Molecular circadian rhythms are robust in marine annelids lacking rhythmic behavior.

The circadian clock controls behavior and metabolism in various organisms. However, the exact timing and strength of rhythmic phenotypes can vary significantly between individuals of the same species. This is highly relevant for rhythmically complex marine environments where organismal rhythmic diversity likely permits the occupation of different microenvironments. When investigating circadian locomotor behavior of Platynereis dumerilii, a model system for marine molecular chronobiology, we found strain-specific, high variability between individual worms. The individual patterns were maintained for several weeks. A diel head transcriptome comparison of behaviorally rhythmic versus arrhythmic wild-type worms showed that 24-h cycling of core circadian clock transcripts is identical between both behavioral phenotypes. While behaviorally arrhythmic worms showed a similar total number of cycling transcripts compared to their behaviorally rhythmic counterparts, the annotation categories of their transcripts, however, differed substantially. Consistent with their locomotor phenotype, behaviorally rhythmic worms exhibit an enrichment of cycling transcripts related to neuronal/behavioral processes. In contrast, behaviorally arrhythmic worms showed significantly increased diel cycling for metabolism- and physiology-related transcripts. The prominent role of the neuropeptide pigment-dispersing factor (PDF) in Drosophila circadian behavior prompted us to test for a possible functional involvement of Platynereis pdf. Differing from its role in Drosophila, loss of pdf impacts overall activity levels but shows only indirect effects on rhythmicity. Our results show that individuals arrhythmic in a given process can show increased rhythmicity in others. Across the Platynereis population, rhythmic phenotypes exist as a continuum, with no distinct "boundaries" between rhythmicity and arrhythmicity. We suggest that such diel rhythm breadth is an important biodiversity resource enabling the species to quickly adapt to heterogeneous or changing marine environments. In times of massive sequencing, our work also emphasizes the importance of time series and functional tests.

Substitution of soybean meal for yellow mealworm meal in the diet of slow-growing chickens provides comparable carcass traits and meat quality.

1. This study investigated the effects of incorporating yellow mealworm (Tenebrio molitor) larval meal as a partial and/or complete substitute for soybean meal on carcass and meat quality in slow-growing chickens.2. A total of 256 one-day-old male broilers were randomly allocated to 1 of 32 experimental units distributed among four treatments (n = 8): the control treatment (C), where soybean (SB) meal was the protein source, and three experimental treatments, in which SB meal was replaced by Tenebrio molitor (TM) larval meal at levels of 50% (T1), 75% (T2) and 100% (T3), respectively. Three different feed phases (1-29; 29-57 and 57-92 d of age) were used for each treatment. All chickens were slaughtered at 92 d of age, with eight animals per treatment randomly selected to assess carcass and meat quality. Near-infrared reflectance spectroscopy (NIR) was used to classify meat quality.3. Carcass traits were not significantly different between treatments, except for head and thigh weight, which were higher in the control group (p < 0.01). In terms of physicochemical characteristics, treatment T2 showed less yellowness (p < 0.05), while water and cooking losses were lower in treatments T1 and T2 (p < 0.01). Meat from both T1 and T2 groups had lower shear forces (p < 0.01), higher moisture content (p < 0.01) and less protein (p < 0.05) compared to treatments C andT3. Birds fed T3 had the highest meat ash content (p < 0.01). Chickens consuming TM had higher monounsaturated fatty acid (MUFA) levels and lower polyunsaturated fatty acid (PUFA) and n-6 acidsPUFA (p < 0.01).4. Substitution of SB with TM is a protein alternative for slow-growing chickens that supports carcass and meat quality comparable to those fed a conventional diet.

The Diversification of Early Emerging Metazoans: A Window into the Evolution of Animal Multicellularity.

The biannual international workshop entitled "The diversification of early emerging metazoans: A window into animal evolution?" took place at the Evangelische Akademie Tutzing, Germany, 11-14. September 2017. It was organized by Thomas Bosch (Kiel), Thomas Holstein (Heidelberg), and Ulrich Technau (Vienna), and it was sponsored by the Deutsche Forschungsgemeinschaft (DFG). The meeting gathered over 140 researchers to discuss the contribution of non-bilaterian metazoan models (Porifera, Ctenophora, Placozoa, and Cnidaria) to our understanding of: a. The evolution of metazoan developmental processes; b. Fundamental molecular mechanisms underlying metazoan features; and c. The complex interactions that animals establish with their environment.

Estimation of somatic cell count levels of hard cheeses using physicochemical composition and artificial neural networks.

This study addresses the prediction of the somatic cell counts of the milk used in the production of sheep cheese using artificial neural networks. To achieve this objective, the neural network was designed using 33 parameters of the physicochemical composition of the cheeses obtained after they have been matured for 12 mo as input data. The physicochemical analysis of the cheeses revealed that the somatic cell count level of the cheese has a significant influence on the amount of protein, fat, dry extract, and fatty acids. When properly set up, the neural network allows the correct classification of the cheeses (100% of correct results in both training and test phases) and therefore their samples in each of the 3 nominal output variables (low, average, and high somatic cell counts). The fatty composition of the cheeses, individual fatty acids, and fat acidity are the variables that most affect the correct operation of the neural network.

The B-cell identity factor Pax5 regulates distinct transcriptional programmes in early and late B lymphopoiesis.

Pax5 controls the identity and development of B cells by repressing lineage-inappropriate genes and activating B-cell-specific genes. Here, we used genome-wide approaches to identify Pax5 target genes in pro-B and mature B cells. In these cell types, Pax5 bound to 40% of the cis-regulatory elements defined by mapping DNase I hypersensitive (DHS) sites, transcription start sites and histone modifications. Although Pax5 bound to 8000 target genes, it regulated only 4% of them in pro-B and mature B cells by inducing enhancers at activated genes and eliminating DHS sites at repressed genes. Pax5-regulated genes in pro-B cells account for 23% of all expression changes occurring between common lymphoid progenitors and committed pro-B cells, which identifies Pax5 as an important regulator of this developmental transition. Regulated Pax5 target genes minimally overlap in pro-B and mature B cells, which reflects massive expression changes between these cell types. Hence, Pax5 controls B-cell identity and function by regulating distinct target genes in early and late B lymphopoiesis.

Antioxidant capacity of different cheeses: Affecting factors and prediction by near infrared spectroscopy.

In this study, we analyzed antioxidant capacity of 224 cheese samples prepared using 16 varied mixtures of milk from cows, ewes, and goats, in 2 manufacturing seasons (winter and summer), and over 6mo of ripening. Antioxidant capacity was evaluated using the spectrophotometric 2,2-azinobis(3-ethylenebenzothiazoline-6-sulfonic acid) (ABTS) method. Total antioxidant capacity was significantly correlated with season of manufacturing and time of ripening but not with animal species providing the milk. Moreover, statistically significant correlations between the total antioxidant capacity and retinol (r=0.399), fat percentage (r=0.308), protein percentage (r=0.366), K (r=0.385), Mg (r=0.312), Na (r=0.432), and P (0.272) were observed. We evaluated the use of near infrared spectroscopy technology, together with the use of a remote reflectance fiber-optic probe, to predict the antioxidant capacity of cheese samples. The model generated allowed us to predict antioxidant capacity in unknown cheeses of different compositions and ripening times.

Tracing the history of cell types.

A study of sea urchin and sea star larvae paves the way for understanding how cell types evolve and give rise to novel morphologies.

Characterization of cephalic and non-cephalic sensory cell types provides insight into joint photo- and mechanoreceptor evolution.

Rhabdomeric opsins (r-opsins) are light sensors in cephalic eye photoreceptors, but also function in additional sensory organs. This has prompted questions on the evolutionary relationship of these cell types, and if ancient r-opsins were non-photosensory. A molecular profiling approach in the marine bristleworm Platynereis dumerilii revealed shared and distinct features of cephalic and non-cephalic r-opsin1-expressing cells. Non-cephalic cells possess a full set of phototransduction components, but also a mechanosensory signature. Prompted by the latter, we investigated Platynereis putative mechanotransducer and found that nompc and pkd2.1 co-expressed with r-opsin1 in TRE cells by HCR RNA-FISH. To further assess the role of r-Opsin1 in these cells, we studied its signaling properties and unraveled that r-Opsin1 is a Gαq-coupled blue light receptor. Profiling of cells from r-opsin1 mutants versus wild-types, and a comparison under different light conditions reveals that in the non-cephalic cells light - mediated by r-Opsin1 - adjusts the expression level of a calcium transporter relevant for auditory mechanosensation in vertebrates. We establish a deep-learning-based quantitative behavioral analysis for animal trunk movements and identify a light- and r-Opsin-1-dependent fine-tuning of the worm's undulatory movements in headless trunks, which are known to require mechanosensory feedback. Our results provide new data on peripheral cell types of likely light sensory/mechanosensory nature. These results point towards a concept in which such a multisensory cell type evolved to allow for fine-tuning of mechanosensation by light. This implies that light-independent mechanosensory roles of r-opsins may have evolved secondarily.

Attitudes of the family toward the mentally ill patient?

INTRODUCTION: In this article the attitudes towards mental illness in those families who have mentally ill members is presented in order to compare them with those found 25 years before in a similar research. METHODOLOGY: In order to carry out the survey we used Struening and Cohen's Opinion about Mental Illness (OMI) questionnaire. adapted for use in Spain by Yllá and Guimón (1979). The population studied were the families associations of the mentally ill persons suffering from sychotic and affective disturbance. alcoholism and Alzheimer's dementia; the control group was made up of a number of Civil Servants and another section of health care workers. The samples were collected at random among the associated members. For the study we undertook a factorial analysis of the answers. RESULTS: The first five factors contain the greater part of the accumulated variation. that is to say 34.2%. 19 factors explain the whole 100% of the variation. The five principle factors are: (i) Mental hygiene (14% of the variation). (ii) therapeutic negativism (7% of the variation). (iii) Social reinsertion (4.87% of the variation). which was also similar to the following factor. (iv) social rehabilitation. and (v) authoritarism (3.62% of the variation). CONCLUSION: The extensive knowledge of these illnesses and a close relationship with those who are ill could be the cause of the large and diverse opinions and that some attitudes are more complex nowadays. According to these five principle factors we can see that the controls are appreciably less authoritarian and restrictive than that of the families of these ill people and that the attitudes vary depending on the age. profession and diagnosis of the illness.

A versatile depigmentation, clearing, and labeling method for exploring nervous system diversity.

Tissue clearing combined with deep imaging has emerged as a powerful alternative to classical histological techniques. Whereas current techniques have been optimized for imaging selected nonpigmented organs such as the mammalian brain, natural pigmentation remains challenging for most other biological specimens of larger volume. We have developed a fast DEpigmEntation-Plus-Clearing method (DEEP-Clear) that is easily incorporated in existing workflows and combines whole system labeling with a spectrum of detection techniques, ranging from immunohistochemistry to RNA in situ hybridization, labeling of proliferative cells (EdU labeling) and visualization of transgenic markers. With light-sheet imaging of whole animals and detailed confocal studies on pigmented organs, we provide unprecedented insight into eyes, whole nervous systems, and subcellular structures in animal models ranging from worms and squids to axolotls and zebrafish. DEEP-Clear thus paves the way for the exploration of species-rich clades and developmental stages that are largely inaccessible by regular imaging approaches.

Establishment of Transgenesis in the Demosponge Suberites domuncula.

Sponges (Porifera) represent one of the most basally branching animal clades with key relevance for evolutionary studies, stem cell biology, and development. Despite a long history of sponges as experimental model systems, however, functional molecular studies are still very difficult to perform in these animals. Here, we report the establishment of transgenic technology as a basic and versatile experimental tool for sponge research. We demonstrate that slice explants of the demosponge Suberites domuncula regenerate functional sponge tissue and can be cultured for extended periods of time, providing easy experimental access under controlled conditions. We further show that an engineered expression construct driving the enhanced green fluorescence protein (egfp) gene under control of the Suberites domuncula ß-actin locus can be transfected into such tissue cultures, and that faithfully spliced transcripts are produced from such transfected DNA. Finally, by combining fluorescence-activated cell sorting (FACS) with quantitative PCR, we validate that transfected cells can be specifically reisolated from tissue based on their fluorescence. Although the number of detected enhanced green fluorescent protein (EGFP)-expressing cells is still limited, our approach represents the first successful introduction and expression of exogenous DNA in a sponge. These results represent a significant advance for the use of transgenic technology in a cornerstone phylum, for instance for the use in lineage tracing experiments.

Influence of somatic cell count and breed on capillary electrophoretic protein profiles of ewes' milk: a chemometric study.

Bulk tank ewe milk from the Assaf, Castellana, and Churra breeds categorized into 3 somatic cell count (SCC) groups (<500,000; 1,000,000 to 1,500,000; and >2,500,000 cells/mL) was used to investigate changes in chemical composition and capillary electrophoresis protein profiles. The results obtained indicated that breed affected fat, protein, and total solids levels, and differences were also observed for the following milk proteins: beta-, beta1-, beta2-, and alpha(s1)-III-casein, alpha-lactalbumin, and beta-lactoglobulin. High SCC affected fat and protein contents and bacterial counts. The level of beta1-, beta2-, and alpha(s1)-I-casein, and alpha-lactalbumin were significantly lower in milk with SCC scores >2,500,000 cells/mL. A preliminary study of the chemical, microbiological, and electrophoretic data was performed by cluster analysis and principal components analysis. Applying discriminant analysis, it was possible to group the milk samples according to breed and level of SCC, obtaining a prediction of 100 and 97% of the samples, respectively.

Fatty acids and fat-soluble vitamins in ewe's milk predicted by near infrared reflectance spectroscopy. Determination of seasonality.

The aim of the present work was to determine the fatty acid and fat-soluble vitamin composition and the season of ewe's milk production using NIR spectroscopy. 219 ewe's milk samples from different breeds and feeding regimes were taken each month over one year. Fatty acids were analyzed by gas chromatography, and retinol and α-, and γ-tocopherol by liquid chromatography. The results showed that the quantification was more accurate for the milk dried on paper, except for vitamins. Calibration statistical descriptors on milk dried on paper were good for capric, lauric, myristic, palmitoleic, stearic and oleic acids, and acceptable for caprilic, undecanoic, 9c, 11tCLA, ΣCLA, PUFA, ω3, ω6, retinol and α-tocopherol. The equations for the discrimination of seasonality was obtained using the partial least squares discriminant analysis (PLSDA) algorithm. 93% of winter samples and 89% of summer samples were correctly classified using the NIR spectra of milk dried on paper.

Pesticide residues in propolis from Spain and Chile. An approach using near infrared spectroscopy.

This study analyses the presence of pesticides in 106 samples of raw propolis from Spain and Chile. The components detected and quantified using gas chromatography mass spectrometry were as follows: triamidefon present in 70.1% of the samples with concentrations between 0.35 and 42.17mg/kg, and dicofol detected in 7.5% of the samples with concentrations between 0.39 and 2.83mg/kg. Diclorofluanid, procymidone, folpet, propham and metazachlor were also detected but only in a small number of samples. Near infrared spectroscopy with a remote reflectance fibre-optic probe was used to detect samples contaminated with triadimefon via partial least squares discriminant, with a R2 value of 0.71. In addition, the presence of the pesticide triadimefon in propolis was quantified using the modified partial least squares (MPLS) regression method. The calibration results showed a coefficient of determination (R2) of 0.81, a root mean square error (RMSE) of 0.36, and a ratio of performance to deviation (RPD) of 2.5.

Effect of breed and ageing time on meat quality and sensory attributes of veal calves of the "Ternera de Aliste" Quality Label.

The effect of breed, sex and ageing time on carcass, meat and eating quality were examined in 32 calves of the "Ternera de Aliste" Spanish Quality Label. Swiss Brown×Alistano-Sanabresa heifers were mated with Charolais or Limousin bulls in order to study the effect of breed. Bull and heifer calves were slaughtered when they were 6-7 months old. Samples of the M. longissimus dorsi were aged for 3, 5 or 7 days and chemical and sensory analyses were done. The results showed no differences due to sex but Charolais sires produced meat which was more tender, juicier and had more odour intensity with a darker colour than meat of the Limousin sired animals. Ageing influenced both sensory and instrumental hardness, colour and odour intensity. After seven days of maturation the meat from such young animals achieved a high quality.

Determination and evaluation of the parameters affecting the choice of veal meat of the "Ternera de Aliste" quality appellation.

The aim of the present work was to determine and assess the parameters affecting the choice of veal under the "Ternera de Aliste" quality appellation. The parameters affecting the choice proved to be colour, taste, odour, hardness and juiciness. Using these parameters, sensory evaluation, both analytical (with trained judges, QDA) and affective (with consumers, the home-use test) was carried out on four veal types, and the relative preferences for the samples assessed. Colour, hardness and losses due to cooking were also analysed instrumentally. The results revealed that the methodology is important for discriminating small differences between samples. The same trend was observed for the results of the panel of judges, consumers, and instrumental analyses regarding both hardness and juiciness. Regarding the determinant parameters in the choice of veal, in raw meat consumers prefer light colours but when expressing their general relative preferences for samples, juiciness, taste and hardness of the cooked meat had the greatest weight.

Developmental gene network analysis.

The developmental process is controlled by the information processing functions executed by the cis-elements that regulate the expression of the participating genes. A model of the network of cis-regulatory interactions that underlies the specification of the endomesoderm of the sea urchin embryo is analyzed here. Although not all the relevant interactions have yet been uncovered, the model shows how the information processing functions executed by the cis-regulatory elements involved can control essential functions of the specification process, such as transforming the localization of maternal factors into a domain-specific program of gene expression; refining the specification pattern; and stabilizing states of specification. The analysis suggests that the progressivity of the developmental process is also controlled by the cis-regulatory interactions unraveled by the network model. Given that evolution occurs by changing the program for development of the body plan, we illustrate the potential of developmental gene network analysis in understanding the process by which morphological features are maintained and diversify. Comparison of the network of cis-regulatory interactions with a portion of that underlying the specification of the endomesoderm of the starfish illustrates how the similarities and differences provide insights into how the programs for development work and how they evolve.

Various applications of liquid chromatography-mass spectrometry to the analysis of phenolic compounds.

Different options of liquid chromatography-mass spectrometry were used to establish the most suitable ion source and conditions to analyse or detect some low-molecular mass phenols, flavan-3-ols, and apply such conditions to a complex sample (wine). Data presented in this work confirm the great utility of atmospheric pressure-ionisation electrospray mass spectrometry coupled to HPLC for analysis of phenolic compounds, under negative mode in the case of low-molecular mass phenols, and under both positive and negative modes in flavan-3-ol compounds. A fragmentor voltage of 60 V could be the most suitable for analysing the compounds under study.

Effect of locust bean/xanthan gum addition and replacement of pork fat with olive oil on the quality characteristics of low-fat frankfurters.

The effects of reducing fat level from 20% to 12% and 9%, substituting pork fat with olive oil and adding locust bean/xanthan gum (0.5% and 0.6%) on emulsion stability, jelly and fat separation, processing yield, cook loss, texture and sensory characteristics of frankfurters were investigated and compared with control samples. Addition of locust bean/xanthan gum produced a significant increase in hydration/binding properties, characterised by lower cook losses, increasing yield, better emulsion stability and lower jelly and fat separation. The substitution of pork fat by olive oil did not affect these parameters. Indeed, results showed that reducing fat levels together with increasing moisture and locust bean/xanthan gum addition do not affect the sensory or textural properties, but olive oil addition produces a decrease in hardness and an increase in adhesiveness, however the overall acceptability was not affected.

Evaluation of the effect of a maternal rearing system on the odour profile of meat from suckling lamb.

The aim was to evaluate the effect of maternal rearing system on the chemical composition and odour profile of meat from suckling lamb. Twenty-four single male suckling lambs raised exclusively on maternal milk came from two rearing systems (extensive and intensive), and from Castellana and Churra breeds. The fatty acid composition of lamb meat from both breeds was affected by the rearing system, and Churra lamb reared under the extensive system had noticeable changes in odour attributes as detected by panellists and an electronic nose. Using partial least squares-discriminant analysis, the electronic nose was able to discriminate accurately between samples of Churra and Castellana meat according to their production system. However, it was not able to correctly classify meat samples from the extensive and intensive systems when both breeds were analysed together. The results suggest a significant effect of the rearing system and breed on suckling lamb meat quality traits.