cell population that contains adult multipotent stem cells. Although it is well established that hADSC have skeletal potential in vivo in adult organisms, in vitro assays suggest further differentiation capacity, such as into glia. Thus, we propose that grafting hADSC into the embryo can provide them with a much more instructive microenvironment, allowing the human cells to adopt diverse fates or niches. Here, hADSC spheroids were grafted into either the presumptive presomitic mesoderm or the first branchial arch (BA1) regions of chick embryos. Cells were identified without previous manipulations via human-specific Alu probes, which allows efficient long-term tracing of heterogeneous primary cultures. When grafted into the trunk, in contrast to previous studies, hADSC were not found in chondrogenic or osteogenic territories up to E8. Surprisingly, 82.5% of the hADSC were associated with HNK1+ tissues, such as peripheral nerves. Human skin fibroblasts showed a smaller tropism for nerves. In line with other studies, hADSC also adopted perivascular locations. When grafted into the presumptive BA1, 74.6% of the cells were in the outflow tract, the final goal of cardiac neural crest cells, and were also associated with peripheral nerves. This is the first study showing that hADSC could adopt a perineural niche in vivo and were able to recognize cues for neural crest cell migration of the host. Therefore, we propose that xenografts of human cells into chick embryos can reveal novel behaviors of heterogeneous cell populations, such as response to migration cues.

5.
Birth Defects Res C Embryo Today ; 105(1): 1-8, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25789860

RESUMO

Many advances have been taken on elucidating embryonic development and tissue homeostasis and repair by the use of experimental strategies that preserve the three-dimensional (3D) organization and allow quantitative analysis of images over time (four-dimensional). Ranging from the understanding about the relationship between blastomeres and the events that take place during gastrulation by the use of time-lapse imaging through 3D cultures that mimic organogenesis, the advances in this area are of critical value. The studies on embryonic development without disrupting the original architecture and the development of 3D organoid cultures pave a new avenue for unprecedented experimental advances that will positively impact the emergence of new treatments applying regenerative principles for both tissue repair and organ transplant.


Assuntos
Desenvolvimento Embrionário/fisiologia , Matriz Extracelular/fisiologia , Homeostase/fisiologia , Imageamento Tridimensional/métodos , Organogênese/fisiologia , Imagem com Lapso de Tempo/métodos , Humanos , Imageamento Tridimensional/tendências , Imagem com Lapso de Tempo/tendências
6.
Int J Dev Biol ; 58(5): 355-362, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25354456

RESUMO

Tiki1 is a Wnt protease and antagonist specifically expressed in the Spemann-Mangold Organizer and is required for head formation in Xenopus embryos. Here we report neighbor-joining phylogenetic analysis of vertebrate Tiki genes and their mRNA expression patterns in chick, mouse, and rabbit embryos. Tiki1 and Tiki2 orthologues are highly conserved, and exhibit similar but also different developmental expression patterns among the vertebrate/mammalian species analyzed. The Tiki1 gene is noticeably absent in the rodent lineage, but is present in lagomorphs and all other vertebrate/mammalian species examined. Expression in Hensen's node, the equivalent of the Xenopus Organizer, was observed for Chick Tiki2 and Rabbit Tiki1 and Tiki2. Mouse Tiki2 was detected at low levels at gastrulation and head fold stages, but not in the node. Mouse Tiki2 and chick Tiki1 display similar expression in the dorsal spinal cord. Chick Tiki1 expression was also detected in the surface ectoderm and maxillary bud, while chick Tiki2 was found in the anterior intestinal portal, head mesenchyme and primitive atrium. Our expression analyses provide evidence that Tiki1 and Tiki2 are evolutionarily conserved among vertebrate species and their expression in the Organizer and other regions suggests contributions of these Wnt inhibitors to embryonic patterning, as well as organogenesis. Our analyses further reveal mis-regulation of TIKI1 and TIKI2 in human cancer and diseases.


Assuntos
Padronização Corporal/genética , Evolução Molecular , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Membrana/genética , Metaloproteases/genética , Filogenia , Animais , Embrião de Galinha , Proteínas de Membrana/metabolismo , Metaloendopeptidases , Metaloproteases/metabolismo , Camundongos , Organizadores Embrionários/embriologia , Organizadores Embrionários/metabolismo , Coelhos
7.
Dev Biol ; 365(2): 350-62, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22426006

RESUMO

Cholesterol-rich membrane microdomains (CRMMs) are specialized structures that have recently gained much attention in cell biology because of their involvement in cell signaling and trafficking. However, few investigations, particularly those addressing embryonic development, have succeeded in manipulating and observing CRMMs in living cells. In this study, we performed a detailed characterization of the CRMMs lipid composition during early frog development. Our data showed that disruption of CRMMs through methyl-ß-cyclodextrin (MßCD) cholesterol depletion at the blastula stage did not affect Spemann's organizer gene expression and inductive properties, but impaired correct head development in frog and chick embryos by affecting the prechordal plate gene expression and cellular morphology. The MßCD anterior defect phenotype was recapitulated in head anlagen (HA) explant cultures. Culture of animal cap expressing Dkk1 combined with MßCD-HA generated a head containing eyes and cement gland. Together, these data show that during Xenopus blastula and gastrula stages, CRMMs have a very dynamic lipid composition and provide evidence that the secreted Wnt antagonist Dkk1 can partially rescue anterior structures in cholesterol-depleted head anlagen.


Assuntos
Padronização Corporal , Colesterol/metabolismo , Microdomínios da Membrana/metabolismo , Prosencéfalo/embriologia , Animais , Embrião de Galinha , Microdomínios da Membrana/efeitos dos fármacos , Organizadores Embrionários/metabolismo , Xenopus laevis , beta-Ciclodextrinas/farmacologia
8.
PLoS One ; 7(1): e30363, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22291942

RESUMO

Population control of the dengue vector mosquito, Aedes aegypti, is difficult due to many reasons, one being the development of resistance to neurotoxic insecticides employed. The biosynthesis of chitin, a major constituent of insect cuticle, is a novel target for population control. Novaluron is a benzoylphenylurea (BPU) that acts as a chitin synthesis inhibitor, already used against mosquitoes. However, information regarding BPU effects on immature mosquito stages and physiological parameters related with mosquito larval development are scarce. A set of physiological parameters were recorded in control developing larvae and novaluron was administered continuously to Ae. aegypti larvae, since early third instar. Larval instar period duration was recorded from third instar until pupation. Chitin content was measured during third and fourth instars. Fourth instars were processed histochemically at the mesothorax region, stained with hematoxylin and eosin (HE) for assessment of internal tissues, and labeled with WGA-FITC to reveal chitinized structures. In control larvae: i) there is a chitin content increase during both third and fourth instars where late third instars contain more chitin than early fourth instars; ii) thoracic organs and a continuous cuticle, closely associated with the underlying epidermis were observed; iii) chitin was continuously present throughout integument cuticle. Novaluron treatment inhibited adult emergence, induced immature mortality, altered adult sex ratio and caused delay in larval development. Moreover, novaluron: i) significantly affected chitin content during larval development; ii) induced a discontinuous and altered cuticle in some regions while epidermis was often thinner or missing; iii) rendered chitin cuticle presence discontinuous and less evident. In both control and novaluron larvae, chitin was present in the peritrophic matrix. This study showed quantitatively and qualitatively evidences of novaluron effects on Ae. aegypti larval development. To our knowledge, this is the first report describing histological alterations produced by a BPU in immature vector mosquitoes.


Assuntos
Aedes/efeitos dos fármacos , Aedes/crescimento & desenvolvimento , Quitina/metabolismo , Compostos de Fenilureia/farmacologia , Aedes/anatomia & histologia , Aedes/fisiologia , Fatores Etários , Animais , Quitina/análise , Quitina Sintase/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Feminino , Inseticidas/farmacologia , Larva/anatomia & histologia , Larva/química , Larva/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Masculino , Controle de Mosquitos , Razão de Masculinidade , Temperatura
9.
Development ; 135(13): 2311-9, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18539924

RESUMO

Previous studies have shown that Sonic Hedgehog (Shh) signaling is crucial for the development of the first branchial arch (BA1) into a lower-jaw in avian and mammalian embryos. We have already shown that if Shh expression is precociously inhibited in pharyngeal endoderm, neural crest cells migrate to BA1 but fail to survive, and Meckel's cartilage and associated structures do not develop. This phenotype can be rescued by addition of an exogenous source of Shh. To decipher the role of Shh, we explored the consequences of providing an extra source of Shh to the presumptive BA1 territory. Grafting quail fibroblasts engineered to produce Shh (QT6-Shh), at the 5- to 8-somite stage, resulted in the induction of mirror-image extra lower jaws, caudolateral to the normal one. It turns out that the oral opening epithelium, in which Shh, Fgf8 and Bmp4 are expressed in a definite pattern, functions as an organizing center for lower-jaw development. In our experimental design, the extra source of Shh activates Fgf8, Bmp4 and Shh genes in caudal BA1 ectoderm in a spatial pattern similar to that of the oral epithelium, and regularly leads to the formation of two extra lower-jaw-organizing centers with opposite rostrocaudal polarities. These results emphasize the similarities between the developmental processes of the limb and mandibular buds, and show that in both cases Shh-producing cells create a zone of polarizing activity for the structures deriving from them.


Assuntos
Padronização Corporal , Proteínas Hedgehog/metabolismo , Mandíbula/embriologia , Mandíbula/metabolismo , Mesoderma/embriologia , Mesoderma/metabolismo , Animais , Osso e Ossos/embriologia , Osso e Ossos/metabolismo , Região Branquial/embriologia , Região Branquial/metabolismo , Cartilagem/embriologia , Cartilagem/metabolismo , Linhagem Celular , Embrião de Galinha , Ectoderma/embriologia , Ectoderma/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Hedgehog/genética , Codorniz
10.
Brain Res Rev ; 55(2): 237-47, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17765317

RESUMO

Since the time of Ramon y Cajal, very significant progress has been accomplished in our knowledge of the fate of the early neural primordium. The origin of the peripheral nervous system from the transient and pluripotent embryonic structure, the neural crest has been fully deciphered using appropriate cell marking techniques. Most of the pioneer work in this field was carried out in lower vertebrates up to 1950 and later on in the avian embryo. New techniques which allow the genetic labelling of embryonic cells by transgenesis are now applied in mammals and fish. One of the highlights of neural crest studies was its paramount role in head and face morphogenesis. Work pursued in our laboratory for the last fifteen years or so has analysed at both cellular and molecular levels the contribution of the NCCs to the construction of the facial and cranial structures. Recently, we have found that the cephalic neural crest plays also a key role in the formation of the fore- and mid-brain.


Assuntos
Encéfalo/embriologia , Encéfalo/crescimento & desenvolvimento , Face/embriologia , Face/fisiologia , Crista Neural/fisiologia , Animais , Desenvolvimento Embrionário , Regulação da Expressão Gênica no Desenvolvimento , Morfogênese , Organogênese
11.
Proc Natl Acad Sci U S A ; 103(31): 11607-12, 2006 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-16868080

RESUMO

We have investigated the role of Sonic hedgehog (Shh) in the development of facial structures by depriving chicken embryos of the most anterior sources of this morphogen, including the prechordal plate and the anterior ventral endoderm of the foregut, before the onset of neural crest cell (NCC) migration to the first branchial arch (BA1). The entire forehead, including the foregut endoderm, was removed at 5- to 10-somite stage (ss), which led to the absence of the lower jaw when the operation was performed before 7-ss. If the embryos were deprived of their forehead at 8- to 10-ss, they were later on endowed with a lower beak. In embryos that were operated on early, the NCCs migrated normally to BA1 but were subjected to massive apoptosis a few hours later. Cell death did not occur when forehead excision was performed at a later stage. In this case, onward expression of Shh in the ventral foregut endoderm extended caudally over the excision limit, and we hypothesized that absence of Shh production by the endoderm in embryos that were operated on early could be responsible for the NCC apoptosis and the failure of BA1 development. We thus provided exogenous Shh to the embryos that were operated on before 7-ss. In this case, the development of the lower jaw was rescued. Therefore, Shh derived from the ventral foregut endoderm ensures the survival of NCCs at a critical stage of BA1 development.


Assuntos
Sobrevivência Celular , Sistema Digestório , Endoderma , Face/embriologia , Arcada Osseodentária/embriologia , Crista Neural/fisiologia , Transativadores/metabolismo , Animais , Padronização Corporal , Região Branquial/anatomia & histologia , Região Branquial/embriologia , Movimento Celular/fisiologia , Embrião de Galinha/anatomia & histologia , Embrião de Galinha/embriologia , Embrião de Galinha/cirurgia , Sistema Digestório/anatomia & histologia , Sistema Digestório/embriologia , Face/anatomia & histologia , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Hedgehog , Hibridização In Situ , Arcada Osseodentária/anatomia & histologia , Crista Neural/citologia , Transdução de Sinais/fisiologia , Transativadores/genética
12.
An. cir. card. cir. vasc ; 12(1): 34-36, ene.-feb. 2006. ilus
Artigo em Es | IBECS | ID: ibc-045517

RESUMO

Se presenta un caso de drenaje venoso pulmonar anómalo parcial izquierdo en una adolescente de 14 años. Se describen los hallazgos clínicos, el ecocardiograma y el cateterismo cardíaco, la indicación quirúrgica y la anatomía atípica hallada intraoperatoriamente (un colector “lejano” a la aurícula izquierda), lo mismo que las maniobras quirúrgicas empleadas en su corrección, y su evolución posterior


We had an interesting case on our hands, in particular that of a 14 year old girl with an unusual form of left partial anomalous pulmonary venous connection. We'd like to show our clinical, echocardiograms and cardiac catheterization results and findings. The patient didn’t have a normal anatomical pattern (a colector far from the left atrial appendage), and we would also like to comment on our postoperative follow up. Because of the unusual anatomy of the lesion and the long distances between the colector and the left atrial appendage, it was necesary to restructure the colector we also had to design a colector flap for the atrial appendage


Assuntos
Feminino , Adolescente , Humanos , Procedimentos Cirúrgicos Cardíacos/métodos , Cardiopatias Congênitas/cirurgia , Veias Pulmonares/cirurgia , Veias Pulmonares/anormalidades , Cateterismo Cardíaco , Átrios do Coração/cirurgia , Resultado do Tratamento , Seguimentos
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