RESUMO
Endoscopic retrograde cholangiopancreatography (ERCP) has become the preferred treatment method for hepatobiliary and pancreatic disease. Despite technological progress this technique continues to account for the greatest morbidity and mortality caused by digestive endoscopic procedures. ERCP carries a risk of pancreatitis, perforation, hemorrhage, cholangitis and cardiopulmonary events occurring in upto 10% of patients in referral centers, implying a mortality of up to 1%, not including therapeutic failures or the need for re-intervention. A greater mortality rate has been demonstrated in prospective studies rather than in retrospective studies, but overall, the number of complications described in the literature is much lower than the number of complications that actually occur.A descriptive prospective study was conducted at San José Hospital from April 1, 2006 to April 30, 2007 in patients who underwent an ERCP and had a 1-month follow-up. A total of 381 patients were included; 9 (2.3%) were excluded, and of the remaining 372 there was an overall success in 79.6% of cases, 8.3% had a second intervention, 7.6% developed complications (pancreatitis, perforation, hemorrhage, cholangitis, pain, intolerance to sedatives, and cardiopulmonary events), and 4.3% were failed ERCP studies. The mortality rate of the ERCP procedure was 0.8%.ERCP-related complications were determined at a teaching center, and this suggests the need to implement centers of excellence in order to improve the efficacy of the procedure.
Assuntos
Colangiopancreatografia Retrógrada Endoscópica/efeitos adversos , Adulto , Fatores Etários , Idoso , Assistência Ambulatorial , Colangiopancreatografia Retrógrada Endoscópica/mortalidade , Colômbia , Coleta de Dados , Interpretação Estatística de Dados , Feminino , Hospitais , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Estudos Retrospectivos , Fatores Sexuais , Fatores de TempoRESUMO
Objetivo: determinar la actividad antimicrobiana del extracto en metanol de Stevia rebaudiana sobre bacterias gram negativas y gram positivas, contaminantes de cavidad oral e importantes en enfermedad periodontal. Materiales y métodos. Estudio experimental (fase I) cuyo objetivo era la Stevia rebaudiana. Se realizó la obtención de plantas y microorganismos, preparación de los extractos y ensayos microbiológicos. Se busca estudiar la actividad antimicrobiana del extracto en metanol de hojas de stevia rebaudiana bertoni pulverizada. Los extractos stevia rebaudiana presentaron, en estudios anteriores, actividad sobre microorganismos cariogénicos (S. mutans y L. acidophilus); se cree que los extractos propuestos tengan una gran actividad sobre los microorganismos relacionados con enfermedad periodontal. Resultados y conclusiones. El solvente metanol indica que por sí sola la stevia rebaudiana no no tuvo acción inhibitoria sobre los dos microorganismos gram-negativos. La acción inhibitoria sobre los microorganismos se inicia a partir de 200 mg/ml.
Assuntos
Bactérias Gram-Negativas , Bactérias Gram-Positivas , Cárie Dentária/tratamento farmacológico , Doenças Periodontais/tratamento farmacológico , Fitoterapia , Stevia/classificação , Meios de Cultura , Cárie Dentária/microbiologia , Cárie Dentária , Doenças Periodontais/microbiologia , Metanol/química , Preparações de Plantas/uso terapêutico , Interpretação Estatística de DadosRESUMO
This work describes the involvement of TRPC proteins in capacitative calcium entry (CCE) induced by 1alpha,25-dihydroxy-vitamin-D3 [1alpha,25(OH)2D3] in chick skeletal muscle and in rat osteoblast-like cells (ROS 17/2.8) and the role of the vitamin D receptor (VDR) in this non-genomic rapid response mediated by the hormone. We propose that an endogenous TRPC3 protein mediates 1alpha,25(OH)2D3 modulation of CCE in these cells, which seems to implicate VDR-TRPC3 association and the participation of an INAD-like scaffold protein.
Assuntos
Calcitriol/metabolismo , Canais de Cálcio/metabolismo , Cálcio/metabolismo , Canais Iônicos/metabolismo , Músculo Esquelético/metabolismo , Osteoblastos/metabolismo , Animais , Embrião de Galinha , Proteínas de Membrana/metabolismo , Ratos , Canais de Cátion TRPCRESUMO
This work describes the involvement of TRPC proteins in capacitative calcium entry (CCE) induced by 1a,25-dihydroxy-vitamin-D3 [1a,25(OH)2D3] in chick skeletal muscle and in rat osteoblast-like cells (ROS 17/2.8) and the role of the vitamin D receptor (VDR) in this non-genomic rapid response mediated by the hormone. We propose that an endogenous TRPC3 protein mediates 1a,25(OH)2D3 modulation of CCE in these cells, which seems to implicate VDR-TRPC3 association and the participation of an INAD-like scaffold protein.
Assuntos
Animais , Ratos , Cálcio/metabolismo , Calcitriol/metabolismo , Canais Iônicos/metabolismo , Músculo Esquelético/metabolismo , Osteoblastos/metabolismo , Embrião de Galinha , Proteínas de Membrana/metabolismoRESUMO
It has been recently shown that the fast non-genomic responses of 1,25(OH)(2)-vitamin D(3) [1,25(OH)(2)D(3)] in skeletal muscle cells involve tyrosine phosphorylation of MAP kinase (ERK1/2), c-Src kinase and the oncoprotein c-myc. In the present work, blockade of vitamin D receptor (VDR) expression (> or =80%) by preincubation of chick embryonic muscle cells with three different antisense oligonucleotides against the VDR mRNA (AS-VDR ODNs) significantly reduced (-94%) 1,25(OH)(2)D(3) stimulation of c-myc tyrosine phosphorylation and inhibited c-Src tyrosine dephosphorylation implying lack of c-Src activation by the hormone. Coimmunoprecipitation experiments revealed that 1,25(OH)(2)D(3) induces the formation of complexes between c-Src and c-myc, in agreement with the above results and previous studies showing hormone-dependent association between c-Src and tyrosine phosphorylated VDR and c-Src mediated c-myc tyrosine phosphorylation. MAPK tyrosine phosphorylation by 1,25(OH)(2)D(3) was affected to a lesser extent (-35%) by transfection with AS-VDR ODNs implying that both VDR-dependent and VDR-independent signalling mediate hormone stimulation of MAPK. These are the first results providing direct evidence on the participation of the VDR in non-genomic 1,25(OH)(2)D(3) signal transduction. Activation of tyrosine phosphorylation cascades through this mechanism may contribute to hormone regulation of muscle growth.
Assuntos
Calcitriol/farmacologia , Proteínas Musculares/metabolismo , Receptores de Calcitriol/metabolismo , Animais , Sequência de Bases , Proteína Tirosina Quinase CSK , Células Cultivadas , Embrião de Galinha , Ativação Enzimática/efeitos dos fármacos , Expressão Gênica , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Proteínas Musculares/química , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Oligodesoxirribonucleotídeos Antissenso/genética , Oligodesoxirribonucleotídeos Antissenso/farmacologia , Fosforilação , Proteínas Tirosina Quinases/química , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-myc/química , Proteínas Proto-Oncogênicas c-myc/metabolismo , RNA Mensageiro/genética , Receptores de Calcitriol/antagonistas & inibidores , Receptores de Calcitriol/genética , Transfecção , Tirosina/metabolismo , Quinases da Família srcRESUMO
In previous work we have demonstrated that the steroid hormone 1,25(OH)(2)-vitamin D(3) [1,25(OH)(2)D(3)] stimulates in skeletal muscle cells the phosphorylation and activity of the extracellular signal-regulated mitogen-activated protein (MAP) kinase isoforms ERK1 and ERK2. In the present study we evaluated the involvement of Ca(2+) and protein kinase C (PKC) on 1,25(OH)(2)D(3)-induced activation of MAP kinase. The hormone response was found to depend on PKC stimulation since it was attenuated by the PKC inhibitors calphostin C (100 nM) and bisindolylmaleimide I (30 nM) and PKC downregulation by prolonged treatment with the phorbol ester TPA (1 microM). Removal of external Ca(2+), chelation of intracellular Ca(2+) with BAPTA (5 microM), inhibition of phosphoinositide-phospholipase C (PLC) by neomycin, the calmodulin antagonist fluphenazine (50 microM) and the specific inhibitor of calmodulin kinase II, KN-62 (10 microM), significantly decreased 1,25(OH)(2)D(3)-activation of MAP kinase. In addition, the Ca(2+)-channel blocker verapamil (5 microM) suppressed hormone-induced MAP kinase activity in these cells. Furthermore, the Ca(2+)-mobilizing agent thapsigargin and the Ca(2+)-inophore A23187 paralleled the phosphorylation of MAP kinase observed with 1,25(OH)(2)D(3). Taken together, these results indicate that PKC and Ca(2+) are two upstream activators mediating the effects of 1,25(OH)(2)D(3) on MAP kinase in skeletal muscle cells.
Assuntos
1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/análogos & derivados , Calcitriol/farmacologia , Cálcio/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Músculo Esquelético/efeitos dos fármacos , Proteína Quinase C/metabolismo , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina/farmacologia , Animais , Calcimicina/farmacologia , Cálcio/antagonistas & inibidores , Calmodulina/antagonistas & inibidores , Embrião de Galinha , Ácido Egtázico/análogos & derivados , Ácido Egtázico/farmacologia , Ativação Enzimática/efeitos dos fármacos , Flufenazina/farmacologia , Indóis/farmacologia , Maleimidas/farmacologia , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Músculo Esquelético/citologia , Músculo Esquelético/enzimologia , Músculo Esquelético/metabolismo , Naftalenos/farmacologia , Neomicina/farmacologia , Fosforilação/efeitos dos fármacos , Proteína Quinase C/antagonistas & inibidores , Acetato de Tetradecanoilforbol/farmacologia , Tapsigargina/farmacologia , Fosfolipases Tipo C/metabolismo , Verapamil/farmacologiaRESUMO
We have recently shown that the hormonal form of vitamin D3, 1,25(OH)2-vitamin D3 (1,25(OH)2D3), stimulates the enzymatic activity of the non-receptor protein tyrosine kinase c-Src in skeletal muscle cells. In this study we show that intracellular and extracellular Ca2+ chelation with BAPTA and EGTA, respectively, blocked hormone stimulation of c-Src activity/dephosphorylation, indicating that the calcium messenger system is an upstream activator of c-Src. Tyrosine phosphorylation and stimulation of the growth-related mitogen-activated protein kinase (MAPK) by 1,25(OH)2D3 was shown to be dependent on activation of c-Src, since pretreatment with the c-Src specific inhibitor PP1 or muscle cell transfection with an antisense oligodeoxynucleotide directed against c-Src mRNA markedly reduced hormone stimulation of MAPK phosphorylation. Evidence was obtained indicating that MAPK is then translocated to the cell nucleus in active phosphorylated form and induces the expression of c-myc oncoprotein, as the MAPK kinase (MEK) inhibitor PD98059 abolished stimulation of c-myc synthesis by 1,25(OH)2D3. In addition, the hormone rapidly stimulated tyrosine phosphorylation of c-myc. In cells pretreated with PP1 (4-amino-5-(4-methylphenyl)-7-(t-butyl)pyrazolo-D3,4-pyrimidine), the 1,25(OH)2D3-induced increase in tyrosine phosphorylation of c-myc was suppressed. Taken together, these results demonstrate that 1,25(OH)2D3 stimulates proliferation-associated signalling pathways in skeletal muscle cells and implicate c-Src kinase as mediator of this response.
Assuntos
Colecalciferol/farmacologia , Ácido Egtázico/análogos & derivados , Músculo Esquelético/metabolismo , Proteínas Tirosina Quinases/metabolismo , Animais , Proteína Tirosina Quinase CSK , Sinalização do Cálcio/efeitos dos fármacos , Núcleo Celular/metabolismo , Células Cultivadas , Embrião de Galinha , Ácido Egtázico/farmacologia , Ativação Enzimática/efeitos dos fármacos , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Fosforilação/efeitos dos fármacos , Proteínas Tirosina Quinases/análise , Proteínas Tirosina Quinases/antagonistas & inibidores , Pirazóis/farmacologia , Pirimidinas/farmacologia , Domínios de Homologia de src , Quinases da Família srcRESUMO
The rapid effect of 1 alpha,25(OH(2))-vitamin D(3) [1 alpha, 25(OH(2))D(3)] on tyrosine kinase Src and its relationship to the vitamin D receptor (VDR) was investigated to further characterize the hormone signaling mechanism in chick muscle cells. Exposure of cultured myotubes to 1 alpha,25(OH(2))D(3) caused a time-dependent increase in Src activity, which was evident at 1 min (one-fold) and reached a maximum at 5 min (15-fold). Immunoblotting with anti-phosphotyrosine antibody of immunoprecipitated Src showed that the hormone decreased Src tyrosine phosphorylation state with maximal effects at 5 min. Using a database for protein consensus motifs we found a putative tyrosine phosphorylation site (amino acids 164-170: KTFDTTY) within the primary sequence of the chick VDR. When the myotube VDR was immunoprecipitated it appeared onto SDS-PAGE gels as a single band of 58 kDa recognized by an anti-phosphotyrosine antibody. Prior treatment of cells with (1)alpha,25(OH(2))D(3) significantly increased tyrosine phosphorylation of the VDR (two- to three-fold above basal levels). In agreement with Src being a SH2-domain containing protein involved in recognition of tyrosine-phosphorylated targets, immunoprecipitation with anti-Src antibody under native conditions followed by blotting with anti-VDR antibody, or using the antibodies in inverse order, showed that the VDR co-precipitates with Src, thus indicating the existence of a VDR/Src complex. Stimulation with the cognate VDR ligand significantly increased formation of the complex with respect to basal conditions. These results altogether provide the first evidence to date for 1 alpha,25(OH(2))D(3) activation involving Src association to tyrosine phosphorylated VDR.
Assuntos
Calcitriol/farmacologia , Músculo Esquelético/efeitos dos fármacos , Receptores de Calcitriol/metabolismo , Tirosina/metabolismo , Quinases da Família src/metabolismo , Animais , Embrião de Galinha , Ativação Enzimática , Músculo Esquelético/citologia , Músculo Esquelético/enzimologia , Fosforilação , Transdução de SinaisRESUMO
In cultured chick skeletal muscle cells loaded with Fura-2, the tyrosine kinase inhibitors herbimycin A and genistein abolished both the fast inositol 1,4,5-trisphosphatedependent Ca(2+) release from internal stores and extracellular Ca(2+) influx induced by 1alpha, 25(OH)(2)-vitamin D(3) (1alpha,25(OH)(2)D(3)). Daidzein, an inactive analog of genistein, was without effects. Tyrosine phosphatase inhibition by orthovanadate increased cytosolic Ca(2+). Anti-phosphotyrosine immunoblot analysis revealed that 1alpha, 25(OH)(2)D(3) rapidly (0.5-10 min) stimulates in a concentrationdependent fashion (0.1-10 nm) tyrosine phosphorylation of several myoblast proteins, among which the major targets of the hormone could be immunochemically identified as phospholipase Cgamma (127 kDa), which mediates intracellular store Ca(2+) mobilization and external Ca(2+) influx, and the growth-related proteins mitogen-activated protein (MAP) kinase (42/44 kDa) and c-myc (65 kDa). Genistein suppressed the increase in phosphorylation and concomitant elevation of MAPK activity elicited by the sterol. Both genistein and the MAPK kinase (MEK) inhibitor PD98059 abolished stimulation of DNA synthesis by 1alpha,25(OH)(2)D(3). The sterol-induced increase in tyrosine phosphorylation of c-myc, a finding not reported before for cell growth regulators, was totally suppressed by the specific Src inhibitor PP1. These results demonstrate that tyrosine phosphorylation is a previously unrecognized mechanism involved in 1alpha,25(OH)(2)D(3) regulation of Ca(2+) homeostasis in hormone target cells. In addition, the data involve tyrosine kinase cascades in the mitogenic effects of 1alpha, 25(OH)(2)D(3) on skeletal muscle cells.
Assuntos
Calcitriol/farmacologia , Músculo Esquelético/efeitos dos fármacos , Proteínas Tirosina Quinases/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Benzoquinonas , Cálcio/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Células Cultivadas , Embrião de Galinha , DNA/biossíntese , Ativação Enzimática/efeitos dos fármacos , Flavonoides/farmacologia , Genisteína/farmacologia , Isoenzimas/metabolismo , Lactamas Macrocíclicas , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Músculo Esquelético/enzimologia , Músculo Esquelético/metabolismo , Fosfolipase C gama , Fosforilação/efeitos dos fármacos , Fosfotirosina/metabolismo , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Proteínas Tirosina Fosfatases/metabolismo , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-myc/metabolismo , Quinonas/farmacologia , Rifabutina/análogos & derivados , Fosfolipases Tipo C/metabolismo , Vanadatos/farmacologiaRESUMO
Estrogens initiate their action by binding to specific intracellular receptors and then acting on gene expression. In addition, there is growing evidence of a direct membrane effect via interaction with a cell surphase receptor. The aim of the present study was to investigate the acute effects of 17beta-estradiol on Ca2+ fluxes through second messenger pathways in rat cardiac muscle. Exposure of rat ventricle to low levels of 17beta-estradiol (10(-12)-10(-8) M) increased 45Ca2+ influx within 1 min (+38%); the response was biphasic, peaking at 2 and 5 min (+60 and +55%, respectively). The effect of the hormone on rat heart seems to be specific since 17alpha-estradiol, dihydrotestosterone, and progesterone were devoid of activity. The effect of 17beta-estradiol (5 min, 10(-10) M) was suppressed by nitrendipine (1 microM) and LaCl3 (10 microM), involving the activation of voltage-dependent Ca2+ channels in the acute increase of rat heart calcium influx by the hormone. 17Beta-estradiol rapidly increased cAMP content and PKA activity of rat cardiac muscle in parallel to the changes in Ca2+ uptake. In addition the cAMP antagonist Rp-cAMPS suppressed 17beta-estradiol-dependent Ca2+ influx. Altogether, the data suggest the involvement of the cAMP/PKA messenger system in the nongenomic modulation of Ca2+ influx in rat cardiac muscle by physiological levels of 17beta-estradiol.
Assuntos
Cálcio/metabolismo , Estradiol/farmacologia , Miocárdio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Feminino , Técnicas In Vitro , Transporte de Íons/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
Un total de 97 adultos, hombres y mujeres, en su mayoría representados en estudiantes de odontología de la Pontificia Universidad Javeriana, completaron un estudio clínico doble ciego de seis (6) semanas diseñado para el efecto sobre la formación de placa de un dentífrico que contenía un 0.3 por ciento de triclosán y un 2 por ciento de gantrez, al compararlo con un dentífrico placebo. Los sujetos fueron clasificados en dos (2) grupos balanceados de acuerdo con los porcentajes de placa iniciales. Durante el estudio los sujetos no utilizaron ningún otro procedimiento de higiene bucal, y se les asignó el uso del dentífrico placebo o del dentífrico T/C durante las siguientes 6 (6) semanas. Después de este tiempo se realizó una evaluación de la formación de placa, en la cual se evidenció que el dentífrico T/C suministró una reducción del 34.5 por ciento de la placa establecida el compararse con el dentífrico placebo y una reducción del 71.1 por ciento según el índice de severidad de placa, igualmente al ser comparada con el placebo. Esta reducción de placa fue estadísticamente significativa al 99 por ciento del nivel de confianza, se concluyó que el uso del dentífrico Tricolosán/Copolímero dos (2) veces al día reduce al índice de severidad de placa en un grado altamente significativo, sin obtenerse ningún efecto colateral adverso como la presencia de manchas extrínsecas
Assuntos
Humanos , Masculino , Feminino , Adulto , Triclosan/farmacologia , Placa Dentária , Dentifrícios/análise , Índice de Placa Dentária , PlacebosRESUMO
Twenty male and female adult subjects were entered into a 7-week, double-blind clinical study to determine the effect on plaque formation of a mouthrinse containing 0.03% [corrected] triclosan and 0.25% of a copolymer of methoxyethylene and maleic acid, as compared to 1) a water placebo mouthrinse, 2) a matching alcohol placebo mouthrinse, and 3) Plax antiplaque pre-brushing dental rinse. The subjects were stratified according to their initial plaque scores and assigned in a type of randomized block design (repeated Latin square), so that each subject received each of the four mouthrinses only once for 1 week during the study. The subjects did not use any other oral hygiene procedure (including use of a toothbrush and dentifrice) during the 1-week period of time when they rinsed with their assigned mouthrinse product. The results indicated that use of the triclosan/copolymer mouthrinse provided a 49.8% reduction in supragingival plaque formation compared to the water placebo mouthrinse. The difference was statistically significant at the 99% level of confidence (P less than .01). Similarly, the use of the triclosan/copolymer mouthrinse provided a 47.6% reduction in supragingival plaque formation compared to Plax antiplaque pre-brushing dental rinse. The difference was statistically significant at the 99% level of confidence (P less than .01). The results further indicated that use of the triclosan/copolymer provided a 31.2% reduction in supragingival plaque formation compared to the alcohol placebo mouthrinse. The difference was statistically significant at the 99% level of confidence (P less than .01).