RESUMO
Nosocomial infections are often induced by the presence of pathogenic organisms on the surface of medical devices or hospital equipment. Chemical modifications of the surface are recognized as efficient strategies to prevent bacterial adhesion but they may have a negative impact on the material's interaction with living tissues. Here we have developed a photoactivated method for the modification of titanium substrates. A photoinduced technique employing a grafting-onto process has been successfully performed to covalently anchor an imidazolium-derivative siloxane onto titanium surfaces. Imidazolium surfaces showed higher bacteria-repellency performances than native titanium substrates, achieving more than 98% anti-adhesion efficiency against Escherichia coli after 24 h of incubation. In addition, these surfaces allowed for the adhesion and viability of osteoblasts cells without evidence of cytotoxicity.
Assuntos
Aderência Bacteriana , Materiais Revestidos Biocompatíveis/química , Escherichia coli/citologia , Imidazóis/química , Siloxanas/química , Titânio/química , Células 3T3 , Animais , Linhagem Celular , Infecções por Escherichia coli/prevenção & controle , Humanos , Hidrólise , Camundongos , Processos Fotoquímicos , Infecções Estafilocócicas/prevenção & controle , Staphylococcus aureus/citologia , Propriedades de SuperfícieRESUMO
P73, a p53-homologue gene, has been studied for its possible role in head and neck squamous epithelium (HNSE) differentiation and carcinogenesis. P73 RNA and protein were analysed in 50 biopsies, including well- and moderately-differentiated carcinomas, and 21 matched normal adjacent tissues. P73 immunohistochemical analyses revealed intense p73 nuclear staining in basal and parabasal cells of normal squamous epithelium, in contrast with complete absence of staining in the more superficial cell layers. Moderately-differentiated carcinomas demonstrated homogeneous and diffuse staining in all tumour cells, while only basal cells were stained in well-differentiated carcinomas as in normal tissue. No correlation was observed between p73 and p53 protein expression. Immunostaining for p63, another p53-related protein previously described as being involved in HNSE morphogenesis and overexpressed in head and neck squamous cell carcinomas (HNSCC), was found to be similar to p73 labelling in carcinomas, but spread to the more differentiated layers in normal epithelium. Biallelic expression of p73 was found in tumours as well as in matched normal tissues. Comparison of p73 transcript levels between tumours and normal tissues showed decreased mRNA expression in 5/17 (30%) tumours independently of the differentiation status. Mutation and loss of heterozygosity analyses of the p73 gene revealed wild type status and no deletion. Our results strongly suggest that: (i) p73 is associated with homeostasis and control of differentiation of head and neck squamous epithelium probably in concert with p53 and p63; (ii) down-regulation of p73 expression could participate in HNSE carcinogenesis.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Proteínas de Ligação a DNA/biossíntese , Células Epiteliais/metabolismo , Neoplasias de Cabeça e Pescoço/metabolismo , Proteínas de Membrana , Proteínas Nucleares/biossíntese , Fosfoproteínas/biossíntese , Transativadores/biossíntese , Proteína Supressora de Tumor p53/biossíntese , Alelos , Diferenciação Celular , Regulação para Baixo , Genes Supressores de Tumor , Heterozigoto , Humanos , Neoplasias Hipofaríngeas/metabolismo , Imuno-Histoquímica , Perda de Heterozigosidade , Modelos Genéticos , Mutação , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição , Proteína Tumoral p73 , Proteínas Supressoras de TumorRESUMO
The p73 gene is a p53 homologue located at 1p36-33, a region submitted to deletions in breast cancer (BC) and putatively imprinted. To study whether p73 was associated with breast carcinogenesis, loss of heterozygosity (LOH), allele expression and transcript levels were assessed in 59 BC, including 39 BC presenting no inflammatory symptoms (NBC) and 20 inflammatory BC (IBC). IBC is a rare but aggressive form of cancer with a very poor prognosis. Normal breast epithelium (BE) and lymphocytes from patients were used as controls. StyI polymorphism generating GC and/or AT alleles was used to select 22 heterozygous patients. p73 LOH was significantly higher in IBC than in NBC [five of eight cases (62%) versus two of 14 cases (14%); Fisher's exact test, P=0.05]. p73 was biallelically expressed in all BE. In contrast, 12 of 16 (75%) BC were monoallelically expressed, showing that allele silencing was significantly associated with breast carcinogenesis (P=0.012), AT being the preferential silent allele (10 out of 12 tumours). p73 mRNA levels in NBC and IBC were two- and threefold lower than in BE, respectively, suggesting that decreased expression could be related to tumour aggressiveness. In conclusion, LOH, allele silencing and decreased expression of the p73 gene may play a role in breast carcinogenesis.
Assuntos
Alelos , Processamento Alternativo , Neoplasias da Mama/genética , Carcinoma Ductal de Mama/genética , Proteínas de Ligação a DNA/genética , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Perda de Heterozigosidade/genética , Proteínas Nucleares/genética , Neoplasias da Mama/epidemiologia , Neoplasias da Mama/imunologia , Carcinoma Ductal de Mama/epidemiologia , Carcinoma Ductal de Mama/imunologia , Feminino , França/epidemiologia , Genes Supressores de Tumor , Humanos , Prevalência , Proteína Tumoral p73 , Proteínas Supressoras de TumorRESUMO
Cells aggregate on an original cellulose substratum (CEL). This influences the signaling programs of adhering cells. CEL thus appears to be a suitable tool for studying the regulation of cell-substratum and cell-cell interactions.
Assuntos
Técnicas de Cultura de Células/métodos , Melanoma , Animais , Carboximetilcelulose Sódica , Adesão Celular , Agregação Celular , Linhagem Celular Tumoral , Humanos , Derivados da Hipromelose , Melanoma/ultraestrutura , Metilcelulose/análogos & derivadosRESUMO
Palindromic rheumatism is characterized by episodes of arthritis or para-arthritis leaving no residual or radiographic changes. Several diseases should be ruled out in the differential diagnosis. Evolution to rheumatoid arthritis is common, especially in patient with positive rheumatoid factor and anticitrullinated peptides. In seronegative patients, palindromic rheumatism could be part of the spectrum of autoinflammatory diseases because of a high frequency of MEFV mutations. Treatment remains discussed. The use of antimalarials could delay the development of rheumatoid arthritis or another connective tissue disease.
Assuntos
Doenças Reumáticas/diagnóstico , Artrite/diagnóstico , Artrite Reumatoide/fisiopatologia , Citrulina/análise , Proteínas do Citoesqueleto/genética , Diagnóstico Diferencial , Progressão da Doença , Doenças Hereditárias Autoinflamatórias/diagnóstico , Humanos , Mutação/genética , Pirina , Doenças Reumáticas/genética , Doenças Reumáticas/fisiopatologia , Fator Reumatoide/análiseRESUMO
The surface of an implant device can be modified by immobilizing biological molecules on it to improve its integration into the host tissue. We have previously demonstrated that enzymatically tailored plant pectins are promising nanocoatings for biomaterials. This study investigates whether a coating of modified hairy region (rhamnogalacturonan-I) from apple pectin (MHR-alpha) which has anti-adhesive properties can inhibit the generation of inflammatory mediators by lipopolysaccharide (LPS)-activated macrophages. For that purpose, J774.2 murine macrophages were cultured for 24h on MHR-alpha-coated Petri dishes and tissue culture polystyrene controls, with and without LPS. Cell morphology, cell growth, nitrite and TNF-alpha secretion were studied. The results indicate that MHR-alpha coating inhibits the LPS-induced activation of macrophages.
Assuntos
Enzimas Imobilizadas/química , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Pectinas/química , Pectinas/farmacologia , Animais , Linhagem Celular , Citocinas , Macrófagos/efeitos dos fármacos , CamundongosRESUMO
There is evidence that the shape of cells and their contact with a matrix direct the growth and the differentiation of both normal and cancer cells. Cells in 3D culture resemble the in vivo situation more closely than do those in conventional 2D cultures. We have studied the interactions and functions of B16F10 mouse melanoma cells, which spread and grow well on tissue culture polystyrene (tPS), when they were made to aggregate on cellulose-coated Petri dishes (CEL). This aggregation of melanoma cells on CEL was Ca2+ dependent and mediated by N-cadherins. The levels of N-cadherin and beta-catenin transcripts in cells cultured on CEL and tPS were similar, but those on CEL contained less beta-catenin protein. Immunoprecipitation and immunostaining showed that both N-cadherins and beta-catenins were present at the membranes of cells on CEL. Cells proliferated significantly more slowly after 48 h on CEL and the cellulose coating caused most of them to arrest in G1. We also compared the melanin contents and tyrosinase activity of cells on CEL and controls grown on tPS. Melanogenesis was induced in cells aggregated on CEL. A cellulose substrate thus appears to be an outstanding tool for studying cell-cell interactions and cell functions in 3D cultures.