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1.
Cell Death Dis ; 15(6): 405, 2024 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-38858390

RESUMO

Genetic mutations causing primary mitochondrial disease (i.e those compromising oxidative phosphorylation [OxPhos]) resulting in reduced bioenergetic output display great variability in their clinical features, but the reason for this is unknown. We hypothesized that disruption of the communication between endoplasmic reticulum (ER) and mitochondria at mitochondria-associated ER membranes (MAM) might play a role in this variability. To test this, we assayed MAM function and ER-mitochondrial communication in OxPhos-deficient cells, including cybrids from patients with selected pathogenic mtDNA mutations. Our results show that each of the various mutations studied indeed altered MAM functions, but notably, each disorder presented with a different MAM "signature". We also found that mitochondrial membrane potential is a key driver of ER-mitochondrial connectivity. Moreover, our findings demonstrate that disruption in ER-mitochondrial communication has consequences for cell survivability that go well beyond that of reduced ATP output. The findings of a "MAM-OxPhos" axis, the role of mitochondrial membrane potential in controlling this process, and the contribution of MAM dysfunction to cell death, reveal a new relationship between mitochondria and the rest of the cell, as well as providing new insights into the diagnosis and treatment of these devastating disorders.


Assuntos
Retículo Endoplasmático , Potencial da Membrana Mitocondrial , Mitocôndrias , Doenças Mitocondriais , Fosforilação Oxidativa , Humanos , Retículo Endoplasmático/metabolismo , Mitocôndrias/metabolismo , Doenças Mitocondriais/metabolismo , Doenças Mitocondriais/genética , Doenças Mitocondriais/patologia , Mutação/genética , DNA Mitocondrial/metabolismo , DNA Mitocondrial/genética
2.
J Allergy Clin Immunol ; 152(4): 1010-1018, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37406823

RESUMO

BACKGROUND: Human infants develop IgG responses to dietary antigens during the first 2 years of life. Yet, the source of these antibodies is unclear. In previous studies we reported on the thymus as a unique functional niche for plasma cells (PCs) specific to environmental antigens. OBJECTIVE: We sought to examine whether PCs specific to dietary antigens are detected in the infant thymus. METHODS: We tested IgG reactivity to 112 food antigens and allergens in the serum of 20 neonates and infants using microarrays. The presence of PC-secreting IgG specific to the most prominent antigens was then assessed among thymocytes in the same cohort. Using an LC-MS proteomics approach, we looked for traces of these antigens in the thymus. RESULTS: Our studies first confirmed that cow's milk proteins are prevalent targets of serum IgG in early life. Subjects with the highest serum IgG titers to cow's milk proteins also harbored IgG-producing PCs specific to the same antigens in the thymic niche. Furthermore, we detected multiple peptide fragments of cow's milk antigens in the thymus. Lastly, we verified that both serum IgG and IgG secreted by thymic PCs recognized the peptide epitopes found in the thymus. CONCLUSIONS: Our studies reveal the presence of antibody-secreting PCs specific to common dietary antigens in the infant thymus. The presence of these antigens in the thymus suggested that activation and differentiation of specific PCs occurred in this organ. Further studies are now warranted to evaluate the possible implication of these cells in tolerance to dietary antigens.


Assuntos
Hipersensibilidade a Leite , Proteínas do Leite , Recém-Nascido , Animais , Feminino , Bovinos , Lactente , Humanos , Formação de Anticorpos , Plasmócitos , Imunoglobulina G , Leite , Alérgenos
4.
Nat Commun ; 12(1): 5761, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34599177

RESUMO

The thymus is a central lymphoid organ primarily responsible for the development of T cells. A small proportion of B cells, however, also reside in the thymus to assist negative selection of self-reactive T cells. Here we show that the thymus of human neonates contains a consistent contingent of CD138+ plasma cells, producing all classes and subclasses of immunoglobulins with the exception of IgD. These antibody-secreting cells are part of a larger subset of B cells that share the expression of signature genes defining mouse B1 cells, yet lack the expression of complement receptors CD21 and CD35. Data from single-cell transcriptomic, clonal correspondence and in vitro differentiation assays support the notion of intrathymic CD138+ plasma cell differentiation, alongside other B cell subsets with distinctive molecular phenotypes. Lastly, neonatal thymic plasma cells also include clones reactive to commensal and pathogenic bacteria that commonly infect children born with antibody deficiency. Thus, our findings point to the thymus as a source of innate humoral immunity in human neonates.


Assuntos
Diferenciação Celular , Plasmócitos/citologia , Timo/citologia , Adulto , Antígenos CD/metabolismo , Linfócitos B/citologia , Linfócitos B/metabolismo , Linfócitos B/ultraestrutura , Sangue Fetal/citologia , Perfilação da Expressão Gênica , Humanos , Imunidade Inata , Imunoglobulina G/metabolismo , Cadeias Pesadas de Imunoglobulinas/imunologia , Região Variável de Imunoglobulina/imunologia , Recém-Nascido , Subpopulações de Linfócitos/citologia , Análise de Componente Principal , RNA-Seq , Análise de Célula Única , Hipermutação Somática de Imunoglobulina/genética , Transcriptoma/genética
5.
J Heart Lung Transplant ; 40(11): 1472-1480, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34420852

RESUMO

BACKGROUND: Antibody mediated rejection (AMR) is an increasingly studied cause of graft failure after heart transplantation. AMR diagnosis previously required the detection of circulating donor specific antibodies (DSA); however, the most recent criteria only require pathological findings. This classification defined a subset of patients with AMR, yet without known antibodies. Here, we sought to evaluate differences in the transcriptome profile associated with different types of AMR. METHODS: RNA sequencing was used on endomyocardial biopsies to analyze and compare transcriptomic profiles associated with different subtypes of AMR defined by immunopathological and histopathological findings, as well as the presence or absence of DSA. Gene expression profiles were characterized for each diagnostic group. RESULTS: The most divergent gene expression profiles were observed between patients with or without DSA. AMR subtypes associated with DSA showed expression of signature genes involved in monocyte activation and response to interferon. There was also substantial difference between the transcriptomic profiles of AMR defined by histopathological and immunopathological findings, the latter being associated with expression of mucin genes. In contrast, there was no differential RNA expression between patients with pAMR1i without DSA and those without AMR. Likewise, no differential expression was observed between patients with pAMR1h with DSA and pAMR2. CONCLUSIONS: Overall, our studies reveal different expression profiles in endomyocardial biopsies in relation to some key criteria used to diagnose AMR. These findings support the view that the diagnosis of AMR encompasses several phenotypes that may rely on distinct mechanisms of injury.


Assuntos
Rejeição de Enxerto/imunologia , Transplante de Coração/efeitos adversos , Isoanticorpos/imunologia , Miocárdio/patologia , Doadores de Tecidos , Transcriptoma/imunologia , Adolescente , Adulto , Biópsia , Criança , Feminino , Rejeição de Enxerto/diagnóstico , Sobrevivência de Enxerto/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Miocárdio/imunologia , Adulto Jovem
6.
Mol Neurobiol ; 58(7): 3270-3289, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33666854

RESUMO

Perturbations in mitochondrial dynamics have been observed in most neurodegenerative diseases. Here, we focus on manganese (Mn)-induced Parkinsonism-like neurodegeneration, a disorder associated with the preferential of Mn in the basal ganglia where the mitochondria are considered an early target. Despite the extensive characterization of the clinical presentation of manganism, the mechanism by which Mn mediated mitochondrial toxicity is unclear. In this study we hypothesized whether Mn exposure alters mitochondrial activity, including axonal transport of mitochondria and mitochondrial dynamics, morphology, and network. Using primary neuron cultures exposed to 100 µM Mn (which is considered the threshold of Mn toxicity in vitro) and intraperitoneal injections of MnCl2 (25mg/kg) in rat, we observed that Mn increased mitochondrial fission mediated by phosphorylation of dynamin-related protein-1 at serine 616 (p-s616-DRP1) and decreased mitochondrial fusion proteins (MFN1 and MFN2) leading to mitochondrial fragmentation, defects in mitochondrial respiratory capacity, and mitochondrial ultrastructural damage in vivo and in vitro. Furthermore, Mn exposure impaired mitochondrial trafficking by decreasing dynactin (DCTN1) and kinesin-1 (KIF5B) motor proteins and increasing destabilization of the cytoskeleton at protein and gene levels. In addition, mitochondrial communication may also be altered by Mn exposure, increasing the length of nanotunnels to reach out distal mitochondria. These findings revealed an unrecognized role of Mn in dysregulation of mitochondrial dynamics providing a potential explanation of early hallmarks of the disorder, as well as a possible common pathway with neurological disorders arising upon chronic Mn exposure.


Assuntos
Corpo Estriado/efeitos dos fármacos , Manganês/toxicidade , Mitocôndrias/efeitos dos fármacos , Dinâmica Mitocondrial/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Animais , Células Cultivadas , Corpo Estriado/metabolismo , Corpo Estriado/patologia , Redes Reguladoras de Genes/efeitos dos fármacos , Redes Reguladoras de Genes/fisiologia , Masculino , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Dinâmica Mitocondrial/fisiologia , Neurônios/metabolismo , Neurônios/patologia , Ratos , Ratos Sprague-Dawley
7.
Cureus ; 12(11): e11510, 2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-33354454

RESUMO

We discuss the case of a three-year-old female patient who presented with a severe episode of immune thrombocytopenia (ITP) and autoimmune hemolytic anemia (AIHA), confirming a diagnosis of Evans syndrome (ES). Over time, she continued to have several episodes of recurrent ITP until, several years later, she experienced a recurrent severe, refractory ES episode. Initially, she responded well to conventional treatment with steroids and intravenous immunoglobulin (IVIG); however, during later episodes, she required anti-CD20 therapy (rituximab). Due to peculiar facies and severe clinical presentation, an underlying immune dysregulation was suspected, which was later confirmed to be 22q11.2 deletion syndrome (22q11.2DS). Over time, her baseline immunoglobulin production decreased significantly. After monthly IVIG replacement, she had a marked reduction in ITP or AIHA events. 22q11.2DS is a frequently underdiagnosed primary immune disorder (PID). Low immunoglobulin production and recurrent ES are infrequent events associated with 22q11.2DS. This condition might cause profound immune dysregulation, predisposing patients to immune-related hematological dyscrasias that still need further research to be fully understood and characterized. We describe a case of 22q11.2DS and recurrent ES episodes, which involves a 13-year history of longitudinal follow-up care.

8.
Bol. méd. postgrado ; 36(1): 32-37, jul.2020. tab
Artigo em Espanhol | LIVECS, LILACS | ID: biblio-1119378

RESUMO

La amigdalectomía es uno de los procedimientos quirúrgicos más frecuentes en la actualidad especialmente en edad pediátrica. Con el objetivo de determinar las complicaciones en amigdalectomía con electrodisección y técnica clásica en pacientes pediátricos que ingresaron al Servicio Desconcentrado Hospital Pediátrico Dr. Agustín Zubillaga en el año 2017, se realizó un estudio de recolección retrospectiva de datos seleccionando 119 historias clínicas, obteniendo que 73,95% de los casos fueron intervenidos por técnica de electrodisección y 26,05% por técnica clásica. Las principales indicaciones para cirugía fueron amigdalitis a repetición e hipertrofia amigdalina. Para ambas técnicas, el tiempo quirúrgico promedio fue entre 21-40 minutos (44,32% y 48,39%, respectivamente) y la estancia hospitalaria menor de un día en 54,55% y 61,29% de los casos, respectivamente. La principal complicación intraoperatoria reportada fue la hemorragia (29,03% para la técnica clásica y 15,91% para la técnica de electrodisección). Entre las complicaciones postoperatorias, el dolor (63,64%) y la fiebre (60,23%) prevaleció en pacientes operados con electrodisección mientras que en los pacientes con técnica clásica la más frecuente fue la fiebre (61,29%). Con estos resultados, se demuestra que ambas técnicas tienen similar tiempo quirúrgico y estancia hospitalaria, pero difieren en en tipo de complicaciones(AU)


Tonsillectomy is one of the most frequent surgical procedures in our times especially in pediatric age. In order to determine the complications of electrodissection and classical tonsillectomy in pediatric patients who were admitted at the Servicio Desconcentrado Hospital Pediátrico Dr. Agustín Zubillaga in 2017, a review of 119 medical charts was performed. The results show that 73.95% of the cases had electrodissection tonsillectomy and 26.05% had classical tonsillectomy. The main indications for surgery were recurrent tonsillitis (electrodissection: 86.09% and classical: 80.65%) and tonsillar hypertrophy (electrodissection: 60.23% and classical: 35.48%). For both techniques, the mean surgical time was between 21-40 minutes (44.32% and 48.39%, respectively) and the hospital stay was less than one day (54.55% and 61.29%, each). The main intraoperative complication reported was hemorrhage (29.03% for classical versus 15.91% for electrodissection). Among the postoperative complications, pain (63.64%) and fever (60.23%) prevailed in patients with electrodissection tonsillectomy while fever was most frequent in patients with classical tonsillectomy (61.29%). These results show that both techniques have similar surgical time and hospital stay but differ in type of complications(AU)


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Complicações Pós-Operatórias , Procedimentos Cirúrgicos Operatórios , Tonsilectomia , Tonsilite/cirurgia , Transtornos de Deglutição , Náusea e Vômito Pós-Operatórios , Hemorragia
9.
PLoS Pathog ; 14(5): e1007105, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29791511

RESUMO

Infection with Burkholderia pseudomallei or B. thailandensis triggers activation of the NLRP3 and NLRC4 inflammasomes leading to release of IL-1ß and IL-18 and death of infected macrophages by pyroptosis, respectively. The non-canonical inflammasome composed of caspase-11 is also activated by these bacteria and provides protection through induction of pyroptosis. The recent generation of bona fide caspase-1-deficient mice allowed us to reexamine in a mouse model of pneumonic melioidosis the role of caspase-1 independently of caspase-11 (that was also absent in previously generated Casp1-/- mice). Mice lacking either caspase-1 or caspase-11 were significantly more susceptible than wild type mice to intranasal infection with B. thailandensis. Absence of caspase-1 completely abolished production of IL-1ß and IL-18 as well as pyroptosis of infected macrophages. In contrast, in mice lacking caspase-11 IL-1ß and IL-18 were produced at normal level and macrophages pyroptosis was only marginally affected. Adoptive transfer of bone marrow indicated that caspase-11 exerted its protective action both in myeloid cells and in radio-resistant cell types. B. thailandensis was shown to readily infect mouse lung epithelial cells triggering pyroptosis in a caspase-11-dependent way in vitro and in vivo. Importantly, we show that lung epithelial cells do not express inflammasomes components or caspase-1 suggesting that this cell type relies exclusively on caspase-11 for undergoing cell death in response to bacterial infection. Finally, we show that IL-18's protective action in melioidosis was completely dependent on its ability to induce IFNγ production. In turn, protection conferred by IFNγ against melioidosis was dependent on generation of ROS through the NADPH oxidase but independent of induction of caspase-11. Altogether, our results identify two non-redundant protective roles for caspase-1 and caspase-11 in melioidosis: Caspase-1 primarily controls pyroptosis of infected macrophages and production of IL-18. In contrast, caspase-11 mediates pyroptosis of infected lung epithelial cells.


Assuntos
Caspase 1/fisiologia , Caspases/fisiologia , Interleucina-18/metabolismo , Pulmão/citologia , Melioidose/prevenção & controle , Piroptose/fisiologia , Animais , Burkholderia/fisiologia , Caspases Iniciadoras , Linhagem Celular , Modelos Animais de Doenças , Feminino , Interferon gama/fisiologia , Macrófagos/microbiologia , Macrófagos/fisiologia , Masculino , Melioidose/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Espécies Reativas de Oxigênio/metabolismo , Mucosa Respiratória/citologia
10.
Mar Pollut Bull ; 128: 324-332, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29571380

RESUMO

Inorganic arsenic (As) is one of the most toxic pollutants in the water. We have studied their effects on the marine teleost European sea bass (Dicentrarchus labrax) at 2 and 10 days of 5 µM of As2O3 (sub-lethal doses) waterborne exposure. Arsenic accumulates in liver and gill tissues. The expression profile of five genes (bax, blc2, casp3, casp8 and casp9) involved in apoptosis cell death confirmed apoptotic effects in liver, slight changes in gill and no effects in skin according with the histopathology findings. Total IgM level and peroxidase activities were increased at 2 and 10 days, respectively. The bactericidal activity was decreased at 2 days after As exposure. A general decrease of cellular immune activities with significant differences in the case of respiratory burst activity was observed after 2 and 10 days of exposure. This work describes for the first time the effects of As exposure on European sea bass.


Assuntos
Apoptose/efeitos dos fármacos , Bass/imunologia , Óxidos/toxicidade , Fagocitose/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Apoptose/genética , Trióxido de Arsênio , Arsenicais , Bass/genética , Expressão Gênica/efeitos dos fármacos , Brânquias/efeitos dos fármacos , Brânquias/patologia , Imunidade Celular/efeitos dos fármacos , Imunoglobulina M/sangue , Fígado/efeitos dos fármacos , Fígado/patologia
11.
Fish Shellfish Immunol ; 75: 291-294, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29448028

RESUMO

Skin lesions contribute to significant rates of morbidity and mortality in fish because they permit the entry of pathogens. Different seric immune parameters (IgM level, protease, antiprotease, peroxidase and lysozyme) and bactericidal activity (against Vibrio harveyi and Photobacterium damsealae) of gilthead seabream (Sparus aurata L.) specimens were evaluated after experimentally wounding fish in two body locations (above or below the lateral line). The results demonstrate that the level of several immune parameters present in fish serum (IgM, proteases, peroxidase and bactericidal activity) showed statistically significant variations depending on the site of the skin wound and the time post-wounding (from 0 to 7 days). However, other parameters (such as antiproteases or lysozyme) remained unaltered during the experiment and did not differs from the values recorded on control fish (non-wounded). The highest activities recorded coincided with the inflammatory healing phase. Moreover, many more significant variations were observed in fish wounded below the lateral line than in those wounded above the lateral line. The present results demonstrate the importance of skin integrity in the maintenance of fish body homeostasis.


Assuntos
Imunidade Humoral/imunologia , Dourada/imunologia , Pele/imunologia , Animais , Dourada/lesões , Pele/lesões
12.
Microb Pathog ; 114: 153-162, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29146500

RESUMO

Disease outbreaks related to waterborne pathogen contamination throughout the world as well as challenges that lie ahead for addressing persistent infection are of renewed interest. In this research, we studied the effects of prolonged exposure of Salmonella enterica serovar Typhimurium to the cues encountered in the extracellular environment particularly in seawater microcosm on bacterial virulence and subsequent infection in Caco-2 cells. Our data show a significant difference in biofilm formation, swimming and swarming motilities between normal and stressed cells of S. Typhimurium under differing NaCl conditions (P < 0.05). Interestingly, adhesion, invasion and apoptotic activity to Caco-2 epithelial cells were determined during infection with normal and stressed Salmonella. Furthermore, we compared the expression of SPI-1 virulence genes (sopA, sopB, sopD, sopE2 and hilA) of normal and stressed S. Typhimurium in response to salt conditions encountered in the extracellular environment in LB broth and after epithelial cell exposure. The interest of the present study is due to the fact that to investigate the bacterial survival strategies during its movement from the natural surroundings to the host cell is fundamental to our understanding of the infection process during the host-pathogen interactions.


Assuntos
Proteínas de Bactérias/genética , Biofilmes , Infecções por Salmonella/microbiologia , Salmonella typhimurium/fisiologia , Fatores de Virulência/genética , Apoptose , Proteínas de Bactérias/metabolismo , Células CACO-2 , Regulação Bacteriana da Expressão Gênica , Interações Hospedeiro-Patógeno , Humanos , Salmonella typhimurium/citologia , Salmonella typhimurium/genética , Água do Mar/análise , Água do Mar/microbiologia , Cloreto de Sódio/análise , Cloreto de Sódio/metabolismo , Fatores de Virulência/metabolismo
13.
BMC Genomics ; 18(1): 939, 2017 Dec 02.
Artigo em Inglês | MEDLINE | ID: mdl-29197330

RESUMO

BACKGROUND: Skin and its mucus are known to be the first barrier of defence against any external stressors. In fish, skin wounds frequently appear as a result of intensive culture and also some diseases have skin ulcers as external clinical signs. However, there is no information about the changes produced by the wounds in the mucosae. In the present paper, we have studied the alterations in the proteome map of skin mucus of gilthead seabream during healing of experimentally produced chronic wounds by 2-DE followed by LC-MS/MS. The corresponding gene expression changes of some identified skin proteins were also investigated through qPCR. RESULTS: Our study has successfully identified 21 differentially expressed proteins involved in immunity and stress processes as well as other metabolic and structural proteins and revealed, for the first time, that all are downregulated in the skin mucus of wounded seabream specimens. At transcript level, we found that four of nine markers (ighm, gst3, actb and krt1) were downregulated after causing the wounds while the rest of them remained unaltered in the wounded fish. Finally, ELISA analysis revealed that IgM levels were significantly lower in wounded fish compared to the control fish. CONCLUSIONS: Our study revealed a decreased-expression at protein and for some transcripts at mRNA levels in wounded fish, which could affect the functionality of these molecules, and therefore, delay the wound healing process and increase the susceptibility to any infection after wounds in the skin of gilthead seabream.


Assuntos
Proteínas de Peixes/metabolismo , Muco/metabolismo , Proteoma/metabolismo , Dourada/metabolismo , Pele/metabolismo , Cicatrização , Animais , Biomarcadores/metabolismo , Proteoma/genética , Proteômica/métodos , Dourada/crescimento & desenvolvimento , Pele/lesões
14.
Fish Shellfish Immunol ; 71: 210-219, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29017945

RESUMO

Skin lesions are very common in fisheries, increasing the risk of pathogens entering through the wounded skin of the fish. In the present assay, the progression of wound healing was studied over a 7 day period in gilthead seabream (Sparus aurata L.) after making experimental wounds in two different locations: above (group A) or below (group B) the lateral line. Macroscopic observation confirmed faster wound healing of the wounds of fish from group B. Furthermore, several immune-related components were studied in the skin mucus of wounded fish to ascertain whether wounding altered the mucus composition compared with the values obtained from non-wounded fish (group C, control). Significant variations were detected depending on both the site of the wound and the studied parameter. At the same time, the gene expression profile of several immune-relevant genes, including pro-inflammatory (il1b,il6, tnfa), anti-inflamamtory (tgfb, il10), immunoglobulins (ighm, ight), involved in oxidative stress (sod, cat) and in skin regeneration (krt1and grhl1) were studied in the three groups of fish (A, B and C). The results throw further light on the complex process of skin wound healing in fish, since substantial changes in the skin mucus and in the skin gene expression originated by the presence of wounds were observed. This work underline some important differences depending on the place of the fish body where the wound is located. Of particular note was the fact that such changes depended on the site of the wound.


Assuntos
Imunidade nas Mucosas , Dourada/imunologia , Pele/imunologia , Transcriptoma , Cicatrização , Animais , Sistema da Linha Lateral/imunologia , Sistema da Linha Lateral/lesões , Dourada/genética , Dourada/lesões , Pele/lesões
15.
PLoS One ; 12(6): e0180438, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28666033

RESUMO

The skin is the first barrier of defence in fish, protecting against any external stressor and preserving the integrity and homeostasis of the fish body. The aim of this study was to characterise gilthead seabream skin by isolating cells and studying the cell cycle by flow cytometry, to study the skin histology by scanning electron microscopy and the transcription level of some immune-relevant genes by RT-PCR. Furthermore, the results obtained from samples taken from the dorsal and the ventral part of the specimens are compared. No differences were observed in the cell cycle of cells isolated from the dorsal and ventral zones of the skin or in the gene expression of the genes studied in both epidermal zones. However, the epidermis thickness of the ventral skin was higher than that of the dorsal skin, as demonstrated by image analysis using light microscopy. Besides, scanning electron microscopy pointed to a greater cell size and area of microridges in the apical part of the dorsal epidermal cells compared with ventral skin epidermal cells. This study represents a step forward in our knowledge of the skin structure of an important farmed teleost, gilthead seabream, one of the most commonly farmed fish worldwide. Furthermore, for functional characterization, experimental wounds were carried out comparing the wound healing rate between the dorsal and ventral regions of skin over the time. The results showed higher ratio of wound healing in the ventral region, whose wounds were closed after 15 days, compared to dorsal region of skin. Taking into account all together, this study represents a step forward in our knowledge of the skin structure and skin regeneration of an important farmed teleost, gilthead seabream, one of the most commonly farmed fish worldwide.


Assuntos
Aquicultura , Dourada/anatomia & histologia , Fenômenos Fisiológicos da Pele , Animais , Ciclo Celular , Perfilação da Expressão Gênica , Microscopia Eletrônica de Varredura , Pele/anatomia & histologia , Pele/citologia , Pele/metabolismo , Pele/ultraestrutura , Cicatrização , Ferimentos e Lesões/patologia
16.
Dev Comp Immunol ; 76: 189-199, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28625747

RESUMO

Haptoglobin (Hp) molecule has been cloned and characterized in two marine teleosts (gilthead seabream and European sea bass), obtaining putative proteins of 319 residues encoded by an ORF of 960 bp in both species. However, the matrix of similarity revealed low identities among bony fish species 78.9% (seabream-sea bass), 43% (seabream/seabass-zebrafish) and lower than 20% with sharks and human. The protein sequences showed a signal peptide from the position 1 to 23, a trypsin domain from 47 to 297, and several predicted disulfide bridges and glycosylation sites. The expression of hp transcript levels during ontogeny showed a progressive increase of expression in seabream whilst remained almost unaltered in sea bass. By tissues, this gene was found constitutively expressed with the highest levels on liver in both species. The main results on hp transcript levels showed the up-regulation in gilthead seabream suffering from naturally occurring lymphocystis disease; and the down-regulation and up-regulation after nodavirus infection in the resistant gilthead seabream and the susceptible European sea bass, respectively. These findings demonstrate for the first time an important role of haptoglobin against viral infections, operating differently in two of the most important marine farmed fish species.


Assuntos
Bass/genética , Proteínas de Peixes/genética , Haptoglobinas/genética , Dourada/genética , Viroses/genética , Animais , Regulação para Baixo/genética , Doenças dos Peixes/genética , Doenças dos Peixes/virologia , Nodaviridae/patogenicidade , Regulação para Cima/genética , Viroses/virologia
17.
Microb Pathog ; 106: 40-49, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-27840222

RESUMO

Pathogenic bacteria such as Salmonella have the ability to respond to a wide variety of environmental stimuli. These responses allow them to survive and withstand insults both of an external location as well as within the host. The aim of this study was to investigate the effect of preadaptation in stressful conditions encountered in seawater microcosms for different periods of time on Salmonella Typhimurium survival, antibiotic susceptibility and interactions with Caco-2 cells. These results showed that the number of bacterial cells depends from the periods of stress in culture medium, highlighting the importance of using the right culture medium for the enumeration of stressed bacteria. The antibiotic resistance of starved cells was modified and their exposure to stressful conditions in seawater during 12 months significantly increased adhesion, invasion and cytotoxic activities on Caco-2 cells. Moreover, cellular cytokines IL-6 and IL-8 secretions were up-regulated. Present results seem to suggest that the preadaptation of S. Typhimurium in seawater microcosms affect the cultural characters by the appearance of the atypical cells that may play a critical role in the intestinal infection and in the systemic spread of the disease. These findings are very important to understand bacterial responses to changing conditions and explain the persistence of these atypical in eukaryotic cells.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Citocinas/metabolismo , Citocinas/farmacologia , Infecções por Salmonella/imunologia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/imunologia , Células CACO-2/citologia , Células CACO-2/efeitos dos fármacos , Meios de Cultura , Farmacorresistência Bacteriana , Meio Ambiente , Interações Hospedeiro-Patógeno/imunologia , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Transmissão , Modelos Biológicos , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/patogenicidade , Água do Mar/microbiologia , Estresse Fisiológico , Fatores de Tempo
18.
Sci Rep ; 6: 39054, 2016 12 14.
Artigo em Inglês | MEDLINE | ID: mdl-27966609

RESUMO

Symbiotic bacteria and mucosal immunoglobulins have co-evolved for millions of years in vertebrate animals. Symbiotic bacteria products are known to modulate different aspects of the host immune system. We recently reported that Flectobacillus major is a predominant species that lives in the gill and skin mucosal surfaces of rainbow trout (Oncorhynchus mykiss). F. major is known to produce sphingolipids of a unique molecular structure. Here we propose a role for F. major and its sphingolipids in the regulation of B cell populations in rainbow trout, as well as an essential role for sphingolipids in trout mucosal homeostasis. We found that F. major-specific IgT titers are confined to the gill and skin mucus, whereas F. major-specific IgM titers are only detected in serum. Live F. major cells are able to stimulate sustained IgT expression and secretion in gills. F. major sphingolipids modulate the growth of trout total skin and gill symbiotic bacteria. In vivo systemic administration of F. major sphingolipids changes the proportion of IgT+ to IgM+ B cells in trout HK. These results demonstrate the key role of the symbiont F. major and its sphingolipids in mucosal homeostasis via the modulation of mucosal and systemic Igs and B cells.


Assuntos
Linfócitos B/imunologia , Cytophagaceae/fisiologia , Mucosa/imunologia , Oncorhynchus mykiss/microbiologia , Esfingolipídeos/imunologia , Animais , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/metabolismo , Cytophagaceae/imunologia , Proteínas de Peixes , Brânquias/imunologia , Brânquias/microbiologia , Homeostase , Imunoglobulinas/sangue , Imunoglobulinas/metabolismo , Mucosa/microbiologia , Oncorhynchus mykiss/imunologia , Pele/imunologia , Pele/microbiologia , Simbiose
19.
Fish Shellfish Immunol ; 58: 500-507, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27697558

RESUMO

Skin mucus is increasingly used as a source for determining immunity-related proteins and enzymes. However, the ability to accurately measure some activities may be modified by inadequate handling and storage of the samples. This study aims to measure the effect of freezing and lyophilization at the time of collection on such activities. Fresh, frozen (immediately after collection at -20 °C and -80 °C) and lyophilized skin mucus samples obtained from the same groups of fish specimens of gilthead seabream (Sparus aurata L.) were analysed in the assays. The amount of total proteins and sugar residues (determined by lectin binding) present in skin mucus samples fell after both freezing and lyophilization of the samples. While no significant differences were exhibited in the levels of some proteins or enzymes (immunoglobulin M, antiprotease, peroxidase, esterase and alkaline phosphatase) determined in fresh or frozen mucus samples, protease and lysozyme activities were lower in frozen mucus samples than in fresh samples. Lyophilization of the mucus samples drastically decreased the total level of proteins obtained, as well as of protease, peroxidase, lysozyme and alkaline phosphatase activities. The results suggest that freezing skin mucus samples is more suitable than lyophilization if samples are stored before determining enzymatic activities.


Assuntos
Liofilização/veterinária , Congelamento , Imunidade Humoral , Dourada/imunologia , Manejo de Espécimes/veterinária , Animais , Muco/imunologia , Pele
20.
Rev. Fac. Nac. Salud Pública ; 34(3): 275-284, set.-dic. 2016. tab
Artigo em Espanhol | LILACS | ID: biblio-957178

RESUMO

RESUMEN Objetivo: analizar el estado de la partería en Jalisco, México, frente al proceso de profesionalización. Metodología: Estudio de caso, realizado desde junio de 2011 a julio de 2013 en tres momentos: I. Entrevistas a profundidad con 20 parteras, II. Cuatro asambleas regionales con 44 parteras, III. Una asamblea estatal con 136 parteras, personal de salud y otros actores sociales, retomando técnicas educativas populares. Los datos cualitativos fueron analizados bajo el modelo actancial semiótico y los datos sociodemográficos de parteras en Epi Info 7, realizando una meta-inferencia con ambos hallazgos. Resultados: 180 parteras informantes, 89% de zona rural, media de edad 56 años, 73% con estudios de secundaria o menos, 95% son parteras tradicionales, adoptando esta práctica desde la adolescencia, capacitándose en instituciones de salud y entre las mismas parteras; 16.7% estudiaron enfermería buscando posicionamiento institucional, sin dejar de ser parteras. Actualmente otorgan atención materno-infantil y a nosologías tradicionales, aludiendo abandono institucional. Existen posturas de rechazo y aceptación de su incorporación al sector salud en médicos y enfermeras, mientras que otros actores sociales, piden su reconocimiento. Conclusiones: la partería en Jalisco, México, es una práctica tradicional vital, con o sin reconocimiento institucional, transformándose en respuesta a necesidades de atención, teniendo que considerar el perfil sociodemográfico de las parteras, su papel actual, la perspectiva del personal de salud y otros actores sociales, junto con la estrecha relación sociocultural y económica del país, frente a su profesionalización.


ABSTRACT Objective: to analyze the status of midwifery in Jalisco, Mexico and its professionalization process. Methodology: a case study conducted from June 2011 to July 2013 in three stages: I. in-depth interviews with 20 midwives, II. four regional assemblies with 44 midwives, and III. a state assembly with 136 midwives, health care staff and other social actors. The three stages employed popular educational techniques. In addition, qualitative data were analyzed using the actantial semiotic model, and the midwives' sociodemographic data were processed in Epi Info 7. Finally, a meta-inference was performed with the findings from both datasets. Results: a total of 180 midwives participated in the study, 89% of which operate in rural areas. The average age was 56, and 73% of the midwives had secondary education or less. Moreover, 95% of them were traditional midwives who adopted this practice since their adolescence and received training from health care providing institutions and from other midwives. In addition, 16.7% studied nursing in order to obtain positions in institutions while remaining midwives. They currently provide care for mothers and children as well as for patients with traditional nosologies. They also mention that they suffer institutional neglect. Among doctors and nurses, there are positions of rejection and acceptance regarding their inclusion in the health sector, while other social actors request their acknowledgement. Conclusions: midwifery in Jalisco, Mexico, is a traditional and vital practice with or without institutional recognition. It is an answer to the need for health care. Likewise, it is necessary to consider the socio-demographic profile of midwives, their current role, the views of the health care staff and other social actors and the close cultural and economic relationship of the Country regarding its professionalization.


RESUMO Objetivo: analisar o estado do partejamento em Jalisco, México, frente ao processo de profissionalização. Métodos: Estudo de caso, realizado desde junho 2011 ao julho 2013 em três momentos: I. Entrevistas em profundidade com 20 parteiras, II. Quatro assembleias regionais com 44 parteiras, III. Uma assembléia estadual com 136 parteiras, funcionários da saúde e outros atores sociais, na retomada das técnicas educacionais populares. Os dados qualitativos foram analisados sob o padrão actancial semiótico e os dados sociais demográficos de parteiras em Epi Info 7, realizando uma meta-inferência com ambos achados. Resultados: 180 parteiras informantes, 89% de zona rural, media de idade 56 anos, 73% com estudos da secundaria ou menos, 95% são parteiras tradicionais, adotando esta prática desde a adolescência, capacitando-se em instituições de saúde e entre as mesmas parteiras; 16.7% estudaram enfermagem procurando posicionamento institucional, sem deixar de ser parteiras. Atualmente outorgam atenção materno-infantil e as nosologías tradicionais, aludindo abandono institucional. Existem posições de rejeito e aceitação da sua incorporação ao setor saúde nos médicos e enfermeiras, entanto que outros atores sociais, pedem o seu reconhecimento. Conclusões: O partejo em Jalisco, México, é uma prática tradicional vital, com ou sem reconhecimento institucional, transformando-se em resposta as necessidades de atenção, tendo que considerar o perfil social demográfico das parteiras, o seu papel atual, a perspectiva dos funcionários da saúde e outros atores sociais, junto com a estreita relação sociocultural e financeira do país, frente a sua profissionalização.

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