Is professional recognition in plastic surgery related to activity in research

OBJECTIVE: To evaluate the relation of medical research, with the participation of prominent plastic surgeon in Congress.METHODS: We reviewed the scientific programs of the last 3 Brazilian Congress of Surgery, were selected 21 Brazilian plástic surgeons invited to serve as panelists or speakers in roundtable sessions in the last 3 congresses (Group 1). We randomly selected and paired by other members (associates) of the Brazilian Society of Plastic Surgery, with no participation in congress as speaker (Group 2). We conducted a search for articles published in journals indexed in Medline, Lilacs and SciELO for all doctors selected during the entire academic career and the last 5 years from March 2007 until March 2012. We assessed the research activity through the simple counting of the number of publications in indexed journals for each professional. The number of publications groups was compared.RESULTS: articles produced throughout career: Group 1- 639 articles (average of 30.42 items each). Group 2- 79 articles (mean 3.95 articles each). Difference between medias: p <0.001.CONCLUSION: The results demonstrate that the Brazilian Society of Plastic Surgery seeking professionals with a greater number of publications and journals of higher impact. This approach encourages new members to pursue a higher qualification, and give security to congressmen, they can rely on the existence of a technical criterion in the choice of speakers.

Evidence for the expression of native Mycobacterium tuberculosis phospholipase C: recognition by immune sera and detection of promoter activity

The genome of Mycobacterium tuberculosis H37Rv contains three contiguous genes (plc-a, plc-b and plc-c) which are similar to the Pseudomonas aeruginosa phospholipase C (PLC) genes. Expression of mycobacterial PLC-a and PLC-b in E. coli and M. smegmatis has been reported, whereas expression of the native proteins in M. tuberculosis H37Rv has not been demonstrated. The objective of the present study was to demonstrate that native PLC-a is expressed in M. tuberculosis H37Rv. Sera from mice immunized with recombinant PLC-a expressed in E. coli were used in immunoblots to evaluate PLC-a expression. The immune serum recognized a 49-kDa protein in immunoblots against M. tuberculosis extracts. No bands were visible in M. tuberculosis culture supernatants or extracts from M. avium, M. bovis and M. smegmatis. A 550-bp DNA fragment upstream of plc-a was cloned in the pJEM12 vector and the existence of a functional promoter was evaluated by detection of ß-galactosidase activity. ß-Galactosidase activity was detected in M. smegmatis transformed with recombinant pJEM12 grown in vitro and inside macrophages. The putative promoter was active both in vitro and in vivo, suggesting that expression is constitutive. In conclusion, expression of non-secreted native PLC-a was demonstrated in M. tuberculosis.

Application and comparison of classification algorithms for recognition of Alzheimer's disease in electrical brain activity (EEG)

The early detection of subjects with probable Alzheimer's disease (AD) is crucial for effective appliance of treatment strategies. Here we explored the ability of a multitude of linear and non-linear classification algorithms to discriminate between the electroencephalograms (EEGs) of patients with varying degree of AD and their age-matched control subjects. Absolute and relative spectral power, distribution of spectral power, and measures of spatial synchronization were calculated from recordings of resting eyes-closed continuous EEGs of 45 healthy controls, 116 patients with mild AD and 81 patients with moderate AD, recruited in two different centers (Stockholm, New York). The applied classification algorithms were: principal component linear discriminant analysis (PC LDA), partial least squares LDA (PLS LDA), principal component logistic regression (PC LR), partial least squares logistic regression (PLS LR), bagging, random forest, support vector machines (SVM) and feed-forward neural network. Based on 10-fold cross-validation runs it could be demonstrated that even tough modern computer-intensive classification algorithms such as random forests, SVM and neural networks show a slight superiority, more classical classification algorithms performed nearly equally well. Using random forests classification a considerable sensitivity of up to 85% and a specificity of 78%, respectively for the test of even only mild AD patients has been reached, whereas for the comparison of moderate AD vs. controls, using SVM and neural networks, values of 89% and 88% for sensitivity and specificity were achieved. Such a remarkable performance proves the value of these classification algorithms for clinical diagnostics.

Prestimulus default mode activity influences depth of processing and recognition in an emotional memory task

Low self-referential thoughts are associated with better concentration, which leads to deeper encoding and increases learning and subsequent retrieval. There is evidence that being engaged in externally rather than internally focused tasks is related to low neural activity in the default mode network (DMN) promoting open mind and the deep elaboration of new information. Thus, reduced DMN activity should lead to enhanced concentration, comprehensive stimulus evaluation including emotional categorization, deeper stimulus processing, and better long-term retention over one whole week. In this fMRI study, we investigated brain activation preceding and during incidental encoding of emotional pictures and on subsequent recognition performance. During fMRI, 24 subjects were exposed to 80 pictures of different emotional valence and subsequently asked to complete an online recognition task one week later. Results indicate that neural activity within the medial temporal lobes during encoding predicts subsequent memory performance. Moreover, a low activity of the default mode network preceding incidental encoding leads to slightly better recognition performance independent of the emotional perception of a picture. The findings indicate that the suppression of internally-oriented thoughts leads to a more comprehensive and thorough evaluation of a stimulus and its emotional valence. Reduced activation of the DMN prior to stimulus onset is associated with deeper encoding and enhanced consolidation and retrieval performance even one week later. Even small prestimulus lapses of attention influence consolidation and subsequent recognition performance. Hum Brain Mapp, 2015. © 2015 Wiley Periodicals, Inc.

Automatic physical activity pattern recognition and meal intake modelling in type 1 diabetes for their inclusion in artificial pancreas systems

La diabetes comprende un conjunto de enfermedades metabólicas que se caracterizan por concentraciones de glucosa en sangre anormalmente altas. En el caso de la diabetes tipo 1 (T1D, por sus siglas en inglés), esta situación es debida a una ausencia total de secreción endógena de insulina, lo que impide a la mayoría de tejidos usar la glucosa. En tales circunstancias, se hace necesario el suministro exógeno de insulina para preservar la vida del paciente; no obstante, siempre con la precaución de evitar caídas agudas de la glucemia por debajo de los niveles recomendados de seguridad. Además de la administración de insulina, las ingestas y la actividad física son factores fundamentales que influyen en la homeostasis de la glucosa. En consecuencia, una gestión apropiada de la T1D debería incorporar estos dos fenómenos fisiológicos, en base a una identificación y un modelado apropiado de los mismos y de sus sorrespondientes efectos en el balance glucosa-insulina. En particular, los sistemas de páncreas artificial –ideados para llevar a cabo un control automático de los niveles de glucemia del paciente– podrían beneficiarse de la integración de esta clase de información. La primera parte de esta tesis doctoral cubre la caracterización del efecto agudo de la actividad física en los perfiles de glucosa. Con este objetivo se ha llevado a cabo una revisión sistemática de la literatura y meta-análisis que determinen las respuestas ante varias modalidades de ejercicio para pacientes con T1D, abordando esta caracterización mediante unas magnitudes que cuantifican las tasas de cambio en la glucemia a lo largo del tiempo. Por otro lado, una identificación fiable de los periodos con actividad física es un requisito imprescindible para poder proveer de esa información a los sistemas de páncreas artificial en condiciones libres y ambulatorias. Por esta razón, la segunda parte de esta tesis está enfocada a la propuesta y evaluación de un sistema automático diseñado para reconocer periodos de actividad física, clasificando su nivel de intensidad (ligera, moderada o vigorosa); así como, en el caso de periodos vigorosos, identificando también la modalidad de ejercicio (aeróbica, mixta o de fuerza). En este sentido, ambos aspectos tienen una influencia específica en el mecanismo metabólico que suministra la energía para llevar a cabo el ejercicio y, por tanto, en las respuestas glucémicas en T1D. En este trabajo se aplican varias combinaciones de técnicas de aprendizaje máquina y reconocimiento de patrones sobre la fusión multimodal de señales de acelerometría y ritmo cardíaco, las cuales describen tanto aspectos mecánicos del movimiento como la respuesta fisiológica del sistema cardiovascular ante el ejercicio. Después del reconocimiento de patrones se incorpora también un módulo de filtrado temporal para sacar partido a la considerable coherencia temporal presente en los datos, una redundancia que se origina en el hecho de que en la práctica, las tendencias en cuanto a actividad física suelen mantenerse estables a lo largo de cierto tiempo, sin fluctuaciones rápidas y repetitivas. El tercer bloque de esta tesis doctoral aborda el tema de las ingestas en el ámbito de la T1D. En concreto, se propone una serie de modelos compartimentales y se evalúan éstos en función de su capacidad para describir matemáticamente el efecto remoto de las concetraciones plasmáticas de insulina exógena sobre las tasas de eleiminación de la glucosa atribuible a la ingesta; un aspecto hasta ahora no incorporado en los principales modelos de paciente para T1D existentes en la literatura. Los datos aquí utilizados se obtuvieron gracias a un experimento realizado por el Institute of Metabolic Science (Universidad de Cambridge, Reino Unido) con 16 pacientes jóvenes. En el experimento, de tipo ‘clamp’ con objetivo variable, se replicaron los perfiles individuales de glucosa, según lo observado durante una visita preliminar tras la ingesta de una cena con o bien alta carga glucémica, o bien baja. Los seis modelos mecanísticos evaluados constaban de: a) submodelos de doble compartimento para las masas de trazadores de glucosa, b) un submodelo de único compartimento para reflejar el efecto remoto de la insulina, c) dos tipos de activación de este mismo efecto remoto (bien lineal, bien con un punto de corte), y d) diversas condiciones iniciales. ABSTRACT Diabetes encompasses a series of metabolic diseases characterized by abnormally high blood glucose concentrations. In the case of type 1 diabetes (T1D), this situation is caused by a total absence of endogenous insulin secretion, which impedes the use of glucose by most tissues. In these circumstances, exogenous insulin supplies are necessary to maintain patient’s life; although caution is always needed to avoid acute decays in glycaemia below safe levels. In addition to insulin administrations, meal intakes and physical activity are fundamental factors influencing glucose homoeostasis. Consequently, a successful management of T1D should incorporate these two physiological phenomena, based on an appropriate identification and modelling of these events and their corresponding effect on the glucose-insulin balance. In particular, artificial pancreas systems –designed to perform an automated control of patient’s glycaemia levels– may benefit from the integration of this type of information. The first part of this PhD thesis covers the characterization of the acute effect of physical activity on glucose profiles. With this aim, a systematic review of literature and metaanalyses are conduced to determine responses to various exercise modalities in patients with T1D, assessed via rates-of-change magnitudes to quantify temporal variations in glycaemia. On the other hand, a reliable identification of physical activity periods is an essential prerequisite to feed artificial pancreas systems with information concerning exercise in ambulatory, free-living conditions. For this reason, the second part of this thesis focuses on the proposal and evaluation of an automatic system devised to recognize physical activity, classifying its intensity level (light, moderate or vigorous) and for vigorous periods, identifying also its exercise modality (aerobic, mixed or resistance); since both aspects have a distinctive influence on the predominant metabolic pathway involved in fuelling exercise, and therefore, in the glycaemic responses in T1D. Various combinations of machine learning and pattern recognition techniques are applied on the fusion of multi-modal signal sources, namely: accelerometry and heart rate measurements, which describe both mechanical aspects of movement and the physiological response of the cardiovascular system to exercise. An additional temporal filtering module is incorporated after recognition in order to exploit the considerable temporal coherence (i.e. redundancy) present in data, which stems from the fact that in practice, physical activity trends are often maintained stable along time, instead of fluctuating rapid and repeatedly. The third block of this PhD thesis addresses meal intakes in the context of T1D. In particular, a number of compartmental models are proposed and compared in terms of their ability to describe mathematically the remote effect of exogenous plasma insulin concentrations on the disposal rates of meal-attributable glucose, an aspect which had not yet been incorporated to the prevailing T1D patient models in literature. Data were acquired in an experiment conduced at the Institute of Metabolic Science (University of Cambridge, UK) on 16 young patients. A variable-target glucose clamp replicated their individual glucose profiles, observed during a preliminary visit after ingesting either a high glycaemic-load or a low glycaemic-load evening meal. The six mechanistic models under evaluation here comprised: a) two-compartmental submodels for glucose tracer masses, b) a single-compartmental submodel for insulin’s remote effect, c) two types of activations for this remote effect (either linear or with a ‘cut-off’ point), and d) diverse forms of initial conditions.

Crystal structure of a Staphylococcus aureus protein A domain complexed with the Fab fragment of a human IgM antibody: Structural basis for recognition of B-cell receptors and superantigen activity

Staphylococcus aureus produces a virulence factor, protein A (SpA), that contains five homologous Ig-binding domains. The interactions of SpA with the Fab region of membrane-anchored Igs can stimulate a large fraction of B cells, contributing to lymphocyte clonal selection. To understand the molecular basis for this activity, we have solved the crystal structure of the complex between domain D of SpA and the Fab fragment of a human IgM antibody to 2.7-Å resolution. In the complex, helices II and III of domain D interact with the variable region of the Fab heavy chain (VH) through framework residues, without the involvement of the hypervariable regions implicated in antigen recognition. The contact residues are highly conserved in human VH3 antibodies but not in other families. The contact residues from domain D also are conserved among all SpA Ig-binding domains, suggesting that each could bind in a similar manner. Features of this interaction parallel those reported for staphylococcal enterotoxins that are superantigens for many T cells. The structural homology between Ig VH regions and the T-cell receptor Vβ regions facilitates their comparison, and both types of interactions involve lymphocyte receptor surface remote from the antigen binding site. However, T-cell superantigens reportedly interact through hydrogen bonds with T-cell receptor Vβ backbone atoms in a primary sequence-independent manner, whereas SpA relies on a sequence-restricted conformational binding with residue side chains, suggesting that this common bacterial pathogen has adopted distinct molecular recognition strategies for affecting large sets of B and T lymphocytes.

Differences in antigen recognition and cytolytic activity of CD8+ and CD8− T cells that express the same antigen-specific receptor

CD8+ and CD8− T cell lines expressing the same antigen-specific receptor [the 2C T cell receptor (TCR)] were compared for ability to bind soluble peptide-MHC and to lyse target cells. The 2C TCR on CD8− cells bound a syngeneic MHC (Kb+)-peptide complex 10–100 times less well than the same TCR on CD8+ cells, and the CD8− 2C cells lysed target cells presenting this complex very poorly. Surprisingly, however, the CD8− cells differed little from CD8+ cells in ability to bind an allogeneic MHC (Ld+)-peptide complex and to lyse target cells presenting this complex. The CD8+/CD8− difference provided an opportunity to estimate how long TCR engagements with peptide-MHC have to persist to initiate the cytolytic T cell response.

SfiI endonuclease activity is strongly influenced by the non-specific sequence in the middle of its recognition site

The SfiI endonuclease cleaves DNA at the sequence GGCCNNNN↓NGGCC, where N is any base and ↓ is the point of cleavage. Proteins that recognise discontinuous sequences in DNA can be affected by the unspecified sequence between the specified base pairs of the target site. To examine whether this applies to SfiI, a series of DNA duplexes were made with identical sequences apart from discrete variations in the 5 bp spacer. The rates at which SfiI cleaved each duplex were measured under steady-state conditions: the steady-state rates were determined by the DNA cleavage step in the reaction pathway. SfiI cleaved some of these substrates at faster rates than other substrates. For example, the change in spacer sequence from AACAA to AAACA caused a 70-fold increase in reaction rate. In general, the extrapolated values for kcat and Km were both higher on substrates with inflexible spacers than those with flexible structures. The dinucleotide at the site of cleavage was largely immaterial. SfiI activity is thus highly dependent on conformational variations in the spacer DNA.

Brain activity evidence for recognition without recollection after early hippocampal damage

Amnesic patients with early and seemingly isolated hippocampal injury show relatively normal recognition memory scores. The cognitive profile of these patients raises the possibility that this recognition performance is maintained mainly by stimulus familiarity in the absence of recollection of contextual information. Here we report electrophysiological data on the status of recognition memory in one of the patients, Jon. Jon's recognition of studied words lacks the event-related potential (ERP) index of recollection, viz., an increase in the late positive component (500–700 ms), under conditions that elicit it reliably in normal subjects. On the other hand, a decrease of the ERP amplitude between 300 and 500 ms, also reliably found in normal subjects, is well preserved. This so-called N400 effect has been linked to stimulus familiarity in previous ERP studies of recognition memory. In Jon, this link is supported by the finding that his recognized and unrecognized studied words evoked topographically distinct ERP effects in the N400 time window. These data suggest that recollection is more dependent on the hippocampal formation than is familiarity, consistent with the view that the hippocampal formation plays a special role in episodic memory, for which recollection is so critical.

D1 cap region involved in the receptor recognition and neural cell survival activity of human ciliary neurotrophic factor.

Human ciliary neurotrophic factor (hCNTF), which promotes the cell survival and differentiation of motor and other neurons, is a protein belonging structurally to the alpha-helical cytokine family. hCNTF was subjected to three-dimensional structure modeling and site-directed mutagenesis to analyze its structure-function relationship. The replacement of Lys-155 with any other amino acid residue resulted in abolishment of neural cell survival activity, and some of the Glu-153 mutant proteins had 5- to 10-fold higher biological activity. The D1 cap region (around the boundary between the CD loop and helix D) of hCNTF, including both Glu-153 and Lys-155, was shown to play a key role in the biological activity of hCNTF as one of the putative receptor-recognition sites. In this article, the D1 cap region of the 4-helix-bundle proteins is proposed to be important in receptor recognition and biological activity common to alpha-helical cytokine proteins reactive with gp130, a component protein of the receptors.

Structural basis for receptor activity-modifying protein-dependent selective peptide recognition by a G protein-coupled receptor

Association of receptor activity-modifying proteins (RAMP1-3) with the G protein-coupled receptor (GPCR) calcitonin receptor-like receptor (CLR) enables selective recognition of the peptides calcitonin gene-related peptide (CGRP) and adrenomedullin (AM) that have diverse functions in the cardiovascular and lymphatic systems. How peptides selectively bind GPCR:RAMP complexes is unknown. We report crystal structures of CGRP analog-bound CLR:RAMP1 and AM-bound CLR:RAMP2 extracellular domain heterodimers at 2.5 and 1.8 Å resolutions, respectively. The peptides similarly occupy a shared binding site on CLR with conformations characterized by a β-turn structure near their C termini rather than the α-helical structure common to peptides that bind related GPCRs. The RAMPs augment the binding site with distinct contacts to the variable C-terminal peptide residues and elicit subtly different CLR conformations. The structures and accompanying pharmacology data reveal how a class of accessory membrane proteins modulate ligand binding of a GPCR and may inform drug development targeting CLR:RAMP complexes.

Structural and chemical basis for anticancer activity of a series of 'beta'-tubulin ligands: molecular modeling and 3D QSAR studies

An important approach to cancer therapy is the design of small molecule modulators that interfere with microtubule dynamics through their specific binding to the ²-subunit of tubulin. In the present work, comparative molecular field analysis (CoMFA) studies were conducted on a series of discodermolide analogs with antimitotic properties. Significant correlation coefficients were obtained (CoMFA(i), q² =0.68, r²=0.94; CoMFA(ii), q² = 0.63, r²= 0.91), indicating the good internal and external consistency of the models generated using two independent structural alignment strategies. The models were externally validated employing a test set, and the predicted values were in good agreement with the experimental results. The final QSAR models and the 3D contour maps provided important insights into the chemical and structural basis involved in the molecular recognition process of this family of discodermolide analogs, and should be useful for the design of new specific ²-tubulin modulators with potent anticancer activity.

Cellular Renewal and Improvement of Local Cell Effector Activity in Peritoneal Cavity in Response to Infectious Stimuli

The peritoneal cavity (PerC) is a singular compartment where many cell populations reside and interact. Despite the widely adopted experimental approach of intraperitoneal (i.p.) inoculation, little is known about the behavior of the different cell populations within the PerC. To evaluate the dynamics of peritoneal macrophage (Mempty set) subsets, namely small peritoneal Mempty set (SPM) and large peritoneal Mempty set (LPM), in response to infectious stimuli, C57BL/6 mice were injected i.p. with zymosan or Trypanosoma cruzi. These conditions resulted in the marked modification of the PerC myelo-monocytic compartment characterized by the disappearance of LPM and the accumulation of SPM and monocytes. In parallel, adherent cells isolated from stimulated PerC displayed reduced staining for beta-galactosidase, a biomarker for senescence. Further, the adherent cells showed increased nitric oxide (NO) and higher frequency of IL-12-producing cells in response to subsequent LPS and IFN-gamma stimulation. Among myelo-monocytic cells, SPM rather than LPM or monocytes, appear to be the central effectors of the activated PerC; they display higher phagocytic activity and are the main source of IL-12. Thus, our data provide a first demonstration of the consequences of the dynamics between peritoneal Mempty set subpopulations by showing that substitution of LPM by a robust SPM and monocytes in response to infectious stimuli greatly improves PerC effector activity.