Hemokinins and endokinins

The mammalian tachykinins are a family of peptides that, until recently, has included substance P (SP), neurokinin A and neurokinin B. Since, the discovery of a third preprotachykinin gene (TAC4), the number of tachykinins has more than doubled to reveal several species-divergent peptides. This group includes hemokinin-1 (HK-1) in mouse and rat, endokinin-1 (EK-1) in rabbit, and EKA, EKB, human HK-1 (hHK-1) and hHK(4-11) in humans. Each exhibits a remarkable selectivity and potency for the tachykinin NK1 receptor similar to SP. Their peripheral expression has led to the proposal that they are the endogenous peripheral SP-like endocrine/paracrine agonists where SP is not expressed. Moreover, their strong cross-reactivity with a specific SP antibody leads us to question many of the proposed locations and roles of SP in the periphery. Additionally, three orphan tachykinin gene-related peptides are identified on TAC4, in rabbit, EK-2 and in humans, EKC and EKD.

Molecular Mechanisms by Which Saturated Fatty Acids Modulate TNF-alpha Expression in Mouse Macrophage Lineage

Many macrophage functions are modulated by fatty acids (FAs), including cytokine release, such as tumor necrosis factor-alpha (TNF-alpha). TNF-alpha is of great interest due to its role in the inflammation process observed in several diseases such as rheumatoid arthritis, atherosclerosis, and obesity. However, the mechanisms by which FA effects occur have not been completely elucidated yet. In this study, we used a mouse monocyte lineage (J774 cells) to evaluate the effect of 50 and 100 mu M of saturated (palmitic and stearic acids), monounsaturated (oleic acid) and polyunsaturated (linoleic acid) FAs on TNF-alpha production. Alterations in gene expression, poly(A) tail length and activation of transcription factors were evaluated. Oleic and linoleic acids, usually known as neutral or pro-inflammatory FA, inhibited LPS-induced TNF-alpha secretion by the cells. Saturated FAs were potent inducers of TNF-alpha expression and secretion under basal and inflammatory conditions (in the presence of LPS). Although the effect of the saturated FA was similar, the mechanism involved in each case seem to be distinct, as palmitic acid increased EGR-1 and CREB binding activity and stearic acid increased mRNA poly(A) tail. These results may contribute to the understanding of the molecular mechanisms by which saturated FAs modulate the inflammatory response and may lead to design of associations of dietary and pharmacological strategies to counteract the pathological effects of TNF-alpha.

Molecular characterization of Arabidopsis thaliana PUF proteins - binding specificity and target candidates

PUF proteins regulate both stability and translation through sequence-specific binding to the 3` UTR of target mRNA transcripts. Binding is mediated by a conserved PUF domain, which contains eight repeats of approximately 36 amino acids each. Found in all eukaryotes, they have been related to several developmental processes. Analysis of the 25 Arabidopsis Pumilio (APUM) proteins presenting PUF repeats reveals that 12 (APUM-1 to APUM-12) have a PUF domain with 50-75% similarity to the Drosophila PUF domain. Through three-hybrid assays, we show that APUM-1 to APUM-6 can bind specifically to the Nanos response element sequence recognized by Drosophila Pumilio. Using an Arabidopsis RNA library in a three-hybrid screening, we were able to identify an APUM-binding consensus sequence. Computational analysis allowed us to identify the APUM-binding element within the 3` UTR in many Arabidopsis transcripts, even in important mRNAs related to shoot stem cell maintenance. We demonstrate that APUM-1 to APUM-6 are able to bind specifically to APUM-binding elements in the 3` UTR of WUSCHEL, CLAVATA-1, PINHEAD/ZWILLE and FASCIATA-2 transcripts. The results obtained in the present study indicate that the APUM proteins may act as regulators in Arabidopsis through an evolutionarily conserved mechanism, which may open up a new approach for investigating mRNA regulation in plants.

Molecular characterization of nitrate reductase gene and its expression in the marine red alga Gracilaria tenuistipitata (Rhodophyta)

The enzyme nitrate reductase (NR) responsible for the conversion of nitrate to nitrite is considered to be the rate-limiting step in nitrogen assimilation. The economically important marine macroalga Gracilaria tenuistipitata presents a circadian oscillation in NR protein content and activity. In order to identify if the regulation of NR in G. tenuistipitata happens at transcriptional levels, the NR cDNA and gene were sequenced and the NR mRNA expression was studied. Analysis of the sequenced gene revealed absence of introns which is unusual for NR genes. The transcriptional profiling revealed a circadian rhythm for NR; furthermore, a rhythm was observed in constant light condition, suggesting a possible regulation by the biological clock at the mRNA levels for NR in G. tenuistipitata.

Molecular masses and sedimentation coefficients of extracellular hemoglobin of Glossoscolex paulistus: Alkaline oligomeric dissociation

The giant extracellular hemoglobin of Glossoscolex paulistus (HbGp) has a molecular mass (M) of 3600 +/- 100 kDa and a standard sedimentation coefficient (s(20.w)(0)) of 58 S. estimated by analytical ultracentrifugation (AUC). In the present work, further AUC studies were developed for HbGp, at pH 10.0, which favors oligomeric dissociation into lower M species. The HbGp oligomer is formed by globin chains a, b, c and d plus the linker chains. The pure monomeric fraction, subunit d, and HbGp at pH 10.0, in the presence of beta-mercaptoethanol, were also studied. Our results indicate that for samples of pure subunit d, besides the monomeric species with s(20.w)(0) of 2.0 S, formation of dimer of subunit d is observed with s(20.w)(0) of around 2.9 S. For the whole HbGp at pH 10.0 contributions from monomers, trimers and linkers are observed. No contribution from 58 S species was observed for the sample of oxy-HbGp at pH 10.0, showing its complete dissociation. For cyanomet-HbGp form a contribution of 17% is observed for the un-dissociated oligomer, consistent with data from other techniques that show the cyanomet-form is more stable as compared to oxy-HbGp. Masses of HbGp subunits, especially trimer abc and monomeric chains a, b, c and d, were also estimated from sedimentation equilibrium data, and are in agreement with the results from MALDI-TOF-MS. (C) 2010 Elsevier B.V. All rights reserved.

A protocol for a discrete choice experiment: understanding preferences of patients with cancer towards their cancer care across metropolitan and rural regions in Australia.

Medical decision-making in oncology is a complicated process and to date there are few studies examining how patients with cancer make choices with respect to different features of their care. It is also unknown whether patient choices vary by geographical location and how location could account for observed rural and metropolitan cancer differences. This paper describes an ongoing study that aims to (1) examine patient and healthcare-related factors that influence choices of patients with cancer; (2) measure and quantify preferences of patients with cancer towards cancer care using a discrete choice experiment (DCE) and (3) explore preference heterogeneity between metropolitan and rural locations.

Firefighter feedback during active cooling: A useful tool for heat stress management?

Monitoring an individual's thermic state in the workplace requires reliable feedback of their core temperature. However, core temperature measurement technology is expensive, invasive and often impractical in operational environments, warranting investigation of surrogate measures which could be used to predict core temperature. This study examines an alternative measure of an individual's thermic state, thermal sensation, which presents a more manageable and practical solution for Australian firefighters operating on the fireground. Across three environmental conditions (cold, warm, hot & humid), 49 Australian volunteer firefighters performed a 20-min fire suppression activity, immediately followed by 20min of active cooling using hand and forearm immersion techniques. Core temperature (Tc) and thermal sensation (TS) were measured across the rehabilitation period at five minute intervals. Despite the decline in Tc and TS throughout the rehabilitation period, there was little similarity in the magnitude or rate of decline between each measure in any of the ambient conditions. Moderate to strong correlations existed between Tc and TS in the cool (0.41, p<0.05) and hot & humid (0.57, p<0.05) conditions, however this was resultant in strong correlation during the earlier stages of rehabilitation (first five minutes), which were not evident in the latter stages. Linear regression revealed TS to be a poor predictor of Tc in all conditions (SEE=0.45-0.54°C) with a strong trend for TS to over-predict Tc (77-80% of the time). There is minimal evidence to suggest that ratings of thermal sensation, which represent a psychophysical assessment of an individual's thermal comfort, are an accurate reflection of the response of an individual's core temperature. Ratings of thermal sensation can be highly variable amongst individuals, likely moderated by local skin temperature. In account of these findings, fire managers require a more reliable source of information to guide decisions of heat stress management.

Relationship between long-term environmental fluctuations and diving effort of female Australian fur seals

For predators foraging within spatially and temporally heterogeneous marine ecosystems, environmental fluctuations can alter prey availability. Using the proportion of time spent diving and foraging trip duration as proxies of foraging effort, a multi-year dataset was used to assess the response of 58 female Australian fur seals Arctocephalus pusillus doriferus to interannual environmental fluctuations. Multiple environmental indices (remotely sensed ocean colour data and numerical weather predictions) were assessed for their influence on inter-annual variations in the proportion of time spent diving and trip duration. Model averaging revealed strong evidence for relationships between 4 indices and the proportion of time spent diving. There was a positive relationship with effort and 2 yr-lagged spring sea-surface temperature, current winter zonal wind and southern oscillation index, while a negative relationship was found with 2 yr-lagged spring zonal wind. Additionally, a positive relationship was found between foraging trip duration and 1 yr-lagged spring surface chlorophyll a. These results suggest that environmental fluctuations may influence prey availability by affecting the survival and recruitment of prey at the larval and post-larval phases while also affecting current distribution of adult prey.

Time-in-area represents foraging activity in a wide-ranging pelagic forager

Successful Marine Spatial Planning depends upon the identification of areas with high importance for particular species, ecosystems or processes. For seabirds, advancements in biologging devices have enabled us to identify these areas through the detailed study of at-sea behaviour. However, in many cases, only positional data are available and the presence of local biological productivity and hence seabird foraging behaviour is inferred from these data alone, under the untested assumption that foraging activity is more likely to occur in areas where seabirds spend more time. We fitted GPS devices and accelerometers to northern gannets Morus bassanus and categorised the behaviour of individuals outside the breeding colony as plunge diving, surface foraging, floating and flying. We then used the locations of foraging events to test the efficiency of 2 approaches: time-in-area and kernel density (KD) analyses, which are widely employed to detect highly-used areas and interpret foraging behaviour from positional data. For KD analyses, the smoothing parameter (h) was calculated using the ad hoc method (KDad hoc), and KDh=9.1, where h = 9.1 km, to designate core foraging areas from location data. A high proportion of foraging events occurred in core foraging areas designated using KDad hoc, KDh=9.1, and time-in-area. Our findings demonstrate that foraging activity occurs in areas where seabirds spend more time, and that both KD analysis and the time-in-area approach are equally efficient methods for this type of analysis. However, the time-in-area approach is advantageous in its simplicity, and in its ability to provide the shapes commonly used in planning. Therefore, the time-in-area approach can be used as a simple way of using seabirds to identify ecologically important locations from both tracking and survey data.

Influence of intrinsic variation on foraging behaviour of adult female Australian fur seals

Phenotypic variation and individual experience can create behavioural and/or dietary variation within a population. This may reduce intra-specific competition, creating a buffer to environmental change. This study examined how intrinsic variation affects foraging behaviour of Australian fur seals. Foraging movements of 29 female Australian fur seals were recorded using FastLoc GPS and dive behaviour recorders. For each individual, body mass, flipper length and axis length were recorded, a tooth was sampled to determine age, and milk was collected for diet analysis. Clustering of fatty acid dietary analysis revealed 5 distinct groups in the population. Behaviour was described using 19 indices, which were then reduced to 7 principal components (>80% of the behavioural variation). Bayesian mixed effect models were developed to describe the relationship between these components and intrinsic variation. No association was found between diet and age or body shape; however, age had a negative relationship with component 1 (27% of variation). Older females spent less time at-sea and foraged nearer to the colony. Age had an effect on component 5 (7% of variation), which represented haul-outs and dive depth; older females made fewer visits to haul-out sites and dived deeper to the benthos. This suggests that as animals age they are able to utilise prior knowledge to exploit nearby foraging sites that younger animals are either unaware of, or have yet to gain the experience required to efficiently utilise. Mass had a negative effect on components representing the directedness of a foraging trip, suggesting heavier individuals were more likely to travel directly to a foraging site.

Exposure mediates transitions between bare and vegetated states in temperate mangrove ecosystems

The resilience of mangroves is dependent on their regeneration capacity. Patchy mid-19th century clearing dramatically affected this capacity, creating stable vegetated and unvegetated states in a fragmented temperate mangrove ecosystem. Mechanisms of mediation between states were tested by monitoring the survival and growth of planted mangrove seedlings and propagules on formerly forested bare mudflats and inside patches of existing forest. Survival (1 to 76%) and growth (-0.83 to 10.45 mm mo-1 increase in plant height) of seedlings was affected by (1) differing levels of exposure found at varying proximities to remnant forest and (2) differing inundation regimes both within and between sites that were randomly selected from locations that varied in aspect relative to prevailing winds. Increases in hydrodynamic energy within and between sites corresponded to a decrease in survival that was much more pronounced at locations that were exposed to prevailing winds. Growth rates were also generally lower at sites in exposed locations, but inundation regime was a more important determinant within sites, where growth was reduced at lower heights on the shore. Results suggest that stability of the bare mudflat state (caused by historical clearance of the mangrove forest) is dependent on level of exposure to hydrodynamic energy, and a return to a forested state is more likely where this exposure is lower. These results have implications for planning and implementing mangrove restoration projects and illustrate the role that physical factors can play in determining the resilience of disturbed temperate mangrove ecosystems.

Recovery pathways from small-scale disturbance in a temperate Australian seagrass

Recovery from disturbance is a key element of ecosystem persistence, and recovery can be influenced by large-scale regional differences and smaller local-scale variations in environmental conditions. Seagrass beds are an important yet threatened nearshore habitat and recover from disturbance by regrowth, vegetative extension and dispersive propagules. We described recovery pathways from small-scale disturbances in the seagrass Zostera nigricaulis in Port Phillip Bay, a large embayment in southeastern Australia, and tested whether these pathways differed between 5 regions with different hydrodynamic conditions and water quality, and between sites within those regions. Recovery pathways were broadly consistent. When aboveground biomass was removed, recovery, defined as the point at which disturbed areas converged with undisturbed controls, took from 2 to 8 mo, but when we removed above-and below-ground biomass, it took between 2 and 13 mo. There was no evidence of recovery resulting from sexual reproduction at any sites regardless of the presence of seeds in the sediment or flower production. We found no differences in recovery at the regional scale, but we found substantial differences between local sites. At some sites, rapid recovery occurred because seagrasses grew quickly, but at others, apparent recovery occurred because regrowth coincided with overall declines in cover of undisturbed areas. Recovery time was unrelated to seagrass canopy height, biomass, percentage cover, stem density, seed bank density, epiphyte cover or sediment organic matter in seagrass adjacent to disturbance experiments. This study highlights the importance of understanding fine-scale variation in local recovery mechanisms, which may override or obscure any regional signal.

Comunicação sinestésica midiática

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Estudos estruturais por dinâmica molecular do peptídeo Polybia-MPI via Replica Exchange

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