RESUMO
Packing biomolecules inside virus capsids has opened new avenues for the study of molecular function in confined environments. These systems not only mimic the highly crowded conditions in nature, but also allow their manipulation at the nanoscale for technological applications. Here, green fluorescent proteins are packed in virus-like particles derived from P22 bacteriophage procapsids. The authors explore individual virus cages to monitor their emission signal with total internal reflection fluorescence microscopy while simultaneously changing the microenvironment with the stylus of atomic force microscopy. The mechanical and electronic quenching can be decoupled by ≈10% each using insulator and conductive tips, respectively. While with conductive tips the fluorescence quenches and recovers regardless of the structural integrity of the capsid, with the insulator tips quenching only occurs if the green fluorescent proteins remain organized inside the capsid. The electronic quenching is associated with the coupling of the protein fluorescence emission with the tip surface plasmon resonance. In turn, the mechanical quenching is a consequence of the unfolding of the aggregated proteins during the mechanical disruption of the capsid.
Assuntos
Imagem Individual de Molécula , Proteínas Virais , Capsídeo/química , Proteínas do Capsídeo/química , Proteínas de Fluorescência Verde , Microscopia de Força Atômica , Proteínas Virais/químicaRESUMO
Microscopes are used to characterize small objects with the help of probes that interact with the specimen, such as photons and electrons in optical and electron microscopies, respectively. In atomic force microscopy (AFM), the probe is a nanometric tip located at the end of a microcantilever which palpates the specimen under study just as a blind person manages a walking stick. In this way, AFM allows obtaining nanometric resolution images of individual protein shells, such as viruses, in a liquid milieu. Beyond imaging, AFM also enables not only the manipulation of single protein cages, but also the characterization of every physicochemical property capable of inducing any measurable mechanical perturbation to the microcantilever that holds the tip. In the present revision, we start revising some recipes for adsorbing protein shells on surfaces. Then, we describe several AFM approaches to study individual protein cages, ranging from imaging to spectroscopic methodologies devoted to extracting physical information, such as mechanical and electrostatic properties. We also explain how a convenient combination of AFM and fluorescence methodologies entails monitoring genome release from individual viral shells during mechanical unpacking.
Assuntos
Proteínas Virais/metabolismo , Vírus/ultraestrutura , Adsorção , Fenômenos Biomecânicos , Genoma Viral , Microscopia de Força Atômica/métodos , Vírus/genéticaRESUMO
In clinical practice, it is not rare to encounter situations in which parents and families are asked to leave the child alone with the health care team in rooms full of devices throughout the performance of procedures, which at times may give rise not only to conflicts but, more importantly, emotional sequelae in children or adolescents. We conducted a narrative review of the literature by searching the digital library of the public health care system of Andalusia for articles concerning the experiences of health care professionals and families with the accompaniment of paediatric patients during health care procedures. We restricted the search to studies published in Spanish or English and conducted in humans. The review evinced the need to humanise care in order to improve care quality. The need to accompany minors is supported by the evidence from works that have analysed the factors involved in the persistence of these behaviours and attitudes in both professionals and parents. We consider it necessary to develop institutional policies and appoint mediators to compile the statements of different national and international societies, taking into account legal aspects but, above all, the pertinent values from a health care ethics perspective, and in pursuit of the best interests of the child.
Assuntos
Atenção à Saúde , Pais , Adolescente , Humanos , Criança , Pais/psicologia , Qualidade da Assistência à SaúdeAssuntos
Transtornos de Deglutição/etiologia , Esôfago/anormalidades , Esôfago/diagnóstico por imagem , Pré-Escolar , Meios de Contraste , Transtornos de Deglutição/complicações , Transtornos de Deglutição/diagnóstico , Transtornos de Deglutição/diagnóstico por imagem , Diagnóstico Diferencial , Acalasia Esofágica/diagnóstico , Junção Esofagogástrica/diagnóstico por imagem , Feminino , Gastroscopia , Humanos , Músculo Liso/anormalidades , Músculo Liso/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Vômito/etiologiaRESUMO
The organization of virus-like particles (VLPs) on surfaces is a relevant matter for both fundamental and biomedical sciences. In this work, the authors have tailored surfaces with different surface tension components aiming at finding a relationship with the affinity of the different geometric/surface features of icosahedral P22 VLPs. The surfaces have been prepared by titanate assisted organosilanization with glycidyloxy, amino, and perfluoro silanes. Vibrational and photoelectron spectroscopies have allowed identifying the different functional groups of the organosilanes on the surfaces. Atomic force microscopy (AFM) showed that, irrespective of the organosilane used, the final root mean square roughness remains below 1 nm. Contact angle analyses confirm the effective formation of a set of surface chemistries exhibiting different balance among surface tension components. The study of the adsorption of P22 VLPs has involved the analysis of the dynamics of virus immobilization by fluorescence microscopy and the interpretation of the final VLP orientation by AFM. These analyses give rise to statistical distributions pointing to a higher affinity of VLPs toward perfluorinated surfaces, with a dominant fivefold conformation on this hydrophobic surface, but threefold and twofold symmetries dominating on hydrophilic surfaces. These results can be explained in terms of a reinforced hydrophobic interaction between the perfluorinated surface and the dominating hydrophobic residues present at the P22 pentons.
Assuntos
Adsorção , Bacteriófago P22/metabolismo , Silanos/metabolismo , Virossomos/metabolismo , Microscopia de Força Atômica , Análise EspectralRESUMO
Microscopes are used to characterize small objects with the help of probes that interact with the specimen, such as photons and electrons in optical and electron microscopies, respectively. In atomic force microscopy (AFM) the probe is a nanometric tip located at the end of a micro cantilever which palpates the specimen under study as a blind person manages a walking stick. In this way AFM allows obtaining nanometric resolution images of individual protein shells, such as viruses, in liquid milieu. Beyond imaging, AFM also enables not only the manipulation of single protein cages, but also the characterization of every physicochemical property able of inducing any measurable mechanical perturbation to the microcantilever that holds the tip. In this chapter we start revising some recipes for adsorbing protein shells on surfaces. Then we describe several AFM approaches to study individual protein cages, ranging from imaging to spectroscopic methodologies devoted for extracting physical information, such as mechanical and electrostatic properties. We also explain how a convenient combination of AFM and fluorescence methodologies entails monitoring genome release from individual viral shells during mechanical unpacking.
Assuntos
Microscopia de Força Atômica , Vírus/ultraestrutura , Capsídeo , Análise de Dados , Humanos , Processamento de Imagem Assistida por Computador/métodos , Fenômenos Mecânicos , Microscopia de Força Atômica/métodos , Eletricidade Estática , Proteínas Virais/químicaRESUMO
En la práctica clínica no es infrecuente observar situaciones en las cuales se invita a los progenitores y la familia a dejar a los menores en soledad junto al equipo asistencial en estancias repletas de tecnología durante la realización de procedimientos, dando lugar en ocasiones a conflictos, pero sobre todo con consecuencias emocionales en los niños o adolescentes.Se ha realizado una revisión narrativa de la literatura mediante búsqueda bibliográfica en la biblioteca virtual del sistema sanitario público de Andalucía, siendo los criterios de inclusión utilizados, estudios que conciernen a las experiencias de profesionales sanitarios y familiares sobre el acompañamiento de la población pediátrica en los procedimientos asistenciales. El resultado de la búsqueda se limitó a trabajos en humanos en español e inglés.Esta revisión pone de manifiesto la necesidad de humanizar la asistencia sanitaria para mejorar la calidad de la atención. Se justifica la necesidad de acompañamiento de los menores, a través de trabajos que han analizado los factores que intervienen en la permanencia de estas conductas y actitudes tanto por profesionales como padres. Se recomienda la necesidad de políticas institucionales y figuras mediadoras que recojan las declaraciones de algunas sociedades nacionales e internacionales teniendo en cuenta aspectos legales, pero sobre todo los valores en juego desde una ética del cuidado y búsqueda del interés superior del menor.(AU)
In clinical practice, it is not rare to encounter situations in which parents and families are asked to leave the child alone with the health care team in rooms full of devices throughout the performance of procedures, which at times may give rise not only to conflicts but, more importantly, emotional sequelae in children or adolescents.We conducted a narrative review of the literature by searching the digital library of the public health care system of Andalusia for articles concerning the experiences of health care professionals and families with the accompaniment of paediatric patients during health care procedures. We restricted the search to studies published in Spanish or English and conducted in humans.The review evinced the need to humanise care in order to improve care quality. The need to accompany minors is supported by the evidence from works that have analysed the factors involved in the persistence of these behaviours and attitudes in both professionals and parents. We consider it necessary to develop institutional policies and appoint mediators to compile the statements of different national and international societies, taking into account legal aspects but, above all, the pertinent values from a health care ethics perspective, and in pursuit of the best interests of the child.(AU)
Assuntos
Humanos , Masculino , Feminino , Criança , Serviço de Acompanhamento de Pacientes , Preceptoria , Qualidade da Assistência à Saúde , Assistência ao Paciente , Reanimação Cardiopulmonar , Ética em Enfermagem , Espanha , Pediatria , Enfermagem Pediátrica , Família , Sistemas de SaúdeRESUMO
Ureterocele is a relatively common malformation of the urinary system in children; however, the ureterocele prolapse through the urethra is an extremely rare presentation. Urological nuclear magnetic resonance is the test of choice for definitive diagnosis. Initial treatment of a prolapsed ureterocele involves decompression of the mass. We report the case of a 7-month-old infant with duplication of the collecting system and right ectopic ureterocele appearing as a vulvar mass.
El ureterocele es una malformación del sistema urinario relativamente frecuente en niños; sin embargo, el prolapso del ureterocele a través de la uretra es una presentación, en extremo rara. La urorresonancia magnética es la prueba de elección para el diagnóstico definitivo. El tratamiento inicial de un ureterocele prolapsado consiste en la descompresión de la masa. Presentamos el caso clínico de una lactante de 7 meses con duplicación del sistema colector y ureterocele ectópico derecho, que apareció como una masa vulvar.
Assuntos
Ureterocele/diagnóstico , Feminino , Humanos , Lactente , Prolapso , Ureterocele/complicações , VulvaRESUMO
RATIONALE: Attention deficit with hyperactivity disorder is a neurodevelopmental disorder associated with alterations in the prefrontal cortex via dopaminergic and noradrenergic neurotransmission. Neurosteroids (e.g. allopregnanolone and dehydroepiandrosterone) modulate the release of multiple neurotransmitters. OBJECTIVE: This study aims to determine the baseline concentrations and daily variations in allopregnanolone and dehydroepiandrosterone in children with attention deficit hyperactivity disorder (ADHD) and to determine the effect of chronic administration of methylphenidate on clinical symptoms and on the concentrations of these two neurosteroids. METHODS: We included 148 children aged 5 to 14 years, subdivided into two groups: ADHD group (n = 107, with a diagnosis of ADHD (DSM-IV-TR criteria), further classified in subtypes by an "attention deficit and hyperactivity scale" and subgroups by the "Children's Depression Inventory") and a control group (n = 41). The clinical workup included blood samples that were drawn at 20:00 and 09:00 hours, at inclusion in both groups, and after 4.61 ± 2.29 months of treatment only in the ADHD group, for measurements for allopregnanolone and dehydroepiandrosterone. Factorial analysis, adjusted for age and gender, was performed by using Stata 12.0. RESULTS: Methylphenidate induced the doubling of allopregnanolone levels in the predominantly inattentive ADHD patients without depressive symptoms (27.26 ± 12.90 vs. 12.67 ± 6.22 ng/ml, morning values). Although without statistical differences, baseline dehydroepiandrosterone levels were higher and slightly increased after methylphenidate in the ADHD subtype with depressive symptoms (7.74 ± 11.46 vs. 6.18 ± 5.99 ng/ml, in the morning), opposite to the lower baseline levels, and further decrease after methylphenidate in the inattentive subtype with depressive symptoms. CONCLUSIONS: Different neurosteroids may have different baseline concentrations and differential responses to methylphenidate treatment as a function of ADHD subtype and subgroup. These differential responses may be a clinical marker of ADHD subtype and/or co-morbidities.
Assuntos
Transtorno do Deficit de Atenção com Hiperatividade/tratamento farmacológico , Estimulantes do Sistema Nervoso Central/uso terapêutico , Transtorno Depressivo/psicologia , Metilfenidato/uso terapêutico , Neurotransmissores/sangue , Adolescente , Atenção/efeitos dos fármacos , Transtorno do Deficit de Atenção com Hiperatividade/complicações , Transtorno do Deficit de Atenção com Hiperatividade/psicologia , Biomarcadores/sangue , Criança , Pré-Escolar , Desidroepiandrosterona/sangue , Transtorno Depressivo/complicações , Manual Diagnóstico e Estatístico de Transtornos Mentais , Feminino , Humanos , Masculino , Pregnanolona/sangueRESUMO
El ureterocele es una malformación del sistema urinario relativamente frecuente en niños; sin embargo, el prolapso del ureterocele a través de la uretra es una presentación, en extremo rara. La urorresonancia magnética es la prueba de elección para el diagnóstico definitivo. El tratamiento inicial de un ureterocele prolapsado consiste en la descompresión de la masa. Presentamos el caso clínico de una lactante de 7 meses con duplicación del sistema colector y ureterocele ectópico derecho, que apareció como una masa vulvar.
Ureterocele is a relatively common malformation of the urinary system in children; however, the ureterocele prolapse through the urethra is an extremely rare presentation. Urological nuclear magnetic resonance is the test of choice for definitive diagnosis. Initial treatment of a prolapsed ureterocele involves decompression of the mass. We report the case of a 7-month-old infant with duplication of the collecting system and right ectopic ureterocele appearing as a vulvar mass.
Assuntos
Humanos , Feminino , Lactente , Ureterocele/diagnóstico , Prolapso , Ureterocele/complicações , VulvaRESUMO
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