RESUMO
The purpose of this study was to investigate whether dental injury diagnoses may predict adverse outcomes occurring 102 weeks after trauma, and to evaluate whether the severity of adverse outcome is related to laser Doppler flowmetry (LDF) measurements of blood flow from teeth. In 309 trauma patients, 404 permanent maxillary incisors and the respective contralateral homologous control teeth were investigated clinically and radiographically, and by LDF to assess local blood flow values. Dental displacement injuries were classified as grade I (subluxation), grade II (lateral or extrusive luxation), and grade III (avulsion or intrusive luxation). Dental fracture injuries were classified as uncomplicated crown fractures, complicated crown fractures, and root fracture. An adverse outcome was defined as the presence of 'periapical radiolucency and/or grey discolouration'. Significant increase in risk of an adverse outcome occurred with a grade II dental displacement injury (15.07 odds ratio; P = 0.000), a grade III dental displacement injury (28.33 odds ratio; P = 0.000), and a root fracture (106.25 odds ratio; P = 0.000). Blood flow measurements that were significantly associated with more severe outcome were blood flow levels of < or =3 perfusion units (PU; 170.72 odds ratio; P = 0.000), and those of >3 PU and < or =6 PU (76.71 odds ratio; P = 0.000). Diagnoses of displaced and root fractured teeth predicted dental injury patients who went on to show adverse treatment outcomes of splinting. Blood flow measurements from teeth were related to the severity of adverse outcome.
Assuntos
Polpa Dentária/irrigação sanguínea , Polpa Dentária/lesões , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Fluxometria por Laser-Doppler , Modelos Logísticos , Masculino , Fluxo Sanguíneo Regional , Avulsão Dentária/fisiopatologia , Avulsão Dentária/terapia , Fraturas dos Dentes/fisiopatologia , Fraturas dos Dentes/terapia , Reimplante Dentário , Raiz Dentária/lesões , Resultado do TratamentoRESUMO
Splinting of traumatically displaced permanent teeth has been described as an effective modality in the treatment of patients with dental injuries. The purpose of this study was to (i) investigate whether dental injury diagnosis may predict adverse outcomes occurring 96 weeks after splint removal, and (ii) evaluate whether the severity of adverse outcome is related to laser Doppler flowmetry (LDF) measurements of pulpal blood flow (PBF). In 206 trauma patients, 273 permanent maxillary incisors treated by repositioning and splinting, and the respective contralateral homologous control teeth were investigated clinically and radiographically, and by LDF to assess local PBF values. Dental displacement injuries were classified as grade I (subluxation), grade II (lateral or extrusive luxation) and grade III (avulsion or intrusive luxation). Outcomes were classified as 'absence of loss of sensitivity, periapical radiolucency, and/or grey discolouration of crown', type I (loss of sensitivity), type II (loss of sensitivity and periapical radiolucency or grey discoloration of crown) and type III (loss of sensitivity, periapical radiolucency and grey discoloration of crown). An adverse outcome was defined as the presence of 'periapical radiolucency and/or grey discoloration'. A multiple logistic regression analysis was used to compute the odds ratio (OR) for dental displacement injury for adverse outcome (n = 69) vs non-adverse outcome (n = 168). An ordinal stepwise regresssion was completed to assess the degree of association between PBF measurements and outcome groups. Significant increase in risk of an adverse outcome occurred with a grade II dental displacement injury (OR 14.3) (P = 0.000) and a grade III dental displacement injury (OR 19.9) (P = 0.000). PBF measurements that were significantly associated with more severe outcome were PBF levels of < or =3 perfusion units (PU) (OR 399.4) (P = 0.000), those of >3 PU and < or =6 PU (OR 100.5) (P = 0.000), and those of >6 PU and < or =9PU (OR 6.2) (P = 0.000). Diagnoses of displaced teeth predicted dental injury patients who went on to show adverse treatment outcomes of splinting. PBF measurements were related to the severity of adverse outcome.
Assuntos
Polpa Dentária/irrigação sanguínea , Incisivo/lesões , Contenções , Avulsão Dentária/terapia , Adolescente , Adulto , Criança , Pré-Escolar , Colagem Dentária/métodos , Doenças da Polpa Dentária/etiologia , Feminino , Seguimentos , Previsões , Humanos , Incisivo/irrigação sanguínea , Fluxometria por Laser-Doppler , Masculino , Maxila , Pessoa de Meia-Idade , Fios Ortodônticos , Doenças Periapicais/etiologia , Fluxo Sanguíneo Regional/fisiologia , Fatores de Risco , Avulsão Dentária/classificação , Avulsão Dentária/patologia , Descoloração de Dente/etiologia , Resultado do TratamentoRESUMO
Laser Doppler flowmetry (LDF) is a non-invasive method to assess pulpal blood-flow (PBF). Dental injury has been associated with significant PBF reduction. The purpose of this study was to assess whether (i) the type of luxation trauma may affect PBF measurements and (ii) whether luxation type-related measurements may show short- and long-term changes of PBF values. In 41 trauma patients, 69 maxillary incisor treated by repositioning and splinting, and the respective contralateral homologous tooth were investigated by LDF to assess local PBF values. Perfusion units were recorded in four sessions, on the day of splint removal, and 12, 24 and 36 weeks after splint removal. Statistical analysis consisted of univariate analysis of variance for repeated measurements. For the LDF measurements, the main effect of the variable 'session' was not significant (P = 0.119). However, there was a significant 'session'/'luxation type' interaction (P = 0.000). Analysis of simple session-within-luxation type effects revealed intrusive luxations to be associated with a significant decrease in PBF values (P = 0.000), while subluxations (P = 0.568), lateral luxations (P = 0.980), extrusive luxations (P = 0.910), and avulsions (P = 0.996) showed no significant difference between session-related values. The PBF measurements did not change over time for the contralateral incisors (P = 0.996). The LDF may become useful in the detection of pulpal ischaemic episodes in luxated maxillary incisors after repositioning and splinting. Further studies are warranted to assess the validity of the diagnosis of post-traumatic 'ischaemic episodes' by comparing it with histological tooth pulp changes, and by determining how well it may predict course and response to treatments in clinical trials.
Assuntos
Polpa Dentária/irrigação sanguínea , Incisivo/lesões , Avulsão Dentária/fisiopatologia , Adolescente , Adulto , Velocidade do Fluxo Sanguíneo/fisiologia , Criança , Feminino , Seguimentos , Humanos , Fluxometria por Laser-Doppler/métodos , Masculino , Maxila , Pessoa de Meia-Idade , Ortodontia Corretiva , Contenções Periodontais , Avulsão Dentária/cirurgiaRESUMO
Laser Doppler flowmetry (LDF) is a non-invasive method to assess pulpal blood-flow (PBF). Dental luxation injuries have been associated with significant PBF reduction. The purpose of this study was to describe diagnostic characteristics for different session-related threshold PBF values for detection of specific adverse outcomes. In 61 trauma patients, a single maxillary incisor treated by repositioning and splinting, and the respective contralateral homologous control tooth were investigated by LDF to assess local PBF values. Perfusion units (PU) were taken 12 weeks (session I) and 24 weeks (session II) after splint removal. The ability of session I-related PBF measurements at 2.8, 6.4 and 10.1 PU levels, and of session II-related ameasurements at 2.4, 6.3 and 10.2 PU levels to identify adverse outcomes occurring 36 weeks after splint removal was investigated. Adverse outcomes were classified as type I (periapical radiolucency), type II (grey discoloration), and type III (periapical radiolucency and grey discoloration of crown). Receiver operator characteristic curves were used to evaluate the sensitivity and specificity of PBF assessments. There was no significant difference in PBF values between session I and session II (P > 0.05) for teeth without an adverse outcome, and those with a type I, II or III outcome (P > 0.05). The PBF measurements did not change over time for the contralateral incisors (P > 0.05). A type I, II and III outcome occurred in 36, 21 and 12% of the incisors, respectively. The best likelihood ratio was found for the PBF 6.4 PU level at session I and incisors associated with a type I (20.8) and type II outcome (15.2). The PBV of 6.4 PU demonstrated a sensitivity of 96% and a specificity of 59% for type I outcomes, and a sensitivity of 100% and a specificity of 50% for type II outcomes. The data suggest the LDF test to be a valuable diagnostic adjunct for luxated teeth showing signs of adverse outcomes.
Assuntos
Polpa Dentária/irrigação sanguínea , Incisivo/irrigação sanguínea , Fluxometria por Laser-Doppler/métodos , Curva ROC , Adolescente , Adulto , Criança , Feminino , Humanos , Incisivo/lesões , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Bucais/métodos , Complicações Pós-Operatórias/etiologia , Prognóstico , Fluxo Sanguíneo Regional/fisiologia , Sensibilidade e Especificidade , Avulsão Dentária/fisiopatologia , Descoloração de Dente/fisiopatologiaRESUMO
Laser Doppler flowmetry (LDF) is a non-invasive method to assess pulpal blood flow (PBF). Dental injury has been associated with significant PBF reduction. The purpose of this study was (i) to describe PBF characteristics of teeth with specific clinical outcomes, and (ii) to demonstrate diagnostic characteristics for different threshold PBF values for detection of specific multiple adverse outcomes. In 80 trauma patients, a single maxillary incisor treated by repositioning and splinting, and the respective contralateral homologous control tooth were investigated by LDF to assess local PBF values. Perfusion units (PU) were taken in two sessions, on the day of splint removal (session I), and 12 weeks after splint removal (session II). The ability of session II-related PBF measurements at 2.9, 6.4 and 9.9 PU levels to identify adverse outcomes occurring 36 weeks after splint removal was investigated. Adverse outcomes were classified as type I (loss of sensitivity), type II (periapical radiolucency), type III (grey discolouration), type IV (loss of sensitivity and periapical radiolucency), and type V (loss of sensitivity, periapical radiolucency and grey discolouration of crown). Receiver-operator characteristic (ROC) curves were used to evaluate the sensitivity and specificity of PBF assessments. There was a significant increase in PBF values from session I to session II (P=0.0001) for teeth without an adverse outcome, while teeth affected by a type II-V outcome showed a significant decrease in PBF values (P <0.05). PBF measurements did not change over time for the contralateral incisors (P >0.05). A type IV and V outcome occurred in 21 and 24% of the instances, respectively. The PBF of 2.9 PU demonstrated a sensitivity of 70% and a specificity of 93% for type V outcomes. The best likelihood ratio was found for the PBF 2.9 PU level and incisors associated with a type V outcome. The data suggest that the LDF test to be a valuable diagnostic adjunct for luxated teeth showing signs of adverse outcomes including grey discolouration or a combination of other signs. However, it may also become necessary to apply clinical decision-making methods in order to correctly evaluate the value of information gathered. The clinical implication is that LDF may become useful in the prediction of adverse outcomes at a much earlier time period than may be accomplished by standard sensitivity tests.
Assuntos
Polpa Dentária/irrigação sanguínea , Incisivo/lesões , Avulsão Dentária/fisiopatologia , Adolescente , Adulto , Criança , Pré-Escolar , Polpa Dentária/fisiopatologia , Feminino , Seguimentos , Humanos , Hipestesia/etiologia , Incisivo/irrigação sanguínea , Fluxometria por Laser-Doppler , Funções Verossimilhança , Masculino , Maxila , Pessoa de Meia-Idade , Doenças Periapicais/etiologia , Curva ROC , Fluxo Sanguíneo Regional/fisiologia , Sensibilidade e Especificidade , Contenções , Avulsão Dentária/complicações , Avulsão Dentária/terapia , Coroa do Dente/patologia , Descoloração de Dente/etiologia , Resultado do TratamentoRESUMO
AIM: To evaluate laser Doppler flowmetry (LDF) measurements of pulpal blood flow (PBF) and severity of dental injury. METHODOLOGY: The relationship between adverse outcomes and PBF measurements was analysed in 94 permanent maxillary incisors of 71 consecutive dental trauma patients. The diagnostic adverse outcome group was comprised of 72 incisors in 52 patients with a type I (loss of sensitivity), type II (loss of sensitivity and periapical radiolucency), or type III (loss of sensitivity, periapical radiolucency and grey discoloration of crown) diagnosis. The nonadverse outcome group consisted of 22 incisors in 19 patients with the finding of an absence of an adverse outcome. At each session, when an injured permanent maxillary incisor was recorded, a contralateral homologous tooth was used as a control. An ordinal stepwise regression was completed to assess the degree of association between PBF measurements and adverse outcomes RESULTS: Using chi-square analysis for pairwise comparison, a significant relationship between PBF measurements and types of adverse outcomes (chi(2) =119.635, d.f. = 12, P = 0.000) was observed. PBF measurements that were significantly associated with more severe outcome were PBF levels of =3 perfusion units (PU) (119.1 odds ratio) (P = 0.000), and those of >3 PU and =6 PU (12.7 odds ratio) (P = 0.000). CONCLUSIONS: PBF measurements were related to the severity of adverse outcomes. Further studies are required to evaluate whether PBF measurements can predict dental injuries that progress to adverse treatment outcomes.
Assuntos
Polpa Dentária/irrigação sanguínea , Incisivo/lesões , Avulsão Dentária/fisiopatologia , Adolescente , Adulto , Análise de Variância , Criança , Pré-Escolar , Feminino , Humanos , Incisivo/irrigação sanguínea , Fluxometria por Laser-Doppler , Masculino , Pessoa de Meia-Idade , Avaliação de Resultados em Cuidados de Saúde/métodos , Análise de Regressão , Distúrbios Somatossensoriais/diagnóstico , Distúrbios Somatossensoriais/etiologia , Avulsão Dentária/complicações , Índices de Gravidade do TraumaRESUMO
The purpose of this study was to assess the prevalences of magnetic resonance (MR) imaging findings of internal derangement (ID) in temporomandibular joints (TMJs) without a specific clinical diagnosis of temporomandibular disorder (TMD), and to investigate whether in this TMJ group the variable of pain may be linked to MR imaging findings of ID. The study comprised 109 patients, who were assigned a clinical uni- or bilateral TMJ-related diagnosis of 'absence of TMD'. Bilateral sagittal and coronal MR images were obtained subsequently to establish the prevalence of TMJ ID. An MR imaging diagnosis of ID was found in 99 (55.9%) of the 177 TMJs investigated. About 30.3% of the closed mouth-related TMJ positions characterized by disc displacement presented with anterior disc displacement, while 27.3% had anterolateral and 25.3% anteromedial disc displacement. Analysis of the data revealed the presence of TMJ pain to be associated with significantly more MR imaging diagnoses of disc displacement without reduction than disc displacement with reduction (P < 0.05), while there was no significant difference in the prevalences of ID and those of absence of ID (P > 0.05). Using chi-square analysis, no significant relationship was found between the presence of TMJ pain and the MR imaging diagnosis of TMJ ID (P=0.93). Use of the kappa statistical test indicated poor diagnostic agreement between the presence of TMJ pain and the MR imaging diagnosis of ID (kappa=0.01). The results suggest TMJs with a clinical diagnosis of 'absence of TMD' to be associated with a high rate of IDs, while in these instances the clinical variable of TMJ pain may have no effect on prevalences of MR imaging diagnoses TMJ ID. The data confirm the aspect of clinical diagnostic criteria as an unreliable instrument in predicting MR imaging diagnoses of TMJ ID.
Assuntos
Disco da Articulação Temporomandibular/patologia , Transtornos da Articulação Temporomandibular/patologia , Adolescente , Adulto , Idoso , Dor Facial/etiologia , Feminino , Humanos , Luxações Articulares/patologia , Imageamento por Ressonância Magnética , Masculino , Côndilo Mandibular/patologia , Pessoa de Meia-Idade , Amplitude de Movimento Articular , Reprodutibilidade dos Testes , Transtornos da Articulação Temporomandibular/complicaçõesRESUMO
Cystic fibrosis (CF) is characterized by impaired Cl(-) secretion and increased Na(+) reabsorption in several tissues including respiratory epithelium. Many CFTR mutations have been identified over the past years. However, only a poor correlation between the genotype and lung phenotype was found suggesting additional factors influencing the phenotype and course of the disease. The serine/threonine kinase SGK1 has recently been shown to stimulate the activity of the epithelial Na(+) channel ENaC. A variety of stimuli such as aldosterone, cell shrinkage, insulin or TGF-beta1 stimulate transcription and activate the SGK1 kinase. Here we further examined the effects of SGK1 on ENaC and CFTR which have mutual interactions and we analyzed sgk1 mRNA abundance in lung tissue from CF patients. Coexpression of CFTR and h-SGK1 in Xenopus oocytes increased ENaC currents as previously described. In addition CFTR mediated currents were also stimulated. h-SGK1 accelerated the expression of the amiloride sensitive Na(+)- current in Xenopus oocytes paralleled by increased ENaC-protein abundance in the oocyte membrane, an effect which was reversed by a h-SGK1(K127R) mutation lacking the ATP-binding site. The cation selectivity or Na(+) affinity were not affected. However, coexpression of h-SGK1 with ENaC altered the sensitivity of the Na(+)-channel to the inhibitors amiloride and triamterene. The inhibitory effect of CFTR expression on ENaC current was not affected by coexpression of h-SGK1 in Xenopus oocytes. Lung tissue from CF patients strongly expressed the serine/threonine kinase h-sgk1 which was not the case for non-CF lung tissue. Loss of CFTR function itself in a CF lung epithelial cell line did not increase SGK1 expression. In summary, enhanced expression of h-SGK1 in epithelial cells of CF-lung tissue may be a novel pathophysiological factor contributing to increased Na(+) channel activity and thus to increased Na(+) transport in CF.
Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Fibrose Cística/metabolismo , Pulmão/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Alvéolos Pulmonares/metabolismo , Canais de Sódio/metabolismo , 1-Metil-3-Isobutilxantina/farmacologia , Substituição de Aminoácidos , Animais , Brônquios/citologia , Brônquios/metabolismo , Linhagem Celular , Fibrose Cística/genética , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Células Epiteliais/metabolismo , Canais Epiteliais de Sódio , Humanos , Hibridização In Situ , Pulmão/citologia , Macrófagos Alveolares/metabolismo , Mutação , Oócitos/metabolismo , Técnicas de Patch-Clamp , Proteínas Serina-Treonina Quinases/genética , Alvéolos Pulmonares/citologia , RNA Complementar , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sódio/metabolismo , Canais de Sódio/genética , Xenopus laevisRESUMO
Extracellular pH has been shown previously to influence transport via type-II Na+/phosphate (NaPi) transporters by modifying the affinity of the carrier for Na+ and by altering the availability of divalent and monovalent phosphate. As the transport of monovalent phosphate would be expected to acidify, and that of divalent phosphate to alkalinize the cell interior, the effect of phosphate transport on cytosolic pH was studied using ion selective microelectrodes in Xenopus oocytes expressing NaPi-3 or NaPi-5. At an alkaline extracellular pH (pHe) of 8.0, addition of phosphate elicited a strong inward current, depolarization of the cell membrane and cytosolic alkalinization. At pHe 6.0 the phosphate-induced inward current and depolarization were reduced and the alkalinization completely abolished. In conclusion, at alkaline pHe phosphate transport is enhanced and the transport of divalent phosphate prevails. At pHe 6.0, phosphate transport is attenuated and is accomplished by transport of both divalent and monovalent phosphate.
Assuntos
Proteínas de Transporte/metabolismo , Concentração de Íons de Hidrogênio , Fosfatos/farmacocinética , Simportadores , Animais , Transporte Biológico/fisiologia , Proteínas de Transporte/genética , Expressão Gênica , Humanos , Túbulos Renais Proximais/metabolismo , Oócitos/metabolismo , Técnicas de Patch-Clamp , Proteínas Cotransportadoras de Sódio-Fosfato , Proteínas Cotransportadoras de Sódio-Fosfato Tipo II , Xenopus laevisRESUMO
The SLC26 gene family (solute carrier family 26) comprises five mammalian genes that encode anion transporter-related proteins. In addition to sat-1 and prestin, which were cloned from rat and gerbil, respectively, three human members have been identified and associated with specific genetic diseases (DTD, diastrophic dysplasia; CLD, congenital chloride diarrhea; PDS, Pendred syndrome). In this study we used a homology approach combined with RACE PCR to identify human SLC26A6, the sixth member of this gene family. Northern blot analysis showed the highest SLC26A6 transcript levels in kidney and pancreas. Expression in MDCK cells and in Xenopus oocytes demonstrated trafficking of the SLC26A6 protein to the cell membrane but did not reveal anion transport activity with tracer uptake or intracellular pH measurements. We determined the genomic structure of the SLC26A6 gene and excluded mutations in the 21 coding exons as the cause of DFNB6 and USH2B, which closely map to the SLC26A6 chromosomal locus (3p21).
Assuntos
Proteínas de Transporte/química , Proteínas de Transporte/genética , Sequência de Aminoácidos , Animais , Proteínas de Transporte de Ânions , Linhagem Celular , Membrana Celular/metabolismo , Mapeamento Cromossômico , Cromossomos Humanos Par 3/genética , Clonagem Molecular , Surdez/genética , Expressão Gênica , Testes Genéticos , Perda Auditiva Neurossensorial/genética , Humanos , Rim/metabolismo , Dados de Sequência Molecular , Família Multigênica , Mutação , Especificidade de Órgãos , Pâncreas/metabolismo , Reação em Cadeia da Polimerase , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Homologia de Sequência de Aminoácidos , SíndromeRESUMO
Ample pharmacological evidence points to a role of kinases in the regulation of cell volume. Given the limited selectivity of most inhibitors, however, the specific molecules involved have remained largely elusive. The search for cell volume regulated genes in liver HepG2 cells led to the discovery of the human serum- and glucocorticoid-dependent serine/threonine kinase hsgk1. Transcription and expression of hsgk1 is markedly and rapidly upregulated by osmotic and isotonic cell shrinkage. The effect of osmotic cell shrinkage on hsgk1 is mediated by p38 kinase. Further stimuli of hsgk1 transcription include glucocorticoids, aldosterone, TGF-beta1, serum, increase of intracellular Ca2+ and phorbolesters, whereas cAMP downregulates hsgk1 transcription. The hsgk1 protein is expressed in several epithelial tissues including human pancreas, intestine, kidney, and shark rectal gland. Co-expression of hsgk1 with the renal epithelial Na+-channel ENaC or the Na+/K+/2Cl(-)-cotransporter NKCC2 (BSC1) in Xenopus oocytes, accelerates insertion of the transport proteins into the cell membrane and thus, stimulates channel or transport activity. Thus, hsgk1 participates in the regulation of transport by steroids and secretagogues increasing intracellular Ca2+-activity. The stimulation of hsgk1 transcription by TGF-beta1 may further bear pathophysiological relevance.
Assuntos
Tamanho Celular , Epitélio/metabolismo , Proteínas Nucleares , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Transporte Biológico , Proteínas Imediatamente PrecocesRESUMO
Besides uptake of Na(+) and Cl(-), mammalian cells counteract osmotic cell shrinkage also by Na(+)-coupled uptake of osmolytes, e. g., myo-inositol, taurine or betaine. The expression of the corresponding transporters is transcriptionally regulated by the ambient pH and osmolarity and is increased upon cell shrinkage, a process requiring hours. The present study has been performed to disclose rapid regulation by pH of osmolyte transport via BGT-1. Transport of GABA was investigated by using the two-electrode voltage-clamp technique with BGT-1 expressing Xenopus oocytes. GABA was used as a substrate, because of the low oocyte endogenous transport activity. Extracellular acidification to pH 5.5 reversibly decreased and extracellular alkalinization to pH 8.5 increased GABA-induced currents. Kinetic analysis revealed that extracellular alkalinization increases the affinity for Cl(-) as reflected by a decrease of the apparent K(m)-value for Cl(-) from >500 mM to 55.8 +/- 4.7 mM upon an increase of the pH from 7.0 to 8.5. The apparent K(m)- values for Na(+) and GABA remained unaltered in the pH range from 6.0 to 8.5. Instead, alkalinization increased the maximal current induced by saturating Na(+) and GABA concentrations. The results are compatible with a model of interference of H(+) ions with Cl(-) binding and a pH-dependent reduction of V(max) for Na(+) and GABA.
Assuntos
Betaína/metabolismo , Proteínas de Transporte/biossíntese , Ácido gama-Aminobutírico/metabolismo , Animais , Cães , Proteínas da Membrana Plasmática de Transporte de GABA , Concentração de Íons de Hidrogênio , Cinética , Oócitos/metabolismo , Técnicas de Patch-Clamp , Xenopus , Ácido gama-Aminobutírico/farmacologiaRESUMO
At moderate cell shrinkage, activation of Na(+) channels is the most prominent mechanism of regulatory cell volume increase in rat hepatocytes. The amiloride sensitivity of these channels suggests a relation to the family of epithelial Na(+) channels (ENaCs). The present study was performed to determine the pharmacological profile of shrinkage-activated Na(+) channels and to test for ENaC expression in primary cultures of rat hepatocytes; in addition, the influence of the cell volume regulated serine/threonine kinase hSGK on activity and pharmacological profile of rENaC was examined in Xenopus oocytes. Conventional electrophysiology in hepatocytes reveals that the shrinkage-activated Na(+) channel is inhibited by amiloride and EIPA with IC(50) values of 6.0 and 0.12 micromol/l, respectively. Western blots and RT-PCR demonstrate that rat hepatocytes do express all three subunits (alpha, beta, gamma) of ENaC. Coexpression of hSGK with rENaC in Xenopus oocytes reveals that the kinase stimulates ENaC by a factor of 4. Moreover, hSGK decreases the affinity to amiloride (increase of IC(50) from 0.12 to 0.26 micromol/l) and increases the affinity to EIPA (decrease of IC(50) from 250 to 50 micromol/l). In conclusion, rat hepatocytes express ENaC, which is activated by the cell volume-sensitive kinase hSGK. ENaC may contribute to the Na(+) channels activated by osmotic cell shrinkage in hepatocytes, whereby the relatively low amiloride and high EIPA sensitivity of the channel could at least be partially due to modification by SGK, which decreases the amiloride and increases the EIPA sensitivity of ENaC.
Assuntos
Hepatócitos/citologia , Hepatócitos/metabolismo , Ativação do Canal Iônico , Proteínas Nucleares , Canais de Sódio/metabolismo , Sódio/metabolismo , Amilorida/análogos & derivados , Amilorida/farmacologia , Animais , Tamanho Celular , Células Cultivadas , DNA Complementar/genética , Canais Epiteliais de Sódio , Hepatócitos/efeitos dos fármacos , Humanos , Proteínas Imediatamente Precoces , Immunoblotting , Concentração Inibidora 50 , Microinjeções , Oócitos/metabolismo , Concentração Osmolar , Pressão Osmótica , Técnicas de Patch-Clamp , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Subunidades Proteicas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Canais de Sódio/química , Canais de Sódio/genética , XenopusRESUMO
L-Carnitine is essential for the translocation of acyl-carnitine into the mitochondria for beta-oxidation of long-chain fatty acids. It is taken up into the cells by the recently cloned Na(+)-driven carnitine organic cation transporter OCTN2. Here we expressed hOCTN2 in Xenopus laevis oocytes and investigated with two-electrode voltage- clamp and flux measurements its functional and pharmacological properties as a Na(+)-carnitine cotransporter. L-carnitine transport was electrogenic. The L-carnitine-induced currents were voltage and Na(+) dependent, with half-maximal currents at 0.3 +/- 0.1 mM Na(+) at -60 mV. Furthermore, L-carnitine-induced currents were pH dependent, decreasing with acidification. In contrast to other members of the organic cation transporter family, hOCTN2 functions as a Na(+)-coupled carnitine transporter. Carnitine transport was stereoselective, with an apparent Michaelis-Menten constant (K(m)) of 4.8 +/- 0.3 microM for L-carnitine and 98.3 +/- 38.0 microM for D-carnitine. The substrate specificity of hOCTN2 differs from rOCT-1 and hOCT-2 as hOCTN2 showed only small currents with classic OCT substrates such as choline or tetraethylammonium; by contrast hOCTN2 mediated transport of betaine. hOCTN2 was inhibited by several drugs known to induce secondary carnitine deficiency. Most potent blockers were the antibiotic emetine and the ion channel blockers quinidine and verapamil. The apparent IC(50) for emetine was 4.2 +/- 1.2 microM. The anticonvulsant valproic acid did not induce a significant inhibition of carnitine transport, pointing to a different mode of action. In summary, hOCTN2 mediates electrogenic Na(+)-dependent stereoselective high-affinity transport of L-carnitine and Na(+). hOCTN2 displays transport properties distinct from other members of the OCT family and is directly inhibited by several substances known to induce systemic carnitine deficiency.
Assuntos
Carnitina/farmacocinética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas de Transporte de Cátions Orgânicos , Sódio/metabolismo , Aminoácidos/química , Aminoácidos/farmacologia , Animais , Betaína/química , Betaína/farmacologia , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/fisiologia , Carnitina/química , Carnitina/deficiência , Proteínas de Transporte/química , Emetina/química , Emetina/farmacologia , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Humanos , Concentração de Íons de Hidrogênio , Lipotrópicos/química , Lipotrópicos/farmacologia , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Proteínas de Membrana/química , Microinjeções , Oócitos/fisiologia , Técnicas de Patch-Clamp , Ácidos Pentanoicos/química , Ácidos Pentanoicos/farmacologia , Inibidores da Síntese de Proteínas/química , Inibidores da Síntese de Proteínas/farmacologia , RNA Complementar/farmacologia , Membro 5 da Família 22 de Carreadores de Soluto , Tetraetilamônio/química , Tetraetilamônio/farmacologia , Trítio , Ácido Valproico/química , Ácido Valproico/farmacologia , Xenopus laevisRESUMO
Transforming growth factor beta (TGF-beta) has been shown to participate in the pathophysiology of diabetic complications. As shown most recently, TGF-beta stimulates the expression of a distinct serine/threonine kinase (hSGK) which had previously been cloned as an early gene transcriptionally regulated by cell volume alterations. The present study was performed to elucidate transcription and function of hSGK in diabetic nephropathy. As shown by Northern blotting, an increase of extracellular glucose concentration increased hSGK mRNA levels in cultured cells, an effect qualitatively mimicked by osmotic cell shrinkage or treatment with TGF-beta (2 microgram/liter), phorbol 12,13-didecanoate (1 microM), or the Ca(2+) ionophore ionomycin (1 microM) and blunted by high concentrations of nifedipine (10 and 100 microM). In situ hybridization revealed that hSGK transcription was markedly enhanced in diabetic nephropathy, with particularly high expression in mesangial cells, interstitial cells, and cells in thick ascending limbs of Henle's loop and distal tubules. According to voltage clamp and tracer flux studies in Xenopus oocytes expressing the renal epithelial Na(+) channel ENaC or the mouse thick ascending limb Na(+),K(+),2Cl(-) cotransporter BSC-1, coexpression with hSGK stimulated ENaC and BSC-1 11-fold and 6-fold, respectively, effects reversed by kinase inhibitors staurosporine (1 microM) and chelerythrine (1 microM) and not elicited by inactive hSGK. In conclusion, excessive extracellular glucose concentrations enhance hSGK transcription, which in turn stimulates renal tubular Na(+) transport. These observations disclose an additional element in the pathophysiology of diabetic nephropathy.
Assuntos
Nefropatias Diabéticas/genética , Proteínas Serina-Treonina Quinases/genética , Células 3T3 , Animais , Proteínas de Transporte/biossíntese , Proteínas de Transporte/genética , Tamanho Celular , Canais Epiteliais de Sódio , Regulação da Expressão Gênica , Humanos , Hibridização In Situ , Rim/patologia , Camundongos , Técnicas de Patch-Clamp , Proteínas Serina-Treonina Quinases/biossíntese , Ratos , Proteínas Recombinantes/biossíntese , Sódio/metabolismo , Canais de Sódio/biossíntese , Canais de Sódio/genética , Simportadores de Cloreto de Sódio-Potássio , Distribuição Tecidual , Transcrição GênicaRESUMO
STATEMENT OF PROBLEM: A precise preparation is required to develop resistance form resulting in mechanical stability of the framework for resin-bonded prostheses (RBPs). PURPOSE: The effects of 4 methods of tooth preparation (freehand, guiding pin, extraoral parallelometer, and intraoral parallelometer) on the deviation of proximal grooves from a preestablished path of insertion (guide planes) were investigated under clinical conditions. MATERIAL AND METHODS: Tooth preparation of proximal grooves was performed by 32 dentists on resin substitutes of posterior segments intraorally with a single test patient. A Latin-square randomized cross-over design was selected as the experimental design. RESULTS: The significant least angular deviation of proximal grooves from path of insertion was achieved with an intraoral parallelometer (mean +/- SD 3.15 +/- 1.67 degrees). Compared with freehand tooth preparations (4.37 +/- 2. 11 degrees), neither use of a guiding pin (4.10 +/- 1.62 degrees) nor an extraoral parallelometer (5.06 +/- 2.33 degrees) improved the results. CONCLUSION: Divergence of guiding grooves from path of insertion was reduced with the use of an intra-oral parallelometer. This should improve mechanical stability of posterior RBPs.
Assuntos
Prótese Adesiva/normas , Preparo Prostodôntico do Dente/métodos , Resinas Acrílicas , Adulto , Análise de Variância , Estudos Cross-Over , Planejamento de Dentadura/instrumentação , Planejamento de Dentadura/métodos , Planejamento de Dentadura/normas , Prótese Adesiva/estatística & dados numéricos , Humanos , Masculino , Mandíbula , Maxila , Modelos Dentários , Distribuição Aleatória , Preparo Prostodôntico do Dente/normas , Preparo Prostodôntico do Dente/estatística & dados numéricosRESUMO
The incline and length of guiding elements, i.e. marginal ridges and lingual surfaces of front teeth, marginal ridges and internal cusp slopes of premolars and molars, play an important role in dentistry. Since the so far reported values differ considerably, it was the purpose of the present investigation to replicate the measurements, including all the occlusal landmarks proposed and defined by previous investigators. The measurements were performed on 34 pairs of mounted casts from a selected group of untreated, naturally grown dentitions from adolescents of mean age 14 years. The upper casts were mounted with a face bow, the kinematic hinge axis and the left incisura infraorbitalis representing the posterior and anterior reference points. The lower pinned casts were mounted joint related. All measurements were carried out with a computer-aided, three-dimensional digitizer. The inclines were expressed as angles related to the axis-orbital-plane. Taking the proposed occlusal landmarks as a basis, the inclines of guiding elements were found to be in agreement with previously reported values, despite ethnic and racial differences of the various study-populations. The values, however, differed markedly when measurements based on individual, functional relevant landmarks were compared to measurements based on anatomical, easy identifiable or mathematically constructed landmarks. The successive decrease of the inclines of the guiding elements from the central incisors to the second molars could be confirmed, the molars displaying very flat inclines. Interestingly, 9% of the first molars and 21% of the second molars showed negative values, pointing to a functional arrangement characterized by a buccally oriented occlusal surface of those teeth. Combined with the finding that the length of the guiding elements of the anterior teeth was almost twice as long as that of the posterior teeth, the results corroborate the occlusal concept of an anterior-posterior sequence of the guiding elements, or a so-called sequential guidance with front-canine-dominance.
Assuntos
Oclusão Dentária , Dente/anatomia & histologia , Adolescente , Dente Pré-Molar/anatomia & histologia , Criança , Estudos Transversais , Dente Canino/anatomia & histologia , Articuladores Dentários , Oclusão Dentária Central , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Incisivo/anatomia & histologia , Masculino , Modelos Dentários , Dente Molar/anatomia & histologia , Odontometria , Radiografia , Dente/diagnóstico por imagem , Dente/fisiologiaRESUMO
This case report describes the treatment of an external root resorption with extensive loss of tooth structure and bone at the labial surface of an upper left central incisor. The area of bone loss and root resorption was surgically exposed and an impression was taken using curing silicone. An individual ceramic insert was fabricated, allowing endodontic retreatment through an artificial root canal. The insert was incorporated using a dentin bonding system and a dual curing luting composite. Following endodontic retreatment and internal bleaching, a ceramic veneer was bonded to the tooth to obtain good esthetics and to improve stability. Twenty months after surgical treatment no further root resorption could be detected radiographically. A shallow residual pocket but no bleeding on probing was found.
Assuntos
Restauração Dentária Permanente/métodos , Incisivo , Reabsorção da Raiz/terapia , Adulto , Perda do Osso Alveolar/terapia , Cerâmica , Resinas Compostas , Colagem Dentária , Facetas Dentárias , Adesivos Dentinários , Feminino , Seguimentos , Bolsa Gengival/patologia , Humanos , Restaurações Intracoronárias , Maxila , Tratamento do Canal Radicular , Clareamento DentalRESUMO
Levels of specific salivary IgA antibodies against mycobacterial heat shock protein (hsp) 65 are significantly increased in patients with gingivitis when compared to clinically healthy subjects. The process of identifying the hsp65 epitopes recognized by the salivary antibodies, binding to overlapping 15-mer-hsp65 peptides, was assessed. Time-resolved fluorescence immunoassays using 15-mer overlapping peptides spanning the whole hsp65 molecule revealed six distinct sequences recognized by anti-hsp65 IgA antibodies. Due to the high degree of sequence homology between mycobacterial hsp65, cognates of the hsp60 family of oral bacterial flora and human hsp60, these six epitopes may serve as cross-reactive autoantigens in certain circumstances in vivo and could incite an autoimmune response that contributes to the initiation of gingivitis.