What do oysters smell? Electrophysiological evidence that the bivalve osphradium is a chemosensory organ in the oyster, Magallana gigas.

The sensing of chemical cues is essential for several aspects of bivalve biology, such as the detection of food and pheromones. However, little is known about chemical communication systems in bivalves or the possible role of the osphradium as a chemosensory organ. To address this, we adapted an electrophysiological technique extensively used in vertebrates-the electro-olfactogram-to record from the osphradium in the Pacific oyster, Magallana gigas. This technique was validated using amino acids as stimulants. The osphradium proved to be sensitive to most proteinogenic L-amino acids tested, evoking tonic, negative, concentration-dependent 'electro-osphradiogram' (EOsG) voltage responses, with thresholds of detection in the range of 10- 6 to 10- 5 M. Conversely, it was insensitive to L-arginine and L-glutamic acid. The current study supports the hypothesis that the osphradium is, indeed, a chemosensory organ. The 'electro-osphradiogram' may prove to be a powerful tool in the isolation and characterization of pheromones and other important chemical cues in bivalve biology.

Applicability of metabolomics to improve sustainable grapevine production.

Metabolites represent the end product of gene expression, protein interaction and other regulatory mechanisms. The metabolome reflects a biological system's response to genetic and environmental changes, providing a more accurate description of plants' phenotype than the transcriptome or the proteome. Grapevine (Vitis vinifera L.), established for the production of wine grapes, table grapes, and raisins, holds immense agronomical and economic significance not only in the Mediterranean region but worldwide. As all plants, grapevines face the adverse impact of biotic and abiotic stresses that negatively affect multiple stages of grape and wine industry, including plant and berry development pre- and post-harvest, fresh grapes processing and consequently wine quality. In the present review we highlight the applicability of metabolome analysis in the understanding of the mechanisms involved in grapevine response and acclimatization upon the main biotic and abiotic constrains. The metabolome of induced morphogenic processes such as adventitious rooting and somatic embryogenesis is also explored, as it adds knowledge on the physiological and molecular phenomena occurring in the explants used, and on the successfully propagation of grapevines with desired traits. Finally, the microbiome-induced metabolites in grapevine are discussed in view of beneficial applications derived from the plant symbioses.

The Influence of (Poly)phenol Intake in Saliva Proteome: Short- and Medium-Term Effects of Apple.

The relationship between salivary proteome and dietary habits was studied in previous works, where a relationship between salivary proteins like cystatins and polyphenol/tannin levels in diet was observed. However, it remains to be elucidated if this association results from an effect of polyphenol-rich food ingestion on saliva composition. The aim of this work was to test the effects of apple intake on the saliva proteome, both in the short and medium term (after 4 days of continuous intake). By incubating saliva samples with apple phenolic-rich extract, protein bands containing α-amylase, S-type cystatins, and proline-rich proteins (PRPs) appeared in the fraction that precipitated, showing the potential of these (poly)phenols to precipitate salivary proteins. Among these, it was salivary cystatins that presented changes in their levels both in the saliva samples collected immediately after apple intake and in the ones collected after 4 days of intake of an extra amount of apple. These results support the thought that intake is reflected in the salivary proteome. The effect of a polyphenol-rich food, like the apple, on salivary cystatin levels is in line with results observed in animal models and, due to the involvement of these proteins in oral food perception, it would be interesting to explore in future studies the effect of these changes on sensory perception and acceptance of polyphenol-rich food.

Exploring the Applicability of Calorespirometry to Assess Seed Metabolic Stability Upon Temperature Stress Conditions-Pisum sativum L. Used as a Case Study.

The availability of phenotyping tools to assist breeding programs in the selection of high-quality crop seeds is of obvious interest with consequences for both seed producers and consumers. Seed germination involves the activation of several metabolic pathways, such as cellular respiration to provide the required ATP and reducing power. This work tested the applicability of calorespirometry, the simultaneous measurement of heat and CO2 rates, as a phenotyping tool to assess seed respiratory properties as a function of temperature. The effect of temperature on seed germination was evaluated after 16 h of seed imbibition by calorespirometric experiments performed in isothermal mode at 15, 20, 25, and 28°C on the seeds of three cultivars of peas (Pisum sativum L.) commonly used in conventional agriculture (cvs. 'Rondo', 'Torta de Quebrar', and 'Maravilha d'América'). Significant differences in metabolic heat rate and CO2 production rate (R CO2 ) as well as in the temperature responses of these parameters were found among the three cultivars. A seed germination trial was conducted during the 6 days of imbibition to evaluate the predictive power of the parameters derived from the calorespirometric measurements. The germination trial showed that the optimal germination temperature was 20°C and low germination rates were observed at extreme temperatures (15 or 28°C). The cv. 'Torta de Quebrar' showed significantly higher germination in comparison with the other two cultivars at all three temperatures. In comparison with the other two cultivars, 'Torta de Quebrar' has the lowest metabolic heat and CO2 rates and the smallest temperature dependence of these measured parameters. Additionally, 'Torta de Quebrar' has the lowest values of growth rate and carbon use efficiency calculated from the measured variables. These data suggest that calorespirometry is a useful tool for phenotyping physiologic efficiency at different temperatures during early germination stages, and can determine the seeds with the highest resilience to temperature variation, in this case 'Torta de Quebrar'.

Assessment of Total Fat and Fatty Acids in Walnuts Using Near-Infrared Hyperspectral Imaging.

"Persian" walnut (Juglans Regia L.) is one of the most consumed tree nuts in the world. It is rich in several bioactive compounds, with polyunsaturated and monounsaturated fatty acids (PUFA and MUFA) appearing at high concentrations. Walnut consumption protects against cardiovascular, carcinogenic, and neurological disorders. The fatty acid profile has usually been determined by gas chromatography, a reliable and robust tool, but also complex, polluting, and time consuming. In this study, near infrared hyperspectral imaging has been used for the screening of total fat, MUFA, PUFA, saturated, and individual fatty acids in walnuts. Five different walnuts varieties have been considered and modified partial least square (MPLS) regressions have been performed. The SEs of prediction (SEP) in external validation (ranged from 2.12% for PUFA to 13.08% for MUFA) suggest that hyperspectral imaging can be a reliable tool for controlling these parameters in a simple, non-destructive and environmentally friendly way.

Comparative study on the use of three different near infrared spectroscopy recording methodologies for varietal discrimination of walnuts.

Walnut fruit (Juglans regia L.) is an internationally well-known product with an important tradition of consumption. Its health benefits and economic importance in the food industry make this nut an interesting research topic. In this feasibility study, 200 walnut samples of 5 different varieties were collected and their near infrared (NIR) spectra were recorded with 3 different devices: a benchtop Fourier transform near infrared (FT-NIR) spectrograph, a dispersive hyperspectral imaging camera and a portable NIR dispersive spectrograph. Discriminant analyses were applied and different methods for the varietal discrimination of walnuts were obtained and compared. Up to 96 and 84% of correct identification were respectively obtained in internal (training set) and external validations. Better results were obtained covering the entire shell surface than collecting a unique random spectrum per sample. Moreover, FT-NIR and hyperspectral tools produced classification models with a lower classification error in internal and external validations than the portable NIR one.

Response of Mycorrhizal 'Touriga Nacional' Variety Grapevines to High Temperatures Measured by Calorespirometry and Near-Infrared Spectroscopy.

Heat stress negatively affects several physiological and biochemical processes in grapevine plants. In this work, two new methods, calorespirometry, which has been used to determine temperature adaptation in plants, and near-infrared (NIR) spectroscopy, which has been used to determine several grapevine-related traits and to discriminate among varieties, were tested to evaluate grapevine response to high temperatures. 'Touriga Nacional' variety grapevines, inoculated or not with Rhizoglomus irregulare or Funneliformis mosseae, were used in this study. Calorespirometric parameters and NIR spectra, as well as other parameters commonly used to assess heat injury in plants, were measured before and after high temperature exposure. Growth rate and substrate carbon conversion efficiency, calculated from calorespirometric measurements, and stomatal conductance, were the most sensitive parameters for discriminating among high temperature responses of control and inoculated grapevines. The results revealed that, although this vine variety can adapt its physiology to temperatures up to 40 °C, inoculation with R. irregulare could additionally help to sustain its growth, especially after heat shocks. Therefore, the combination of calorespirometry together with gas exchange measurements is a promising strategy for screening grapevine heat tolerance under controlled conditions and has high potential to be implemented in initial phases of plant breeding programs.

Predicting calcium in grape must and base wine by FT-NIR spectroscopy.

Calcium content in sparkling wines may not exceed 80 mg/L due to the risk of aggregation with alginate capsules. The high calcium content usually found in wine and must emphasizes the need to develop alternative and appropriate techniques faster and cleaner than atomic absorption spectrometry (AAS). To obtain a robust model to predict calcium content, FT-NIR spectroscopy was used in 98 base white wine samples and 60 must samples from an Alentejo winery. The reference method for calcium determination was AAS technique, with a dry ashing sample procedure, as a prior treatment. Results confirmed the ability of FT-NIR as an alternative technique to AAS, to predict calcium content in grape must and base wine. Advance knowledge of the calcium content in the grape must will help avoid obtaining a mixture of musts with a high calcium content in the same container.

Fatty Acid Profile of Pacific Oyster, Crassostrea gigas, Fed Different Ratios of Dietary Seaweed and Microalgae during Broodstock Conditioning.

The fatty acid (FA) profile of oysters generally reflects the dietary FA composition. Moreover, incorporation of FA into tissues is modulated by various metabolic factors, and final composition will depend upon the dietary sources, cumulative intake, and oysters' development stage. Thus, the aim of this study was to assess the impact of dietary incorporation of seaweed (SW) Ulva rigida, in replacement of traditional microalgae diet, on the FA composition of Pacific oysters Crassostrea gigas, during broodstock conditioning. The dietary conditioning consisted of direct replacement of microalgae (33% Tisochrysis lutea, 50.25% Skeletonema costatum, and 16.75% Chaetoceros calcitrans) by SW at four different substitution levels (0%, 25%, 50%, and 100% diet). The dietary docosahexaenoic acid (DHA) (22:6n-3) and eicosapentaenoic acid (EPA) (20:5n-3) contents showed a positive correlation with the dietary microalgae level. During the trial, oysters fed with higher percentages of microalgae revealed a depletion of DHA and accumulation of EPA. The 100% SW caused a significant reduction in oxygen consumption and, consequently, in the standard metabolic rate. Based on these results, a partial substitution of up to 25% of dietary microalgae seems to be a suitable alternative, because it elicited similar results to the commercial 100% microalgae diet.

Elemental composition and bioaccessibility of farmed oysters (Crassostrea gigas) fed different ratios of dietary seaweed and microalgae during broodstock conditioning.

The Pacific oyster (Crassostrea gigas) culture has been expanding, thereby leading to a greater importance of hatcheries. Broodstock conditioning is very important in the hatchery process, in which diet composition may have a strong influence on the offspring production and quality. Therefore, the current study evaluated elemental composition and bioaccessibility of oysters fed different ratios of dietary seaweed (SW) and microalgae. The dietary conditioning consisted of direct replacement of microalgae by SW at four substitution levels (0%, 25%, 50%, and 100% diet). It was observed that oysters fed 100% SW had the highest levels of Be, Cu, Zn, Sr, and Cd. The most important trend was a concentration decline of most elements with progressively lower levels of SW substitution for microalgae in the feeds. No Cd or Pb hazard (contents below 1.0 mg/kg for Cd and 1.5 mg/kg for Pb) was found in oyster meat. Regarding elemental bioaccessibility, values were similar, near 100% in the cases of Cu, Br, and I. Only for Mn and Pb, bioaccessibility percentages deviated more from 100%. Indeed, the value for Pb was 50% ± 7% (initial group), and for Mn, all values were equal or lower than 29% ± 2% (final group of oysters fed microalgae). It was observed that Mn, Cd, and Pb bioaccessibility increased with a growing share of microalgal biomass in the feed. Therefore, this study showed that SW incorporation into the feed influences elemental composition and bioaccessibility of the oysters.

Comparison of butterfly volumetric modulated arc therapy to full arc with or without deep inspiration breath hold for the treatment of mediastinal lymphoma.

PURPOSE: Radiotherapy is an effective treatment for mediastinal lymphoma but induces late effects including cardiac toxicity and secondary breast and lung cancer. Therefore reducing the dose to these organs is vital. We compared full arc volumetric modulated arc therapy (F-VMAT) against limited angle 'Butterfly' VMAT (B-VMAT) on free breathing (FB) and deep inspiration breath-hold (DIBH) computed tomography scans. The aim was to assess the benefits of B-VMAT over F-VMAT and to establish if the addition of DIBH results is a cumulative benefit. MATERIALS AND METHODS: F-VMAT and B-VMAT plans were calculated for 20 consecutive patients (15 females) with mediastinal lymphoma on both FB and DIBH scans. The planning target volume V95% was kept comparable between all plans while reducing organ doses as much as possible. RESULTS: B-VMAT significantly reduced low lung doses (V5-10), while F-VMAT was better for higher lung doses (V20-30). DIBH further improved lung doses for both types of plans. DIBH B-VMAT produced the lowest mean lung dose. With FB, heart doses were slightly higher for B-VMAT but the maximum difference was small (0.8% for V20) and only statistically significant for V10-20. The mean heart dose increased by only 0.1 Gy. The addition of DIBH however significantly reduced heart doses. While DIBH F-VMAT had the lowest heart doses, the difference was small compared with DIBH B-VMAT. B-VMAT significantly reduced breast V4 while DIBH reduced the V10. CONCLUSION: B-VMAT and DIBH are both effective in reducing organ doses and the dosimetric benefit is additive for some parameters and complementary for others.

Viability of dietary substitution of live microalgae with dry Ulva rigida in broodstock conditioning of the Pacific oyster (Crassostrea gigas).

The current study evaluated the microalgae replacement by dry macroalgae (Ulva rigida) in the reproductive success and biochemical composition of the Pacific oyster (Crassostrea gigas) during broodstock conditioning. Five nutritional regimes were tested: 100% macroalgae (diet 1), 50% macroalgae+50% microalgae (diet 2), 25% macroalgae+75% microalgae (diet 3) and 100% microalgae (diet 4). An unfed group was used as a negative control. The microalgae blend was composed of 33% Isochrysis galbana and 67% diatoms (75% Skeletonema costatum+25% Chaetoceros calcitrans). Gonadal maturation was reflected in the physiological condition of the individuals. All treatments, except diet 1, showed an increase in condition index and were fully matured at the end of the trial, with the best physiological condition observed in oysters fed diet 3 and diet 4. Protein and total lipid content increased during the conditioning period, whereas glycogen content decreased. Oysters conditioned with diet 3 had higher protein and total lipid content and lower glycogen content than the other treatments. In addition, diet 3 showed the highest percentage of viable veliger larvae. The current study demonstrated that it is possible to replace 25% of microalgae with macroalgae in the broodstock conditioning, minimizing the operative cost in bivalve hatcheries.This article has an associated First Person interview with the first author of the paper.

Chemical diplomacy in male tilapia: urinary signal increases sex hormone and decreases aggression.

Androgens, namely 11-ketotestosterone (11KT), have a central role in male fish reproductive physiology and are thought to be involved in both aggression and social signalling. Aggressive encounters occur frequently in social species, and fights may cause energy depletion, injury and loss of social status. Signalling for social dominance and fighting ability in an agonistic context can minimize these costs. Here, we test the hypothesis of a 'chemical diplomacy' mechanism through urinary signals that avoids aggression and evokes an androgen response in receiver males of Mozambique tilapia (Oreochromis mossambicus). We show a decoupling between aggression and the androgen response; males fighting their mirror image experience an unresolved interaction and a severe drop in urinary 11KT. However, if concurrently exposed to dominant male urine, aggression drops but urinary 11KT levels remain high. Furthermore, 11KT increases in males exposed to dominant male urine in the absence of a visual stimulus. The use of a urinary signal to lower aggression may be an adaptive mechanism to resolve disputes and avoid the costs of fighting. As dominance is linked to nest building and mating with females, the 11KT response of subordinate males suggests chemical eavesdropping, possibly in preparation for parasitic fertilizations.

Identity of a tilapia pheromone released by dominant males that primes females for reproduction.

Knowledge of the chemical identity and role of urinary pheromones in fish is scarce, yet it is necessary in order to understand the integration of multiple senses in adaptive responses and the evolution of chemical communication [1]. In nature, Mozambique tilapia (Oreochromis mossambicus) males form hierarchies, and females mate preferentially with dominant territorial males, which they visit in aggregations or leks [2]. Dominant males have thicker urinary bladder muscular walls than subordinates or females and store large volumes of urine, which they release at increased frequency in the presence of subordinate males or preovulatory, but not postspawned, females [3-5]. Females exposed to dominant-male urine augment their release of the oocyte maturation-inducing steroid 17α,20ß-dihydroxypregn-4-en-3-one (17,20ß-P) [6]. Here we isolate and identify a male Mozambique tilapia urinary sex pheromone as two epimeric (20α- and 20ß-) pregnanetriol 3-glucuronates. We show that both males and females have high olfactory sensitivity to the two steroids, which cross-adapt upon stimulation. Females exposed to both steroids show a rapid, 10-fold increase in production of 17,20ß-P. Thus, the identified urinary steroids prime the female endocrine system to accelerate oocyte maturation and possibly promote spawning synchrony. Tilapia are globally important as a food source but are also invasive species, with devastating impact on local freshwater ecosystems [7, 8]. Identifying the chemical cues that mediate reproduction may lead to the development of tools for population control [9-11].

[Beliefs and attitudes of professionals about marital violence: studies with health professionals, policemen and teachers]. / Crenças e atitudes dos profissionais face à violência conjugal: estudos com profissionais de saúde, polícias e profesores.

This article presents a set of three research projects that aimed to characterize the beliefs about marital violence of the professionals most directly implied in the response and prevention of this problem. A research questionnaire that evaluates beliefs about marital violence was administered to a sample of 226 health professionals, 85 law enforcement agents and 280 teachers. On the global results show a tendency to moderate disagreement with beliefs that legitimize this form of abuse. Some myths, however, persist in these samples, namely those related to the privacy of the problem, its rarity and attribution to external causes. Male and older subjects show, in general, higher levels of violence legitimization.