Broodstock nutritional programming differentially affects the hepatic transcriptome and genome-wide DNA methylome of farmed gilthead sea bream (Sparus aurata) depending on genetic background.

BACKGROUND: Broodstock nutritional programming improves the offspring utilization of plant-based diets in gilthead sea bream through changes in hepatic metabolism. Attention was initially focused on fatty acid desaturases, but it can involve a wide range of processes that remain largely unexplored. How all this can be driven by a different genetic background is hardly underlined, and the present study aimed to assess how broodstock nutrition affects differentially the transcriptome and genome-wide DNA methylome of reference and genetically selected fish within the PROGENSA® selection program. RESULTS: After the stimulus phase with a low fish oil diet, two offspring subsets of each genetic background received a control or a FUTURE-based diet. This highlighted a different hepatic transcriptome (RNA-seq) and genome-wide DNA methylation (MBD-seq) pattern depending on the genetic background. The number of differentially expressed transcripts following the challenge phase varied from 323 in reference fish to 2,009 in genetically selected fish. The number of discriminant transcripts, and associated enriched functions, were also markedly higher in selected fish. Moreover, correlation analysis depicted a hyper-methylated and down-regulated gene expression state in selected fish with the FUTURE diet, whereas the opposite pattern appeared in reference fish. After filtering for highly represented functions in selected fish, 115 epigenetic markers were retrieved in this group. Among them, lipid metabolism genes (23) were the most reactive following ordering by fold-change in expression, rendering a final list of 10 top markers with a key role on hepatic lipogenesis and fatty acid metabolism (cd36, pitpna, cidea, fasn, g6pd, lipt1, scd1a, acsbg2, acsl14, acsbg2). CONCLUSIONS: Gene expression profiles and methylation signatures were dependent on genetic background in our experimental model. Such assumption affected the magnitude, but also the type and direction of change. Thus, the resulting epigenetic clock of reference fish might depict an older phenotype with a lower methylation for the epigenetically responsive genes with a negative methylation-expression pattern. Therefore, epigenetic markers will be specific of each genetic lineage, serving the broodstock programming in our selected fish to prevent and mitigate later in life the risk of hepatic steatosis through changes in hepatic lipogenesis and fatty acid metabolism.

Going to work with COVID-19 symptoms among non-sanitary (or socio-sanitary) workers: an issue of social inequality.

OBJECIVES: To describe the characteristics of the workers of activity sectors other than sanitary and socio-sanitary, who go to work with COVID-19 symptoms (GWC19S) during the lockdown or first phase of the lockdown de-escalation in Spain. STUDY DESIGN: An observational cross-sectional study based on a convenience sample selected from the COTS online survey. METHODS: A cross-sectional study based on a sample of n = 9601 workers. Descriptive analyses were performed calculating GWC19S prevalences and fitting robust Poisson regressions to obtain crude and adjusted prevalence ratios. RESULTS: The overall GWC19S prevalence is 5.6%, greater in young people (8.7%), manual workers (8.7%), workers with low salaries (9.5%), and workers of essential sectors (7.4%). Among those workers who went to work regularly to their workplaces, the GWC19S prevalence is 10.0%, greater in young (15.1%), workers with low salaries (14.2%), and women (13.2%). CONCLUSIONS: The axes of inequality of the labor market are clearly represented in the GWC19S phenomenon.

Functional insights into the infective larval stage of Anisakis simplex s.s., Anisakis pegreffii and their hybrids based on gene expression patterns.

BACKGROUND: Anisakis simplex sensu stricto and Anisakis pegreffii are sibling species of nematodes parasitic on marine mammals. Zoonotic human infection with third stage infective larvae causes anisakiasis, a debilitating and potentially fatal disease. These 2 species show evidence of hybridisation in geographical areas where they are sympatric. How the species and their hybrids differ is still poorly understood. RESULTS: Third stage larvae of Anisakis simplex s.s., Anisakis pegreffii and hybrids were sampled from Merluccius merluccius (Teleosti) hosts captured in waters of the FAO 27 geographical area. Specimens of each species and hybrids were distinguished with a diagnostic genetic marker (ITS). RNA was extracted from pools of 10 individuals of each taxon. Transcriptomes were generated using Illumina RNA-Seq, and assembled de novo. A joint assembly (here called merged transcriptome) of all 3 samples was also generated. The inferred transcript sets were functionally annotated and compared globally and also on subsets of secreted proteins and putative allergen families. While intermediary metabolism appeared to be typical for nematodes in the 3 evaluated taxa, their transcriptomes present strong levels of differential expression and enrichment, mainly of transcripts related to metabolic pathways and gene ontologies associated to energy metabolism and other pathways, with significant presence of excreted/secreted proteins, most of them allergens. The allergome of the 2 species and their hybrids has also been thoroughly studied; at least 74 different allergen families were identified in the transcriptomes. CONCLUSIONS: A. simplex s.s., A. pegreffi and their hybrids differ in gene expression patterns in the L3 stage. Strong parent-of-origin effects were observed: A. pegreffi alleles dominate in the expression patterns of hybrids albeit the latter, and A. pegreffii also display significant differences indicating that hybrids are intermediate biological entities among their parental species, and thus of outstanding interest in the study of speciation in nematodes. Analyses of differential expression based on genes coding for secreted proteins suggests that co-infections presents different repertoires of released protein to the host environment. Both species and their hybrids, share more allergen genes than previously thought and are likely to induce overlapping disease responses.

Intensification and isolation: psychosocial work environment changes in Spain 2005-10.

BACKGROUND: Work organization in Spain has traditionally been based on a high proportion of passive work. Changes in the labour market in Europe and the economic crisis that began in 2008 may have had an impact on the pace of work in Spain. AIMS: To estimate the prevalence of exposure to high-strain work and passive work in 2010 compared with 2005 and to analyse the distribution by gender, age and occupation of workers exposed to high strain and iso-strain in 2010 compared with 2005. METHODS: Two representative samples of the Spanish working population were compared. Unweighted and weighted prevalences in 2010 were calculated and compared with those in 2005. RESULTS: In the 2010 sample of 5110 workers, 29% (95% CI 27.8; 30.7) were exposed to high strain (of whom 83% had low social support). There was an increase of 6% (95% CI 3.8; 7.1) in high strain, and of 7% (95% CI 5.2; 8.3) to iso-strain, compared with 2005 (n = 7612). In 2010, as in 2005, the proportion of manual workers exposed to strain and iso-strain was more than double the corresponding proportion in non-manual workers. CONCLUSIONS: There has been an intensification of work, reduction in social support and a notable increase in exposure to high strain and iso-strain. The class inequalities reflect the segmentation of the Spanish labour market.

Simulating the impact of non-pharmaceutical interventions limiting transmission in COVID-19 epidemics using a membrane computing model.

Epidemics caused by microbial organisms are part of the natural phenomena of increasing biological complexity. The heterogeneity and constant variability of hosts, in terms of age, immunological status, family structure, lifestyle, work activities, social and leisure habits, daily division of time and other demographic characteristics make it extremely difficult to predict the evolution of epidemics. Such prediction is, however, critical for implementing intervention measures in due time and with appropriate intensity. General conclusions should be precluded, given that local parameters dominate the flow of local epidemics. Membrane computing models allows us to reproduce the objects (viruses and hosts) and their interactions (stochastic but also with defined probabilities) with an unprecedented level of detail. Our LOIMOS model helps reproduce the demographics and social aspects of a hypothetical town of 10 320 inhabitants in an average European country where COVID-19 is imported from the outside. The above-mentioned characteristics of hosts and their lifestyle are minutely considered. For the data in the Hospital and the ICU we took advantage of the observations at the Nursery Intensive Care Unit of the Consortium University General Hospital, Valencia, Spain (included as author). The dynamics of the epidemics are reproduced and include the effects on viral transmission of innate and acquired immunity at various ages. The model predicts the consequences of delaying the adoption of non-pharmaceutical interventions (between 15 and 45 days after the first reported cases) and the effect of those interventions on infection and mortality rates (reducing transmission by 20, 50 and 80%) in immunological response groups. The lockdown for the elderly population as a single intervention appears to be effective. This modeling exercise exemplifies the application of membrane computing for designing appropriate multilateral interventions in epidemic situations.

The Gypsy Database (GyDB) of mobile genetic elements.

In this article, we introduce the Gypsy Database (GyDB) of mobile genetic elements, an in-progress database devoted to the non-redundant analysis and evolutionary-based classification of mobile genetic elements. In this first version, we contemplate eukaryotic Ty3/Gypsy and Retroviridae long terminal repeats (LTR) retroelements. Phylogenetic analyses based on the gag-pro-pol internal region commonly presented by these two groups strongly support a certain number of previously described Ty3/Gypsy lineages originally reported from reverse-transcriptase (RT) analyses. Vertebrate retroviruses (Retroviridae) are also constituted in several monophyletic groups consistent with genera proposed by the ICTV nomenclature, as well as with the current tendency to classify both endogenous and exogenous retroviruses by three major classes (I, II and III). Our inference indicates that all protein domains codified by the gag-pro-pol internal region of these two groups agree in a collective presentation of a particular evolutionary history, which may be used as a main criterion to differentiate their molecular diversity in a comprehensive collection of phylogenies and non-redundant molecular profiles useful in the identification of new Ty3/Gypsy and Retroviridae species. The GyDB project is available at http://gydb.uv.es.

Total phallic reconstruction using radial forearm free flap after iatrogenic penile amputation. / Reconstrucción fálica total con colgajo libre antebraquial radial tras amputación peneana iatrogénica.

INTRODUCTION: The iatrogenic loss of the penis is a rare situation. We present a challenging case of deferred total penile reconstruction in a genetic male. MATERIAL AND METHODS: A 57-year-old man with the loss of the penis due to a penile abscess and necrosis secondary to penile curvature surgery. The reconstruction was performed over several operations using a radial forearm free flap (RFFF) and placement of a customised inflatable prosthesis a year later. RESULTS: During the first operation, the penile abscess was drained, the necrotic residues were debrided and placement of hypogastric drainage. Seven weeks later, phalloplasty was performed with RFFF and a tube-in-tube neourethra was constructed. Multiple microsurgical anastomosis was performed, and the donor site was coated with a skin graft from the thigh of partial thickness. The surgery lasted 10hours and had the complication of hair growth in the neourethra, which required mechanical endoscopic depilation on repeated occasions. The patient regained penile sensitivity. Eighteen months after the phalloplasty, a Zephyr single-body inflatable prosthesis (Geneva, Switzerland) was implanted, using the tunica albuginea of the proximal corpus cavernosum. The patient was satisfied with the aesthetics and urinary and sensory function. Four months later, the patient is gaining confidence to consider penetration. CONCLUSIONS: Despite the risk of postoperative complications and the need for multiple operations, phallic reconstruction with RFFF and the placement of a customised prosthetic implant can improve urinary and sexual function secondary to the loss of the penis.

The fern cavitation catapult: mechanism and design principles.

Leptosporangiate ferns have evolved an ingenious cavitation catapult to disperse their spores. The mechanism relies almost entirely on the annulus, a row of 12-25 cells, which successively: (i) stores energy by evaporation of the cells' content, (ii) triggers the catapult by internal cavitation, and (iii) controls the time scales of energy release to ensure efficient spore ejection. The confluence of these three biomechanical functions within the confines of a single structure suggests a level of sophistication that goes beyond most man-made devices where specific structures or parts rarely serve more than one function. Here, we study in detail the three phases of spore ejection in the sporangia of the fern Polypodium aureum. For each of these phases, we have written the governing equations and measured the key parameters. For the opening of the sporangium, we show that the structural design of the annulus is particularly well suited to inducing bending deformations in response to osmotic volume changes. Moreover, the measured parameters for the osmoelastic design lead to a near-optimal speed of spore ejection (approx. 10 m s(-1)). Our analysis of the trigger mechanism by cavitation points to a critical cavitation pressure of approximately -100 ± 14 bar, a value that matches the most negative pressures recorded in the xylem of plants. Finally, using high-speed imaging, we elucidated the physics leading to the sharp separation of time scales (30 versus 5000 µs) in the closing dynamics. Our results highlight the importance of the precise tuning of the parameters without which the function of the leptosporangium as a catapult would be severely compromised.

Enkephalinase activity in rat peripheral organs.

Hydrolysis of ([3H]Leu5)-enkephalin into the tripeptide [3H]Tyr-Gly-Gly can be detected using particulate fractions from a variety of peripheral organs. In some tissues, e.g. exocrine and endocrine glands, this dipeptidyl carboxypeptidase ("enkephalinase") activity is even higher than in brain.

Regional distribution of a high-affinity enkephalin-degrading peptidase ('enkephalinase') and effects of lesions suggest localization in the vicinity of opiate receptors in brain.

The distribution of a high affinity enkephalin-dipeptidylcarboxypeptidase between regions of mouse brain is markedly heterogenous and parallels that of opiate receptors. Furthermore intrastriatal administration of kainic acid as well as interruption of the nigrostriatal dopaminergic pathway by 6-hydroxydopamine (6-OHDA) lead to similar decreases in this peptidase activity and in the number of opiate receptors. On the contrary, no correlation was found between low affinity enkephalin degrading enzymes and opiate receptors.

Differential recognition of "enkephalinase" and angiotensin-converting enzyme by new carboxyalkyl inhibitors.

New carboxylalkyl compounds derived from Phe-Leu and corresponding to the general formula C6H5-CH2-CH(R)CO-L.Leu with R = -COOH, 3, R = -CH2-COOH, 4, R = -NH-CH2-COOH, 5, R = -NH-(CH2)2-COOH, 6, have been found to inhibit the breakdown of the Gly3-Phe4 bond of [3H] Leu-enkephalin or [3H]D.Ala2-Leu-enkephalin resulting from the action of the mouse striatal metallopeptidases: "enkephalinase" or angiotensin-converting enzyme (A.C.E.). The carboxyl coordinating ability of the Zn atom seems to be significantly higher in ACE than in "enkephalinase". Moreover, IC50 values against "enkephalinase" were found in the same range whatever the length of the chain bearing the carboxyl group whereas a well-defined position of this group with respect to the Zn atom is required for strong ACE inhibition. These features suggest a larger degree of freedom of the carboxyalkyl moieties within the active site of "enkephalinase". Therefore the differential recognition of active sites of both peptidases leads to: i) N-(carboxymethyl)-L-Phe-L-Leu, 5, a competitive inhibitor of "enkephalinase" (KI = 0.7 microM) and ACE (KI = 1.2 microM) which could be used as mixed inhibitor for both enzymes; ii) N-[(R,S)-2-carboxy, 3-benzylpropanoyl]-L-Leucine, 3, a full competitive inhibitor of "enkephalinase" (KI = 0.34 microM) which does not interact with ACE (IC50 greater than 10,000 microM). This compound can be considered as the first example of a new series of highly potent and specific "enkephalinase" inhibitors.

Fragmentation pathways of organoarsenical compounds by electrospray ion trap multiple mass spectrometry (MS6).

With its detection limit well below 30 pg microl(-1) LC-MS-MS has become a sensitive and thus popular analytical technique for organoarsenical compounds. Collision induced dissociation (CID) is a valuable tool for speciation and facilitates a positive identification of the species detected. However, it is not straightforward to understand the fragmentation pathways of organoarsenical compounds when only CID-MS-MS data is available. In the present paper we have investigated multiple mass spectrometry (MSn, n=1, 2, 3, 4, 5, 6) with electrospray CID fragmentation for a number of organoarsenical compounds likely to occur in the environment. The investigated compounds were tetramethylarsonium, trimethylarsinoxide, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, arsenocholine, and dimethylarsinoylethanol. By CID of (protonated) organoarsenical cations mostly even-electron fragments are produced after neutral loss processes such as elimination of H2, H2O, CH4, C2H2, C2H4, C2H6, HCHO, CH3OH, C2H5OH, C2H4O, and CH2CO. However, abundant odd-electron fragments are also formed after elimination of radical species. Evidence for reduction of As(V) to As(III) as a driving force in the odd-electron ion formation is obtained.

Fibronectin levels in stimulated whole-saliva and their relationship with cariogenic oral bacteria.

AIM: Fibronectin is a multifunctional glycoprotein that plays a fundamental role in mechanism of cellular union and in bacterial adhesion in the oral cavity. The aim of this study was to test the clinical relationship between fibronectin concentration in stimulated whole-saliva and the number of Mutans streptococci bacteria. PARTICIPANTS: The sample consisted of 167 children aged 12 years. METHODS: Saliva samples were collected from the subjects. Bacterial quantification was carried out using a semiquantitative method (Dentocult SM, Vivadent). Fibronectin was measured by the ELISA method with modifications. RESULTS: An inverse clinical relation was found between the levels of soluble fibronectin and the number of SM colony forming units. CONCLUSIONS: Bacterial counts sufficiently high to destroy the protein films increase bacterial binding to dental surfaces, thereby contributing to the regulation of bacterial plaque composition and its pathogenic potential. This capacity of glycoproteins to reduce the adhesion of microorganisms to the oral tissues and favour macrophage action supports the hypothesis that such proteins contribute to the first line of immune defence together with secretory antibodies.

[Intraparenchymal pulmonary inflammatory pseudotumors]. / Seudotumor inflamatorio pulmonar de localización intraparenquimatosa.

Inflammatory pseudotumors are rare entities which occasionally affect the pulmonary parenchyma. Their clinical diagnosis arises in a variety of ways and half the time they are detected by chance. Deciding on a therapeutic approach is difficult because a firm diagnosis is established only after studying the excised tissue. We describe our experience with the diagnosis, surgical treatment, and follow up of 4 cases of inflammatory pseudotumor located in the pulmonary parenchyma and treated by surgical resection (by thoracotomy or video assisted thoracoscopic surgery) with good outcomes following complete resection.

The fern sporangium: a unique catapult.

Various plants and fungi have evolved ingenious devices to disperse their spores. One such mechanism is the cavitation-triggered catapult of fern sporangia. The spherical sporangia enclosing the spores are equipped with a row of 12 to 13 specialized cells, the annulus. When dehydrating, these cells induce a dramatic change of curvature in the sporangium, which is released abruptly after the cavitation of the annulus cells. The entire ejection process is reminiscent of human-made catapults with one notable exception: The sporangia lack the crossbar that arrests the catapult arm in its returning motion. We show that much of the sophistication and efficiency of the ejection mechanism lies in the two very different time scales associated with the annulus closure.

Reconstrucción fálica total con colgajo libre antebraquial radial tras amputación peneana iatrogénica / Total phallic reconstruction using radial forearm free flap after iatrogenic penile amputation

Introducción: La pérdida del pene de causa iatrogénica es una circunstancia muy infrecuente. Se presenta un caso desafiante de reconstrucción peneana total diferida en varón genético. Material y métodos: Varón de 57 años con pérdida del falo por absceso peneano y necrosis secundaria a cirugía de incurvadura peneana. La reconstrucción se realizó en varios tiempos empleando colgajo libre de antebrazo radial (CLAR) y colocación de prótesis inflable personalizada un año después. Resultados: En un primer tiempo quirúrgico se llevó a cabo drenaje de absceso peneano, desbridamiento de restos necróticos y colocación de talla hipogástrica. Siete semanas después se llevó a cabo faloplastia con CLAR y construcción de neouretra tubo-en-tubo, anastomosis microquirúrgica múltiple y recubrimiento del sitio donante con injerto de piel del muslo de espesor parcial. La duración de esta cirugía fue 10 horas y tuvo como complicación crecimiento de vello en la neouretra, que obligó a depilación mecánica endoscópica en repetidas ocasiones. El paciente recuperó sensibilidad peneana, y 18 meses tras la faloplastia se le implantó prótesis Zephyr (Ginebra, Suiza) inflable de un solo cuerpo, utilizando la propia albugínea del cuerpo cavernoso proximal. El paciente se encuentra satisfecho desde el punto de vista cosmético, miccional y sensitivo. Cuatro meses después se encuentra ganando confianza para abordar la penetración. Conclusiones: A pesar del riesgo de complicaciones postoperatorias y de la necesidad de operaciones múltiples, la reconstrucción fálica con CLAR y colocación de implante protésico personalizado puede mejorar el impacto en la función urinaria y sexual secundario a la pérdida del pene

Introduction: The iatrogenic loss of the penis is a rare situation. We present a challenging case of deferred total penile reconstruction in a genetic male. Material and methods: A 57-year-old man with the loss of the penis due to a penile abscess and necrosis secondary to penile curvature surgery. The reconstruction was performed over several operations using a radial forearm free flap (RFFF) and placement of a customised inflatable prosthesis a year later. Results: During the first operation, the penile abscess was drained, the necrotic residues were debrided and placement of hypogastric drainage. Seven weeks later, phalloplasty was performed with RFFF and a tube-in-tube neourethra was constructed. Multiple microsurgical anastomosis was performed, and the donor site was coated with a skin graft from the thigh of partial thickness. The surgery lasted 10 hours and had the complication of hair growth in the neourethra, which required mechanical endoscopic depilation on repeated occasions. The patient regained penile sensitivity. Eighteen months after the phalloplasty, a Zephyr single-body inflatable prosthesis (Geneva, Switzerland) was implanted, using the tunica albuginea of the proximal corpus cavernosum. The patient was satisfied with the aesthetics and urinary and sensory function. Four months later, the patient is gaining confidence to consider penetration. Conclusions: Despite the risk of postoperative complications and the need for multiple operations, phallic reconstruction with RFFF and the placement of a customised prosthetic implant can improve urinary and sexual function secondary to the loss of the penis

Ty3/Gypsy retrotransposons: description of new Arabidopsis thaliana elements and evolutionary perspectives derived from comparative genomic data.

We performed a comprehensive analysis of the evolution of the Ty3/GYPSY: group of long-terminal-repeat retrotransposons (also known as METAVIRIDAE:). Exhaustive database searches allowed us to detect novel elements of this group. In particular, the Arabidopsis thaliana and Drosophila melanogaster genome sequencing projects have recently disclosed a large number of new Ty3/GYPSY: sequences. So far, elements of three different Ty3/GYPSY: lineages had been described for A. thaliana. Here, we describe six new lineages, which we have called Tit-for-tat1, Tit-for-tat2, Gimli, Gloin, Legolas, and Little Athila. We confirm that plant Ty3/GYPSY: elements form two main monophyletic groups. Moreover, our results suggest that at least four independent ancestral lineages existed before the monocot-dicot split, about 200 MYA. Twelve sequences from D. melanogaster that may correspond to new elements are also described. Some of these sequences are similar to those of OSVALDO: and Ulysses, two elements of the OSVALDO: clade that had never before been described for D. melanogaster. Comparative analyses of multiple organisms, some of them with completely sequenced genomes, show that the number of lineages of Ty3/GYPSY: elements is very variable. Thus, while only 1 lineage is present in Saccharomyces cerevisiae, at least 6 exist in Caenorhabditis elegans, at least 9 are present in the A. thaliana, and perhaps 20 are present in D. melanogaster. Finally, we suggest that the presence of a chromodomain-containing integrase, a feature of some closely related Ty3/GYPSY: elements of fungi, plants, and animals, may be used to define a new METAVIRIDAE: genus.