RESUMO
A new electron beam-based approach for the direct functionalization of polyethersulfone, polyvinylidene fluoride, polysulfone as well as polyacrylonitrile membranes in a one-step procedure is presented. Aqueous solutions of functional molecules were immobilized on the membrane surface by electron beam treatment. The resulting membranes show significantly increased flux and water wettability accompanied by decreased protein adsorption. Stability tests demonstrated the permanence of the modification. This new method neither requires any preceding surface functionalization nor the use of catalysts/photoinitiators or other toxic reagents. In addition, it avoids the synthesis of hydrophilic monomers/polymers, thus avoiding additional synthetic and purification steps as well as the use of organic solvents.
Assuntos
Elétrons , Membranas Artificiais , Polímeros/química , Adsorção , Estrutura Molecular , Permeabilidade , Proteínas/química , Água/química , Poluentes Químicos da Água/químicaRESUMO
INTRODUCTION: Over the last decade, proprioceptive function gained increasing attention in joint disorders such as instability of the shoulder. Common tests for evaluation of proprioception are limited by their complexity and high technical demands. Thus, they are hardly applicable during daily routine. Our hypothesis was that the simplified "laser-pointer assisted angle reproduction test" (LP-ART) presented here allows for clinically feasible assessment of proprioceptive shoulder function. METHODS: Active angle reproduction capability as an aspect of sensorimotor function was evaluated with the new method in patients with shoulder instability (n = 24) and healthy controls (n = 24). 15 patients had traumatic, 9 non-traumatic anterior instability (6 bilateral), 17 were treated surgically, 13 non-operatively. Tests were performed in flexion and abduction in different angles (55°, 90°, 125°) in randomized order. RESULTS: Angle reproduction capability was worst below shoulder level (55°) in all groups. Best results were achieved at shoulder level (90°). Healthy controls showed overall better results than patients with instability. Patients after surgical stabilization had better results in 55° and 90° abduction compared to instability patients before surgery. CONCLUSIONS: The new LP-ART presented here is a technically simple, yet effective instrument for evaluation of the proprioceptive function of the shoulder. In contrast to former test setups it is feasible in daily routine. Compared to healthy controls, patients with unstable shoulder joints show significant proprioceptive disorders that can be quantified by the LP-ART.
Assuntos
Instabilidade Articular/fisiopatologia , Lasers , Propriocepção/fisiologia , Articulação do Ombro/fisiopatologia , Adulto , Estudos de Viabilidade , Feminino , Humanos , MasculinoRESUMO
The treatment of traumatic anterior glenohumeral instability has been a topic of debate over the last couple of decades. However, a consensus exists regarding the necessity of an individualized treatment plan based on the type and degree of instability and the patient's functional demands. Various open and arthroscopic techniques are among the currently used surgical procedures. Open reconstruction of the capsulolabral complex had been considered the treatment of choice for many years, but the latest results for arthroscopic anterior shoulder stabilization demonstrate its increasing use. The current literature suggests that with the introduction of suture anchors, it is possible to mirror the principles of open anterior shoulder stabilization and thus combine the general advantages of arthroscopic surgery with a low recurrence rate.
Assuntos
Artroplastia/métodos , Artroscopia/métodos , Instabilidade Articular/patologia , Instabilidade Articular/cirurgia , Luxação do Ombro/patologia , Luxação do Ombro/cirurgia , Articulação do Ombro/cirurgia , Alemanha , Humanos , Articulação do Ombro/patologia , Técnicas de SuturaRESUMO
BACKGROUND: Paired helical filaments (PHFs) are a characteristic pathological feature of Alzheimer's disease; their principal component is the microtubule-associated protein tau. The tau in PHFs (PHF-tau) is hyperphosphorylated, but the cellular mechanisms responsible for this hyperphosphorylation have yet to be elucidated. A number of kinases, including mitogen-activated protein (MAP) kinase, glycogen synthase kinase (GSK)-3 alpha, GSK-3 beta and cyclin-dependent kinase-5, phosphorylate recombinant tau in vitro so that it resembles PHF-tau as judged by its reactivity with a panel of antibodies capable of discriminating between normal tau and PHF-tau, and by a reduced electrophoretic mobility that is characteristic of PHF-tau. To determine whether MAP kinase, GSK-3 alpha and GSK-3 beta can also induce Alzheimer's disease-like phosphorylation of tau in mammalian cells, we studied the phosphorylation status of tau in primary neuronal cultures and transfected COS cells following changes in the activities of MAP kinase and GSK-3. RESULTS: Activating MAP kinase in cultures of primary neurons or transfected COS cells expressing tau isoforms did not increase the level of phosphorylation for any PHF-tau epitope investigated. But elevating GSK-3 activity in the COS cells by co-transfection with GSK-3 alpha or GSK-3 beta decreased the electrophoretic mobility of tau so that it resembled that of PHF-tau, and induced reactivity with eight PHF-tau-selective monoclonal antibodies. CONCLUSIONS: Our data indicate that GSK-3 alpha and/or GSK-3 beta, but not MAP kinase, are good candidates for generating PHF-type phosphorylation of tau in Alzheimer's disease. The involvement of other kinases in the generation of PHFs cannot, however, be eliminated. Our results suggest that aberrant regulation of GSK-3 may be a pathogenic mechanism in Alzheimer's disease.
Assuntos
Doença de Alzheimer/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas Quinases Ativadas por Mitógeno , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , Proteínas tau/metabolismo , Doença de Alzheimer/enzimologia , Animais , Encéfalo/citologia , Encéfalo/embriologia , Encéfalo/enzimologia , Linhagem Celular , Quinase 3 da Glicogênio Sintase , Quinases da Glicogênio Sintase , Humanos , Mamíferos , Proteína Quinase 1 Ativada por Mitógeno , Proteína Quinase 3 Ativada por Mitógeno , Neurônios/metabolismo , Fosforilação , Ratos , TransfecçãoRESUMO
The phorbol ester PMA/TPA (phorbol 12-myristate 13-acetate) is a potent tumor promoter which mimics distinct intracellular signalling events triggered by activated growth factor receptors, e.g. the activation of MAP kinases. The largest known family of TPA-binding proteins comprise members of the protein kinase C (PKC) family although other TPA-binding proteins outside the PKC family have recently been identified. In this report we addressed the mechanism and the pathway by which TPA induces the activation of MAPkinases. Using recombinant proteins and in vitro phosphorylation reactions we identified the components in the signal transduction pathway from TPA to MAPkinase and we show that the activation of MAPkinase by TPA requires the presence of protein kinase C, c-raf and the MAPkinase activator MEK. We also find that the activation of raf autophosphorylation in vitro correlates with the ability of Raf to signal to MAPkinase. Thus the activation of Raf by PKC apparently can trigger the same signalling pathway as oncogenic Raf or Raf activation by ras in combination with tyrosine phosphorylation.
Assuntos
Quinases de Proteína Quinase Ativadas por Mitógeno , Proteínas Quinases Ativadas por Mitógeno , Proteína Quinase C/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Transdução de Sinais , Acetato de Tetradecanoilforbol/metabolismo , Animais , Baculoviridae/genética , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Humanos , MAP Quinase Quinase 1 , Proteína Quinase 1 Ativada por Mitógeno , Proteína Quinase 3 Ativada por Mitógeno , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Proteínas Tirosina Quinases/genética , Proteínas Proto-Oncogênicas c-raf , Ratos , SpodopteraRESUMO
The Mos protein kinase is a serine-/threonine-specific protein kinase with a crucial role in meiotic cell divisions in vertebrates. Several oncogenic derivatives of the c-Mos protein have been discovered in murine retroviruses. These proteins have acquired mutations and exhibit different degrees of protein kinase activity in vitro. In an attempt to understand the factors governing Mos protein kinase activity we have compared the kinase activities of the wild-type c-Mos protein and two v-Mos proteins (strain HT1 and MSV124) after expression in insect cells. Only the 124 v-Mos protein showed kinase activity in vitro as measured by autophosphorylation, vimentin phosphorylation or by phosphorylation and activation of MAP kinase kinase. By domain swapping and site-directed mutagenesis we identified a single point mutation in the 124 v-Mos protein (Arg145-->Gly) which is responsible for its constitutive activity. This residue is located in the alpha-helix C of the kinase domain close to the ATP binding fold and is conserved in all known c-Mos proteins. Introduction of the corresponding mutation into HT1 v-Mos and into murine c-Mos activated both proteins for autophosphorylation, vimentin phosphorylation and for signalling via MAP kinase kinase in vitro. We hypothesize that the Arg145-->Gly mutation found in 124 v-Mos mimicks a conformational change which might be an obligatory step in the activation of c-Mos in vivo.
Assuntos
Proteínas Oncogênicas v-mos/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas c-mos/metabolismo , Sequência de Aminoácidos , Animais , Arginina/química , Sequência de Bases , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Primers do DNA/química , Glicina/química , Camundongos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fosforilação , Conformação Proteica , Proteínas Recombinantes , Transdução de Sinais , Relação Estrutura-Atividade , Vimentina/metabolismoRESUMO
Raman and near-infrared (NIR) spectroscopy have been evaluated for determining fatty acid composition and contents of main constituents in a complex food model system. A model system consisting of 70 different mixtures of protein, water, and oil blends was developed in order to create a rough chemical imitation of typical fish and meat samples, showing variation both in fatty acid composition and in contents of main constituents. The model samples as well as the pure oil mixtures were measured using Raman and NIR techniques. Partial least squares regression was utilized for prediction, and fatty acid features were expressed in terms of the iodine value and as contents of saturated, monounsaturated, and polyunsaturated fatty acids. Raman spectroscopy provided the best results for predicting iodine values of the model samples, giving validated estimation errors accounting for 2.8% of the total iodine value range. Both techniques provided good results for predicting the content of saturated, monounsaturated, and polyunsaturated fatty acids in the model samples, yielding validated estimation errors in the range of 2.4-6.1% of the total range of fatty acid content. Prediction results for determining fatty acid features of the pure oil mixtures were similar for the two techniques. NIR was clearly the best technique for modeling content of main constituents in the model samples.
Assuntos
Misturas Complexas/análise , Ácidos Graxos/análise , Produtos Pesqueiros/análise , Análise de Alimentos/métodos , Carne/análise , Espectrofotometria Infravermelho/métodos , Análise Espectral Raman/métodos , Algoritmos , Proteínas/análise , Água/análiseRESUMO
Mitogen-activated protein (MAP) kinases are cytoplasmic and/or nuclear protein kinases which are activated by one or several signal transduction pathways from the cell surface into the nucleus. Their activity is regulated by phosphorylation on Tyr as well as on Ser/Thr residues. A cDNA encoding the rat ERK1 member of the MAP kinase family was isolated and sequenced. The longest cDNA consisted of 1875 nucleotides and coded for a polypeptide of 380 amino acids with a predicted M(r) of 42987.
Assuntos
DNA/genética , Proteínas Quinases/genética , Proteínas/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas Quinases Dependentes de Cálcio-Calmodulina , Clonagem Molecular , DNA/isolamento & purificação , Humanos , Dados de Sequência Molecular , Família Multigênica , Ratos , Homologia de Sequência de AminoácidosRESUMO
A proportion of the microtubule-associated protein, tau, is in an elevated state of phosphorylation in foetal and adult brain whereas all of the tau in paired helical filaments, which are characteristic of Alzheimer's disease is hyperphosphorylated; it is important therefore to elucidate the mechanisms that regulate tau phosphorylation. Here we describe results that show that although MAP kinase can hyperphosphorylate tau in vitro, activation of MAP kinase in transformed fibroblasts does not result in hyperphosphorylation of transfected tau, whereas glycogen synthase kinase-3 beta (GSK-3 beta) when co-transfected with tau does result in tau hyperphosphorylation. The findings imply that GSK-3 beta may be a stronger candidate than MAP kinase for inducing tau hyperphosphorylation in vivo.
Assuntos
Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Proteínas Quinases Ativadas por Mitógeno , Proteínas Oncogênicas de Retroviridae/metabolismo , Proteínas tau/metabolismo , Células 3T3 , Animais , Western Blotting , Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Ativação Enzimática , Quinase 3 da Glicogênio Sintase , Quinases da Glicogênio Sintase , Camundongos , Proteína Quinase 3 Ativada por Mitógeno , Proteínas Oncogênicas v-raf , Fosforilação , Proteínas Oncogênicas de Retroviridae/genética , Transformação Genética , Proteínas tau/genéticaRESUMO
Despite widespread use of radiofrequency (RF)-shrinkage, there have been no studies on the influence of RF-energy on neural elements of collagenous tissue. The purpose of this study was to examine the effect of RF-shrinkage on neural structures of capsuloligamentous tissue and the recovery of neural elements under different postoperative treatment protocols. One patellar tendon of 46 New-Zealand-White rabbits was shrunk. Six rabbits were sacrificed immediately postoperative. Twenty rabbits were not immobilized, 10 were immobilized for 3 and 10 were immobilized for 6 weeks. A monoclonal antibody, specific against a neurofilament protein, was used to detect nerves and neural structures. Staining pattern of nerve fibres was significantly altered immediately postoperative. After 3 weeks the number of nerve fibres and bundles decreased significantly in immobilized and non-immobilized limbs. The loss of nerve fibres was significantly less in immobilized limbs. At 6 weeks the number of neural elements in immobilized limbs increased to the level of untreated control tissue. In non-immobilized limbs we found no recovery of neural elements 9 weeks postoperatively. At this time the number of nerve fibres and bundles was still significantly less compared to the untreated control limbs. RF-shrinkage causes significant alteration of neural elements. Under immobilization nerve fibres and bundles reach the level of normal untreated tissue. Careful rehabilitation is important after RF-shrinkage. Not only for biomechanical reasons, but also to allow the neural elements to recover, thermally modified tissue should be protected from normal physiologic loads.
Assuntos
Ablação por Cateter , Imobilização , Regeneração Nervosa/efeitos da radiação , Tendões/inervação , Tendões/cirurgia , Animais , Hipertermia Induzida , Articulação do Joelho/inervação , Articulação do Joelho/cirurgia , Fibras Nervosas/fisiologia , Patela , Complicações Pós-Operatórias/prevenção & controle , Propriocepção , CoelhosRESUMO
The angiographic diagnostic features of concordant tumors of the posterior fossa in monozygotic twins are described. In both cotwins cranio-spinal subependymomas were found which had grown through the foramen magnum and reached from the fourth ventricle down to the second cervical vertebra. Thereby, the conformity of the clinical mainifestation, symptomotology and the course of the disease, the localisation and extent of the tumors as well as the neuroradiologic findings and the histologic features of the tumors were particularly striking. After a review and critical summary of the few cases of concordant affections with brain tumors in identical twins published in the literature at present our couple is discussed under the question of the possible influence of genetic factors or a hereditary disposition on the development of cerebral tumors.
Assuntos
Neoplasias Encefálicas/diagnóstico por imagem , Doenças em Gêmeos , Ependimoma/diagnóstico por imagem , Adulto , Angiografia , Neoplasias Encefálicas/irrigação sanguínea , Neoplasias Encefálicas/genética , Fossa Craniana Posterior , Ependimoma/irrigação sanguínea , Ependimoma/genética , Feminino , Humanos , Gravidez , Gêmeos MonozigóticosRESUMO
In a 3-year-old child, a rare combination of a Dandy-Walker syndrome, a primitive trigeminal artery and a facial haemangioma was found. These multiple dysplasias originate from different germinal layers and result from disturbances of different periods of gestation.
Assuntos
Encéfalo/irrigação sanguínea , Síndrome de Dandy-Walker/complicações , Neoplasias Faciais/complicações , Hemangioma/complicações , Hidrocefalia/complicações , Artérias/anormalidades , Pré-Escolar , Síndrome de Dandy-Walker/diagnóstico , Neoplasias Faciais/diagnóstico , Feminino , Hemangioma/diagnóstico , HumanosRESUMO
Despite widespread use of radiofrequency (RF) shrinkage, there have been no animal studies on the effects of post-operative immobilisation on the histological properties of the shrunken tissue. We have therefore examined the role of post-operative immobilisation after RF shrinkage with special emphasis on the histological properties of collagenous tissue. One patellar tendon of 66 New Zealand White rabbits was shrunk. Six rabbits were killed immediately after the operation. Twenty rabbits were not immobilised, 20 were immobilised for three weeks and 20 for six weeks. Fibroblasts, collagen and vascular quality and density were evaluated on sections, stained by haematoxylin and eosin. Nine weeks after operation the histological properties were inferior to those of the contralateral control tendons. Shrunk tendons did not return to normal at any time after operation irrespective of whether the animals had been immobilised or not. All the parameters improved significantly between zero and three weeks after operation. Immobilised tendons tended to have a better and faster recovery. Careful rehabilitation is imperative after RF shrinkage. Immobilisation aids recovery of the histological properties. Our findings in this animal model support a period of immobilisation of more than three weeks.
Assuntos
Ablação por Cateter/efeitos adversos , Imobilização , Tendões/cirurgia , Animais , Colágeno , Fibroblastos/patologia , Membro Posterior , Articulações , Microscopia/métodos , Período Pós-Operatório , Coelhos , Tendões/patologiaRESUMO
INTRODUCTION: The discussion of diagnostic pre- and intraoperative procedures of the common bile duct (CBD) for laparoscopic cholecystectomy is still open. METHODS: Results of a prospective study of 4000 laparoscopic cholecystectomies were evaluated, adhering to the concept of selective common bile duct examination. In 1999 an inquiry was performed on 1197 patients an average of 38.7 months postoperatively to judge this concept. RESULTS: Three hundred and nineteen patients (8%) examined preoperatively by ERC revealing a CBD stone in 126 cases. The morbidity of this procedure was 3.1%. In 0.18% a lesion of the common bile duct was documented following laparoscopic cholecysTectomy. A intraoperative cholangiography was performed in 67 patients (1.7%) because of unclear anatomy or to exclude a CBD stone. Perioperatively a CBD stone was found in 0.5% of cases. Additionally, in 0.3% the same problem occurred in the long-term follow-up, so that the risk for unknown CBD stones following the selective diagnostic concept is 0.8%. CONCLUSION: The concept of selective diagnostic procedures of the common bile duct shows a low morbidity with sufficient efficiency in avoiding CBD stones as well as duct lesions. Therefore it is recommended for routine use in laparoscopic cholecystectomy.
Assuntos
Colangiografia , Colangiopancreatografia Retrógrada Endoscópica , Colecistectomia Laparoscópica , Cálculos Biliares/diagnóstico , Complicações Intraoperatórias/diagnóstico , Testes de Função Hepática , Complicações Pós-Operatórias/diagnóstico , Seguimentos , Cálculos Biliares/cirurgia , Humanos , Complicações Intraoperatórias/cirurgia , Complicações Pós-Operatórias/cirurgia , Estudos Prospectivos , Reoperação , Sensibilidade e EspecificidadeRESUMO
Self-assembled quantum dots (QDs) are prominent candidates for solid-state quantum information processing. For these systems, great progress has been made in addressing spin states by optical means. In this study, we introduce an all-electrical measurement technique to prepare and detect non-equilibrium many-particle spin states in an ensemble of self-assembled QDs at liquid helium temperature. The excitation spectra of the one- (QD hydrogen), two- (QD helium) and three- (QD lithium) electron configuration are shown and compared with calculations using the exact diagonalization method. An exchange splitting of 10 meV between the excited triplet and singlet spin states is observed in the QD helium spectrum. These experiments are a starting point for an all-electrical control of electron spin states in self-assembled QDs above liquid helium temperature.