RESUMEN
The aim of this study was to evaluate the barrier function of platelet-induced epithelial sheets on titanium surfaces. The lack of functional peri-implant epithelial sealing with basal lamina (BL) attachment at the interface of the implant and the adjacent epithelium allows for bacterial invasion, which may lead to peri-implantitis. Although various approaches have been reported to combat bacterial infection by surface modifications to titanium, none of these have been successful in a clinical application. In our previous study, surface modification with protease-activated receptor 4-activating peptide (PAR4-AP), which induced platelet activation and aggregation, was successful in demonstrating epithelial attachment via BL and epithelial sheet formation on the titanium surface. We hypothesized that the platelet-induced epithelial sheet on PAR4-AP-modified titanium surfaces would reduce bacterial attachment, penetration, and invasion. Titanium surface was modified with PAR4-AP and incubated with platelet-rich plasma (PRP). The aggregated platelets released collagen IV, a critical BL component, onto the PAR4-AP-modified titanium surface. Then, human gingival epithelial cells were seeded on the modified titanium surface and formed epithelial sheets. Green fluorescent protein (GFP)-expressing Escherichia coli was cultured onto PAR4-AP-modified titanium with and without epithelial sheet formation. While Escherichia coli accumulated densely onto the PAR4-AP titanium lacking epithelial sheet, few Escherichia coli were observed on the epithelial sheet on the PAR4-AP surface. No bacterial invasion into the interface of the epithelial sheet and the titanium surface was observed. These in vitro results indicate the efficacy of a platelet-induced epithelial barrier that functions to prevent bacterial attachment, penetration, and invasion on PAR4-AP-modified titanium.
Asunto(s)
Plaquetas/fisiología , Implantes Dentales , Materiales Dentales/química , Inserción Epitelial , Periimplantitis/prevención & control , Receptores de Trombina/química , Titanio/química , Adhesión Bacteriana/efectos de los fármacos , Pilares Dentales , Escherichia coli , Humanos , Técnicas In Vitro , Periimplantitis/etiología , Plasma Rico en Plaquetas , Propiedades de Superficie , Cicatrización de HeridasRESUMEN
The objectives of this research were to examine the current status of perioperative treatment among foreigners, to elucidate the health status/outcome disparities that contribute to ethnic differences, and to recommend counter-measures to rectify these ethnic disparities. The authors identified 36 non-Japanese and 111 Japanese females who underwent gynecological surgery from 2004 to 2009 at a single institution. Electronic medical records were reviewed and telephone survey was conducted in order to obtain patient background, preoperative, operative, and postoperative data. The non-Japanese group showed significantly larger number of uninsured, shorter length of stay (LOS), higher rate of emergency surgery, and higher cases of spinal anesthesia. There were significant differences in length of residency in Japan and LOS among four foreign countries. Seventy-nine percent of patients contacted by phone understood informed consent from doctors, 73.7% understood explanation in operating room (OR), and 84.2% understood explanation from anesthesiologists. This research was the first survey of the ethnic disparities in perioperative management among foreign patients treated in Osaka. The authors have demonstrated differences in operative method, emergency surgery, anesthesia, and American Society of Anesthesiologists physical status (ASA-PS) due to the difference in disease structure, language, and culture. It is recommended that the barriers between non-Japanese patients and medical staff are rectified during the perioperative period when mutual understanding is needed the most.
Asunto(s)
Parto Obstétrico/estadística & datos numéricos , Emigrantes e Inmigrantes/estadística & datos numéricos , Disparidades en el Estado de Salud , Adulto , Anestesia Obstétrica/estadística & datos numéricos , Anestesia Raquidea/estadística & datos numéricos , Barreras de Comunicación , Femenino , Encuestas de Atención de la Salud , Humanos , Lenguaje , Tiempo de Internación , Atención Perinatal , Adulto JovenRESUMEN
In this study, we examined the role of nitric oxide (NO) in controlling vascular integrity mediated by vascular endothelial (VE)-cadherin in chronic inflammation. Periapical granulomas were analysed for the expression of inducible NO synthase (iNOS) and VE-cadherin, and more iNOS expression than VE-cadherin was shown. Human umbilical vein endothelial cells (HUVECs) were stimulated with proinflammatory cytokines and lipopolysaccharide extracted from Porphyromonas gingivalis and it induced iNOS expression, whereas it reduced VE-cadherin expression, compared with negative controls. On the other hand, pre-incubation with 1400W, an iNOS-specific inhibitor, markedly reduced iNOS expression in stimulated HUVECs and restored VE-cadherin expression to its control level, suggesting that vascular integrity was modulated in conjunction with the reduction of NO. Immunocytochemistry confirmed the functional role of NO in cultured HUVEC monolayers with or without 1400W. These data are consistent with a hypothesis suggesting that NO could attenuate VE-cadherin-mediated vascular integrity in human chronic inflammation.
Asunto(s)
Antígenos CD/metabolismo , Cadherinas/metabolismo , Endotelio Vascular/metabolismo , Óxido Nítrico/fisiología , Granuloma Periapical/metabolismo , Adulto , Anciano , Antígenos CD/genética , Cadherinas/genética , Células Cultivadas , Enfermedad Crónica , Citocinas/inmunología , Endotelio Vascular/inmunología , Endotelio Vascular/patología , Regulación de la Expresión Génica/inmunología , Humanos , Lipopolisacáridos/inmunología , Persona de Mediana Edad , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Granuloma Periapical/inmunología , Granuloma Periapical/patología , ARN Mensajero/genética , Venas Umbilicales/citología , Venas Umbilicales/metabolismo , Adulto JovenRESUMEN
BACKGROUND/AIMS: The cell surface protein antigen (PAg) and glucosyltransferases (GTFs) produced by Streptococcus sobrinus are considered to be major colonization factors of the organism. METHODS: We constructed a fusion gene encoding a protein composed of the alanine-rich region of PAg (PAgA) and the glucan-binding domain (GB) of GTF-I, which catalyzes the synthesis of water-insoluble glucan in S. sobrinus. The fusion protein PAgA-GB was purified from cell extracts of Escherichia coli harboring the fusion gene, and antibodies against the fusion protein were prepared in rabbits. RESULTS: In the presence of sucrose, the antibody against PAgA-GB significantly inhibited the adhesion of both S. sobrinus MT8145 and Streptococcus mutans Xc to saliva-coated hydroxyapatite beads, and the inhibitory effect on S. sobrinus was stronger than that on S. mutans. In the absence of sucrose, the antibody against PAgA-GB significantly inhibited the adhesion of both S. sobrinus and S. mutans, however the inhibitory effect on S. sobrinus was unexpectedly weaker than that on S. mutans. A similar result was observed with the antibody against the intact recombinant PAg protein (rPAg), while the same antibody reacted more strongly against S. sobrinus than against S. mutans cells. CONCLUSION: Taken together, these results show that the antibody against S. sobrinus GTF-I may be useful for effective inhibition of the sucrose-dependent adhesion of S. sobrinus. However, PAg of S. sobrinus may not function primarily as a receptor for acquired pellicles, and other cell surface proteins may be involved in the sucrose-independent adhesion of S. sobrinus.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Antígenos Bacterianos/inmunología , Antígenos de Superficie/inmunología , Adhesión Bacteriana/inmunología , Proteínas Bacterianas/inmunología , Glicosiltransferasas/inmunología , Proteínas de la Membrana/inmunología , Streptococcus mutans/inmunología , Streptococcus sobrinus/inmunología , Adulto , Animales , Durapatita/química , Electroforesis en Gel de Poliacrilamida , Humanos , Immunoblotting , Conejos , Proteínas Recombinantes , Saliva/fisiología , Sacarosa/farmacología , Propiedades de SuperficieRESUMEN
OBJECTIVE: To evaluate the clinical importance of mixed mutans streptococci colonization in predicting caries in preschool children. METHODS: Caries prevalence was examined twice, with a 6-month interval, in 410 preschool children aged 3-4 years at baseline. A commercial strip method was used to evaluate the mutans streptococci score in plaque collected from eight selected interdental spaces and in saliva. Mutans streptococci typing polymerase chain reaction (PCR) assays (Streptococcus sobrinus and Streptococcus mutans, including serotypes c, e, and f) were performed using colonies on the strips as template. RESULTS: Twenty variables were examined in a univariate analysis to predict caries development: questionnaire variables, results of clinical examination, mutans streptococci scores, and PCR detection of S. sobrinus and S. mutans (including serotypes c, e, and f). Sixteen variables showed statistically significant associations (P < 0.04) in the univariate analysis. However, when entered into a logistic regression, only five variables remained significant (P < 0.05): caries experience at baseline; mixed colonization of S. sobrinus and S. mutans including S. mutans serotypes; high plaque mutans streptococci score; habitual use of sweet drinks; and nonuse of fluoride toothpaste. CONCLUSION: 'Mixed mutans streptococci colonization' is a novel measure correlated with caries development in their primary dentition.
Asunto(s)
Caries Dental/microbiología , Streptococcus mutans/fisiología , Streptococcus sobrinus/fisiología , Bebidas , Preescolar , Recuento de Colonia Microbiana , Índice CPO , Susceptibilidad a Caries Dentarias/fisiología , Placa Dental/microbiología , Carbohidratos de la Dieta/administración & dosificación , Femenino , Estudios de Seguimiento , Predicción , Encía/microbiología , Humanos , Masculino , Tiras Reactivas , Saliva/microbiología , Serotipificación , Streptococcus mutans/clasificación , Streptococcus sobrinus/clasificación , Diente Primario/microbiología , Pastas de Dientes/análisisRESUMEN
The clinical feasibility of both dual-isotope single photon emission tomography (SPET) and gated SPET have been described. The present study evaluates the feasibility of combining gated SPET with exercise 201Tl/rest 99mTc-tetrofosmin dual-isotope SPET corrected for scatter. Ninety-one patients with known or suspected coronary artery disease underwent cardiac catheterization and coronary angiography. Twenty-nine of them underwent exercise 201Tl/rest 99mTc-tetrofosmin dual-isotope SPET with a second 201Tl injection 3 h after the initial 201Tl injection (protocol 1). We then segregated a Bull's eye polar map into three coronary artery territories and quantified the relative regional uptake. The remaining 62 patients underwent exercise 201Tl/rest 99mTc-tetrofosmin dual-isotope SPET combined with gated SPET. We visually evaluated exercise and rest images from the three coronary artery territories. Left ventricular (LV) function was assessed globally by means of the LV ejection fraction and regionally by means of visual scoring analysis, compared with left ventriculography (LVG). The correlation between rest 99mTc-tetrofosmin and 201Tl reinjection images in 87 areas of coronary artery territory (r=0.89, P<0.01) and in 13 infarcted areas (r =0.94, P<0.01) was very close in protocol 1. The overall values for vessel-related sensitivity, specificity and accuracy were 88%, 79% and 82%, respectively, in protocol 2. The correlation between gated SPET and LVG was significant and linear with respect to the LV ejection fraction (r=0.77, P<0.01). The wall motion score from visual evaluation in gated SPET revealed a close overall agreement with LVG (concordance rate, 88%; kappa, 0.670). Exercise 201Tl/rest 99mTc-tetrofosmin dual-isotope SPET with scatter correction for assessing the coronary artery disease offers excellent diagnostic accuracy and the additional gated SPET provides useful information about LV function similar to that for LVG. This sequential protocol requires only 2 h to generate much useful clinical information.
Asunto(s)
Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Corazón/diagnóstico por imagen , Anciano , Cateterismo Cardíaco , Circulación Coronaria/fisiología , Estudios de Factibilidad , Femenino , Imagen de Acumulación Sanguínea de Compuerta , Humanos , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/diagnóstico por imagen , Compuestos Organofosforados , Compuestos de Organotecnecio , Perfusión , Radiofármacos , Volumen Sistólico/fisiología , Radioisótopos de Talio , Función Ventricular Izquierda/fisiologíaRESUMEN
A 39-year-old woman had exercise-induced ST segment depression associated with chest pain. Cardiac evaluation revealed moderate aortic stenosis (AS), related to the bicuspid valves, with an aortic mean pressure gradient of 22 mmHg, a calculated aortic valve area of 1.3 cm2 and normal left ventricular (LV) peak systolic and end-diastolic pressures, but no LV hypertrophy, resulting in normal LV wall stress. Although the coronary arteries were angiographically normal, rapid atrial pacing and an intracoronary papaverine injection revealed a significantly decreased coronary flow reserve (CFR), which may have played an important role in the pathogenesis of angina pectoris in this patient. Though the CFR is usually decreased in patients with AS, as well as in microvascular angina, in this particular case, it appeared to have decreased as a consequence of microvascular dysfunction rather than of AS-related mechanisms.
Asunto(s)
Estenosis de la Válvula Aórtica/patología , Angina Microvascular/etiología , Adulto , Estenosis de la Válvula Aórtica/fisiopatología , Vasos Coronarios/patología , Vasos Coronarios/fisiopatología , Electrocardiografía , Ejercicio Físico , Femenino , Humanos , Angina Microvascular/patología , Angina Microvascular/fisiopatología , Papaverina/administración & dosificación , Flujo Sanguíneo Regional , Vasodilatadores/administración & dosificaciónRESUMEN
Primitive blood cells differentiate from the ventral mesoderm blood islands in Xenopus embryos. In order to determine the tissue interactions that propagate blood formation in early embryogenesis, we used embryos that had the ventral cytoplasm removed. These embryos gastrulated normally, formed a mesodermal layer and lacked axial structures, but displayed a marked enhancement of alpha-globin expression. Early ventral markers, such as msx-1, vent-1 and vent-2 were highly expressed at the gastrula stage, while a dorsal marker, goosecoid, was diminished. Several lines of experimental evidence demonstrate the critical role of animal pole-derived ectoderm in blood cell formation: 1) Mesoderm derived from dorsal blastomeres injected with beta-galactosidase mRNA (as a lineage tracer) expressed alpha-globin when interfaced with an animal pole-derived ectodermal layer; 2) Embryos in which the animal pole tissue had been removed by dissection at the blastula stage failed to express alpha-globin; 3) Exogastrulated embryos that lacked an interaction between the mesodermal and ectodermal layers failed to form blood cells, while muscle cells were observed in these embryos. Using dominant-negative forms of the BMP-4 and ALK-4 receptors, we showed that activin and BMP-4 signaling is necessary for blood cell differentiation in ventral marginal zone explants, while FGF signaling is not essential. In ventralized embryos, inactivation of the BMP-4 signal within a localized area of the ectoderm led to suppression of globin expression in the adjacent mesoderm layer, but inactivation of the activin signal did not have this effect. These observations suggest that mesodermal cells, derived from a default pathway that is induced by the activin signal, need an additional BMP-4-dependent factor from the overlying ectoderm for further differentiation into a blood cell lineage.
Asunto(s)
Diferenciación Celular , Eritrocitos/citología , Eritropoyesis/fisiología , Estratos Germinativos/metabolismo , Transducción de Señal , Xenopus laevis/embriología , Receptores de Activinas Tipo I , Activinas , Animales , Northern Blotting , Proteína Morfogenética Ósea 4 , Proteínas Morfogenéticas Óseas/metabolismo , Embrión no Mamífero , Eritrocitos/metabolismo , Factores de Crecimiento de Fibroblastos/metabolismo , Globinas/metabolismo , Hibridación in Situ , Inhibinas/metabolismo , Morfogénesis , ARN Mensajero/análisis , Receptores de Factores de Crecimiento/metabolismo , Proteínas de Xenopus , Xenopus laevis/metabolismoRESUMEN
We carried out stress 201Tl (Tl) and rest 99mTc-tetrofosmin (TF) myocardial scintigraphy with dual energy acquisition in 24 patients with suspected ischemic heart disease performed coronary arteriography and elucidated the sensitivity of this method. One hour after light meal eating, TF (555 MBq) was injected intravenously at rest and after 3 minutes from injection of TF exercise or pharmacologic stress was performed. During stress Tl (111 MBq) was injected intravenously before end-point or at adequate point of pharmacologic stress. Dual energy acquisition using triple energy windows (TEW) was started after 5 minutes (early) and 4 hours (delayed) from stress. The sensitivity (Sn), specificity (Sp) and accuracy (Ac) in diagnosis of non-infarcted branches by using Tl (early)-TF (rest) and Tl (early)-Tl (delayed) were 79% vs. 53% (Sn), 78% vs. 96% (Sp) and 79% vs. 71% (Ac) respectively. Accordance of uptake score of infarcted region between TF (rest) and Tl (delayed) was 70%. In conclusion, this protocol is seemed to be useful as usual protocol for detection of myocardial ischemia and viability during about only 1 hour.
Asunto(s)
Corazón/diagnóstico por imagen , Isquemia Miocárdica/diagnóstico por imagen , Compuestos Organofosforados , Compuestos de Organotecnecio , Radiofármacos , Radioisótopos de Talio , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Descanso , Sensibilidad y Especificidad , Tomografía Computarizada de Emisión de Fotón ÚnicoRESUMEN
BACKGROUND: Although enamel matrix-derived protein (EMD) can stimulate attachment of human periodontal ligament (HPDL) cells to the root surface, the biological mechanism of this phenomenon is unclear. The purpose of this study was to determine which molecules in EMD are involved in the attachment of HPDL cells, and which types of integrins on the cell surface mediate the interaction between the cells and EMD. METHODS: HPDL explants were obtained from tooth surfaces extracted from 4 individuals, and cells taken from the individual explants were separately harvested and subcultured through as many as 5 passages. Cells were incubated on EMD-coated culture plates with and without neutral antibodies for integrins or RGD-sequence blocking peptides and stained with toluidine blue. Proteins in EMD that were able to induce cell attachment were identified by incubating sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) replicas with HPDL cells; the cell-binding regions were detected by staining the cells with toluidine blue. Characteristics of the cell-binding proteins in the EMD were identified by Western blot analysis. RESULTS: It was shown that anti-alpha(v)beta3 antibody and GRGDSP peptide markedly reduced attachment of HPDL cells to EMD. When the cells were incubated with SDS-PAGE replicas, distinct cell attachment was observed at a molecular mass of approximately 55 kDa. The cell-binding ability of this protein was completely blocked by treatment with anti-alpha(v)beta3 antibody or GRGDSP peptide. In the Western blot analysis, the 55 kDa protein was recognized by anti-bone sialoprotein (BSP) antibody as a single band. CONCLUSIONS: Our study provides the first evidence that the attachment of HPDL cells to EMD can be mediated by interaction between a BSP-like molecule and integrin alpha(v)beta3 on the cell surface.
Asunto(s)
Proteínas del Esmalte Dental/fisiología , Ligamento Periodontal/citología , Receptores de Vitronectina/fisiología , Sialoglicoproteínas/fisiología , Secuencia de Aminoácidos , Anticuerpos Monoclonales , Western Blotting , Adhesión Celular , Técnicas de Cultivo de Célula , Colorantes , Proteínas del Esmalte Dental/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida , Humanos , Sialoproteína de Unión a Integrina , Peso Molecular , Oligopéptidos/antagonistas & inhibidores , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Vitronectina/aislamiento & purificación , Sialoglicoproteínas/aislamiento & purificación , Estadística como Asunto , Cloruro de TolonioRESUMEN
To ascertain whether periodontal fibroblasts could be involved in the pathogenesis of periodontal pocket formation, the chemotactic activity of periodontal ligament fibroblast-conditioned medium (PLF-CM) and gingival fibroblast-conditioned medium (GF-CM) for gingival epithelial cells was examined using a modified Boyden chamber assay. Both PLF-CM and GF-CM possessed significant chemotactic activity, which was decreased markedly by treatment with anti-human hepatocyte growth factor (HGF) neutralizing antibody. Furthermore, the chemotactic activity of PLF-CM and GF-CM was well correlated with HGF content. These results show that PLF and GF secrete an HGF-like factor, and suggest that such a factor derived from periodontal fibroblasts might play a role in epithelial apical migration in periodontitis.
Asunto(s)
Factores Quimiotácticos/fisiología , Células Epiteliales/fisiología , Encía/metabolismo , Factor de Crecimiento de Hepatocito/fisiología , Ligamento Periodontal/metabolismo , Línea Celular , Movimiento Celular/efectos de los fármacos , Factores Quimiotácticos/metabolismo , Factores Quimiotácticos/farmacología , Medios de Cultivo Condicionados/farmacología , Células Epiteliales/efectos de los fármacos , Fibroblastos/metabolismo , Encía/citología , Factor de Crecimiento de Hepatocito/metabolismo , Factor de Crecimiento de Hepatocito/farmacología , Humanos , Ligamento Periodontal/citología , Estadísticas no ParamétricasRESUMEN
The msx homeodomain protein is a downstream transcription factor of the bone morphogenetic protein (BMP)-4 signal and a key regulator for neural tissue differentiation. Xmsx-1 antagonizes the dorsal expression of noggin and cerberus, as revealed by in situ hybridization and reverse transcription-polymerase chain reaction assays. In animal cap explants, Xmsx-1 and BMP-4 inhibit the neural tissue differentiation induced by noggin or cerberus. A loss-of-function study using the Xmsx-1/VP-16 fusion construct indicated that neural tissue formation was directly induced by the injection of fusion ribonucleic acid, although the expression of neural cell adhesion molecule (N-CAM) in the cap was less than that in the cap injected with tBR or noggin. In contrast to the single cap assay, unexpectedly, both BMP-4 and Xmsx-1 failed to inhibit neurulation in the ectodermal explants to which the organizer mesoderm was attached. The results of cell-lineage tracing experiments indicated that the neural cells were differentiated from the animal pole tissue where the excess RNA of either BMP-4 or Xmsx-1 was injected, whereas notochord was differentiated from the organizer mesoderm. Neural tissue differentiated from BMP-4-injected ectodermal cells strongly expressed posterior neural markers, such as hoxB9 and krox20, suggesting that the posterior neural cells differentiated regardless of the existence of the BMP signal. The introduction of a dominant-negative form of the fibroblast growth factor (FGF) receptor (XFD) into the ectodermal cells drastically reduced the expression of pan and posterior neural markers (N-CAM and hoxB-9) if co-injected with BMP-4 RNA, although XFD alone at the same dose did not shut down the expression of N-CAM in the combination explants. Therefore, it is proposed that an FGF-related molecule was involved in the direct induction of posterior neural tissue in the inducing signals from the organizer mesoderm in vivo.
Asunto(s)
Proteínas Morfogenéticas Óseas/fisiología , Embrión no Mamífero/inervación , Inducción Embrionaria/fisiología , Factores de Crecimiento de Fibroblastos/fisiología , Proteínas de Homeodominio/fisiología , Sistema Nervioso/embriología , Transducción de Señal/fisiología , Factores de Transcripción , Xenopus laevis/embriología , Animales , Tipificación del Cuerpo , Proteína Morfogenética Ósea 4 , Proteínas Portadoras , Diferenciación Celular , Linaje de la Célula , Cartilla de ADN/química , Ectodermo/fisiología , Inhibidores Enzimáticos/farmacología , Etopósido/metabolismo , Hibridación in Situ , Factor de Transcripción MSX1 , Morfogénesis , Mutación , Sistema Nervioso/metabolismo , Proteínas/metabolismo , ARN Mensajero/análisis , ARN Mensajero/genética , Proteínas Recombinantes de Fusión/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas de Xenopus , Xenopus laevis/fisiologíaRESUMEN
To clarify whether fibroblasts could be involved in the pathogenesis of periodontal pocket formation, the chemotactic activity of radicular cyst-derived fibroblast-like cell (RCF)-conditioned medium (RCF-CM) for gingival epithelial cells was examined using a modified Boyden chamber assay. RCF-CM possessed significant chemotactic activity, which was decreased markedly by treatment with anti-human hepatocyte growth factor (HGF) antibody. Furthermore, the chemotactic activity of RCF-CM was well correlated with HGF content. These results show that the RCF secrete an HGF-like factor, and suggest that such a factor derived from periodontal fibroblasts might play a role in epithelial apical migration in periodontitis.
Asunto(s)
Factores Quimiotácticos/fisiología , Células Epiteliales/fisiología , Fibroblastos/fisiología , Encía/citología , Factor de Crecimiento de Hepatocito/fisiología , Quiste Radicular/patología , Línea Celular , Movimiento Celular/fisiología , Factores Quimiotácticos/farmacología , Medios de Cultivo Condicionados , Cámaras de Difusión de Cultivos , Células Epiteliales/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/patología , Encía/efectos de los fármacos , Factor de Crecimiento de Hepatocito/farmacología , Humanos , Bolsa Periodontal/etiología , Bolsa Periodontal/patología , Periodontitis/etiología , Periodontitis/patologíaRESUMEN
We demonstrated previously that Xmsx-1 is involved in mesoderm patterning along the dorso-ventral axis, under the regulation of BMP-4 signaling. When Xmsx-1 RNA was injected into the dorsal blastomeres, a mass of muscle tissue formed instead of notochord. This activity was similar to that of Xwnt-8 reported previously. In this study, we investigated whether the activity of Xmsx-1 is related to the ventralizing signal and myogenesis promoting factor, Xwnt-8. Whole-mount in situ hybridization showed that Xmsx-1, Xwnt-8, and XmyoD were expressed in overlapping areas, including the ventro-lateral marginal zone at mid-gastrula stage. The expression of XmyoD was induced by the ectopic expression of either Xmsx-1 or Xwnt-8 in dorsal blastomeres, and Xwnt-8 was induced by the ectopic expression of Xmsx-1. On the other hand, the expression of Xmsx-1 was not affected by the loading of pCSKA-Xwnt-8 or dominant-negative Xwnt-8 (DN-Xwnt-8) RNA. In addition, Xmsx-1 RNA did not abrogate the formation of notochord if coinjected with DN-Xwnt-8 RNA. These results suggest that Xmsx-1 functions upstream of the Xwnt-8 signal. Furthermore, the antagonistic function of Xmsx-1 to the expression of organizer genes, such as Xlim-1 and goosecoid, was shown by in situ hybridization analysis and luciferase reporter assay using the goosecoid promoter construct. Finally if Xmsx-1/VP-16 fusion RNA, which was expected to function as a dominant-negative Xmsx-1, was injected into ventral blastomeres, a partial secondary axis formed in a significant number of embryos. In such embryos, the activity of luciferase, under the control of goosecoid promoter sequence, was significantly elevated at gastrula stage. These results led us to conclude that Xmsx-1 plays a central role in establishing dorso-ventral axis in gastrulating embryo, by suppressing the expression of organizer genes.
Asunto(s)
Tipificación del Cuerpo/genética , Regulación del Desarrollo de la Expresión Génica , Glicoproteínas , Proteínas de Homeodominio/metabolismo , Péptidos y Proteínas de Señalización Intercelular , Proteínas Represoras , Factores de Transcripción , Xenopus laevis/embriología , Xenopus laevis/genética , Actinas/genética , Animales , Tipificación del Cuerpo/fisiología , Proteínas del Citoesqueleto , Femenino , Proteína Goosecoide , Proteínas de Homeodominio/genética , Hibridación in Situ , Proteínas con Homeodominio LIM , Factor de Transcripción MSX1 , Microinyecciones , Proteína MioD/genética , Organizadores Embrionarios , Regiones Promotoras Genéticas , Proteínas/genética , ARN/genética , Proteínas Recombinantes de Fusión/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Eliminación de Secuencia , Proteínas Wnt , Proteínas de Xenopus , Proteínas de Pez CebraRESUMEN
Levels of messenger RNA (mRNA) extracted from five samples of radicular cyst-lining epithelium were analyzed for cytokines, growth factors and epithelial cell growth-related receptors by RT-PCR. All five samples expressed IL-1alpha, -1beta, IL-6, IL-8, IL-11, TGF-beta1, PDGF-A and aFGF, and receptors for EGF (c-erbB), KGF, HGF (c-met) and IL-6. Some of the specimens expressed MIP-1alpha, RANTES, GM-CSF, M-CSF, TNF-alpha, PDGF-B and bFGF, but no expression of IL-2, IL-4, IFN-gamma, IGF-I, EGF and KGF was detected. These results indicate that radicular cyst-lining epithelium, which is considered to be identical to the cell rests of Malassez, may play a role in periodontal pocket formation or apical cyst formation by interaction with surrounding connective tissue or hematopoietic cells through the expression of various cytokines.
Asunto(s)
Citocinas/biosíntesis , Células Epiteliales/metabolismo , Sustancias de Crecimiento/biosíntesis , Quiste Radicular/metabolismo , Humanos , ARN Mensajero/análisis , Receptores de Factores de Crecimiento/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Msx is a homeodomain-containing transcriptional factor that plays an essential role in pattern formation in vertebrata and invertebrata embryos. In Xenopus laevis, two msx genes have been identified (Xmsx-1 and Xmsx-2). In the present study, we attempted to elucidate the expression and function of Xmsx-2B (formerly designated as Xhox7.1') in early embryogenesis. Whole mount in situ hybridization analyses showed that the expression pattern of Xmsx-2B at gastrula and neurula stages was very similar to that of Xmsx-1: the transcript of Xmsx-2B was observed in ventral and lateral sides of the embryo. At the tailbud stage, however, the expression pattern of Xmsx-2B in neural tissues was distinct from that of Xmsx-1. An RNA injection experiment revealed that, like BMP-4, Xmsx-2B has a strong ventralizing activity. In the Xmsx-2B -injected embryos, differentiation of axial structures such as the notochord, muscle, and neural tissue was completely suppressed, whereas alpha-globin mRNA, a blood cell marker, was highly expressed. Simultaneous injection of Xmsx-1 and Xmsx-2B RNAs showed that they function in an additive manner. It was also shown that coinjection of Xmsx-2B with a dominant-negative BMP-4 receptor (tBR), which can induce formation of secondary axis when injected alone in ventral blastomeres, suppressed secondary axis formation. Furthermore, Xmsx-2B also suppressed secondary axis formation, which was induced by a dominant-negative form of Xmsx-1 (VP16/msx-1). Therefore, like Xmsx-1, Xmsx-2B is a downstream nuclear factor of the BMP-4-derived ventralizing signal, and these two factors probably share the same target molecules. In conclusion, Xmsx-1 and Xmsx-2B function in dorso-ventral axis formation in early Xenopus laevis development.
RESUMEN
Whey peptides in a yogurt-like product fermented by Lactobacillus helveticus CPN4 were fractionated by a Sep-pak C-18 cartridge followed by two-step reverse-phase HPLC. The antihypertensive activity was measured by systolic blood pressure in spontaneously hypertensive rats after oral administration of each fraction. Five major peptides in the final fraction were further purified by reverse-phase HPLC and were measured for these antihypertensive activities in spontaneously hypertensive rats. The only peptide in the final fraction that showed strong antihypertensive activity had a sequence of Tyr-Pro, which is found in alpha s1-casein (CN), beta-CN, and kappa-CN. The synthetic peptide Tyr-Pro yielded significant antihypertensive activity from 2 to 8 h after oral administration of 1 mg of peptide/kg of body weight, and the effect was maximal at 6 h after oral administration. The antihypertensive effect of the peptide was dependent on the peptide dosage from 0.1 to 10 mg of peptide/kg of body weight. The concentration of Tyr-Pro peptide increased during fermentation and reached about 8.1 micrograms/ml of whey in the pH 4.3 yogurt-like product. The antihypertensive peptide had a low inhibitory activity against angiotensin I-converting enzyme. The inhibition of 50% of the angiotensin I-converting enzyme (IC50) was 720 microM.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/aislamiento & purificación , Antihipertensivos/aislamiento & purificación , Dipéptidos/aislamiento & purificación , Sístole/efectos de los fármacos , Yogur/análisis , Administración Oral , Inhibidores de la Enzima Convertidora de Angiotensina/química , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Antihipertensivos/química , Antihipertensivos/farmacología , Cromatografía Líquida de Alta Presión , Dipéptidos/química , Dipéptidos/farmacología , Fermentación , Manipulación de Alimentos , Masculino , Proteínas de la Leche , Ratas , Ratas Endogámicas SHR , Proteína de Suero de LecheRESUMEN
In order to analyze biochemical properties of Xenopus bone morphogenetic protein-1 (XBMP-1), rabbit antiserum (alpha-B1) was raised against a synthetic peptide (P1) corresponding to a hydrophilic N-terminal region. XBMP-1B (Xtld) synthesized in the reticulocyte lysate was successfully immunoprecipitated by this antiserum. This precipitation was completely blocked when P1 was added to the reaction, indicating that alpha-B1 recognized XBMP-1B specifically. In Western blot analysis, two distinct sizes of protein (107 and 34 kD) were detected in hind limbs in metamorphosing animals. Both proteins were detected in various adult tissues such as lung, liver, kidney, heart, muscle, intestine, brain, and testis. The mixing of the liver and muscle extracts, and the following detection of immunoreactive proteins suggested that the 34 kD band was a proteolytic product of the 107 kD protein. In the embryonic extracts from the unfertilized egg (stage 0) to swimming tadpoles (stage 40), a 63 kD protein was detected in addition to the 107 kD protein. We also showed that the 107 kD protein was much more expressed in the animal half of the unfertilized eggs than in the vegetal half, but that it was ubiquitously expressed in the gastrula embryos. We suggest that the 63 and 107 kD proteins correspond to full-length proteins encoded by XBMP-1A and XBMP-1B genes, and these proteins are expressed in embryo and in various adult tissues.
Asunto(s)
Anticuerpos/metabolismo , Proteínas Morfogenéticas Óseas , Metaloendopeptidasas/inmunología , Proteínas de Xenopus , Xenopus/embriología , Xenopus/metabolismo , Animales , Proteína Morfogenética Ósea 1 , Sueros Inmunes , Immunoblotting , Larva/inmunología , Metaloendopeptidasas/metabolismo , Pruebas de Precipitina , Factores de Tiempo , Distribución TisularRESUMEN
A 77-year-old man with malignant lymphoma presented with dizziness and exertional dyspnea. Physical examination revealed marked bradycardia (36 beats/min). Twelve-lead electrocardiography showed complete atrioventricular block with narrow QRS escape beats. Gallium scintigraphy demonstrated significant abnormal uptake in the heart. Transesophageal echocardiography showed a thick interatrial septum with increased echogenecity. He underwent chemotherapy under external temporary pacing with a suspected diagnosis of complete atrioventricular block secondary to cardiac invasion of malignant lymphoma. Atrioventricular conduction progressively improved and the complete atrioventricular block disappeared. He is currently well and has required no cardiac pacing for 6 months. We conclude that complete atrioventricular block may be reversible in some patients with malignant lymphoma, even in the elderly.