RESUMO
IFNß (recombinant interferon Beta) has been widely used for the treatment of Multiple sclerosis for the last four decades. Despite the human origin of the IFNß sequence, IFNß is immunogenic, and unwanted immune responses in IFNß-treated patients may compromise its efficacy and safety in the clinic. In this study, we applied the DeFT (De-immunization of Functional Therapeutics) approach to producing functional, de-immunized versions of IFNß-1a. Two de-immunized versions of IFNß-1a were produced in CHO cells and designated as IFNß-1a(VAR1) and IFNß-1a(VAR2). First, the secondary and tertiary protein structures were analyzed by circular dichroism spectroscopy. Then, the variants were also tested for functionality. While IFNß-1a(VAR2) showed similar in vitro antiviral activity to the original protein, IFNß-1a(VAR1) exhibited 40% more biological potency. Finally, in vivo assays using HLA-DR transgenic mice revealed that the de-immunized variants showed a markedly reduced immunogenicity when compared to the originator.
Assuntos
Esclerose Múltipla , Animais , Camundongos , Cricetinae , Humanos , Esclerose Múltipla/tratamento farmacológico , Interferon beta , Interferon beta-1a/uso terapêutico , Cricetulus , Recidiva Local de Neoplasia , Adjuvantes ImunológicosRESUMO
Endo-ß-1,3-glucanases from several organisms have attracted much attention in recent years because of their capability for in vitro degrading ß-1,3-glucan as a critical step for both biofuels production and short-chain oligosaccharides synthesis. In this study, we biochemically characterized a putative endo-ß-1,3-glucanase (EgrGH64) belonging to the family GH64 from the single-cell protist Euglena gracilis. The gene coding for the enzyme was heterologously expressed in a prokaryotic expression system supplemented with 3% (v/v) ethanol to optimize the recombinant protein right folding. Thus, the produced enzyme was highly purified by immobilized-metal affinity and gel filtration chromatography. The enzymatic study demonstrated that EgrGH64 could hydrolyze laminarin (KM 23.5 mg ml-1,kcat 1.20 s-1) and also, but with less enzymatic efficiency, paramylon (KM 20.2 mg ml-1,kcat 0.23 ml mg-1 s-1). The major product of the hydrolysis of both substrates was laminaripentaose. The enzyme could also use ramified ß-glucan from the baker's yeast cell wall as a substrate (KM 2.10 mg ml-1, kcat 0.88 ml mg-1 s-1). This latter result, combined with interfacial kinetic analysis evidenced a protein's greater efficiency for the yeast polysaccharide, and a higher number of hydrolysis sites in the ß-1,3/ß-1,6-glucan. Concurrently, the enzyme efficiently inhibited the fungal growth when used at 1.0 mg/mL (15.4 µM). This study contributes to assigning a correct function and determining the enzymatic specificity of EgrGH64, which emerges as a relevant biotechnological tool for processing ß-glucans.
Assuntos
Euglena gracilis , Cinética , Polissacarídeos/metabolismo , Hidrólise , Saccharomyces cerevisiae/metabolismo , Especificidade por SubstratoRESUMO
Se presenta el caso de un hombre de 57 años que consulta por parálisis alta del nervio radial, con dolor y prueba de Tinel positiva en la cara lateral del brazo dominante, de inicio súbito, luego de grandes esfuerzos musculares repetitivos, sin mejoría clínica al tercer mes de evolución. Se realizó un tratamiento quirúrgico descompresivo. El paciente tuvo una rápida recuperación a partir del séptimo día, y remisión completa a los 25 días de la cirugía. Conclusión: El atrapamiento del nervio radial en el brazo es un cuadro poco frecuente. Según los estudios publicados, la evolución clínica es variada, pero si no hay remisión o la evolución de la parálisis no es favorable en 3 meses, creemos que la cirugía es el tratamiento de elección. Nivel de Evidencia: IV
We present the case of a 57-year-old male patient who consulted for high radial nerve palsy, with pain and positive Tinel test on the lateral side of the dominant arm, of sudden onset after great repetitive muscular efforts, without clinical improvement after three months of evolution. A decompressive surgical treatment was performed, presenting a rapid recovery since the 7th day and full recovery after 25 postoperative days. Conclusion: The entrapment of the radial nerve in the arm is a rare pathology and its clinical presentation may vary. We consider that in the face of no remission or favorable evolution of paralysis within the first three months of conservative treatment, surgery should be performed. Level of Evidence: IV
Assuntos
Pessoa de Meia-Idade , Apraxias , Braço , Neuropatia RadialRESUMO
Objetivo: Comunicar los resultados funcionales y radiográficos de pacientes tratados con prótesis reversa por fracturas complejas. El objetivo secundario fue determinar la relación entre rangos de movilidad y puntaje ASES con la evolución radiográfica del troquíter. Materiales y Métodos: Se incluyeron 16 pacientes >65 años, tratados con prótesis reversa y reinserción del troquíter, entre 2013 y 2017, operados antes de las 4 semanas del trauma y con un seguimiento mínimo de 2 años. Se consignaron el puntaje ASES y el rango de movilidad activa. En las radiografías, se evaluaron la posición y la consolidación del troquíter, y se registraron las complicaciones y su tratamiento. Resultados: La media de la edad fue 74.5 años (RIC 66-78.5), 11 (69%) eran mujeres. Once fracturas (69%) eran a 4 fragmentos y 5, luxofracturas a 4 fragmentos. La media entre el trauma y la cirugía fue 9.4 días y el seguimiento, 29.5 meses. En 9 casos (56%), el troquíter presentó consolidación. Rotación interna: 5 pacientes alcanzaron la región glútea con el pulgar; 4, la vértebra T12; 4, la vértebra L3; 3, la T7. Las medianas de rotación externa y flexión anterior fueron 30° (RIC 17,5-40) y 100° (RIC 87,5-160). El puntaje ASES promedio fue 78,3 (RIC 63,3-87,4). No hubo una asociación estadísticamente significativa entre la evolución del troquíter y la flexión anterior y el puntaje (p = 0,24 y 0,52, respectivamente). Conclusión: La prótesis reversa en fracturas agudas con reinserción de las tuberosidades puede llevar a buenos resultados funcionales. No se encontró relación entre la consolidación del troquíter y el puntaje ASES. Nivel de Evidencia: IV
Objective: To report functional and radiologic outcomes of reverse shoulder arthroplasty (RSA) in patients with complex proximal humeral fractures. A second objective was to assess the relation between the greater tuberosity healing and the range of motion (ROM) and the American Shoulder and Elbow Surgeons (ASES) score. Materials and Methods: Sixteen patients treated between 2013 and 2017, older than 65 years old, operated before 4 weeks after the trauma, and with a minimum of 2-year follow-up were included. ASES scores and active ROMs were recorded. Greater tuberosity and the prosthesis position and healing were radiologically evaluated, and the complications and treatment were recorded. Results: The median age was of 74.5 years (IQR 66-78.5), 11 patients were females (69%). According to Neer classification, 11 cases were four-part fractures and 5 were four-part fracture-dislocations. The average time between trauma and surgery was 9.4 days, and the average follow-up was of 29.5 months. The greater tuberosity was healed in 9 cases (56%). Internal rotation: 5 patients (31.25%) were able to reach up with their thumbs to gluteal level, 4 (25%) to T12, 3 (18.75%) to T7, and 4 (25%) to L3. The medians for external rotation and forward flexion were 30° (IQR 17.5°-40°) and 100° (IQR 87.5°-160°). The average ASES score was of 78.3 (IQR 63.3-87.4). There was no significant statistical relation between greater tuberosity healing and forward flexion or ASES score (P=0.24 and P=0.52, respectively). Conclusion: The use of reverse prostheses for complex fractures with greater tuberosity reattachment could lead to good functional outcomes, low complication rates and reoperations. There was no significant statistical relation between ASES score and greater tuberosity healing or failure to heal. Level of Evidence: IV
Assuntos
Idoso , Fraturas do Ombro/cirurgia , Amplitude de Movimento Articular , Resultado do Tratamento , Artroplastia do Ombro , Úmero/lesõesRESUMO
Objetivo: Presentar una nueva técnica quirúrgica y los resultados clínicos comparativos de pacientes con roturas de tenorrafia primaria o lesiones crónicas del flexor profundo en las zonas I y II de Verdan, tratados con avance tendinoso no convencional. Ma-teriales y Métodos: Se incluyó a pacientes >18 años, con antecedente de lesiones en las zonas I y II de Verdan y un seguimiento mínimo de 36 meses. La serie estaba formada por 13 pacientes (edad promedio 29 años) que fueron divididos en dos grupos según el tipo de lesión (7 con roturas de tenorrafia primaria y 6 con lesiones crónicas del flexor profundo) y la técnica quirúrgica utilizada (alargamiento en Z más sutura término-terminal y sutura lateral del tendón alargado a un flexor vecino indemne en la zona V, respectivamente). Se empleó la Clasificación de Strickland para la evaluación. Resultados: El seguimiento promedio fue de 51 meses. El intervalo promedio entre la lesión y la cirugía fue de 11.7 semanas. Según la Clasificación de Strickland, 8 pa-cientes tuvieron resultados excelentes; 4, buenos y uno, pobre. El avance tendinoso promedio fue de 20,5 mm en ambos grupos.Conclusiones: El avance tendinoso no convencional para lesiones en las zonas I y II de Verdan, sea en roturas de tenorrafia primaria o lesiones crónicas del flexor profundo, resultó un tratamiento reproducible y eficaz. Nivel de Evidencia: III
Objective: To report a new surgical technique and the comparative clinical outcomes in patients with primary tenorrhaphy rupture or chronic flexor digitorum profundus (FDP) injuries in Verdan zones I and II, treated with non-conventional tendon advancement. Materials and Methods: The study population consisted of 13 patients over 18 years (averaging 29 years) who had had injuries in Verdan zones I and II and at least a 36-month follow-up. Patients were divided into two groups according to injury type (7 cases of primary tenorrhaphy rupture and 6 cases of chronic FDP injuries) and surgical technique (Z-plasty lengthening plus end-to-end suture or lateral suture to an adjacent undamaged FDP tendon at zone-V level, respectively). The results were evaluated according to Strickland's scoring system. Results: The average follow-up was 51 months. Time period between injury and surgery averaged 11.7 weeks (range, 2-24 weeks). Strickland scoring system results: excellent in 8 patients, good in 4 patients, and poor in 1 patient. Study average tendon advancement was 20.5mm. Conclusions: Non-conventional tendon advancement for primary tenorrhaphy ruptures or chronic FDP injuries in Verdan zones I and II proved to be a reproducible and effective treatment. Level of evidence: III
Assuntos
Traumatismos dos Tendões , MãosRESUMO
BACKGROUND: A poor ability of recommended risk scores for predicting in-hospital bleeding has been reported in elderly patients with acute coronary syndromes (ACS). No study assessed the prediction of post-discharge bleeding in the elderly. The new BleeMACS score (Bleeding complications in a Multicenter registry of patients discharged with diagnosis of Acute Coronary Syndrome), was designed to predict post-discharge bleeding in ACS patients. We aimed to assess the predictive ability of the BleeMACS score in elderly patients. METHODS: We assessed the incidence and characteristics of severe bleeding after discharge in ACS patients aged ≥ 75 years. Bleeding was defined as any intracranial bleeding or bleeding leading to hospitalization and/or red blood transfusion, occurring within the first year after discharge. We assessed the predictive ability of the BleeMACS score according to age by Fine-Gray proportional hazards regression analysis, calculating receiver-operating characteristic (ROC) curves and the area under the ROC curves (AUC). RESULTS: The BleeMACS registry included 15,401 patients of whom 3,376/15,401 (21.9%) were aged ≥ 75 years. Elderly patients were more commonly treated with clopidogrel and less often treated with ticagrelor or prasugrel. Of 3,376 elderly patients, 190 (5.6%) experienced post-discharge bleeding. The incidence of bleeding was moderately higher in elderly patients (hazard ratio [HR], 2.31, 95% confidence interval [CI], 1.92-2.77). The predictive ability of the BleeMACS score was moderately lower in elderly patients (AUC, 0.652 vs. 0.691, p = 0.001). CONCLUSION: Elderly patients with ACS had a significantly higher incidence of post-discharge bleeding. Despite a lower predictive ability in older patients, the BleeMACS score exhibited an acceptable performance in these patients.
Assuntos
Síndrome Coronariana Aguda/cirurgia , Clopidogrel/efeitos adversos , Técnicas de Apoio para a Decisão , Hemorragias Intracranianas/induzido quimicamente , Alta do Paciente , Intervenção Coronária Percutânea , Inibidores da Agregação Plaquetária/efeitos adversos , Síndrome Coronariana Aguda/diagnóstico , Fatores Etários , Idoso , Ásia/epidemiologia , Brasil/epidemiologia , Canadá/epidemiologia , Transfusão de Eritrócitos , Europa (Continente)/epidemiologia , Feminino , Humanos , Incidência , Hemorragias Intracranianas/diagnóstico , Hemorragias Intracranianas/epidemiologia , Hemorragias Intracranianas/terapia , Masculino , Pessoa de Meia-Idade , Readmissão do Paciente , Intervenção Coronária Percutânea/efeitos adversos , Cloridrato de Prasugrel/efeitos adversos , Valor Preditivo dos Testes , Sistema de Registros , Medição de Risco , Fatores de Risco , Ticagrelor/efeitos adversos , Fatores de Tempo , Resultado do TratamentoRESUMO
BACKGROUND: The prevalence and outcome of patients with cancer that experience acute coronary syndrome (ACS) have to be determined. METHODS AND RESULTS: The BleeMACS project is a multicentre observational registry enrolling patients with acute coronary syndrome undergoing percutaneous coronary intervention worldwide in 15 hospitals. The primary endpoint was a composite event of death and re-infarction after one year of follow-up. Bleedings were the secondary endpoint. 15,401 patients were enrolled, 926 (6.4%) in the cancer group and 14,475 (93.6%) in the group of patients without cancer. Patients with cancer were older (70.8±10.3 vs. 62.8±12.1 years, P<0.001) with more severe comorbidities and presented more frequently with non-ST-segment elevation myocardial infarction compared with patients without cancer. After one year, patients with cancer more often experienced the composite endpoint (15.2% vs. 5.3%, P<0.001) and bleedings (6.5% vs. 3%, P<0.001). At multiple regression analysis the presence of cancer was the strongest independent predictor for the primary endpoint (hazard ratio (HR) 2.1, 1.8-2.5, P<0.001) and bleedings (HR 1.5, 1.1-2.1, P=0.015). Despite patients with cancer generally being undertreated, beta-blockers (relative risk (RR) 0.6, 0.4-0.9, P=0.05), angiotensin-converting enzyme inhibitors/angiotensin receptor blockers (RR 0.5, 0.3-0.8, P=0.02), statins (RR 0.3, 0.2-0.5, P<0.001) and dual antiplatelet therapy (RR 0.5, 0.3-0.9, P=0.05) were shown to be protective factors, while proton pump inhibitors (RR 1, 0.6-1.5, P=0.9) were neutral. CONCLUSION: Cancer has a non-negligible prevalence in patients with acute coronary syndrome undergoing percutaneous coronary intervention, with a major risk of cardiovascular events and bleedings. Moreover, these patients are often undertreated from clinical despite medical therapy seems to be protective. Registration:The BleeMACS project (NCT02466854).
Assuntos
Síndrome Coronariana Aguda/epidemiologia , Neoplasias/epidemiologia , Intervenção Coronária Percutânea , Complicações Pós-Operatórias/epidemiologia , Sistema de Registros , Medição de Risco , Síndrome Coronariana Aguda/complicações , Síndrome Coronariana Aguda/cirurgia , Idoso , Ásia/epidemiologia , Europa (Continente)/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/complicações , América do Norte/epidemiologia , Prevalência , América do Sul/epidemiologia , Taxa de Sobrevida/tendências , Resultado do TratamentoRESUMO
Introducción: No existe consenso en la elección de los implantes pero, sin duda, la aparición de las placas conbloqueo angular fijo y luego variable o poliaxial constituye un hito en el tratamiento de las fracturas del húmero proximal. El objetivo de este estudio es la evaluación comparativa de los resultados funcionales obtenidos con dos tipos de bloqueo cefálico. Materiales y métodos: Se evaluaron, en forma retrospectiva, 29 pacientes divididos en dos grupos: uno comprendió 17 pacientes (bloqueo fijo Philos, AxSos, LCP) y otro, 12 pacientes (bloqueo poliaxial Numelock), que fueron clasificados según los criterios de Neer. Se usó el puntaje de Constant y radiografías a los 3 y los 6 meses. En todos los casos se usó abordaje deltopectoral. Resultados: En el grupo 1 hubo consolidación radiológica en 16 casos y 1 seudoartrosis que fue reoperada. El promedio de Constant fue de 77,37 puntos a los 6 meses. En el grupo 2 la consolidación fue de 100 por ciento, hubo 1 caso de desplazamiento al varo y 1 caso de necrosis aséptica que representó el puntaje más bajo (50). El promedio de Constant fue de 75 puntos. Conclusiones: En ambos grupos el principal progreso en el puntaje a los 6 meses fue a expensas de la movilidad. El resultado en los dos grupos se encuentra en el rango bueno, comparable con las casuísticas internacionales. No se encontraron diferencias clínicas significativas.(AU)
Assuntos
Humanos , Adulto , Fraturas do Ombro/cirurgia , Articulação do Ombro/cirurgia , Placas Ósseas , Estudos Retrospectivos , Resultado do Tratamento , Articulação do Ombro/diagnóstico por imagem , Fraturas do Ombro/classificaçãoRESUMO
Introducción: No existe consenso en la elección de los implantes pero, sin duda, la aparición de las placas conbloqueo angular fijo y luego variable o poliaxial constituye un hito en el tratamiento de las fracturas del húmero proximal. El objetivo de este estudio es la evaluación comparativa de los resultados funcionales obtenidos con dos tipos de bloqueo cefálico. Materiales y métodos: Se evaluaron, en forma retrospectiva, 29 pacientes divididos en dos grupos: uno comprendió 17 pacientes (bloqueo fijo Philos, AxSos, LCP) y otro, 12 pacientes (bloqueo poliaxial Numelock), que fueron clasificados según los criterios de Neer. Se usó el puntaje de Constant y radiografías a los 3 y los 6 meses. En todos los casos se usó abordaje deltopectoral. Resultados: En el grupo 1 hubo consolidación radiológica en 16 casos y 1 seudoartrosis que fue reoperada. El promedio de Constant fue de 77,37 puntos a los 6 meses. En el grupo 2 la consolidación fue de 100 por ciento, hubo 1 caso de desplazamiento al varo y 1 caso de necrosis aséptica que representó el puntaje más bajo (50). El promedio de Constant fue de 75 puntos. Conclusiones: En ambos grupos el principal progreso en el puntaje a los 6 meses fue a expensas de la movilidad. El resultado en los dos grupos se encuentra en el rango bueno, comparable con las casuísticas internacionales. No se encontraron diferencias clínicas significativas.
Assuntos
Humanos , Adulto , Articulação do Ombro/cirurgia , Fraturas do Ombro/cirurgia , Articulação do Ombro , Estudos Retrospectivos , Fraturas do Ombro/classificação , Placas Ósseas , Resultado do TratamentoRESUMO
We have studied the effect of the insertion of spin-labeled molecules n-doxyl-stearic acid (n-SASL, n = 5, 12, 16) on the structure and dynamics of a model lipid bilayer in gel-like phases using molecular dynamics simulations. We have studied the atomic density depth profiles and configurations of the labeled molecules in a host hydrated stearic acid bilayer system. We have found that the 5-SASL label positions its paramagnetic group at the water-lipid interface, and its polar head builds H bonds to neighboring lipids and to the solvent. 16-SASL positions its paramagnetic group at the lipid-lipid interface. The 12-SASL label presents two configurations at high lateral pressure. In one configuration, the doxyl ring lays at the lipid-lipid interface, shifting its polar head toward the bilayer center. The other equilibrium configuration of 12-SASL presents its paramagnetic group laying in the center of the compact hydrophobic region of the layer (erected configuration). It was determined that the coexistence of these two configurations is governed by the polar head-water interaction. We have found that the insertion of the labeled molecules at the concentrations used in the present work (0.36 mol %) do not perturb global properties like area per lipid, tilt angle, or order parameters. Nevertheless, there are local perturbations of the host system that are confined to a 10 angstroms neighboring shell around the spin label molecule. To study the interactions that determine the position of the labeled molecules in the bilayer, we performed simulations at different lateral pressures, which allowed us to extract important conclusions.
Assuntos
Bicamadas Lipídicas , Marcadores de Spin , Ácidos Esteáricos/química , Géis , Estrutura MolecularRESUMO
The effects of the insertion of a spin-labeled molecule (10-doxyl-stearic acid) on the structure and dynamics of model lipid bilayers in gel-like as well as in liquid-ordered-like phases are studied using molecular dynamic simulations. The perturbing effects of the labeled molecule on the structure of the bilayers are analyzed. We have also studied the relationship between the structural and dynamic properties of the bilayer phase and those of the labeled molecule. We found that the insertion of the labeled molecule in the bilayer at the concentration considered here (1:70) produces local and global perturbations in the gel-like phase. There is an increase of the area associated with the lipid molecules that produces a larger tilting angle of this condensed phase. In this gel-like phase, we also found that the z component of the order parameter of the labeled molecule associated with the electron paramagnetic resonance (EPR) spectra has the same temperature dependence as the axial correlation times of the lipid molecules. The mechanism by which the doxyl reorientation senses the dynamics of the layers is determined by the correlation between the gauche defect transitions of the labeled alkyl chain and its environment. For the liquid-ordered-like phase, we found that cholesterol molecules play the role of wedges that open free spaces in the lipid structure below the ring position and order the alkyl chains at the depths of the rings, leading to small inclination angles. The doxyl ring of the labeled molecule is located just below the cholesterol ring moiety, having fewer gauche defects than in the case of the gel-like phase. The change in depth of the doxyl ring causes a reorientation of this group that leads to an increase of the order parameter as the temperature rises.
Assuntos
Colesterol/química , Óxidos N-Cíclicos/química , Bicamadas Lipídicas/química , Ácidos Esteáricos/química , Simulação por Computador , Modelos Moleculares , Conformação Molecular , Marcadores de Spin , Água/químicaRESUMO
Se presentan 27 casos de mallet finger operados entre 1991 y 1995. Veinte de ellos no habían recibido tratamiento quirúrgico previo. Fueron tratados por un procedimiento simple con el que no hemos tenido recidivas. Describimos, además, la técnica que empleamos en siete casos inveterados que habían sido operados en una o más oportunidades
Assuntos
Argentina , MãosRESUMO
Se presentan 27 casos de mallet finger operados entre 1991 y 1995. Veinte de ellos no habían recibido tratamiento quirúrgico previo. Fueron tratados por un procedimiento simple con el que no hemos tenido recidivas. Describimos, además, la técnica que empleamos en siete casos inveterados que habían sido operados en una o más oportunidades