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2.
Rev. Costarric. psicol ; 40(2)dic. 2021.
Artigo em Espanhol | LILACS, SaludCR | ID: biblio-1387262

RESUMO

Resumen La utilización de pruebas psicológicas validadas y de confiabilidad comprobadas aportan evidencias valiosas al profesional en una multiplicidad de actividades, que comprenden desde el diagnóstico, el plan y el seguimiento de tratamiento hasta la selección laboral, la orientación vocacional, las pericias judiciales e investigación; sin embargo, no siempre se disponen de baremos adaptados a la región en la que se pretende aplicar un instrumento y, en ocasiones, siquiera corresponden a datos normativos del país. En Argentina, la WISC-IV fue adaptada considerando población perteneciente al Área Metropolitana de Buenos Aires, no se incluye muestra de otras regiones del extenso país. Por lo tanto, se realizó una investigación cuantitativa no experimental, exploratoria- descriptiva, con el objetivo de determinar la importancia de contar con baremos adaptados de la WISC-IV para diferentes regiones de un mismo país; para ello, se analizaron comparativamente los índices obtenidos por 520 escolares de 6 a 14 años (agrupados según los rangos de edad 6-8, 9-11 y 12-14) al aplicar dos baremos argentinos (Buenos Aires y Resistencia) para su corrección mediante un análisis de ANOVA mixto. Los resultados evidenciaron diferencias significativas según el baremo sea de Buenos Aires o de Resistencia, en 4 de los 5 índices de la WISC-IV, al abarcar tanto en aspectos relacionados con el funcionamiento cognitivo general (i.e., CIT), como en campos más específicos (i.e., memoria de trabajo - IMO, velocidad de procesamiento - IVP y razonamiento perceptivo - IRP); además, se observaron diferencias entre baremos en CIT, ICV, IRP e IMO y sugerir comportamientos diferentes en los distintos índices según el tipo de baremo aplicado en los diferentes grupos de edad. Los resultados sugieren que la corrección de la escala a un/a estudiante, según las normativas establecidas para una región con características sociodemográficas distintas a la que pertenece el individuo, podría derivar en errores interpretativos de sus aptitudes cognitivas, por lo que se determina la importancia de contar con adaptaciones de las pruebas psicológicas para arribar a interpretaciones que eviten infra o sobrevalorar sus puntuaciones.


Abstract: The use of validated psychological tests provides valuable evidence to specialists in a multiplicity of activities (e.g., predicting diagnoses, guiding treatment and follow-up plans, guiding the job selection process and vocational orientation, determining disability levels for medico-legal purposes). However, a psychological instrument is not generally adapted and standardized in the country in which it is intended to be applied. In Argentina, the Wechsler Intelligence Scale for Children-Fourth Edition (WISC-IV) was standardized in the Buenos Aires Metropolitan Area; but did not include samples from other regions across the vast country. This situation is problematic, because Argentina presents disparities in its socio-demographic characteristics across the territory, and an accurate interpre- tation of intelligence test performance, depends on the use of appropriately standardized data. Hence, a quantitative non-experimental, exploratory-descriptive study was conducted to determine the importance of obtaining locally standardized data for WISC-IV. To this end, the indices obtained from 520 children and adolescents aged 6 to 14 years (grouped according to age ranges 6-8, 9-11 and 12-14) were comparatively analyzed, by analyzing two Argentine adaptations (Buenos Aires and Resistencia). Results revealed differences between normative data from the country's different regions in four indices, which included both aspects of general cognitive functioning (i.e., CIT) and more specific processes (i.e., working memory - IMO, processing speed - IVP, and perceptual reasoning - IRP). These results suggest that the assessment of students, according to non-locally established standardized data, could lead to interpretative errors regarding their cognitive abilities. Thus, the study contributes to knowledge about the importance of using contextually appropriate standardized data in the implementation of intelligence tests, to arrive at evaluations that avoid over- or under- estimating students' abilities.


Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Padrões de Referência , Escalas de Wechsler , Avaliação Educacional , Argentina
3.
ESC Heart Fail ; 8(6): 5204-5212, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34514741

RESUMO

AIMS: There are conflicting data on the benefit of cardiac resynchronization therapy (CRT) in heart failure (HF) patients with permanent atrial fibrillation (AF). We aimed to compare patient outcomes according to the presence or absence of permanent AF at device implantation. METHODS AND RESULTS: We retrospectively analysed remote monitoring data from 1141 CRT defibrillators. Propensity score with inverse-probability weighting method was used to balance AF and sinus rhythm (SR) groups. Analysis endpoints included total mortality, appropriate defibrillation shocks, and CRT percentage. There were 229 patients (20.1%) in the AF group and 912 patients (79.9%) in the SR group. Compared with SR patients, AF patients were older (median age, 77 vs. 72 years, P < 0.001), more frequently male (82.5% vs. 75.5%, P = 0.02), and had higher heart rate (75.7 vs. 71.0 b.p.m., P < 0.001). Of the 229 AF patients, 162 (70.7%) received suboptimal CRT (<98%) and 67 (29.3%) had adequate CRT (≥98%). During a median follow-up of 24 months, total mortality did not differ between AF and SR groups (propensity-score-weighted hazard ratio, HR 1.32 [95% confidence interval, 0.82-2.15], P = 0.25). The risk of appropriate shocks was significantly higher in the AF group with <98% CRT than in the SR group (weighted-HR, 1.99 [1.21-3.26], P = 0.006) and was similar in the AF group with ≥98% CRT versus the SR group (1.29 [0.66-2.53], P = 0.45). During follow-up, sinus rhythm was recovered in 23 patients in the AF group (10%) after a median time of 106 (42-256) days. The rate of sinus rhythm recovery in the AF group was 4.5 (95% CI, 2.8-6.7) per 100 patient-years; the rate of permanent AF occurrence in the SR group was 2.5 (95% CI, 1.9-3.3) per 100 patient-years. CONCLUSIONS: Although mortality was similar across patient groups, patients with permanent AF and suboptimal CRT had twofold higher risk of appropriate shocks than SR patients or AF patients with CRT ≥ 98%.


Assuntos
Fibrilação Atrial , Terapia de Ressincronização Cardíaca , Idoso , Fibrilação Atrial/epidemiologia , Fibrilação Atrial/terapia , Terapia de Ressincronização Cardíaca/métodos , Desfibriladores , Humanos , Masculino , Estudos Retrospectivos , Resultado do Tratamento
4.
Interdisciplinaria ; 38(3): 83-102, jun. 2021. tab, graf
Artigo em Espanhol | LILACS-Express | LILACS | ID: biblio-1356329

RESUMO

Resumen El ámbito educativo, indudablemente, puede beneficiarse de los aportes de las neurociencias, enfoque desde el cual, en este estudio, se plantearon los objetivos de explorar la relación existente entre la capacidad intelectual (CI), las funciones ejecutivas (FE) y el rendimiento académico (RA) en adolescentes; analizar si la CI y las FE constituyen variables predictoras del RA, e indagar el funcionamiento ejecutivo de la muestra según la CI fuera baja o alta. Para ello, se realizó un estudio no experimental, transversal, descriptivo-correlacional, en el que participaron 36 sujetos de 13-14 años. Los instrumentos de evaluación utilizados fueron WISC-IV, BANFE-2 y el registro de calificaciones provisto por la institución educativa. Los datos obtenidos fueron sometidos a análisis descriptivos, de correlación y de regresión. Los resultados arrojaron correlación positiva significativa entre las variables CI y FE, y entre la prueba WISC-IV y RA, determinándose el CI como variable predictiva del RA, mientras que en relación con las FE evaluadas con BANFE-2 solo se encontró asociación estadísticamente significativa con las subpruebas correspondientes al área prefrontal anterior. Además se observaron diferencias significativas entre los grupos de alta y baja CI en las FE de planificación, memoria de trabajo (MT) y fluidez verbal, en favor del grupo de alto desempeño intelectual. La conclusión se centra en la importancia de involucrar los hallazgos sobre los diferentes procesos cognitivos en el diseño de las estrategias pedagógicas para favorecer al máximo el RA del estudiante. Finalmente, las limitaciones versaron en relación con el tamaño muestral, y se proponen futuras líneas de estudio que posibiliten generalizar los resultados.


Abstract The study of the relationship between executive functions, intellectual abilities, and academic achievement plays an important role in the field of cognitive neuroscience, particularly for their implications in the educational field. Intelligence has been linked to executive functions. In fact, core aspects of intelligence, such as reasoning, problem-solving and planning, are often included in the wide range of frontal lobe functions and over laps with the so-called higher-level executive functions. In addition, executive functioning abilities are associated with skills put in play when participating in learning activities at school. In this sense, different studies have demonstrated that a lower performance on executive function measures typically is associated with a lower performance across different academic areas. The aim of this work was focused on the relationship between intelligence and executive functions, and their ability to predict academic achievement. In the present study, a non-experimental, transversal and descriptive-correlational design was implemented to explore the relationship between intellectual abilities, executive functions, and academic performance in a sample of 37 adolescents (17 girls) aged 13-14 years. The information was collected through the school reports of participants, and their performance on tests BANFE-2 (neuropsychological battery of the executive functions and frontal lobes) and WISC-IV (Wechsler intelligence scale for children 4th Ed.). Also, adolescents were classified into two groups (high- and low performers) based on their performance on the general intelligence score. Participants with IQ scores below the median were assigned to the low-performing group, while participants with IQ scores above the median were assigned to the high-performing group. The data obtained were subjected to descriptive, correlation and regression analyses. Results showed a pattern of positive and significant correlations between executive functions variables (BANFE-2) and intelligence scores (WISC-IV). The 69 % of significant correlations indicated a moderate - strong degree of association (i. e., 49

5.
Pacing Clin Electrophysiol ; 44(6): 995-1003, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33908052

RESUMO

BACKGROUND: During the COVID-19 pandemic in-person visits for patients with cardiac implantable electronic devices should be replaced by remote monitoring (RM), in order to prevent viral transmission. A direct home-delivery service of the RM communicator has been implemented at 49 Italian arrhythmia centers. METHODS: According to individual patient preference or the organizational decision of the center, patients were assigned to the home-delivery group or the standard in-clinic delivery group. In the former case, patients received telephone training on the activation process and use of the communicator. In June 2020, the centers were asked to reply to an ad hoc questionnaire to describe and evaluate their experience in the previous 3 months. RESULTS: RM was activated in 1324 patients: 821 (62%) received the communicator at home and the communicator was activated remotely. Activation required one additional call in 49% of cases, and the median time needed to complete the activation process was 15 min [25th-75th percentile: 10-20]. 753 (92%) patients were able to complete the correct activation of the system. At the time when the questionnaire was completed, 743 (90%) communicators were regularly transmitting data. The service was generally deemed useful (96% of respondents) in facilitating the activation of RM during the COVID-19 pandemic and possibly beyond. CONCLUSIONS: Home delivery of the communicator proved to be a successful approach to system activation, and received positive feedback from clinicians. The increased use of a RM protocol will reduce risks for both providers and patients, while maintaining high-quality care.


Assuntos
Arritmias Cardíacas/terapia , COVID-19/epidemiologia , Desfibriladores Implantáveis , Serviços de Assistência Domiciliar , Distanciamento Físico , Pneumonia Viral/epidemiologia , Tecnologia de Sensoriamento Remoto/instrumentação , Feminino , Humanos , Incidência , Itália , Masculino , Marca-Passo Artificial , Pandemias , Pneumonia Viral/virologia , SARS-CoV-2
6.
Rev. iberoam. psicol. (En línea) ; 12(2): 13-23, 2019. graf, tab
Artigo em Espanhol | LILACS | ID: biblio-1253278

RESUMO

Durante el nivel inicial, los infantes suelen expresar respuestas emocionales y conductuales de manera impulsiva, dado que carecen de la aptitud de abstenerse de efectuar conductas que no son apropiadas a la ocasión. En este punto, las Habilidades Sociales (HHSS) y las Funciones Ejecutivas (FE) serán facilitadoras de la interacción asertiva con otros, favoreciendo en control comportamental y evitando los Problemas de Conducta (PC). El objetivo de este estudio fue analizar la correlación entre las FE, la percepción parental en relación a las emociones, y la percepción de docentes respecto de las HHSS y la conducta en niños del nivel inicial de educación; a tal fin se realizó un estudio cuantitativo, transversal, descriptivo-correlacional, en base a un diseño no experimental. La muestra estuvo integrada por 36 niños/as de nivel inicial, a quienes se les administró una batería neuropsicológica, además se incluyeron dos cuestionarios, uno para padres y otro para docentes. Se encontró correlación entre HHSS y PC; entre Planeación y Emociones; y entre Atención y Concentración y Memoria de Trabajo. Se concluye que los niños de nivel inicial poseen una conducta socialmente habilidosa, y manejan sus emociones sin dificultad indistintamente del sexo, según la perspectiva parental y docente


During the initial level, the children usually express emotional and behavioral responses impulsively, since they lack the ability to refrain from behaviors that are not appropriate to the occasion. At this point, Social Skills (SS) and Executive Functions (EF) will facilitate assertive interaction with others, favoring behavioral control and avoiding Conduct Problems (CP). The aim of this study was to analyze the correlation between EFs, parental perception in relation to emotions, and teachers' perception of SS and behavior in children at the initial level of education. To this end, a quantitative, cross-sectional, descriptive-correlational study was carried out, based on a non-experimental design. The sample consisted of 36 children of initial level, who were administered a neuropsychological battery, and two questionnaires were included, one for parents and another for teachers. Correlation was found between SS and CP; between Planning and Emotions; and between Attention and Concentration and Work Memory. It is concluded that the children of initial level have a socially skilled behavior, and handle their emotions without difficulty regardless of sex, according to the parental and teaching perspective


Assuntos
Humanos , Comportamento Social , Comportamento , Controle Comportamental , Função Executiva , Pais , Percepção , Aptidão , Atenção , Ensino , Inquéritos e Questionários , Educação , Emoções , Docentes , Habilidades Sociais , Memória
7.
New Microbiol ; 38(4): 565-70, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26485015

RESUMO

On November 25, 2014, an Italian physician infected by Ebola virus in Sierra Leone was admitted to the "Lazzaro Spallanzani" National Institute for Infectious Diseases in Rome, Italy. He was the first Italian case and was successfully cured in 38 days. The staff responsible for communication had a critical role ensuring that this challenging mission went smoothly. The Institutional Press Office working together with the press offices of the Ministry of Health was able to provide the high level of expertise necessary within both medical and communication contexts. Communication strategy, tools and procedures adopted before and after the arrival of the patient are summarized.


Assuntos
Comunicação , Doença pelo Vírus Ebola/psicologia , Adulto , Ebolavirus/genética , Ebolavirus/isolamento & purificação , Doença pelo Vírus Ebola/diagnóstico , Doença pelo Vírus Ebola/tratamento farmacológico , Doença pelo Vírus Ebola/virologia , Hospitais , Humanos , Itália , Masculino
8.
Mol Immunol ; 57(2): 317-22, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24240062

RESUMO

Generation of human recombinant antibody libraries displayed on the surface of the filamentous phage and selection of specific antibodies against desirable targets allows production of fully human antibodies usable for repeated administration in humans. Various lymphoid tissues from immunized donors, such as lymph nodes or peripheral blood lymphocytes from individuals with tumor or lymphocytes infiltrating tumor masses may serve as a source of specific anti-tumor antibody repertoire for generation of tumor-focused phage display libraries. In the case of lack of tumor-associated antigens in the purified form, high affinity anti-tumor antibodies can be isolated through library panning on whole cells expressing these antigens. However, affinity selection against cell surface specific antigens within highly heterogeneous population of molecules is not a very efficient process that often results in the selection of unspecific antibodies or antibodies against intracellular antigens that are generally useless for targeted immunotherapy. In this work, we developed a new cell-based antibody selection protocol that, by eliminating the contamination of dead cells from the cell suspension, dramatically improves the selection frequency of anti-tumor antibodies recognizing cell surface antigens.


Assuntos
Anticorpos Antineoplásicos/isolamento & purificação , Biblioteca de Peptídeos , Proteínas Recombinantes/isolamento & purificação , Anticorpos de Cadeia Única/imunologia , Anticorpos Anti-Idiotípicos/genética , Anticorpos Anti-Idiotípicos/imunologia , Anticorpos Antineoplásicos/genética , Anticorpos Antineoplásicos/imunologia , Antígenos de Neoplasias/imunologia , Antígenos de Superfície/imunologia , Linhagem Celular Tumoral , Feminino , Humanos , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/imunologia , Células MCF-7 , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia
9.
BMC Biotechnol ; 13: 79, 2013 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-24073829

RESUMO

BACKGROUND: Consistent progress in the development of bacteriophage lambda display platform as an alternative to filamentous phage display system was achieved in the recent years. The lambda phage has been engineered to display efficiently multiple copies of peptides or even large protein domains providing a powerful tool for screening libraries of peptides, proteins and cDNA. RESULTS: In the present work we describe an original method for dual display of large proteins on the surface of lambda particles. An anti-CEA single-chain antibody fragment and green fluorescent protein or alkaline phosphatase were simultaneously displayed by engineering both gpD and gpV lambda proteins. CONCLUSIONS: Here we show that such modified phage particles can be used for the detection of target molecules in vitro and in vivo. Dual expression of functional moieties on the surface of the lambda phage might open the way to generation of a new class of diagnostic and therapeutic targeted nanoparticles.


Assuntos
Bacteriófago lambda/genética , Proteínas/química , Proteínas/genética , Fosfatase Alcalina/metabolismo , Animais , Proteínas do Capsídeo/química , Proteínas do Capsídeo/genética , Linhagem Celular , Clonagem Molecular , DNA Complementar/química , DNA Complementar/genética , Escherichia coli/genética , Vetores Genéticos , Glicoproteínas/química , Glicoproteínas/genética , Proteínas de Fluorescência Verde/genética , Células HT29 , Humanos , Camundongos , Camundongos Nus , Microscopia Confocal , Nanopartículas/química , Biblioteca de Peptídeos , Plasmídeos/genética , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
10.
J Cardiothorac Vasc Anesth ; 26(4): 604-7, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22445181

RESUMO

OBJECTIVE: An intra-aortic balloon pump (IABP) is used routinely in high-risk patients undergoing cardiac surgery to prevent or treat low-cardiac-output syndrome and to reduce perioperative mortality. The insertion and management of IABP carry the risk of major vascular complications. The authors reviewed their database to ascertain the incidence of IABP-related severe complications. DESIGN: A retrospective study. SETTING: A teaching hospital. PARTICIPANTS: Ten thousand three hundred sixty-five patients scheduled for elective or emergency cardiac surgery over a 12-year period at a single center. INTERVENTIONS: Four hundred twenty-three patients received an IABP perioperatively. Careful preoperative screening for peripheral arterial disease, strict postoperative control, and the sheathless insertion technique to spare the arterial flow to the lower limb were performed routinely. MEASUREMENTS AND MAIN RESULTS: The use of a perioperative IABP was 0.7% at the beginning of the observation period in 1999 and 7.3% in 2010, showing a fluctuating trend. Two patients (0.47%) died of direct complications, arterial wall damage and bleeding. Immediate surgical exploration and control of bleeding were followed by multiple-organ failure and death. Vascular complications, leading to lower-limb ischemia, occurred in 4 of 423 patients (0.94%). All of them underwent urgent vascular surgery and survived. Local sepsis occurred in 2 other patients (0.47%). CONCLUSIONS: These data indicate that an IABP is a valuable option in high-risk patients undergoing cardiac surgery even if not devoid of intrinsic risks for vascular complications (0.94%), septic complications (0.47%) and mortality (0.47%).


Assuntos
Procedimentos Cirúrgicos Cardíacos , Balão Intra-Aórtico/efeitos adversos , Complicações Pós-Operatórias/epidemiologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco
11.
Cancer Detect Prev ; 30(3): 248-56, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16876336

RESUMO

OBJECTIVE: In this article we provide evidence of a significant spontaneous humoral response in cancer patients. METHODS: A panel of tumor-associated antigens, previously identified through serological screening of phage-displayed cDNA libraries from solid human tumors, breast carcinoma cell lines and human testis by employing breast cancer patient sera, was used in this study to survey sera from 182 patients with known disease histories and clinical stages. RESULTS: This analysis reveals a statistically significant association between tumor disease and presence in peripheral blood of IgG antibodies against four autoantigens. One of these antigens (D7-1) is particularly interesting in that the antibody response against it grows with cancer progression from stages I through IV, with an incidence of 13.2, 13.5, 18.2 and 27%, respectively. The significance of this stage-dependent increase in the incidence is confirmed by the Mantel-Haenszel Chi-squared test (P=0.001). CONCLUSIONS: Our data confirm association between breast cancer diagnosis of patients and presence in their peripheral blood of antibodies against several autoantigens identified by phage display.


Assuntos
Antígenos de Neoplasias/biossíntese , Neoplasias da Mama/sangue , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Neoplasias/sangue , Adulto , Formação de Anticorpos , Antígenos de Neoplasias/metabolismo , Autoantígenos/química , Neoplasias da Mama/imunologia , Clonagem Molecular , DNA Complementar/metabolismo , Feminino , Humanos , Imunoglobulina G/química , Masculino , Pessoa de Meia-Idade , Biblioteca de Peptídeos , Plasmídeos/metabolismo
12.
BMC Cancer ; 6: 41, 2006 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-16504122

RESUMO

BACKGROUND: CEA is a tumor-associated antigen abundantly expressed on several cancer types, including those naturally refractory to chemotherapy. The selection and characterization of human anti-CEA single-chain antibody fragments (scFv) is a first step toward the construction of new anticancer monoclonal antibodies designed for optimal blood clearance and tumor penetration. METHODS: The human MA39 scFv, selected for its ability to recognize a CEA epitope expressed on human colon carcinomas, was first isolated from a large semi-synthetic ETH-2 antibody phage library, panned on human purified CEA protein. Subsequently, by in vitro mutagenesis of a gene encoding for the scFv MA39, a new library was established, and new scFv antibodies with improved affinity towards the CEA cognate epitope were selected and characterized. RESULTS: The scFv MA39 antibody was affinity-maturated by in vitro mutagenesis and the new scFv clone, E8, was isolated, typed for CEA family member recognition and its CEACAM1, 3 and 5 shared epitope characterized for expression in a large panel of human normal and tumor tissues and cells. CONCLUSION: The binding affinity of the scFv E8 is in a range for efficient, in vivo, antigen capture in tumor cells expressing a shared epitope of the CEACAM1, 3 and 5 proteins. This new immunoreagent meets all criteria for a potential anticancer compound: it is human, hence poorly or not at all immunogenic, and it binds selectively and with good affinity to the CEA epitope expressed by metastatic melanoma and colon and lung carcinomas. Furthermore, its small molecular size should provide for efficient tissue penetration, yet give rapid plasma clearance.


Assuntos
Antígeno Carcinoembrionário/química , Região Variável de Imunoglobulina/química , Sequência de Aminoácidos , Anticorpos Monoclonais/química , Especificidade de Anticorpos , Antineoplásicos/farmacologia , Bactérias/metabolismo , Biotinilação , Western Blotting , Linhagem Celular Tumoral , Neoplasias do Colo/metabolismo , DNA/química , Ensaio de Imunoadsorção Enzimática , Epitopos/química , Citometria de Fluxo , Biblioteca Gênica , Humanos , Fragmentos de Imunoglobulinas/química , Imuno-Histoquímica , Cinética , Neoplasias Pulmonares/metabolismo , Melanoma/metabolismo , Microscopia Confocal , Dados de Sequência Molecular , Mutagênese , Metástase Neoplásica , Biblioteca de Peptídeos , Estrutura Terciária de Proteína , Sensibilidade e Especificidade , Ressonância de Plasmônio de Superfície , Transfecção
13.
J Immunol Methods ; 310(1-2): 149-58, 2006 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-16497320

RESUMO

In the present work we demonstrate the efficient display of functional scFv antibodies on the bacteriophage lambda capsid. A single-chain (scFv) anti-CEA antibody gene was cloned in two different vectors to obtain fusion of the scFv antibody to the N- or C-terminus of the bacteriophage lambda capsid protein D (gpD). Lambda bacteriophage assembly occurs in the reducing environment of the cytoplasm; despite this the lambda-displayed anti-CEA antibody fragments retain the capacity to recognize the antigen, indicating correct single-chain antibody folding. Efficient production of functional scFv exposed on lambda capsid with viable antigen binding specificity allowed us to study and compare the capacity of display, the stability of recombinant antibody expression and the assembly efficiency of bacteriophage particles decorated with recombinant antibody fused to the amino- or carboxy-terminus of lambda D protein.


Assuntos
Bacteriófago lambda/imunologia , Fragmentos de Imunoglobulinas/imunologia , Western Blotting , Proteínas do Capsídeo/imunologia , Antígeno Carcinoembrionário/imunologia , Cromatografia de Afinidade , Humanos , Região Variável de Imunoglobulina/imunologia , Oligopeptídeos , Biblioteca de Peptídeos , Peptídeos/imunologia , Proteínas Recombinantes/imunologia
14.
BMC Cancer ; 4: 78, 2004 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-15541172

RESUMO

BACKGROUND: Tumor-associated antigens recognized by humoral effectors of the immune system are a very attractive target for human cancer diagnostics and therapy. Recent advances in molecular techniques have led to molecular definition of immunogenic tumor proteins based on their reactivity with autologous patient sera (SEREX). METHODS: Several high complexity phage-displayed cDNA libraries from breast carcinomas, human testis and breast carcinoma cell lines MCF-7, MDA-MB-468 were constructed. The cDNAs were expressed in the libraries as fusion to bacteriophage lambda protein D. Lambda-displayed libraries were efficiently screened with sera from patients with breast cancer. RESULTS: A panel of 21 clones representing 18 different antigens, including eight proteins of unknown function, was identified. Three of these antigens (T7-1, T11-3 and T11-9) were found to be overexpressed in tumors as compared to normal breast. A serological analysis of the 21 different antigens revealed a strong cancer-related profile for at least five clones (T6-2, T6-7, T7-1, T9-21 and T9-27). CONCLUSIONS: Preliminary results indicate that patient serum reactivity against five of the antigens is associated with tumor disease. The novel T7-1 antigen, which is overexpressed in breast tumors and recognized specifically by breast cancer patient sera, is potentially useful in cancer diagnosis.


Assuntos
Antígenos Glicosídicos Associados a Tumores/genética , Bacteriófago lambda/genética , Neoplasias da Mama/genética , Biblioteca Gênica , Neoplasias/genética , Testículo/química , Testículo/metabolismo , Sequência de Aminoácidos , Antígenos Glicosídicos Associados a Tumores/sangue , Antígenos Glicosídicos Associados a Tumores/química , Antígenos Glicosídicos Associados a Tumores/imunologia , Neoplasias da Mama/sangue , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica/genética , Vetores Genéticos/genética , Humanos , Masculino , Análise de Sequência de DNA/métodos , Testes Sorológicos/métodos
15.
J Biol Chem ; 279(42): 43760-8, 2004 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-15265865

RESUMO

Synapsins, a family of neuron-specific phosphoproteins, have been demonstrated to regulate the availability of synaptic vesicles for exocytosis by binding to both synaptic vesicles and the actin cytoskeleton in a phosphorylation-dependent manner. Although the above-mentioned observations strongly support a pre-docking role of the synapsins in the assembly and maintenance of a reserve pool of synaptic vesicles, recent results suggest that the synapsins may also be involved in some later step of exocytosis. In order to investigate additional interactions of the synapsins with nerve terminal proteins, we have employed phage display library technology to select peptide sequences binding with high affinity to synapsin I. Antibodies raised against the peptide YQYIETSMQ (syn21) specifically recognized Rab3A, a synaptic vesicle-specific small G protein implicated in multiple steps of exocytosis. The interaction between synapsin I and Rab3A was confirmed by photoaffinity labeling experiments on purified synaptic vesicles and by the formation of a chemically cross-linked complex between synapsin I and Rab3A in intact nerve terminals. Synapsin I could be effectively co-precipitated from synaptosomal extracts by immobilized recombinant Rab3A in a GTP-dependent fashion. In vitro binding assays using purified proteins confirmed the binding preference of synapsin I for Rab3A-GTP and revealed that the COOH-terminal regions of synapsin I and the Rab3A effector domain are required for the interaction with Rab3A to occur. The data indicate that synapsin I is a novel Rab3 interactor on synaptic vesicles and suggest that the synapsin-Rab3 interaction may participate in the regulation of synaptic vesicle trafficking within the nerve terminals.


Assuntos
Sinapsinas/metabolismo , Vesículas Sinápticas/fisiologia , Proteínas rab3 de Ligação ao GTP/metabolismo , Sequência de Aminoácidos , Animais , Química Encefálica , Bovinos , Clonagem Molecular , Escherichia coli , Biblioteca Gênica , Terminações Nervosas/fisiologia , Fragmentos de Peptídeos/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Sinapsinas/isolamento & purificação , Proteínas rab3 de Ligação ao GTP/isolamento & purificação
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