RESUMEN
Triple-negative breast cancer (TNBC) can be divided into six subtypes. Among these subtypes, the basal-like 2 (BL2) subtype shows the lowest five-year survival rate and highest risk of metastasis. Alpha-crystallin B chains (αB-crystallin), a small heat shock protein that is known to be involved in breast cancer metastasis, is highly expressed in the basal-like subtype but not in the other non-basal subtypes. Thus, we hypothesized that αB-crystallin may be an important factor involved in the worse prognosis of the BL2 subtype compared with those of the other TNBC subtypes. Here, we examined the role of αB-crystallin in cell motility in two TNBC cell lines: HCC1806 (BL2 subtype) and, as control, MDA-MB-436 (mesenchymal stem-like subtype). HCC1806 showed greater cell migration capacity and a higher expression level of the gene encoding αB-crystallin (CRYAB) than did MDA-MB-436. Short interfering RNA-mediated silencing of CRYAB expression significantly reduced the cell migration capacity of HCC1806 cells, whereas it had no effect in MDA-MB-436 cells, indicating that αB-crystallin is essential for the migration of HCC1806 cells. Thus, high αB-crystallin expression may be a contributing factor to the poor prognosis of BL2 TNBC.
Asunto(s)
Movimiento Celular , Neoplasias de la Mama Triple Negativas , Cadena B de alfa-Cristalina , Humanos , Pronóstico , Neoplasias de la Mama Triple Negativas/genética , Neoplasias de la Mama Triple Negativas/metabolismo , Cadena B de alfa-Cristalina/genética , Cadena B de alfa-Cristalina/metabolismoRESUMEN
Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) provide a favorable treatment outcome in patients with EGFR mutation-positive non-small cell lung cancer. However, most of such patients become resistant to EGFR-TKIs within a year. Thus, clarifying the mechanism of acquired resistance to EGFR-TKIs has been a research focus. Here, we demonstrated that the expression of progesterone receptor membrane component 2 (PGRMC2) was upregulated in an erlotinib-resistant cell line, PC9/ER, compared with the parental PC9 lung cancer cells. Our previous study showed that PGRMC1 is responsible for acquired resistance to erlotinib; however, PGRMC2 has not been discussed yet. Thus, the aim of this study was to determine the role of PGRMC2 in acquired resistance to erlotinib. Transfection with PGRMC2 siRNA significantly enhanced the sensitivity to erlotinib in PC9/ER cells. Furthermore, knockdown of PGRMC2 reduced the expression of p21, which is known as cell-cycle inhibitor and antiproliferative effector. These results suggest that PGRMC2 partially contributes to erlotinib resistance in PC9/ER cells, and that investigation into the effect of PGRMC2 on apoptosis and the cell cycle are warranted.
Asunto(s)
Adenocarcinoma del Pulmón , Carcinoma de Pulmón de Células no Pequeñas , Neoplasias Pulmonares , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/genética , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular Tumoral , Resistencia a Antineoplásicos/genética , Receptores ErbB , Clorhidrato de Erlotinib/farmacología , Humanos , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Proteínas de la Membrana/genética , Mutación , Inhibidores de Proteínas Quinasas/farmacología , Receptores de Progesterona/genética , Transducción de SeñalRESUMEN
Exosomes are potent players in the development of metastases and they play an important role in cancer angiogenesis and exacerbation. However, it is unclear how proteins on exosomes affect development of blood vessel networks. In this study, we focused on relationships between membrane proteins on exosomes and angiogenesis using human umbilical vein endothelial cells (HUVEC). Lung tumor cell-derived exosomes induced tube formation and growth of endothelial cells in vitro in a dose-dependent manner involving MAPK activation, but this was not seen in normal lung epithelial cells. Ephrin type-A receptor 2 (EphA2) was identified by proteomic analysis and an inhibition assays showed it is a major MAPK activator on exosomes. Thus EphA2 on exosomes participates in angiogenesis as a ligand of the ephrin signaling pathway. These results support the development of novel therapeutic strategies such as blockade of remote cancer communications through exosomes.
Asunto(s)
Efrina-A2/metabolismo , Exosomas/metabolismo , Neoplasias Pulmonares/irrigación sanguínea , Sistema de Señalización de MAP Quinasas , Inductores de la Angiogénesis , Línea Celular Tumoral , Células Epiteliales/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Neoplasias Pulmonares/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Cultivo Primario de Células , Receptor EphA2 , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Fimbriae are protein-based filamentous appendages that protrude from the bacterial cell surface and facilitate host adhesion. Two types of fimbriae, FimA and Mfa1, of the periodontal pathogen Porphyromonas gingivalis are responsible for adherence to other bacteria and to host cells in the oral cavity. Both fimbrial forms are composed of 5 proteins, but there is limited information about their polymerization mechanisms. Here, the authors evaluated the function of Mfa5, one of the Mfa1 fimbrial accessory proteins. Using mfa5 gene disruption and complementation studies, the authors revealed that Mfa5 affects the incorporation of other accessory proteins, Mfa3 and Mfa4, into fibers and the expression of fimbriae on the cell surface. Mfa5 is predicted to have a C-terminal domain (CTD) that uses the type IX secretion system (T9SS), which is limited to this organism and related Bacteroidetes species, for translocation across the outer membrane. To determine the relationship between the putative Mfa5 CTD and the T9SS, mutants were constructed with in-frame deletion of the CTD and deletion of porU, a C-terminal signal peptidase linked to T9SS-mediated secretion. The ∆CTD-expressing strain presented a similar phenotype to the mfa5 disruption mutant with reduced expression of fimbriae lacking all accessory proteins. The ∆porU mutants and the ∆CTD-expressing strain showed intracellular accumulation of Mfa5. These results indicate that Mfa5 function requires T9SS-mediated translocation across the outer membrane, which is dependent on the CTD, and subsequent incorporation into fibers. These findings suggest the presence of a novel polymerization mechanism of the P. gingivalis fimbriae.
Asunto(s)
Proteínas Fimbrias/fisiología , Porphyromonas gingivalis/fisiología , Adhesión Bacteriana/fisiología , Western Blotting , Electroforesis en Gel de Poliacrilamida , Proteínas Fimbrias/genética , Proteínas Fimbrias/aislamiento & purificación , Regulación Bacteriana de la Expresión Génica/fisiología , Mutación/genética , Porphyromonas gingivalis/genéticaRESUMEN
Tumor necrosis factor (TNF)/TNF receptors (TNFR1/TNFR2) are considered to be potential drug targets to treat refractory diseases, including autoimmune diseases and malignant tumors. However, their specific functions, especially in the case of TNFR2, are poorly understood. In this study, we constructed a mouse TNFR2 (mTNFR2)-mediated biological assay system that shows no effects of mouse TNFR1 (mTNFR1) in order to screen mTNFR2-selective stimulating agents. Mouse TNFR1(-/-)R2(-/-) preadipocytes were transfected with the gene encoding the mTNFR2/mouse Fas (mFas) chimeric receptor in which the extracellular and transmembrane domains of mTNFR2 were fused to the intracellular domain of mFas. Our results demonstrated that this cell line exhibits highly sensitive mTNFR2-mediated cytotoxic effects. We propose that this mTNFR2-mediated biological assay system would be a useful tool to screen for mTNFR2-selective stimulating agents.
Asunto(s)
Adipocitos/citología , Receptores Tipo II del Factor de Necrosis Tumoral/genética , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Receptor fas/genética , Animales , Bioensayo/métodos , Línea Celular , Ratones , Ratones Noqueados , Receptores Tipo I de Factores de Necrosis Tumoral/efectos de los fármacos , Receptores Tipo II del Factor de Necrosis Tumoral/efectos de los fármacos , TransfecciónRESUMEN
BACKGROUND: The relationship among natural allergen exposure, induction of blocking antibody and the occurrence of atopic allergy-particularly in the presence of IgE production-is debatable. OBJECTIVE: To clarify the relationship between the dose of cutaneous exposure to dust mite allergen and susceptibility to the IgE-mediated allergic response in relation to IgG production. METHODS: NC/Nga mice were epicutaneously exposed to various doses of Dermatophagoides pteronyssinus allergen to induce atopic dermatitis-like skin lesions. We then evaluated the skin lesions, induction of mite-specific immune responses, and susceptibility to anaphylaxis. RESULTS: Dose-dependent exacerbation of atopic dermatitis-like skin lesions and increases in mite-specific IgG and IgE production were observed. However, mice exposed to relatively low doses of mite allergen showed hypersusceptibility to mite allergen-specific anaphylaxis. We also showed that adoptive transfer of total IgG from Dp-sensitized mice rescued mice from the hypersusceptibility seen in those exposed to low doses of mite allergen. CONCLUSIONS AND CLINICAL RELEVANCE: High-dose cutaneous exposure to dust mites induced effective blocking IgG production, even if accompanied by IgE production. Our data might support the concept that an increase in IgG titre, not a decrease in IgE titre, is a marker of clinical improvement in allergen-specific immunotherapy.
Asunto(s)
Alérgenos/administración & dosificación , Alérgenos/inmunología , Anafilaxia/prevención & control , Anticuerpos Bloqueadores/inmunología , Antígenos Dermatofagoides/administración & dosificación , Antígenos Dermatofagoides/inmunología , Inmunoglobulina G/inmunología , Anafilaxia/inmunología , Anafilaxia/metabolismo , Animales , Especificidad de Anticuerpos/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Inmunización , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Inmunoglobulina G/sangre , RatonesRESUMEN
The periodontal pathogen Porphyromonas gingivalis is known to express 2 distinct types of fimbriae: FimA and Mfa1 fimbriae. However, we previously reported that fimbria-like structures were found in a P. gingivalis strain in which neither FimA nor Mfa1 fimbriae were detected. In this study, we identified a major protein in the bacterial lysates of the strain, which has been reported as the 53-kDa major outer membrane protein of P. gingivalis (53K protein) and subsequently reported as a major fimbrilin of a novel-type fimbria. Sequencing of the chromosomal DNA of the strain showed that the 53k gene (encoding the 53K protein) was located at a locus corresponding to the mfa1 gene (encoding the Mfa1 protein, which is a major fimbrilin of Mfa1 fimbriae) of the ATCC 33277 type strain. However, the 53K and Mfa1 proteins showed a low amino acid sequence homology and different antigenicity. The 53K protein was detected in 34 of 84 (41%) P. gingivalis strains, while the Mfa1 protein was detected in 44% of the strains. No strain expressed both 53K and Mfa1 proteins. Additionally, fimbriae were normally expressed in mutants in which the 53k and mfa1 genes were interchanged. These results indicate that the 53K protein is another major fimbrilin of Mfa1 fimbriae in P. gingivalis.
Asunto(s)
Proteínas Fimbrias/genética , Fimbrias Bacterianas/fisiología , Porphyromonas gingivalis/fisiología , Secuencia de Aminoácidos , Proteínas Bacterianas , Western Blotting , Electroforesis en Gel de Poliacrilamida , Fimbrias Bacterianas/química , Eliminación de Gen , Proteínas de la Membrana/genética , Microscopía Electrónica de Transmisión , Reacción en Cadena de la Polimerasa , Porphyromonas gingivalis/química , Porphyromonas gingivalis/genéticaRESUMEN
Periodontal diseases are semi-ubiquitous and caused by chronic, plaque-induced inflammation. The 55-kDa immunodominant RagB outer membrane protein of Porphyromonas gingivalis, a keystone periodontal pathogen, has been proposed to facilitate nutrient transport. However, potential interactions between RagB and the innate response have not been examined. We determined that RagB exposure led to the differential and dose-related expression of multiple genes encoding proinflammatory mediators [interleukin-1α (IL-1α), IL-1ß, IL-6, IL-8 and CCL2; all P < 0.05] in primary human monocytes and to the secretion of tumor necrosis factor and IL-8, but not interferon-γ or IL-12. RagB was shown to be a Toll-like receptor 2 (TLR2) and TLR4 agonist that activated signal transducer and activator of transcription 4 and nuclear factor-κB signaling, as determined by a combination of blocking antibodies, pharmaceutical inhibitors and gene silencing. In keeping, a ΔragB mutant similarly exhibited reduced inflammatory capacity, which was rescued by ragB complementation. These results suggest that RagB elicits a major pro-inflammatory response in primary human monocytes and, therefore, could play an important role in the etiology of periodontitis and systemic sequelae.
Asunto(s)
Proteínas Bacterianas/inmunología , Porphyromonas gingivalis/genética , Factor de Transcripción STAT4/agonistas , Receptor Toll-Like 2/agonistas , Receptor Toll-Like 4/agonistas , Proteínas Bacterianas/genética , Citocinas/inmunología , Encía/inmunología , Humanos , Lipopolisacáridos/inmunología , Monocitos/inmunología , FN-kappa B/inmunología , Periodontitis/inmunología , Fosforilación , Transducción de Señal/inmunologíaRESUMEN
OBJECTIVE: This study aimed to clarify the influence of a low-temperature environment on muscle atrophy and apoptosis. METHODS: Wistar rats were divided into four groups: two groups of hindlimb-unloading rats maintained in a normal (25°C, HU) or low-temperature (10°C, HU+LT) environment for 3 weeks and two corresponding control groups (CON; normal temperature, CON+LT; low-temperature). RESULTS: The soleus muscle wet weight and muscle-to-body mass ratio were lower in the experimental groups than in the control groups. The cross-sectional areas of myofibers in the HU+LT and HU groups were significantly decreased than those in the CON and CON+LT groups. Ubiquitin ladder levels from soleus muscle lysates were significantly increased in the HU+LT group. Caspase-3-activated myofibers were observed only in the HU+LT group. Decreased cytochrome c levels were present in these caspase-3-activated myofibers. Meanwhile, cytochrome c levels were increased significantly in CON+LT rats but unchanged in HU+LT rats. CONCLUSION: Our results suggest that apoptosis caused by hindlimb unloading at low temperatures is associated with a lack of cytochrome c in myofibers. This indicates that long-term hindlimb unloading at low temperatures did not suppress muscle atrophy. We conclude that low-temperature stimulation should not be used as a long-term treatment for preventing disuse atrophy.
Asunto(s)
Apoptosis/fisiología , Frío , Citocromos c/metabolismo , Suspensión Trasera/fisiología , Atrofia Muscular/fisiopatología , Animales , Técnica del Anticuerpo Fluorescente , Masculino , Fibras Musculares Esqueléticas/enzimología , Fibras Musculares Esqueléticas/patología , Músculo Esquelético/enzimología , Músculo Esquelético/fisiopatología , Atrofia Muscular/enzimología , Ratas , Ratas WistarRESUMEN
Oral malignancy is rare in chimpanzees. A 34-year-old female chimpanzee (Pan troglodytes) at Kumamoto Sanctuary, Japan, had developed it. Treatment is technically difficult for chimpanzees while malignant neoplasm is seemingly rising in captive populations. Widespread expert discussion, guidelines for treatment, especially for great apes in terminal stages is urgently needed.
Asunto(s)
Animales de Zoológico , Enfermedades del Simio Antropoideo/diagnóstico , Neoplasias de la Boca/veterinaria , Pan troglodytes , Sarcoma/veterinaria , Animales , Enfermedades del Simio Antropoideo/patología , Enfermedades del Simio Antropoideo/terapia , Resultado Fatal , Femenino , Hepacivirus/aislamiento & purificación , Japón , Neoplasias de la Boca/diagnóstico , Neoplasias de la Boca/terapia , Sarcoma/diagnóstico , Sarcoma/terapiaRESUMEN
The fimbriae of Porphyromonas gingivalis, the causative agent of periodontitis, have been implicated in various aspects of pathogenicity, such as colonization, adhesion and aggregation. Porphyromonas gingivalis ATCC 33277 has two adhesins comprised of the FimA and Mfa1 fimbriae. We characterized the PGN0289 (Mfa3) protein, which is one of the three accessory proteins of Mfa1 fimbriae in P. gingivalis. The Mfa3 protein was present in two different sizes, 40 and 43 kDa, in the cell. The 43-kDa and 40-kDa Mfa3 were detected largely in the inner membrane and the outer membrane, respectively. Purified Mfa1 fimbriae contained the 40-kDa Mfa3 alone. Furthermore, the 40-kDa Mfa3 started with the Ala(44) residue of the deduced amino acid sequence, indicating that the N-terminal region of the nascent protein expressed from the mfa3 gene is processed in the transport step from the inner membrane into fimbriae. Immuno-electron microscopy revealed that Mfa3 localized at the tip of the fimbrial shaft. Interestingly, deletion of the mfa3 gene resulted in the absence of other accessory proteins, PGN0290 and PGN0291, in the purified Mfa1 fimbriae, suggesting that Mfa3 is required for integration of PGN0290 and PGN0291 into fimbriae. A double mutant of mfa3 and fimA genes (phenotype Mfa1 plus, FimA minus) showed increased auto-aggregation and biofilm formation similar to a double mutant of mfa1 and fimA genes (phenotype Mfa1(-) , FimA(-) ). These findings suggest that the tip protein Mfa3 of the Mfa1 fimbriae may function in the integration of accessory proteins and in the colonization of P. gingivalis.
Asunto(s)
Proteínas Bacterianas/análisis , Proteínas Fimbrias/análisis , Proteínas Fimbrias/fisiología , Fimbrias Bacterianas/fisiología , Porphyromonas gingivalis/fisiología , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Biopelículas/crecimiento & desarrollo , Proteínas Fimbrias/química , Proteínas Fimbrias/genética , Fimbrias Bacterianas/química , Mutación , Porphyromonas gingivalis/química , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/ultraestructuraRESUMEN
The periodontal pathogen Porphyromonas gingivalis generally expresses two distinct fimbriae, FimA and Mfa1, which play a role in biofilm formation. The fimA gene that encodes FimA fimbrilin is polymorphic, and polymerase chain reaction analysis has identified six genotypes called types I-V and Ib. We found recently that fimbriae exhibit antigenic heterogeneity among the genotypes. In the present study, we analysed the fimA DNA sequences of 84 strains of P. gingivalis and characterized the antigenicity of FimA fimbriae. Strains analysed here comprised 10, 16, 29, 13, 10 and 6 strains of types I, Ib, II, III, IV and V, respectively. DNA sequencing revealed that type Ib does not represent a single cluster and that type II sequences are remarkably diverse. In contrast, the fimA sequences of the other types were relatively homogeneous. Antigenicity was investigated using antisera elicited by pure FimA fimbriae of types I-V. Antigenicity correlated generally with the respective genotype. Type Ib strains were recognized by type I antisera. However, some strains showed cross-reactivity, especially, many type II strains reacted with type III antisera. The levels of fimbrial expression were highly variable, and expression was positively correlated with ability of biofilm formation on a saliva-coated plate. Further, two strains without FimA and Mfa1 fimbriae expressed fimbrial structures, suggesting that the strains produce other types of fimbriae.
Asunto(s)
Antígenos Bacterianos/genética , Proteínas Fimbrias/genética , Pili Sexual/genética , Porphyromonas gingivalis/genética , Variación Antigénica/genética , Antígenos Bacterianos/clasificación , Antígenos Bacterianos/inmunología , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Biopelículas , Reacciones Cruzadas/inmunología , ADN Bacteriano/genética , Película Dental/microbiología , Proteínas Fimbrias/clasificación , Proteínas Fimbrias/inmunología , Genotipo , Humanos , Sistemas de Lectura Abierta/genética , Pili Sexual/inmunología , Porphyromonas gingivalis/inmunología , Saliva/microbiología , Análisis de Secuencia de ADNRESUMEN
The fullerene C60 is used in consumer products such as cosmetics owing to its antioxidative effects and is being developed for nanomedical applications. However, knowledge regarding the safety of fullerene C60, especially after oral administration, is sparse. Here, we examined the safety of fullerene C60 in mice after 7 d of exposure to orally administered polyvinylpyrrolidone (PVP)-wrapped fullerene C60 (PVP-fullerene C60). Mice treated with PVP-fullerene C60 showed few changes in the plasma levels of various markers of kidney and liver injury and experienced no significant hematologic effects. Furthermore, the histology of the colon of PVP-fullerene C60-treated mice was indistinguishable from that of control mice. These results suggest that PVP-fullerene C60 lacks toxicity after high-dose oral administration and indicate that PVP-fullerene C60 can be considered safe for oral medication. These data provide basic information that likely will facilitate the production of safe and effective forms of fullerene C60.
Asunto(s)
Fulerenos/farmacología , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Administración Oral , Animales , Recuento de Células Sanguíneas , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Colitis/inducido químicamente , Colitis/patología , Femenino , Fulerenos/administración & dosificación , Luz , Ratones , Ratones Endogámicos C57BL , Povidona , Dispersión de Radiación , Fijación del TejidoRESUMEN
Detection of drug-target proteins and biomarkers that are expressed in cancer tissue has significant potential for both diagnosis and treatment of cancer. However, current immuno-histochemical and cytogenetic analyses of biopsy specimens for pre-operational diagnosis are highly invasive and often difficult to apply to lung cancer patients. The purpose of this study was to evaluate the possible utility of determining epidermal growth factor receptor (EGFR) expression on exosomal membranes using a targeted ELISA with an anti-CD81 antibody as a capture antibody for lung cancer diagnosis. While soluble EGFR (sEGFR) levels in plasma were not remarkably different between lung cancer patients and normal controls, significantly higher exosomal EGFR expression levels were observed in 5/9 cancer cases compared to normal controls. These results suggest that measurement of exosomal protein levels could be useful for in vitro diagnosis, and that exosomal EGFR is a possible biomarker for characterization of lung cancer.
Asunto(s)
Biomarcadores/análisis , Receptores ErbB/metabolismo , Exosomas/metabolismo , Neoplasias Pulmonares/diagnóstico , Adulto , Anciano , Animales , Western Blotting , Línea Celular Tumoral , Medios de Cultivo Condicionados , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunohistoquímica , Masculino , Membranas/metabolismo , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica , Persona de Mediana Edad , Plásmidos , Tetraspanina 28/metabolismoRESUMEN
Porphyromonas gingivalis has been implicated as a major pathogen associated with chronic periodontitis. To extend our knowledge of post-translational protein glycosylation in P. gingivalis, a proteomic analysis involving two-dimensional polyacrylamide gel electrophoresis combined with carbohydrate staining and mass spectrometry was performed. Four novel glycoproteins, PGN0743, PGN0876, PGN1513 and PGN0729, in P. gingivalis ATCC 33277 were identified. These four identified glycoproteins possess a range of biochemical activities and cellular localization. PGN0743 contains a sequence motif identifying it as a FKBP-type cis-trans isomerase, which has activity usually associated with chaperone functions. PGN0876 and PGN1513 contain tetratricopeptide repeat domains that mediate protein-protein interactions. PGN0729 encodes the outer membrane protein 41 precursor, which was previously identified as Pgm6, and is homologous to the OmpA protein in Escherichia coli. Several different types of glycoprotein were identified, suggesting that P. gingivalis possesses a general mechanism for protein glycosylation. PGN0743-deficient and PGN0876-deficient mutants were constructed to examine the role(s) of the two identified glycoproteins. Both mutants showed a decreased growth rate under nutrient-limited conditions and reduced biofilm formation activity. These results suggest that the novel glycoproteins PGN0743 and PGN0876 play an important role in the growth and colonization of P. gingivalis.
Asunto(s)
Proteínas Bacterianas/aislamiento & purificación , Biopelículas/crecimiento & desarrollo , Glicoproteínas/aislamiento & purificación , Porphyromonas gingivalis/crecimiento & desarrollo , Secuencias de Aminoácidos , Adhesión Bacteriana , Proteínas de la Membrana Bacteriana Externa/aislamiento & purificación , Fraccionamiento Celular , Colorantes , Electroforesis en Gel Bidimensional , Humanos , Espectrometría de Masas , Chaperonas Moleculares/aislamiento & purificación , Mutación/genética , Periplasma/química , Periplasma/ultraestructura , Porphyromonas gingivalis/genética , Porphyromonas gingivalis/fisiología , Proteoma/aislamiento & purificación , Secuencias Repetitivas de Aminoácido , Saliva/microbiología , Proteínas de Unión a Tacrolimus/aislamiento & purificaciónRESUMEN
Since metastasis is one of the most important prognostic factors in colorectal cancer, development of new methods to diagnose and prevent metastasis is highly desirable. However, the molecular mechanisms leading to the metastatic phenotype have not been well elucidated. In this study, a proteomics-based search was carried out for metastasis-related proteins in colorectal cancer by analyzing the differential expression of proteins in primary versus metastasis focus-derived colorectal tumor cells. Protein expression profiles were determined using a tissue microarray (TMA), and the results identified Rho GDP-dissociation inhibitor alpha (Rho GDI) as a metastasis-related protein in colon and prostate cancer patients. Consequently, Rho GDI may be useful as a diagnostic biomarker and/or a therapeutic to prevent colon and prostate cancer metastasis.
Asunto(s)
Neoplasias del Colon/secundario , Inhibidores de Disociación de Guanina Nucleótido/fisiología , Neoplasias de la Próstata/secundario , Anciano , Western Blotting , Línea Celular Tumoral , Cromatografía Líquida de Alta Presión , Electroforesis en Gel Bidimensional , Colorantes Fluorescentes , Geles , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Humanos , Hidrólisis , Inmunohistoquímica , Masculino , Espectrometría de Masas , Análisis por Micromatrices , Persona de Mediana Edad , Tripsina/química , Inhibidores de la Disociación del Nucleótido Guanina rho-EspecíficoRESUMEN
Tannerella forsythia, a gram-negative fusiform rod, is implicated in several types of oral anaerobic infections. Most gram-negative bacteria have OmpA-like proteins that are homologous to the OmpA protein in Escherichia coli. We identified an OmpA-like protein in T. forsythia encoded by the tf1331 gene as one of the major proteins by mass spectrometric analysis. Two-dimensional, diagonal electrophoresis showed that the OmpA-like protein formed a dimeric or trimeric structure via intermolecular disulfide bonds. A biotin labeling experiment revealed that a portion of the protein was exposed on the cell surface, even though T. forsythia possesses an S-layer at the outermost cell surface. Using a tf1331-deletion mutant, we showed that the OmpA-like protein affected cell morphology. The length of the mutant cell was reduced almost by half. Cell swelling was observed in more than 40% of the mutant cells. Moreover, the mutant exhibited decreased adhesion to fibronectin, retarded autoaggregation, and reduced cell surface hydrophobicity. These results suggest that the OmpA-like protein in T. forsythia plays an important role in cellular integrity and adhesive function.
Asunto(s)
Adhesión Bacteriana/fisiología , Proteínas de la Membrana Bacteriana Externa/fisiología , Bacteroides/fisiología , Antibacterianos/farmacología , Proteínas de la Membrana Bacteriana Externa/análisis , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/análisis , Bacteroides/citología , Biopelículas , Western Blotting , Electroforesis en Gel Bidimensional , Electroforesis en Gel de Poliacrilamida , Escherichia coli/genética , Matriz Extracelular/microbiología , Fibronectinas/metabolismo , Vectores Genéticos/genética , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Glicoproteínas de Membrana/análisis , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica , Plásmidos/genética , Eliminación de Secuencia/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
The immune-modulating effect following intradermal injection of various-sized amorphous silica particles was analyzed in terms of induction of ovalbumin-specific CD8+ T cells in vivo. IFN-gamma ELISPOT assays revealed that only nanosilica particles with a diameter of less than 100 nm significantly enhanced CD8+ T cell responses against ovalbumin. These results indicate that the size of nanomaterials is a critical determinant in terms of their safe use.
Asunto(s)
Factores Inmunológicos , Nanopartículas , Dióxido de Silicio/farmacología , Animales , Linfocitos T CD8-positivos/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Femenino , Interferón gamma , Ratones , Ratones Endogámicos C57BL , Ovalbúmina/inmunología , Tamaño de la Partícula , Dióxido de Silicio/química , Bazo/citología , Bazo/inmunologíaRESUMEN
SUMMARYIn December 2006, an outbreak of gastroenteritis occurred involving 372 guests and 72 employees at a hotel after a guest vomited in corridors on the third (F3) and 25th (F25) floors. Norovirus with identical genotype was confirmed by real-time reverse transcription-polymerase chain reaction in faecal samples from guest cases and employees. Spread of the outbreak on F25 was compared with that on F3. The attack rate in the guests who visited F25 alone (15·0%, 106/708 guests) was significantly higher than in those who visited F3 alone (3·5%, 163/4710 guests) (relative risk 4·3, 95% confidence interval 3·4-5·5, P < 0·001). The outbreak on F3 ended within 2 days, while that on F25 extended over 7 days. The environmental ratios of F3 to F25 were 7·4 for volume, 6·9 for floor area and 7·6 for ventilation rate. This outbreak suggests that environmental differences can affect the propagation and persistence of a norovirus outbreak following environmental contamination.