Interaction of severe acute respiratory syndrome-coronavirus and NL63 coronavirus spike proteins with angiotensin converting enzyme-2

Although in different groups, the coronaviruses severe acute respiratory syndrome-coronavirus (SARS-CoV) and NL63 use the same receptor, angiotensin converting enzyme (ACE)-2, for entry into the host cell. Despite this common receptor, the consequence of entry is very different; severe respiratory distress in the case of SARS-CoV but frequently only a mild respiratory infection for NL63. Using a wholly recombinant system, we have investigated the ability of each virus receptor-binding protein, spike or S protein, to bind to ACE-2 in solution and on the cell surface. In both assays, we find that the NL63 S protein has a weaker interaction with ACE-2 than the SARS-CoV S protein, particularly in solution binding, but the residues required for contact are similar. We also confirm that the ACE-2-binding site of NL63 S lies between residues 190 and 739. A lower-affinity interaction with ACE-2 might partly explain the different pathological consequences of infection by SARS-CoV and NL63.

Cleavage and serum reactivity of the severe acute respiratory syndrome coronavirus spike protein

Severe acute respiratory syndrome (SARS) coronavirus (SCoV) spike (S) protein is the major surface antigen of the virus and is responsible for receptor binding and the generation of neutralizing antibody. To investigate SCoV S protein, full-length and individual domains of S protein were expressed on the surface of insect cells and were characterized for cleavability and reactivity with serum samples obtained from patients during the convalescent phase of SARS. S protein could be cleaved by exogenous trypsin but not by coexpressed furin, suggesting that the protein is not normally processed during infection. Reactivity was evident by both flow cytometry and Western blot assays, but the pattern of reactivity varied according to assay and sequence of the antigen. The antibody response to SCoV S protein involves antibodies to both linear and conformational epitopes, with linear epitopes associated with the carboxyl domain and conformational epitopes associated with the amino terminal domain. Recombinant SCoV S protein appears to be a suitable antigen for the development of an efficient and sensitive diagnostic test for SARS, but our data suggest that assay format and choice of S antigen are important considerations.

Theoretical and experimental investigation of wall confluent jets ventilation and comparison with wall displacement ventilation

This paper reports CFD and experimental results of the characteristics of wall confluent jets in a room. The results presented show the behaviour of wall confluent jets in the form of velocity profiles, the spreading rate of jets on the surface, jets decay, etc. The empirical equations derived are compared with other types of air jets. In addition, the flow in wall confluent jets is compared with the flow in displacement ventilation supply, with regards to the vertical and horizontal spreading on the floor. It is concluded that the jet momentum of wall confluent jets can be more conserved than other jets. Thus, wall confluent jets have a greater spread over the floor than displacement flow.

Preliminary observations on the presence of sustained tendon strain and eccentric contractions of the wrist extensors during a common manual task: implications for lateral epicondylitis

Lateral epicondylitis (LE) is hypothesized to occur as a result of repetitive, strenuous and abnormal postural activities of the elbow and wrist. There is still a lack of understanding of how wrist and forearm positions contribute to this condition during common manual tasks. In this study the wrist kinematics and the wrist extensors’ musculotendon patterns were investigated during a manual task believed to elicit LE symptoms in susceptible subjects. A 42-year-old right-handed male, with no history of LE, performed a repetitive movement involving pushing and turning a spring-loaded mechanism. Motion capture data were acquired for the upper limb and an inverse kinematic and dynamic analysis was subsequently carried out. Results illustrated the presence of eccentric contractions sustained by the extensor carpi radialis longus (ECRL), together with an almost constant level of tendon strain of both extensor carpi radialis brevis (ECRB) and extensor digitorum communis lateral (EDCL) branch. It is believed that these factors may partly contribute to the onset of LE as they are both responsible for the creation of microtears at the tendons’ origins. The methodology of this study can be used to explore muscle actions during movements that might cause or exacerbate LE.

Lateral boundary contributions to wave-activity invariants and nonlinear stability theorems for balanced dynamics

The slow advective-timescale dynamics of the atmosphere and oceans is referred to as balanced dynamics. An extensive body of theory for disturbances to basic flows exists for the quasi-geostrophic (QG) model of balanced dynamics, based on wave-activity invariants and nonlinear stability theorems associated with exact symmetry-based conservation laws. In attempting to extend this theory to the semi-geostrophic (SG) model of balanced dynamics, Kushner & Shepherd discovered lateral boundary contributions to the SG wave-activity invariants which are not present in the QG theory, and which affect the stability theorems. However, because of technical difficulties associated with the SG model, the analysis of Kushner & Shepherd was not fully nonlinear. This paper examines the issue of lateral boundary contributions to wave-activity invariants for balanced dynamics in the context of Salmon's nearly geostrophic model of rotating shallow-water flow. Salmon's model has certain similarities with the SG model, but also has important differences that allow the present analysis to be carried to finite amplitude. In the process, the way in which constraints produce boundary contributions to wave-activity invariants, and additional conditions in the associated stability theorems, is clarified. It is shown that Salmon's model possesses two kinds of stability theorems: an analogue of Ripa's small-amplitude stability theorem for shallow-water flow, and a finite-amplitude analogue of Kushner & Shepherd's SG stability theorem in which the ‘subsonic’ condition of Ripa's theorem is replaced by a condition that the flow be cyclonic along lateral boundaries. As with the SG theorem, this last condition has a simple physical interpretation involving the coastal Kelvin waves that exist in both models. Salmon's model has recently emerged as an important prototype for constrained Hamiltonian balanced models. The extent to which the present analysis applies to this general class of models is discussed.

Climatology of Upper Tropospheric–Lower Stratospheric (UTLS) Jets and Tropopauses in MERRA

A global climatology (1979–2012) from the Modern-Era Retrospective Analysis for Research and Applications (MERRA) shows distributions and seasonal evolution of upper tropospheric jets and their relationships to the stratospheric subvortex and multiple tropopauses. The overall climatological patterns of upper tropospheric jets confirm those seen in previous studies, indicating accurate representation of jet stream dynamics in MERRA. The analysis shows a Northern Hemisphere (NH) upper tropospheric jet stretching nearly zonally from the mid-Atlantic across Africa and Asia. In winter–spring, this jet splits over the eastern Pacific, merges again over eastern North America, and then shifts poleward over the North Atlantic. The jets associated with tropical circulations are also captured, with upper tropospheric westerlies demarking cyclonic flow downstream from the Australian and Asian monsoon anticyclones and associated easterly jets. Multiple tropopauses associated with the thermal tropopause “break” commonly extend poleward from the subtropical upper tropospheric jet. In Southern Hemisphere (SH) summer, the tropopause break, along with a poleward-stretching secondary tropopause, often occurs across the tropical westerly jet downstream of the Australian monsoon region. SH high-latitude multiple tropopauses, nearly ubiquitous in June–July, are associated with the unique polar winter thermal structure. High-latitude multiple tropopauses in NH fall–winter are, however, sometimes associated with poleward-shifted upper tropospheric jets. The SH subvortex jet extends down near the level of the subtropical jet core in winter and spring. Most SH subvortex jets merge with an upper tropospheric jet between May and December; although much less persistent than in the SH, merged NH subvortex jets are common between November and April.

NAD Biosynthesis Evolution in Bacteria: Lateral Gene Transfer of Kynurenine Pathway in Xanthomonadales and Flavobacteriales

The biosynthesis of quinolinate, the de novo precursor of nicotinamide adenine dinucleotide (NAD), may be performed by two distinct pathways, namely, the bacterial aspartate (aspartate-to-quinolinate) and the eukaryotic kynurenine (tryptophan-to-quinolinate). Even though the separation into eukaryotic and bacterial routes is long established, recent genomic surveys have challenged this view, because certain bacterial species also carry the genes for the kynurenine pathway. In this work, both quinolinate biosynthetic pathways were investigated in the Bacteria clade and with special attention to Xanthomonadales and Bacteroidetes, from an evolutionary viewpoint. Genomic screening has revealed that a small number of bacterial species possess some of the genes for the kynurenine pathway, which is complete in the genus Xanthomonas and in the order Flavobacteriales, where the aspartate pathway is absent. The opposite pattern (presence of the aspartate pathway and absence of the kynurenine pathway) in close relatives (Xylella ssp. and the order Bacteroidales, respectively) points to the idea of a recent acquisition of the kynurenine pathway through lateral gene transfer in these bacterial groups. In fact, sequence similarity comparison and phylogenetic reconstruction both suggest that at least part of the genes of the kynurenine pathway in Xanthomonas and Flavobacteriales is shared by eukaryotes. These results reinforce the idea of the role that lateral gene transfer plays in the configuration of bacterial genomes, thereby providing alternative metabolic pathways, even with the replacement of primary and essential cell functions, as exemplified by NAD biosynthesis.

Recording from Two Neurons: Second-Order Stimulus Reconstruction from Spike Trains and Population Coding

We study the reconstruction of visual stimuli from spike trains, representing the reconstructed stimulus by a Volterra series up to second order. We illustrate this procedure in a prominent example of spiking neurons, recording simultaneously from the two H1 neurons located in the lobula plate of the fly Chrysomya megacephala. The fly views two types of stimuli, corresponding to rotational and translational displacements. Second-order reconstructions require the manipulation of potentially very large matrices, which obstructs the use of this approach when there are many neurons. We avoid the computation and inversion of these matrices using a convenient set of basis functions to expand our variables in. This requires approximating the spike train four-point functions by combinations of two-point functions similar to relations, which would be true for gaussian stochastic processes. In our test case, this approximation does not reduce the quality of the reconstruction. The overall contribution to stimulus reconstruction of the second-order kernels, measured by the mean squared error, is only about 5% of the first-order contribution. Yet at specific stimulus-dependent instants, the addition of second-order kernels represents up to 100% improvement, but only for rotational stimuli. We present a perturbative scheme to facilitate the application of our method to weakly correlated neurons.

Undersampled Critical Branching Processes on Small-World and Random Networks Fail to Reproduce the Statistics of Spike Avalanches

The power-law size distributions obtained experimentally for neuronal avalanches are an important evidence of criticality in the brain. This evidence is supported by the fact that a critical branching process exhibits the same exponent t~3=2. Models at criticality have been employed to mimic avalanche propagation and explain the statistics observed experimentally. However, a crucial aspect of neuronal recordings has been almost completely neglected in the models: undersampling. While in a typical multielectrode array hundreds of neurons are recorded, in the same area of neuronal tissue tens of thousands of neurons can be found. Here we investigate the consequences of undersampling in models with three different topologies (two-dimensional, small-world and random network) and three different dynamical regimes (subcritical, critical and supercritical). We found that undersampling modifies avalanche size distributions, extinguishing the power laws observed in critical systems. Distributions from subcritical systems are also modified, but the shape of the undersampled distributions is more similar to that of a fully sampled system. Undersampled supercritical systems can recover the general characteristics of the fully sampled version, provided that enough neurons are measured. Undersampling in two-dimensional and small-world networks leads to similar effects, while the random network is insensitive to sampling density due to the lack of a well-defined neighborhood. We conjecture that neuronal avalanches recorded from local field potentials avoid undersampling effects due to the nature of this signal, but the same does not hold for spike avalanches. We conclude that undersampled branching-process-like models in these topologies fail to reproduce the statistics of spike avalanches.

Lateral septal area alpha(1)-and alpha(2)-adrenoceptors differently modulate baroreflex activity in unanaesthetized rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The influence of visual inter-hemispheric connections on spiking, assembly and LFP activities, and their phase relationship during figure-ground stimulation

Desde os descobrimentos pioneiros de Hubel e Wiesel acumulou-se uma vasta literatura descrevendo as respostas neuronais do córtex visual primário (V1) a diferentes estímulos visuais. Estes estímulos consistem principalmente em barras em movimento, pontos ou grades, que são úteis para explorar as respostas dentro do campo receptivo clássico (CRF do inglês classical receptive field) a características básicas dos estímulos visuais como a orientação, direção de movimento, contraste, entre outras. Entretanto, nas últimas duas décadas, tornou-se cada vez mais evidente que a atividade de neurônios em V1 pode ser modulada por estímulos fora do CRF. Desta forma, áreas visuais primárias poderiam estar envolvidas em funções visuais mais complexas como, por exemplo, a separação de um objeto ou figura do seu fundo (segregação figura-fundo) e assume-se que as conexões intrínsecas de longo alcance em V1, assim como as conexões de áreas visuais superiores, estão ativamente envolvidas neste processo. Sua possível função foi inferida a partir da análise das variações das respostas induzidas por um estímulo localizado fora do CRF de neurônios individuais. Mesmo sendo muito provável que estas conexões tenham também um impacto tanto na atividade conjunta de neurônios envolvidos no processamento da figura quanto no potencial de campo, estas questões permanecem pouco estudadas. Visando examinar a modulação do contexto visual nessas atividades, coletamos potenciais de ação e potenciais de campo em paralelo de até 48 eletrodos implantados na área visual primária de gatos anestesiados. Estimulamos com grades compostas e cenas naturais, focando-nos na atividade de neurônios cujo CRF estava situado na figura. Da mesma forma, visando examinar a influência das conexões laterais, o sinal proveniente da área visual isotópica e contralateral foi removido através da desativação reversível por resfriamento. Fizemos isso devido a: i) as conexões laterais intrínsecas não podem ser facilmente manipuladas sem afetar diretamente os sinais que estão sendo medidos, ii) as conexões inter-hemisféricas compartilham as principais características anatômicas com a rede lateral intrínseca e podem ser vistas como uma continuação funcional das mesmas entre os dois hemisférios e iii) o resfriamento desativa as conexões de forma causal e reversível, silenciando temporariamente seu sinal, permitindo conclusões diretas a respeito da sua contribuição. Nossos resultados demonstram que o mecanismo de segmentação figurafundo se reflete nas taxas de disparo de neurônios individuais, assim como na potência do potencial de campo e na relação entre sua fase e os padrões de disparo produzidos pela população. Além disso, as conexões laterais inter-hemisféricas modulam estas variáveis dependendo da estimulação feita fora do CRF. Observamos também uma influência deste circuito lateral na coerência entre potenciais de campo entre eletrodos distantes. Em conclusão, nossos resultados dão suporte à ideia de um mecanismo complexo de segmentação figura-fundo atuando desde as áreas visuais primárias em diferentes escalas de frequência. Esse mecanismo parece envolver grupos de neurônios ativos sincronicamente e dependentes da fase do potencial de campo. Nossos resultados também são compatíveis com a hipótese que conexões laterais de longo alcance também fazem parte deste mecanismo

Development of phantom for radiographic image optimization of standard patient in the lateral view of chest and skull examination

We present the construction of a homogeneous phantom to be used in simulating the scattering and absorption of X-rays by a standard patient chest and skull when irradiated laterally. This phantom consisted of Incite and aluminium plates with their thickness determined by a tomographic exploratory method applied to the anthropomorphic phantom. Using this phantom, an optimized radiographic technique was established for chest and skull of standard sized patient in lateral view. Images generated with this optimized technique demonstrated improved image quality and reduced radiation doses. (c) 2006 Elsevier Ltd. All rights reserved.

Electroacupuncture analgesia in dogs: is there a difference between uni- and bi-lateral stimulation?

Objective To compare the analgesic effect of uni- and bi-lateral electroacupuncture (EA) in response to thermal and mechanical nociceptive stimuli and to investigate the cardiorespiratory, endocrine, and behavioral changes in dogs submitted to EA.Study design Prospective, randomized cross-over experimental study.Animals Eight adult, clinically healthy, cross-breed dogs, weighing 13 +/- 4 kg.Methods Dogs underwent electrostimulation at false acupoints (T-false); bilateral EA at acupoints, stomach 36, gall bladder 34 and spleen 6 (T-EA/bil); unilateral EA at the same points (T-EA/uni) or were untreated (T-control). All animals received acepromazine (0.05 mg kg(-1)) IV; and heart rate, pulse oximetry, indirect arterial blood pressure, respiratory rate, PECO2, rectal temperature, and plasma cortisol concentration were measured before, during, and after EA. Analgesia was tested using thoracic and abdominal cutaneous thermal and mechanical stimuli, and an interdigital thermal stimulus. Behavior was classified as calm or restless. Analysis of variance for repeated measures followed by Tukey's test was used for analysis of the data.Results There were no cardiorespiratory differences among the treatments. The cutaneous pain threshold was higher after EA, compared with false points. The latency period was shorter and analgesia was more intense in T-EA/bil than T-EA/uni, when both were compared with T-false and T-control. Six out of eight animals treated with EA were calm during treatment, and 5/8 and 4/8 of the T-false and T-control animals, respectively, were restless. Latency to interdigital thermal stimulation increased in T-EA/bil compared with the others. There was no difference in plasma cortisol concentrations among the treatments.Conclusions Bilateral EA produced a shorter latency period, a greater intensity, and longer duration of analgesia than unilateral stimulation, without stimulating a stress response.Clinical relevance Bilateral EA produces a better analgesic effect than unilateral EA.

GABA(A) receptor activation in the lateral parabrachial nucleus induces water and hypertonic NaCl intake

Inhibitory serotonergic and cholecystokinergic mechanisms in the lateral parabrachial nucleus and central GABAergic mechanisms are involved in the regulation of water and NaCl intake. In the present study we investigated if the GABA(A) receptors in the lateral parabrachial nucleus are involved in the control of water, NaCl and food intake in rats. Male Holtzman rats with stainless steel cannulas implanted bilaterally into the lateral parabrachial nucleus were used. Bilateral injections of muscimol (0.2 nmol/0.2 mu l) into the lateral parabrachial nucleus strongly increased 0.3 M NaCl (20.3 +/- 7.2 vs. saline: 2.6 +/- 0.9 ml/180 min) without changing water intake induced by the treatment with the diuretic furosemide combined with low dose of the angiotensin converting enzyme inhibitor captopril s.c. In euhydrated and satiated rats, bilateral lateral parabrachial nucleus injections of muscimol (0.2 and 0.5 nmol/0.2 0) induced 0.3 M NaCl intake (12.1 +/- 6.5 and 32.5 +/- 7.3 ml/180 min, respectively, vs. saline: 0.4 +/- 0.2 ml/180 min) and water intake (5.2 +/- 2.0 and 7.6 +/- 2.8 ml/ 180 min, respectively, vs. saline: 0.8 +/- 0.4 ml/180 min), but no food intake (2 +/- 0.4 g/240 min vs. saline: 1 +/- 0.3 g/240 min). Bilateral lateral parabrachial nucleus injections of the GABAA antagonist bicuculline (1.6 nmol/0.2 mu l) abolished the effects of muscimol (0.5 nmol/0.2 mu l) on 0.3 M NaCl and water intake. Muscimol (0.5 nmol/0.2 mu l) into the lateral parabrachial nucleus also induced a slight ingestion of water (4.2 +/- 1.6 ml/240 min vs. saline: 1.1 +/- 0.3 ml/240 min) when only water was available, a long lasting (for at least 2 h) increase on mean arterial pressure (14 +/- 4 mm Hg, vs. saline: -1 +/- 1 mm Hg) and only a tendency to increase urinary volume and Na+ and K+ renal excretion. Therefore the activation of GABAA receptors in the lateral parabrachial nucleus induces strong NaCl intake, a small ingestion of water and pressor responses, without changes on food intake. (c) 2005 Published by Elsevier Ltd on behalf of IBRO.