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1.
JAMA Netw Open ; 5(10): e2238677, 2022 10 03.
Artigo em Inglês | MEDLINE | ID: mdl-36301545

RESUMO

Importance: The health of the US population is decreasing, and novel strategies are needed to reverse the trajectory. The surgical episode may be an opportune time to screen for poor health behaviors and social needs, yet little is known about the feasibility or acceptability of such efforts. Objective: To assess the feasibility and acceptability of a pilot program that engages patients in health behavior change and addresses unmet social needs at the time of surgery. Design, Setting, and, Participants: This quality improvement study was conducted among 10 338 patients seen in a preoperative clinic for elective surgery between February 8 and August 31, 2021. Patients who received a referral for institutional resources were contacted via telephone to complete follow-up surveys 30 to 90 days after surgery and between July 1, 2021, and March 31, 2022. Interventions: Implementation of a tool to screen patients for smoking and food insecurity in a preoperative clinic. Those who screened positive were offered referrals for institutional resources. Telephone surveys were conducted with patients who accepted referrals to understand attitudes toward addressing health behaviors and social needs. Main Outcomes and Measures: Screening and referral rates, patient-perceived acceptability of addressing health behaviors and social needs at the time of surgery, smoking cessation rates, and resolution of food insecurity. Results: A total of 10 338 patients (6052 women [58.5%]; mean [SD] age, 56.5 [17.9] years) were evaluated in the preoperative clinic. Of the 10 338 patients, 7825 (75.7%) were successfully screened. Of the 641 identified smokers, 152 (23.7%) accepted a referral for smoking cessation counseling. Of the 181 identified patients with food insecurity, 121 (66.9%) accepted a referral for nonmedical needs assistance. On follow-up surveys, 64 of 78 smokers (82.1%) agreed that the preoperative appointment was an appropriate time to discuss smoking cessation, and 34 of 78 smokers (43.6%) reported quitting smoking. Similarly, 69 of 84 patients with food insecurity (82.1%) agreed it was a good or very good idea for health systems to address nonmedical needs at the time of surgery, and 27 patients (32.1%) reported no longer being insecure about food since their preoperative visit. Conclusions and Relevance: This study suggests that it is feasible to address patients' foundational health at the time of surgery. Most patients agreed that these interventions were appropriate during the perioperative period. These results support using the surgical episode as an opportunity to address foundational health.


Assuntos
Abandono do Hábito de Fumar , Fumar , Humanos , Feminino , Pessoa de Meia-Idade , Fumar/epidemiologia , Fumar/psicologia , Abandono do Hábito de Fumar/métodos , Aconselhamento/métodos , Fumar Tabaco , Insegurança Alimentar
2.
Beilstein J Nanotechnol ; 10: 975-984, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31165024

RESUMO

We present the successful growth of few-layer graphene on top of AlN-based solidly mounted resonators (SMR) using a low-temperature chemical vapour deposition (CVD) process assisted by Ni catalysts, and its effective bio-functionalization with antibodies. The SMRs are manufactured on top of fully insulating AlN/SiO2 acoustic mirrors able to withstand the temperatures reached during the CVD growth of graphene (up to 650 °C). The active AlN films, purposely grown with the c-axis tilted, effectively excite shear modes displaying excellent in-liquid performance, with electromechanical coupling and quality factors of around 3% and 150, respectively, which barely vary after graphene integration. Raman spectra reveal that the as-grown graphene is composed of less than five weakly coupled layers with a low density of defects. Two functionalization protocols of the graphene are proposed. The first one, based on a covalent binding approach, starts with a low-damage O2 plasma treatment that introduces a controlled density of defects in graphene, including carboxylic groups. After that, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) chemistry is used to covalently bind streptavidin molecules to the surface of the sensors. The second functionalization protocol is based on the non-covalent bonding of streptavidin on hydrophobic graphene surfaces. The two protocols end with the effective bonding of biotinylated anti-IgG antibodies to the streptavidin, which leaves the surface of the devices ready for possible IgG detection.

3.
J Acoust Soc Am ; 145(4): 2718, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31046325

RESUMO

The acoustic diffusion equation model has shown to be a versatile alternative model for certain types of scenarios where the classical geometrical methods have been demonstrated to be inefficient or even inaccurate. In certain scenarios, the transmission loss plays a fundamental role for accounting, i.e., noise levels and sound propagation between rooms. This paper presents an extension of the absorption boundary conditions known as modified that unifies previously proposed boundary conditions in one equation, including transmission of energy between coupled rooms by both openings and enclosures.

4.
Sci Rep ; 7(1): 1367, 2017 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-28465522

RESUMO

Shear mode solidly mounted resonators (SMRs) are fabricated using an inclined c-axis ZnO grown on a rough Al electrode. The roughness of the Al surface is controlled by changing the substrate temperature during the deposition process to promote the growth of inclined ZnO microcrystals. The optimum substrate temperature to obtain homogeneously inclined c-axis grains in ZnO films is achieved by depositing Al at 100 °C with a surface roughness ~9.2 nm, which caused an inclination angle of ~25° of the ZnO c-axis with respect to the surface normal. Shear mode devices with quality-factors at resonance, Q r and effective electromechanical coupling factors, [Formula: see text], as high as 180 and 3.4% are respectively measured. Mass sensitivities, S m of (4.9 ± 0.1) kHz · cm2/ng and temperature coefficient of frequency (TCF) of ~-67 ppm/K are obtained using this shear mode. The performance of the devices as viscosity sensors and biosensors is demonstrated by determining the frequency shifts of water-ethanol mixtures and detection of Rabbit immunoglobin G (IgG) whole molecule (H&L) respectively.

5.
J Acoust Soc Am ; 135(2): 742-53, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25234883

RESUMO

Sound source localization using a two-microphone array is an active area of research, with considerable potential for use with video conferencing, mobile devices, and robotics. Based on the observed time-differences of arrival between sound signals, a probability distribution of the location of the sources is considered to estimate the actual source positions. However, these algorithms assume a given number of sound sources. This paper describes an updated research account on the solution presented in Escolano et al. [J. Acoust. Am. Soc. 132(3), 1257-1260 (2012)], where nested sampling is used to explore a probability distribution of the source position using a Laplacian mixture model, which allows both the number and position of speech sources to be inferred. This paper presents different experimental setups and scenarios to demonstrate the viability of the proposed method, which is compared with some of the most popular sampling methods, demonstrating that nested sampling is an accurate tool for speech localization.

6.
J Acoust Soc Am ; 134(4): 2627-30, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24116399

RESUMO

Source localization using the steered response power (SRP) usually requires a costly grid-search procedure. To address this issue, a modified SRP algorithm was recently introduced, providing improved robustness when using coarser spatial grids. In this letter, an iterative method based on the modified SRP is presented. A coarse spatial grid is initially evaluated with the modified SRP, selecting the point with the highest accumulated value. Then, its corresponding volume is iteratively decomposed by using a finer spatial grid. Experiments have shown that this method provides almost the same accuracy as the fine-grid search with a substantial reduction of functional evaluations.


Assuntos
Acústica , Algoritmos , Processamento de Sinais Assistido por Computador , Som , Acústica/instrumentação , Movimento (Física) , Análise Numérica Assistida por Computador , Razão Sinal-Ruído , Espectrografia do Som , Fatores de Tempo , Transdutores
7.
J Acoust Soc Am ; 133(6): 3975-85, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23742351

RESUMO

Some recent concert hall designs have incorporated coupled reverberation chambers to the main hall that have stimulated a range of research activities in architectural acoustics. The coupling apertures between two or more coupled-volume systems are of central importance for sound propagation and sound energy decays throughout the coupled-volume systems. In addition, positions of sound sources and receivers relative to the aperture also have a profound influence on the sound energy distributions and decays. This work investigates the effect of aperture size on the behavior of coupled-volume systems using both acoustic scale-models and diffusion equation models. In these physical and numerical models, the sound source and receiver positions relative to the aperture are also investigated. Through systematic comparisons between results achieved from both physical scale models and numerical models, this work reveals valid ranges and limitations of the diffusion equation model for room-acoustic modeling.

8.
J Acoust Soc Am ; 133(3): 1218-21, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23463993

RESUMO

The diffusion equation model was used for room acoustic simulations to predict the sound pressure level and the reverberation time. The technical literature states that the diffusion equation method accurately models the late portion of the room impulse response if the energy is sufficiently scattered. This work provides conclusions on the validity of the diffusion equation model for rooms with homogeneous dimensions in relation to the scattering coefficients of the boundaries. A systematic evaluation was conducted out to determine the ranges of the absorption and scattering coefficient values that result in low noticeable differences between the predictions from a geometrical acoustic model and those from the diffusion equation model.


Assuntos
Acústica , Simulação por Computador , Arquitetura de Instituições de Saúde/métodos , Modelos Teóricos , Som , Absorção , Difusão , Movimento (Física) , Análise Numérica Assistida por Computador , Pressão , Espalhamento de Radiação , Fatores de Tempo
9.
J Acoust Soc Am ; 132(3): 1257-60, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22978853

RESUMO

The localization of active speakers with microphone arrays is an active research line with a considerable interest in many acoustic areas. Many algorithms for source localization are based on the computation of the Generalized Cross-Correlation function between microphone pairs employing phase transform weighting. Unfortunately, the performance of these methods is severely reduced when wall reflections and multiple sound sources are present in the acoustic environment. As a result, estimating the number of active sound sources and their actual directions becomes a challenging task. To effectively tackle this problem, a Bayesian inference framework is proposed. Based on a nested sampling algorithm, a mixture model and its parameters are estimated, indicating both the number of sources-model selection-and their angle of arrival-parameter estimation, respectively. A set of measured data demonstrates the accuracy of the proposed model.


Assuntos
Acústica/instrumentação , Teorema de Bayes , Modelos Teóricos , Processamento de Sinais Assistido por Computador , Fala , Algoritmos , Humanos , Movimento (Física) , Razão Sinal-Ruído , Som , Fatores de Tempo , Transdutores
10.
J Acoust Soc Am ; 128(4): 1586-9, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20968330

RESUMO

In recent years a model for predicting sound fields in enclosures has been proposed, based on the mathematical theory of diffusion. This model is held to be valid for predicting the late reverberation component of the impulse response, on the basis that sufficient reflection events must occur [Valeau et al., J. Acoust. Soc. Am. 119, 1504-1513 (2006)]. The present work determines numerically the extent of reflections necessary for the solution of the diffusion equation model to be accurate in quasi-cubic rooms. Some preliminary numerical experiments have been carried out to determine after how many mean-free times of the impulse response, which is obtained by a geometrical-acoustic approach, gives a similar result to the solution obtained from a diffusion equation model.


Assuntos
Acústica , Arquitetura de Instituições de Saúde , Modelos Teóricos , Simulação por Computador , Difusão , Movimento (Física) , Análise Numérica Assistida por Computador , Som , Fatores de Tempo , Vibração
11.
J Acoust Soc Am ; 126(6): 2856-9, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20000897

RESUMO

This work investigates a generalized implementation of directive sources on discrete-time methods. Results based on previous work for sinusoidal sources have been extended to accomplish real life source requirements, which have a frequency-dependent pattern and show gradual transitions between the different directivities at successive frequencies. One of the main advantages of this method lies in its capacity to be applied to any discrete-time method, since it is based on a proper combination of impulse responses radiated from discrete positions, which, in turn, will produce a particular pressure distribution at far-field. Furthermore, some considerations about mesh-dispersion effects on source modeling are addressed using some examples.

12.
Toxicon ; 54(7): 938-48, 2009 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-19539636

RESUMO

Bothrops asper is responsible for the vast majority of snakebite accidents in Central America and several studies have demonstrated that specific toxic and enzymatic activities of its venom vary with the geographic origin and age of the specimens. Variability in venom proteins and enzymes between specimens from the Caribbean and the Pacific versants of Costa Rica has been reported since 1964. Recently, we performed a comparative proteomic characterization of the venoms from one population of each versant. Proteins belonging to several families, including disintegrin, phospholipases A(2), serine proteinases, C-type lectins, CRISP, l-amino acid oxidase, and Zn(2+)-dependent metalloproteinases show a variable degree of relative occurrence in the venoms of both populations. The occurrence of prominent differences in the protein profile between venoms from adults and newborns, and among venom samples from individual specimens of the same region or developmental stage, further demonstrated the existence of geographic, ontogenetic and individual variability in the venom proteome of this species. These findings provide new insights towards understanding the biology of B. asper, contribute to a deeper understanding of the pathology induced by its venom and underscore the importance of the use of venoms pooled from specimens from both regions for producing antivenom exhibiting the broadest cross-reactivity. Furthermore, knowledge of the protein composition of B. asper venom paves the way for detailed future structure-function studies of individual toxins as well as for the development of new protocols to study the reactivity of therapeutic antivenoms.


Assuntos
Bothrops , Venenos de Crotalídeos/química , Proteoma , Animais , América Central , Venenos de Crotalídeos/genética , Eletroforese em Gel de Poliacrilamida , América do Sul
13.
J Proteomics ; 72(2): 227-40, 2009 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-19457355

RESUMO

The taxonomic status of the medically important pitviper of the Bothrops atrox-asper complex endemic to Venezuela, which has been classified as Bothrops colombiensis, remains incertae cedis. To help resolving this question, the venom proteome of B. colombiensis was characterized by reverse-phase HPLC fractionation followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venom contained proteins belonging to 8 types of families. PI Zn(2+)-metalloproteinases and K49 PLA(2) molecules comprise over 65% of the venom proteins. Other venom protein families comprised PIII Zn(2+)-metalloproteinases (11.3%), D49 PLA(2)s (10.2%), l-amino acid oxidase (5.7%), the medium-sized disintegrin colombistatin (5.6%), serine proteinases (1%), bradykinin-potentiating peptides (0.8%), a DC-fragment (0.5%), and a CRISP protein (0.1%). A comparison of the venom proteomes of B. colombiensis and B. atrox did not support the suggested synonymy between these two species. The closest homologues to B. colombiensis venom proteins appeared to be toxins from B. asper. A rough estimation of the similarity between the venoms of B. colombiensis and B. asper indicated that these species share approximately 65-70% of their venom proteomes. The close kinship of B. colombiensis and B. asper points at the ancestor of B. colombiensis as the founding Central American B. asper ancestor. This finding may be relevant for reconstructing the natural history and cladogenesis of Bothrops. Further, the virtually indistinguishable immunological crossreactivity of a Venezuelan ABC antiserum (raised against a mixture of B. colombiensis and Crotalus durissus cumanensis venoms) and the Costa Rican ICP polyvalent antivenom (generated against a mixture of B. asper, Crotalus simus, and Lachesis stenophrys venoms) towards the venoms of B. colombiensis and B. asper, supports this view and suggests the possibility of indistinctly using these antivenoms for the management of snakebites by any of these Bothrops species. However, our analyses also evidenced the limited recognition capability or avidity of these antivenoms towards a number of B. colombiensis and B. asper venom components, most notably medium-size disintegrins, bradykinin-potentiating peptides, PLA(2) proteins, and PI Zn(2+)-metalloproteinases.


Assuntos
Antivenenos/análise , Conotoxinas/análise , Proteômica/métodos , Mordeduras de Serpentes/terapia , Venenos de Serpentes/análise , Animais , Antivenenos/imunologia , Bothrops , Cromatografia Líquida de Alta Pressão , Conotoxinas/química , Eletroforese em Gel de Poliacrilamida , Metaloproteases/química , Fosfolipases A2/química , Estrutura Terciária de Proteína , Especificidade da Espécie , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Zinco/química
14.
J Proteome Res ; 7(10): 4396-408, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18785768

RESUMO

The venom proteomes of the snakes Bothrops caribbaeus and Bothrops lanceolatus, endemic to the Lesser Antillean islands of Saint Lucia and Martinique, respectively, were characterized by reverse-phase HPLC fractionation, followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venoms contain proteins belonging to seven ( B. caribbaeus) and five ( B. lanceolatus) types of toxins. B. caribbaeus and B. lanceolatus venoms contain phospholipases A 2, serine proteinases, l-amino acid oxidases and zinc-dependent metalloproteinases, whereas a long disintegrin, DC-fragments and a CRISP molecule were present only in the venom of B. caribbaeus, and a C-type lectin-like molecule was characterized in the venom of B. lanceolatus. Compositional differences between venoms among closely related species from different geographic regions may be due to evolutionary environmental pressure acting on isolated populations. The venoms of these two species differed in the composition and the relative abundance of their component toxins, but they exhibited similar toxicological and enzymatic profiles in mice, characterized by lethal, hemorrhagic, edema-forming, phospholipase A 2 and proteolytic activities. The venoms of B. caribbaeus and B. lanceolatus are devoid of coagulant and defibrinogenating effects and induce only mild local myotoxicity in mice. The characteristic thrombotic effect described in human envenomings by these species was not reproduced in the mouse model. The toxicological profile observed is consistent with the abundance of metalloproteinases, PLA 2s and serine proteinases in the venoms. A polyvalent (Crotalinae) antivenom produced in Costa Rica was able to immunodeplete approximately 80% of the proteins from both B. caribbaeus and B. lanceolatus venoms, and was effective in neutralizing the lethal, hemorrhagic, phospholipase A 2 and proteolytic activities of these venoms.


Assuntos
Antivenenos , Bothrops/metabolismo , Venenos de Crotalídeos/química , Proteoma/análise , Sequência de Aminoácidos , Animais , Antivenenos/imunologia , Antivenenos/toxicidade , Cromatografia Líquida de Alta Pressão , Venenos de Crotalídeos/metabolismo , Evolução Molecular , Humanos , Martinica , Espectrometria de Massas , Camundongos , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/genética , Peptídeos/metabolismo , Santa Lúcia
15.
J Proteomics ; 71(1): 46-60, 2008 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-18541473

RESUMO

We report the proteomic characterization of the venoms of two closely related pit vipers of the genus Lachesis, L. muta (South American Bushmaster) and L. stenophrys (Central American Bushmaster), and compare the toxin repertoire of the former revealed through a proteomic versus a transcriptomic approach. The protein composition of the venoms of Lachesis muta and L. stenophrys were analyzed by RP-HPLC, N-terminal sequencing, MALDI-TOF peptide mass fingerprinting and CID-MS/MS. Around 30-40 proteins of molecular masses in the range of 13-110 kDa and belonging, respectively, to only 8 and 7 toxin families were identified in L. muta and L. stenophrys venoms. In addition, both venoms contained a large number of bradykinin-potentiating peptides (BPP) and a C-type natriuretic peptide (C-NP). BPPs and C-NP comprised around 15% of the total venom proteins. In both species, the most abundant proteins were Zn(2+)-metalloproteinases (32-38%) and serine proteinases (25-31%), followed by PLA(2)s (9-12%), galactose-specific C-type lectin (4-8%), l-amino acid oxidase (LAO, 3-5%), CRISP (1.8%; found in L. muta but not in L. stenophrys), and NGF (0.6%). On the other hand, only six L. muta venom-secreted proteins matched any of the previously reported 11 partial or full-length venom gland transcripts, and venom proteome and transcriptome depart in their relative abundances of different toxin families. As expected from their close phylogenetic relationship, the venoms of L. muta and L. stenophrys share (or contain highly similar) proteins, in particular BPPs, serine proteinases, a galactose-specific C-type lectin, and LAO. However, they dramatically depart in their respective PLA(2) complement. Intraspecific quantitative and qualitative differences in the expression of PLA(2) molecules were found when the venoms of five L. muta specimens (3 from Bolivia and 2 from Peru) and the venom of the same species purchased from Sigma were compared. These observations indicate that these class of toxins represents a rapidly-evolving gene family, and suggests that functional differences due to structural changes in PLA(2)s molecules among these snakes may have been a hallmark during speciation and adaptation of diverging snake populations to new ecological niches, or competition for resources in existing ones. Our data may contribute to a deeper understanding of the biology and ecology of these snakes, and may also serve as a starting point for studying structure-function correlations of individual toxins.


Assuntos
Venenos de Crotalídeos/química , Venenos de Crotalídeos/genética , Perfilação da Expressão Gênica , Proteômica , Proteínas de Répteis/análise , Viperidae/genética , Viperidae/metabolismo , Sequência de Aminoácidos , Animais , América Central , Dados de Sequência Molecular , Proteoma/metabolismo , Proteínas de Répteis/química , Proteínas de Répteis/isolamento & purificação , América do Sul
16.
J Proteome Res ; 7(8): 3556-71, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18557640

RESUMO

We report the comparative proteomic characterization of the venoms of adult and newborn specimens of the lancehead pitviper Bothrops asper from two geographically isolated populations from the Caribbean and the Pacific versants of Costa Rica. The crude venoms were fractionated by reverse-phase HPLC, followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The two B. asper populations, separated since the late Miocene or early Pliocene (8-5 mya) by the Guanacaste Mountain Range, Central Mountain Range, and Talamanca Mountain Range, contain both identical and different (iso)enzymes from the PLA 2, serine proteinase, and SVMP families. Using a similarity coefficient, we estimate that the similarity of venom proteins between the two B. asper populations may be around 52%. Compositional differences between venoms among different geographic regions may be due to evolutionary environmental pressure acting on isolated populations. To investigate venom variability among specimens from the two B. asper populations, the reverse-phase HPLC protein profiles of 15 venoms from Caribbean specimens and 11 venoms from snakes from Pacific regions were compared. Within each B. asper geographic populations, all major venom protein families appeared to be subjected to individual variations. The occurrence of intraspecific individual allopatric variability highlights the concept that a species, B. asper in our case, should be considered as a group of metapopulations. Analysis of pooled venoms of neonate specimens from Caribbean and Pacific regions with those of adult snakes from the same geographical habitat revealed prominent ontogenetic changes in both geographical populations. Major ontogenetic changes appear to be a shift from a PIII-SVMP-rich to a PI-SVMP-rich venom and the secretion in adults of a distinct set of PLA 2 molecules than in the neonates. In addition, the ontogenetic venom composition shift results in increasing venom complexity, indicating that the requirement for the venom to immobilize prey and initiate digestion may change with the size (age) of the snake. Besides ecological and taxonomical implications, the geographical venom variability reported here may have an impact in the treatment of bite victims and in the selection of specimens for antivenom production. The occurrence of intraspecies variability in the biochemical composition and symptomatology after envenomation by snakes from different geographical location and age has long been appreciated by herpetologist and toxinologists, though detailed comparative proteomic analysis are scarce. Our study represents the first detailed characterization of individual and ontogenetic venom protein profile variations in two geographical isolated B. asper populations, and highlights the necessity of using pooled venoms as a statistically representative venom for antivenom production.


Assuntos
Bothrops , Venenos de Crotalídeos/análise , Proteoma/análise , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos , Cromatografia Líquida de Alta Pressão , Costa Rica , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Especificidade da Espécie
17.
J Proteome Res ; 7(6): 2445-57, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18444672

RESUMO

We report the comparative proteomic characterization of the venoms of two related neotropical arboreal pitvipers from Costa Rica of the genus Bothriechis, B. lateralis (side-striped palm pit viper) and B. schlegelii (eyelash pit viper). The crude venoms were fractionated by reverse-phase HPLC, followed by analysis of each chromatographic fraction by SDS-PAGE, N-terminal sequencing, MALDI-TOF mass fingerprinting, and collision-induced dissociation tandem mass spectrometry of tryptic peptides. The venom proteomes of B. lateralis and B. schlegelii comprise similar number of distinct proteins belonging, respectively, to 8 and 7 protein families. The two Bothriechis venoms contain bradykinin-potentiating peptides (BPPs), and proteins from the phospholipase A 2 (PLA 2), serine proteinase, l-amino acid oxidase (LAO), cysteine-rich secretory protein (CRISP), and Zn (2+)-dependent metalloproteinase (SVMP) families, albeit each species exhibit different relative abundances. Each venom also contains unique components, for example, snake venom vascular endothelial growth factor (svVEGF) and C-type lectin-like molecules in B. lateralis, and Kazal-type serine proteinase inhibitor-like proteins in B. schlegelii. Using a similarity coefficient, we estimate that the similarity of the venom proteins between the two Bothriechis taxa may be <10%, indicating a high divergence in their venom compositions, in spite of the fact that both species have evolved to adapt to arboreal habits. The major toxin families of B. lateralis and B. schlegelii are SVMP (55% of the total venom proteins) and PLA 2 (44%), respectively. Their different venom toxin compositions provide clues for rationalizing the distinct signs of envenomation caused by B. schlegelii and B. lateralis. An antivenomic study of the immunoreactivity of the Instituto Clodomiro Picado (ICP) polyvalent antivenom toward Bothriechis venoms revealed that l-amino acid oxidase and SVMPs represent the major antigenic protein species in both venoms. Our results provide a ground for rationalizing the reported protection of the ICP polyvalent antivenom against the hemorrhagic, coagulant, defibrinating, caseinolytic and fibrin(ogen)olytic activities of Bothriechis ( schlegelii, lateralis) venoms. However, these analyses also evidenced the limited recognition capability of the polyvalent antivenom toward a number of Bothriechis venom components, predominantly BPPs, svVEGF, Kazal-type inhibitors, some PLA 2 proteins, some serine proteinases, and CRISP molecules.


Assuntos
Antivenenos/análise , Venenos de Crotalídeos/metabolismo , Proteoma/análise , Viperidae/metabolismo , Animais , Antivenenos/imunologia , Cromatografia Líquida de Alta Pressão , Venenos de Crotalídeos/química , Venenos de Crotalídeos/imunologia , Eletroforese em Gel de Poliacrilamida , L-Aminoácido Oxidase/análise , L-Aminoácido Oxidase/imunologia , Lectinas Tipo C/análise , Metaloproteases/análise , Metaloproteases/imunologia , Oligopeptídeos/análise , Fosfolipases A2 Secretórias/análise , Proteoma/imunologia , Serina Endopeptidases/análise , Inibidores de Serina Proteinase/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Fator A de Crescimento do Endotélio Vascular/análise
18.
J Proteome Res ; 7(2): 708-19, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18161938

RESUMO

We report the proteomic characterization of the Central American pitvipers Atropoides nummifer and Atropoides picadoi. The crude venoms were fractionated by reverse-phase high-performance liquid chromatography (HPLC), followed by analysis of each chromatographic fraction by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), N-terminal sequencing, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass fingerprinting, and collision-induced dissociation-tandem mass spectrometry (CID-MS/MS) of tryptic peptides. Each venom contained a number of bradykinin-potentiating peptides and around 25-27 proteins of molecular masses in the range of 7-112 kDa, belonging to only nine different toxin families (disintegrin, DC fragment, snake venom vascular endothelial growth factor, phospholipases A2, serine protease, cysteine-rich secretory proteins, C-type lectins, L-amino acid oxidase, and Zn2+-dependent metalloproteases), albeit distinctly distributed among the two Atropoides species. In addition, A. nummifer expresses low amounts of a three-finger toxin not detected in the venom of A. picadoi. The major toxins of A. nummifer belong to the PLA2 (relative abundance, 36.5%) and the serine proteinase (22%) families, whereas the most abundant A. picadoi toxins are Zn2+-dependent metalloproteinases (66.4%). We estimate that the similarity of venom proteins between the two Atropoides taxa may be around 14-16%. The high degree of differentiation in the venom proteome among congeneric taxa emphasizes unique aspects of venom composition of related species of Atropoides snakes and points to a strong role for adaptive diversification via natural selection as a cause of this distinctiveness. On the other hand, their distinct venom toxin compositions provide clues for rationalizing the low hemorrhagic, coagulant, and defibrinating activities and the high myotoxic and proteolytic effects evoked by A. nummifer snakebite in comparison to other crotaline snake venoms and the high hemorrhagic activity of A. picadoi.


Assuntos
Venenos de Crotalídeos/química , Proteômica , Proteínas de Répteis/química , Viperidae/metabolismo , Sequência de Aminoácidos , Animais , Costa Rica , Venenos de Crotalídeos/biossíntese , Venenos de Crotalídeos/toxicidade , Dados de Sequência Molecular , Proteínas de Répteis/biossíntese , Proteínas de Répteis/toxicidade
19.
J Proteome Res ; 6(7): 2732-45, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17559253

RESUMO

The protein composition of the venoms of the West African Gaboon viper (Bitis gabonica rhinoceros), the rhinoceros viper (Bitis nasicornis), and the horned puff adder (Bitis caudalis) were analyzed by RP-HPLC, N-terminal sequencing, SDS-PAGE, MALDI-TOF peptide mass fingerprinting, and CID-MS/MS. In line with previous proteomic and transcriptomic analyses showing that snake venom proteins belong to only a few major protein families, the venom proteomes of Bitis gabonica rhinoceros, Bitis nasicornis, and Bitis caudalis comprise, respectively, toxins from 11, 9, and 8 toxin families. Dimeric disintegrins, PLA2 molecules, serine proteinases, a CRISP, C-type lectin-like proteins, L-amino acid oxidases, and snake venom metalloproteases are present in the three Bitis snake venoms, though they depart from each other in the composition and the relative abundance of their toxins. The venom composition appears to keep information on the evolutionary history of congeneric taxa. Protein similarity coefficients used to estimate the similarity of venom proteins of the Bitis taxa sampled here and in previous studies (eg. Bitis arietans and Bitis gabonica gabonica) support the monophyly of the three West African taxa (B.g. gabonica, B.g. rhinoceros, and B. nasicornis) based on genetic distance reconstructions, the lack of alliances between B. arietans and any other Bitis species, and are consistent with the taxonomic association of Bitis caudalis within the differentiated group of small Bitis species. The low level of venom toxin composition similarity between the two conventionally recognized subspecies of Bitis gabonica, B. g. gabonica and B. g. rhinoceros, supports the consideration by some authors of B. g. rhinoceros as a separate species, Bitis rhinoceros. Moreover, our proteomic data fit better to a weighted phylogram based on overall genetic distances than to an unweighted maximum-parsimony tree.


Assuntos
Proteínas/química , Proteoma/química , Proteômica , Venenos de Víboras/química , Viperidae/classificação , Sequência de Aminoácidos , Animais , Cromatografia Líquida de Alta Pressão , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Filogenia , Proteínas/análise , Proteoma/análise , Análise de Sequência de Proteína , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Viperidae/metabolismo
20.
J Acoust Soc Am ; 121(6): EL256-62, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17552578

RESUMO

Discrete-time domain methods provide a simple and flexible way to solve initial boundary value problems. With regard to the sources in such methods, only monopoles or dipoles can be considered. However, in many problems such as room acoustics, the radiation of realistic sources is directional-dependent and their directivity patterns have a clear influence on the total sound field. In this letter, a method to synthesize the directivity of sources is proposed, especially in cases where the knowledge is only based on discrete values of the directivity diagram. Some examples have been carried out in order to show the behavior and accuracy of the proposed method.


Assuntos
Localização de Som , Simulação por Computador , Humanos , Matemática , Modelos Teóricos
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