RESUMEN
Polyphosphate [Poly(P)] has positive effects on osteoblast mineralization; however, the underlying mechanism remains unclear. In addition, it is unknown whether Poly(P) promotes mineralization in soft tissues. We investigated this by using various cells. Poly(P) concentrations of 1 and 0.5 mg/mL yielded high levels of mineralization in ROS17/2.8 osteoblast cells. Similarly, Poly(P) induced mineralization in cell types expressing alkaline phosphatase (ALP), namely, ATDC5 and MC3T3-E1, but not in CHO, C3H10T1/2, C2C12, and 3T3-L1 cells. Furthermore, forced expression of ALP caused Poly(P)-induced mineralization in CHO cells. These results suggest that ALP determines Poly(P)-induced mineralization in a cell-type independent manner.
Asunto(s)
Fosfatasa Alcalina/biosíntesis , Calcificación Fisiológica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Osteoblastos/enzimología , Polifosfatos/farmacología , Células 3T3-L1 , Fosfatasa Alcalina/genética , Animales , Células CHO , Calcificación Fisiológica/genética , Cricetinae , Cricetulus , Regulación Enzimológica de la Expresión Génica/fisiología , Ratones , Especificidad de Órganos/efectos de los fármacos , Especificidad de Órganos/fisiología , Osteoblastos/citología , RatasRESUMEN
We examined cell-to-cell interaction between pre-osteoblasts and osteocytes using MC3T3-E1 and MLO-Y4, respectively. First, GFP expressing MC3T3-E1 (E1-GFP) cells were generated to isolate the cells from co-culture with MLO-Y4. No changes were observed in the expression of osteogenic transcription factors Runx2, Osterix, Dlx5 and Msx2, but expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP) in E1-GFP co-cultured with MLO-Y4 was 300-400-fold greater than that in mono-cultured E1-GFP. In addition, mineralized nodule formation was drastically increased in co-cultured E1-GFP cells compared to mono-cultured cells. Patch clamp assay showed the presence of gap junctions between E1-GFP and MLO-Y4. Furthermore, when the gap junction inhibitor carbenoxolone (CBX) was added to the culture, increased expression of ALP and BSP in E1-GFP co-cultured with MLO-Y4 was suppressed. These results suggest that gap junction detected between pre-osteoblasts and osteocytes plays an important role on the terminal differentiation of pre-osteoblasts.
Asunto(s)
Uniones Comunicantes/fisiología , Regulación de la Expresión Génica , Osteoblastos/citología , Osteocitos/citología , Células 3T3 , Fosfatasa Alcalina/metabolismo , Animales , Carbenoxolona/química , Proteínas Portadoras/metabolismo , Comunicación Celular , Ciclo Celular , Diferenciación Celular , Técnicas de Cocultivo , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Sialoproteína de Unión a Integrina/metabolismo , Ratones , Osteocitos/metabolismo , Técnicas de Placa-Clamp , ARN Mensajero/metabolismoRESUMEN
The effects of dissolved elements from metal dental restorations are a major concern in lesions of the oral mucosa, and the evaluation of accumulated metal elements, especially their distribution and chemical state, is essential for determining the precise effects of trace metals. In this study, X-ray fluorescence with synchrotron radiation (SR-XRF) and particle-induced X-ray emission (PIXE) were applied for distribution analysis of the trace metal elements contained in the oral mucosa, and the chemical states of the elements were estimated using X-ray absorption fine structure (XAFS) analysis. Appropriate combination of these analysis techniques, particularly SR-XRF and PIXE, to visualize the distributions of the elements in the oral mucosa allowed for the observation and evaluation of accumulated metal ions and debris. Importantly, the analyses in this study could be carried out using conventional histopathological specimens without damaging the specimens. Therefore, this method would be applicable for the detection of accumulated trace metal elements in biopsy specimens from the oral mucosa.
Asunto(s)
Mucosa Bucal/metabolismo , Espectrometría por Rayos X/métodos , Oligoelementos/análisis , Adulto , Femenino , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana EdadRESUMEN
The 75 kDa transmembrane protein, p75(NTR), is a marker of mesenchymal stem cells (MSCs). Isolated MSCs are capable of differentiating into osteoblasts, but the molecular function of p75(NTR) in MSCs and osteoblasts is poorly understood. The aim of this study was to examine the function of p75(NTR) in the human MG63 osteoblast cell line compared to the murine MC3T3E-1 pre-osteoblast cell line. MG63 cells and MC3T3-E1 cells expressing exogenous p75(NTR) protein (denoted as p75-MG63 and p75GFP-E1, respectively) were generated to compare osteogenic differentiation and cell proliferation abilities. Overexpression of p75(NTR) induced alkaline phosphatase activity and the mRNA expression of osteoblast-related genes such as osterix and bone sialoprotein in both p75-MG63 and p75GFP-E1. Interestingly, exogenous p75(NTR) stimulated cell proliferation and cell cycle progression in p75GFP-E1, but not in p75-MG63. To elucidate any different effects of p75(NTR) expression on osteogenic differentiation and cell proliferation, we examined the mRNA expression of tropomyosin receptor kinase (trk) genes (trkA, trkB, trkC) and Nogo receptor (NgR), which are binding partners of p75(NTR). Although trkA, trkB, and trkC were detected in both p75-MG63 and p75GFP-E1, only NgR was detected in p75-MG63. We then used the K252a inhibitor of the trks to identify the signaling pathway for osteogenic differentiation and cell proliferation. Inhibition of trks by K252a suppressed p75(NTR)-mediated osteogenic differentiation of p75GFP-E1, whereas deletion of the GDI domain in P75(NTR) from the p75-MG63 produced enhanced cell proliferation compared to p75-MG63. These results suggest that p75(NTR) signaling associated with trk receptors promotes both cell proliferation and osteoblast differentiation, but that p75(NTR)-mediated proliferation may be suppressed by signaling from the p75(NTR)/NgR complex.
Asunto(s)
Proliferación Celular , Osteoblastos/metabolismo , Receptor de Factor de Crecimiento Nervioso/metabolismo , Fosfatasa Alcalina/metabolismo , Carbazoles/farmacología , Línea Celular , Humanos , Alcaloides Indólicos/farmacología , Osteoblastos/citología , Osteogénesis , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Proteínas Tirosina Quinasas Receptoras/metabolismo , Receptor de Factor de Crecimiento Nervioso/genética , Sialoglicoproteínas/genética , Sialoglicoproteínas/metabolismo , Transducción de Señal , Factor de Transcripción Sp7 , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Defensins, a major family of cationic antimicrobial peptides, play important roles in innate immunity. In the present study, we investigated whether double-stranded RNA (dsRNA), a by-product of RNA virus replication, can induce human ß-defensins-2 (hBD-2) expression in oral epithelial cells (OECs). We also examined the hBD-2-inducible activity of acid-electrolyzed functional water (FW). The results indicated that both dsRNA- and FW-induced hBD-2 expression in OECs. The induction efficiency was much higher for FW than for dsRNA. FW-induced production of hBD-2 was clearly observed by immunofluorescence staining. A luciferase assay was performed with 1.2 kb of the 5'-untranslated region (5'-UTR) of the hBD-2 gene. The results indicated that the nuclear factor-kappa B (NF-κB)-binding site proximal to the translation initiation site was indispensable for dsRNA-stimulated hBD-2 expression, but not in the case of FW. Moreover, FW-stimulated hBD-2 expression did not depend on NF-κB activity; instead, FW inhibited NF-κB activity. Pretreatment of the cells with specific inhibitors against NF-κB further confirmed NF-κB-independent hBD-2 induction by FW. In analogy to the results for intestinal epithelial cells (IECs), the dsRNA signal, but not FW, was sensed by toll-like receptor 3 (TLR3) in OECs. These results suggested that hBD-2 expression induced by dsRNA and FW is regulated by distinct mechanisms in OECs.
Asunto(s)
Carcinoma de Células Escamosas/metabolismo , Peróxido de Hidrógeno/farmacología , Neoplasias de la Boca/metabolismo , ARN Bicatenario/genética , Transducción de Señal , beta-Defensinas/metabolismo , Western Blotting , Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/genética , Células Cultivadas , Técnica del Anticuerpo Fluorescente , Humanos , Neoplasias de la Boca/tratamiento farmacológico , Neoplasias de la Boca/genética , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , beta-Defensinas/genéticaRESUMEN
Acinar cell regeneration from tubular structures has been reported to occur in duct-deligated salivary glands. However, the detailed process of acinar cell regeneration has not been clarified. We have developed a mouse duct ligation model to clarify the mechanisms underlying acinar cell regeneration, and we analyzed the epidermal growth factor receptor (EGFR) and epidermal growth factor (EGF) ligands using the model. We studied these ligands expressions in the course of acinar cell regeneration using immunohistochemistry and RT-PCR methods. In the duct-ligated portion of the submandibular gland (SMG) that underwent atrophy, newly formed acinar cells were observed arising from the tubular structures after the release of the duct obstruction. The constitutive expression of EGFR was observed by immunohistochemistry in both the duct-ligated and duct-deligated animals as well as in normal controls. The EGFR phosphorylation detected on the tubular structures after duct ligation paralleled the acinar cell regeneration. RT-PCR showed an increase in the epiregulin and heparin-binding EGF levels from day 0 to day 3 after the release of the duct obstruction. The EGF level was increased only after day 7. In vitro, cultured cells isolated from ligated SMGs proliferated and produced EGF ligands following the addition of epiregulin to the culture medium. These findings suggest that the tubular structures localized in an atrophic gland are the source of acinar cell regeneration of the salivary gland. The induction of EGF ligands, in particular epiregulin, may play an important role in acinar cell regeneration in this model.
Asunto(s)
Células Acinares/fisiología , Epirregulina/análisis , Regeneración/fisiología , Conductos Salivales/metabolismo , Enfermedades de la Glándula Submandibular/metabolismo , Glándula Submandibular/metabolismo , Anfirregulina , Animales , Atrofia , Betacelulina/análisis , Técnicas de Cultivo de Célula , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Modelos Animales de Enfermedad , Familia de Proteínas EGF/análisis , Factor de Crecimiento Epidérmico/análisis , Factor de Crecimiento Epidérmico/efectos de los fármacos , Epigen/análisis , Epirregulina/farmacología , Receptores ErbB/análisis , Receptores ErbB/efectos de los fármacos , Femenino , Factor de Crecimiento Similar a EGF de Unión a Heparina/análisis , Calicreínas/análisis , Calicreínas/efectos de los fármacos , Ligadura , Ratones , Ratones Endogámicos C57BL , Isomerasa de Peptidilprolil/análisis , Antígeno Nuclear de Célula en Proliferación/análisis , Conductos Salivales/efectos de los fármacos , Conductos Salivales/patología , Glándula Submandibular/patología , Enfermedades de la Glándula Submandibular/patología , Factor de Crecimiento Transformador alfa/análisis , Factor de Crecimiento Transformador alfa/efectos de los fármacosRESUMEN
OBJECTIVES: The pathogenesis of denture-induced fibrous hyperplasias has not been examined in detail to explain how tissue injury results in fibrous hyperplasia of the oral mucosa. PATIENTS AND METHODS: We examined the presence of mast cells and myofibroblasts in 33 denture-induced fibrous hyperplasias (DIFH) compared with 10 healthy gingival tissues. The parameters examined included mast cell numbers, tissue distribution, degranulation, and cell subtypes using immunohistochemistry. The presence of myofibroblasts and their likely origin was also examined by double immunofluorescense staining. Furthermore, we investigated the synthesis of osteopontin and TGF-ß, considered to be involved in the transformation of a fibroblast to a myofibroblast. RESULTS: The results demonstrated that the mast cell numbers are significantly increased in the DIFH compared with non-disease controls. The mast cell localization in lesions was higher in the superficial areas with inflammatory cell infiltration compared with the deep fibrotic area (P < 0.01). The number of tryptase-positive mast cells was significantly higher compared with chymase-positive ones. The TGF-ß- or osteopontin-positive cell infiltration into the lesion was found in high numbers. The presence of myofibroblasts was identified in 14 of 33 cases (42%), and some of these cells showed apoptosis when assessed by the TUNEL assay. On the survey of the origin of myofibroblasts, results showed αSMA and vimentin positivity indicating these transformed from fibroblasts. CONCLUSION: These results are the first to show that mast cells and myofibroblasts can be detected in DIFH, indicating important roles of these cells in the pathogenesis of this lesion.
Asunto(s)
Dentaduras/efectos adversos , Mastocitos/patología , Mucosa Bucal/patología , Miofibroblastos/patología , Actinas/análisis , Anciano , Anciano de 80 o más Años , Apoptosis/fisiología , Recuento de Células , Degranulación de la Célula/fisiología , Transdiferenciación Celular/fisiología , Quimasas/análisis , Femenino , Fibroblastos/patología , Fibrosis , Encía/patología , Humanos , Hiperplasia , Masculino , Mastocitos/fisiología , Persona de Mediana Edad , Osteopontina/análisis , Factor de Crecimiento Transformador beta/análisis , Triptasas/análisis , Vimentina/análisisRESUMEN
BACKGROUND: Patch tests are often used in the clinical diagnosis of metal allergies. In currently available patch tests, high concentrations of metal salt solutions are used. However, diagnosis accuracy can be influenced not only by acute skin reactions to high concentrations of metal salt, but also by skin reactions to other components present in the patch or to pH changes. In this study, we developed Ni nanoparticles (termed "nanoballs") for use in patch-test solutions. FINDINGS: Highly soluble, spherical Ni nanoballs were prepared using plasma electrolysis. The Ni released from the nanoballs permeated through a dialysis membrane, and the nanoball-containing solution's pH was maintained constant. Ni ions were released slowly at low concentrations in a time-dependent manner, which contrasted the rapid release observed in the case of a commercial patch test. Consequently, in the new test system, reactions caused by high concentrations of metal salts were avoided. CONCLUSIONS: By exploiting the high specific surface area of Ni nanoballs, we obtained an effective dissolution of Ni ions that triggered Ni allergy in the absence of direct contact between the nanoballs and mouse skin. This novel patch system can be applied to other metals and alloys for diagnosing various types of metal-induced contact dermatitis.
Asunto(s)
Nanopartículas/química , Níquel/química , Níquel/inmunología , Pruebas del Parche/instrumentación , Pruebas del Parche/métodos , Animales , Usos Diagnósticos de Compuestos Químicos , Concentración de Iones de Hidrógeno , Ratones Endogámicos C57BL , Nanotecnología/métodos , Níquel/farmacocinética , Piel/inmunología , Espectrometría por Rayos X/métodosRESUMEN
Transfection is a powerful method for investigating variable biological functions of desired genes. However, the efficiency of transfection into primary cultures of dental pulp-derived cells (DPDC) is low. Therefore, using a recombinant vaccinia virus (vTF7-3), which contains T7 RNA polymerase, we have established a transient protein expression system in DPDCs. In this study, we used the human polymeric immunoglobulin receptor (pIgR) cDNA as a model gene. pIgR expression by the vTF7-3 expression system was confirmed by flow cytometry analysis and Western blotting. Furthermore, exogenous pIgR protein localized at the cell surface in DPDCs and formed a secretory component (SC). This suggests that exogenous pIgR protein expressed by the vTF7-3 expression system acts like endogenous pIgR protein. These results indicate the applicability of the method for cells outgrown from dental pulp tissue. In addition, as protein expression could be detected shortly after transfection (approximately 5h), this experimental system has been used intensely for experiments examining very early steps in protein exocytosis.
Asunto(s)
Biotecnología/métodos , Pulpa Dental/citología , Pulpa Dental/metabolismo , Proteínas/química , Receptores de Inmunoglobulina Polimérica/química , Animales , Western Blotting , Línea Celular , Núcleo Celular , Células Cultivadas , Cricetinae , Fibroblastos , Citometría de Flujo , Células HeLa , Humanos , Microscopía Fluorescente , Biosíntesis de Proteínas , Proteínas/metabolismo , Receptores de Inmunoglobulina Polimérica/metabolismo , Transfección , Virus VacciniaRESUMEN
BACKGROUND: With the rapid worldwide increase in the oldest old population, considerable concern has arisen about the social and economic burden of diseases and disability in this age group. Understanding of multidimensional structure of health and its life-course trajectory is an essential prerequisite for effective health care delivery. Therefore, we organized an interdisciplinary research team consisting of geriatricians, dentists, psychologists, sociologists, and epidemiologists to conduct a longitudinal observational study. METHODS/DESIGN: For the Tokyo Oldest Old Survey on Total Health (TOOTH) study, a random sample of inhabitants of the city of Tokyo, aged 85 years or older, was drawn from the basic city registry. The baseline comprehensive assessment consists of an in-home interview, a self-administered questionnaire, and a medical/dental examination. To perform a wide variety of biomedical measurements, including carotid ultrasonography and a detailed dental examination, participants were invited to our study center at Keio University Hospital. For those who were not able to visit the study center, we provided the option of a home-based examination, in which participants were simultaneously visited by a geriatrician and a dentist. Of 2875 eligible individuals, a total of 1152 people were recruited, of which 542 completed both the in-home interview and the medical/dental examination, with 442 completed the in-home interview only, and another 168 completed self or proxy-administered data collection only. Carotid ultrasonography was completed in 458 subjects, which was 99.6% of the clinic visitors (n = 460). Masticatory assessment using a colour-changeable chewing gum was completed in 421 subjects, a 91.5% of the clinic visitors. DISCUSSION: Our results demonstrated the feasibility of a new comprehensive study that incorporated non-invasive measurements of subclinical diseases and a detailed dental examination aiming at community-dwelling individuals aged 85 years or older. The bimodal recruitment strategy is critically important to capture a broad range of health profiles among the oldest old. Results form the TOOTH study will help develop new models of health promotion, which are expected to contribute to an improvement in lifelong health and well-being.
Asunto(s)
Envejecimiento/fisiología , Envejecimiento/psicología , Estado de Salud , Encuestas Epidemiológicas , Anciano de 80 o más Años , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Humanos , Estudios Longitudinales , Masculino , Estudios Prospectivos , Sistema de Registros/estadística & datos numéricos , Tokio/epidemiologíaRESUMEN
To elucidate the underlying mechanism of secretory leukocyte protease inhibitor (SLPI)-induced cell migration, we compared SLPI-deleted human gingival carcinoma Ca9-22 (ΔSLPI) cells and original (wild-type: wt) Ca9-22 cells using several microscopic imaging methods and gene expression analysis. Our results indicated reduced migration of ΔSLPI cells compared to wtCa9-22 cells. The lamellipodia/dorsal ruffles were smaller and moved slower in ΔSLPI cells compared to wtCa9-22 cells. Furthermore, well-developed intermediate filament bundles were observed at the desmosome junction of ΔSLPI cells. In addition, Galectin4 was strongly expressed in ΔSLPI cells, and its forced expression suppressed migration of wtCa9-22 cells. Taken together, SLPI facilitates cell migration by regulating lamellipodia/ruffles and desmosomes, in which Galectin4 plays an important role.
Asunto(s)
Movimiento Celular , Desmosomas/metabolismo , Galectina 4/metabolismo , Seudópodos/metabolismo , Inhibidor Secretorio de Peptidasas Leucocitarias/metabolismo , Línea Celular Tumoral , Proliferación Celular , Desmosomas/ultraestructura , Galectina 4/genética , Humanos , Seudópodos/ultraestructura , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Dendritic cells (DC) play a crucial role in the pathogenesis of oral lichen planus (OLP) with respect to antigens presented to T cells. We performed immunohistochemical analysis to elucidate the process of activation of DC in OLP. METHODS: Thirty biopsy specimens were obtained from the patients with OLP. The expressions of CD1a, Langerin, S-100, fascin, chemokine receptor-7 (CCR-7), D2-40, cyclooxygenase-2 (COX-2), and microsomal prostaglandin E synthase-1 (mPGES-1) in DC from OLP and disease free control were investigated using specific antibodies. The distribution and number (1 mm(2)) of DC were assessed in the intra-epithelium and the submucosa specimens. Correlation between the number of DC and epithelium thickness was also determined. RESULT: Immature DC (Langerin(+), CD1a(+), and S-100(+)) were identified in the epithelia from OLP patients and control, though the numbers of Langerin(+) and CD1a(+) positive cells were decreased in the OLP samples as compared to the control. Mature DC (fascin(+)) were identified in the submucosa specimens, not found in the epithelium from OLP or control. Double immunostaining revealed DC positive for fascin and CCR-7 in the submucosa, which had migrated into D2-40(+) lymph vessels. Furthermore, keratinocytes expressed both Prostaglandin E(2) (PGE(2)) converting enzymes, COX-2, and mPGES-1, indicating PGE(2) synthesis in the epithelial layer of the OLP specimens. CONCLUSION: Our results indicate that DC change from immature to mature in the epithelium and are then drawn out to the submucosa. We demonstrate that mature DC localized in the submucosa, it consequently migrates into lymph vessels. This maturation process of DC is an important immunopathological feature of OLP.
Asunto(s)
Liquen Plano Oral/inmunología , Liquen Plano Oral/patología , Adulto , Anciano , Anciano de 80 o más Años , Antígenos CD/análisis , Proteínas Portadoras/análisis , Proteínas Portadoras/biosíntesis , Ciclooxigenasa 2/biosíntesis , Células Dendríticas/química , Células Dendríticas/inmunología , Células Dendríticas/patología , Dinoprostona/análisis , Dinoprostona/biosíntesis , Epitelio/patología , Femenino , Humanos , Inmunohistoquímica , Oxidorreductasas Intramoleculares/biosíntesis , Lectinas Tipo C/análisis , Liquen Plano Oral/metabolismo , Vasos Linfáticos/patología , Masculino , Lectinas de Unión a Manosa/análisis , Proteínas de Microfilamentos/análisis , Proteínas de Microfilamentos/biosíntesis , Persona de Mediana Edad , Mucosa Bucal/patología , Prostaglandina-E Sintasas , Receptores CCR7/análisis , Receptores CCR7/biosíntesis , Estudios Retrospectivos , Adulto JovenRESUMEN
BACKGROUND/AIMS: Hepatitis E virus (HEV) in Bangladesh has not been adequately documented. We report HEV RNA and genotype detection in Bangladesh. METHODS: In total, 82 samples were used; 36 sporadic acute hepatitis (AH), 12 fulminant hepatitis (FH), 14 chronic liver disease (CLD) and 20 from an apparently healthy population (HP) positive for both immunoglobulin (Ig) M and IgG specific anti-HEV antibodies (anti-HEV). The male/female ratio was 61/21, ages 12-67 (mean 30.4) years. RNA was extracted, transcribed to cDNA and amplified in nt 6345-6490 (ORF2) of HEV. Nucleic and amino acid sequences were determined. Homology comparison between Bangladesh clones and other representative HEV clones and phylogenetic tree analyses were done. Relations between HEV RNA-positivity and clinical factors were analyzed. RESULTS: HEV RNA was positive in 9/36 (25.0%) of AH cases, 4/12 (33.3%) FH, 3/14 (21.4%) CLD and 0/20 (0%) HP samples; total 16/82 (19.5%). Four factors correlated significantly with HEV RNA-positivity (Mann-Whitney U test); alanine aminotransferase (ALT) (P = 0.0229), aspartate aminotransferase (AST) (P = 0.0448), and titers of IgG (P = 0.0208) and IgM (P = 0.0095) specific anti-HEV. The 16 HEV clones were divided mainly into two groups, A and B, including six different cDNA sub-groups. CONCLUSION: HEV RNA was found in sporadic AH and FH and sub-clinical CLD cases, but not in HP. HEV RNA-positivity was significantly related to values of ALT and AST and titers of IgG and IgM specific anti-HEV, with IgM specific anti-HEV showing the most significant relationship. All clones were genotype I, which is prevalent in South Asia.
Asunto(s)
Virus de la Hepatitis E/genética , Hepatitis E/diagnóstico , ARN Viral/sangre , Enfermedad Aguda , Adolescente , Adulto , Anciano , Secuencia de Aminoácidos , Bangladesh/epidemiología , Secuencia de Bases , Niño , Enfermedad Crónica , Femenino , Genotipo , Anticuerpos Antihepatitis/sangre , Hepatitis E/epidemiología , Hepatitis E/virología , Virus de la Hepatitis E/inmunología , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Filogenia , Adulto JovenRESUMEN
1,25-Dihydroxyvitamin D(3) [1,25(OH)(2)D(3); 1,25-dihydroxycholecalciferol or calcitriol] is the active form of vitamin D(3), a lipid-soluble vitamin that plays a role in calcium and bone metabolism. Recently, vitamin D(3) has been shown to function in cancer prevention, immunity and cardiovascular regulation. 1,25(OH)(2)D(3) exhibits physiological and pharmacological effects by activating the vitamin D receptor (VDR), a transcription factor of the nuclear receptor superfamily. 1,25(OH)(2)D(3) plays a role in maintaining oral health through its effects on bone and mineral metabolism and innate immunity, and several VDR gene polymorphisms have been reported to be associated with periodontal disease. VDR ligands should prove to be useful in the treatment and prevention of periodontal disease.
Asunto(s)
Calcitriol/uso terapéutico , Agonistas de los Canales de Calcio/uso terapéutico , Enfermedades Periodontales/tratamiento farmacológico , Huesos/metabolismo , Calcitriol/inmunología , Agonistas de los Canales de Calcio/inmunología , Humanos , Inmunidad Innata/efectos de los fármacos , Minerales/metabolismo , Enfermedades Periodontales/genética , Enfermedades Periodontales/prevención & control , Polimorfismo Genético/genética , Receptores de Calcitriol/efectos de los fármacos , Receptores de Calcitriol/genéticaRESUMEN
BACKGROUND/AIM: Epidermal growth factor receptor (EGFR) is a target of molecular therapeutics for non-small cell lung cancer. EGFR gene mutations at codons 746-753 promote constitutive EGFR activation and result in worst prognosis. However, these mutations augment the therapeutic effect of EGFR-tyrosine kinase inhibitor. Therefore, the detection of EGFR gene mutations is important for determining treatment planning. The aim of the study was to establish a method to detect EGFR gene mutations at codons 746-753. MATERIALS AND METHODS: EGFR gene mutation at codons 746-753 in six cancer cell lines were investigated. A loop-mediated isothermal amplification (LAMP)-based procedure was developed, that employed peptide nucleic acid to suppress amplification of the wild-type allele. RESULTS: This mutation-oriented LAMP can amplify the DNA fragment of the EGFR gene with codons 746-753 mutations within 30 min. Moreover, boiled cells can work as template resources. CONCLUSION: Mutation oriented-LAMP assay for EGFR gene mutation is sensitive on extracted DNA. This procedure would be capable of detecting EGFR gene mutation in sputum, pleural effusion, broncho-alveolar lavage fluid or trans-bronchial lung biopsy by chair side.
Asunto(s)
Análisis Mutacional de ADN/métodos , Receptores ErbB/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/genética , Biopsia , Líquido del Lavado Bronquioalveolar/química , Línea Celular Tumoral , Células HT29 , Humanos , Pulmón/metabolismo , Pulmón/patología , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Mutación , Ácidos Nucleicos de Péptidos/química , Derrame Pleural/genética , Derrame Pleural/metabolismo , Reacción en Cadena de la Polimerasa/métodos , Esputo/química , Esputo/metabolismoRESUMEN
Although surgical resection is the first choice for oral cancer, the development of new anti-cancer drugs is of great interest. The effect of the histone deacetylation inhibitor, sodium butyrate (NaBu) on oral cancer cell (OCC) HSC-3 and HSC-4 proliferation in vitro was investigated. The synthesis of rate-limiting enzymes such as sPLA2 (-IIA, -V, -X) and COX-2 was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, as well as PGE2 by ELISA. NaBu acted in a concentration-dependent manner. Over 3 mM, it inhibited OCC proliferation, due to increased p21 expression and cell cycle arrest in the G2/M-phase. At low concentration (< or = 1 mM), NaBu showed no effects or enhanced cell proliferation. NaBu also regulated COX-2 and sPLA2-X expression, and augmented PGE2 synthesis in OCC. These results indicate that NaBu is a novel candidate agent for the treatment of oral cancer. The treatment efficacy must be investigated in additional experiments considering NaBu concentration and tumor cell heterogeneity.
Asunto(s)
Antineoplásicos/farmacología , Butiratos/farmacología , Ciclooxigenasa 2/metabolismo , Proteínas de la Membrana/metabolismo , Neoplasias de la Boca/enzimología , Fosfolipasas A/metabolismo , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Ciclooxigenasa 2/análisis , Ciclooxigenasa 2/genética , Dinoprostona/análisis , Dinoprostona/metabolismo , Fosfolipasas A2 Grupo II , Fosfolipasas A2 Grupo V , Fosfolipasas A2 Grupo X , Histonas/metabolismo , Humanos , Proteínas de la Membrana/análisis , Proteínas de la Membrana/genética , Fosfolipasas A/análisis , Fosfolipasas A/genética , Fosfolipasas A2RESUMEN
Secretory leukocyte protease inhibitor (SLPI) is a serine protease inhibitor whose expression level is positively correlated with tumor aggressiveness and metastatic potential. However, the mechanism underlying SLPI-induced enhancement of malignant phenotype is not completely understood. The malignancy of cancer cells is highly dependent on cell migration activity. Our previous study revealed that gingival carcinoma Ca9-22 cells, but not colorectal adenocarcinoma HT-29 cells, expressed SLPI. Therefore, we investigated the migration activity of these two cell types to understand the nature of SLPI-mediated tumor aggressiveness and metastatic potential. In vitro wound healing assay indicated that HT-29 cells and SLPI-deleted Ca9-22 cells showed lower migration activity than wild-type Ca9-22 cells, suggesting that SLPI-induced cell migration plays an important role in tumor aggressiveness and metastatic potential. In addition, our gene expression profiling study based on microarray data for the three cell types identified a number of candidates, including LCP1 and GLI, that could be key molecules in the mechanism of SLPI-induced cell migration.
Asunto(s)
Adenocarcinoma/genética , Movimiento Celular/fisiología , Neoplasias Colorrectales/genética , Perfilación de la Expresión Génica , Neoplasias Gingivales/genética , Inhibidor Secretorio de Peptidasas Leucocitarias/fisiología , Adenocarcinoma/patología , Neoplasias Colorrectales/patología , Neoplasias Gingivales/patología , Células HT29 , Humanos , Metástasis de la Neoplasia , Inhibidor Secretorio de Peptidasas Leucocitarias/genéticaRESUMEN
Age-related deterioration in physical and oral health reduces healthy life expectancy and is thus an important problem for very elderly people. We investigated the effects of satisfaction with dietary life (SDL) in everyday life on oral health-related quality of life (OHRQoL) and subjective well-being and examined associations between these factors. We evaluated 426 elders aged 85 years or older. All participants completed a questionnaire that inquired about age, gender, drinking status, body mass index, cognitive function, disability, and comorbidities, among other covariates. Oral, physical, and mental health conditions were also examined. Associations of questionnaire results for SDL with items on subjective well-being (Philadelphia Geriatric Center Morale Scale [PGC] and World Health Organization-5 [WHO-5]) and OHRQoL (Geriatric Oral Health Assessment Index [GOHAI]) were confirmed with multiple logistic regression analysis. In a multivariate model adjusted for various confounders, participants with self-reported "enjoyable" SDL had significantly lower risks for having the lowest scores on the GOHAI, PGC, and WHO-5 (odds ratio [OR] = 0.460, 95% confidence interval [CI] = 0.277-0.762; OR = 0.589, 95% CI = 0.348-0.996; and OR = 0.452, 95% CI = 0.263-0.775, respectively). These associations remained after further adjustment for number of teeth.
Asunto(s)
Dieta , Salud Bucal , Satisfacción Personal , Calidad de Vida , Anciano , Anciano de 80 o más Años , Femenino , Humanos , MasculinoRESUMEN
Although a number of sPLA2 (secretory phospholipase A2) enzymes have been identified in mammals, the localization and functions of individual enzymes in human pathologic tissues still remain obscure. In the present study, we have examined the expression and function of sPLA2s in human lung-derived cells and in human lungs with pneumonia. Group IID, V and X sPLA2s were expressed in cultured human bronchial epithelial cells (BEAS-2B) and normal human pulmonary fibroblasts with distinct requirement for cytokines (interleukin-1b, tumour necrosis factor a and interferon-g). Lentivirus- or adenovirus-mediated transfection of various sPLA2s into BEAS-2B or normal human pulmonary fibroblast cells revealed that group V and X sPLA2s increased arachidonate release and prostaglandin production in both cell types, whereas group IIA and IID sPLA2s failed to do so. Immunohistochemistry of human lungs with pneumonia demonstrated that group V and X sPLA2s were widely expressed in the airway epithelium, interstitium and alveolar macrophages, in which group IID sPLA2 was also positive, whereas group IIA sPLA2 was restricted to the pulmonary arterial smooth muscle layers and bronchial chondrocytes, and group IIE and IIF sPLA2s were minimally detected. These results suggest that group V and X sPLA2s affect lung pathogenesis by facilitating arachidonate metabolism or possibly through other functions.
Asunto(s)
Bronconeumonía/enzimología , Pulmón/enzimología , Fosfolipasas A/biosíntesis , Fosfolipasas A/metabolismo , Anciano , Ácido Araquidónico/metabolismo , Bronquios/citología , Bronquios/enzimología , Bronconeumonía/patología , Línea Celular , Citosol/enzimología , Citosol/metabolismo , Dinoprostona/biosíntesis , Retículo Endoplásmico/química , Retículo Endoplásmico/metabolismo , Células Epiteliales/enzimología , Células Epiteliales/metabolismo , Fibroblastos/química , Fibroblastos/enzimología , Fibroblastos/metabolismo , Humanos , Inmunohistoquímica/métodos , Riñón/química , Riñón/citología , Riñón/embriología , Riñón/metabolismo , Pulmón/citología , Macrófagos Alveolares/enzimología , Macrófagos Alveolares/metabolismo , Persona de Mediana Edad , Músculo Liso Vascular/enzimología , Músculo Liso Vascular/patología , Fosfolipasas A2 , Arteria Pulmonar/enzimología , Arteria Pulmonar/patología , Proteínas Recombinantes/biosíntesisRESUMEN
This study evaluated the effects of different sizes of beta-TCP particles on bone augmentation within a titanium cap. In 20 rabbits, the calvarium was exposed and a circular groove was prepared. After marrow penetration, a standardized hemispherical titanium cap was placed in the circular grove. The cap was filled with small-sized (100-250 microm) or medium-sized (250-500 microm) beta-TCP particles for the experimental site and without beta-TCP for the control site. After one and three months of healing, the animals were euthanized and examined histologically. There was a statistically significant difference in the amount of mineralized bone generated between the experimental and control groups in the three-month specimens. Furthermore, the medium-sized particles showed significantly more mineralized bone than did the small-sized particles. Based on these findings, we suggested that beta-TCP might be effective for bone formation and that medium-sized particles are more useful than small-sized particles in bone maturation.