Most animals, including sponges (Porifera), have species‐specific microbiomes. Which genetic or environmental factors play major roles structuring the microbial community at the intraspecific level ...in sponges is, however, largely unknown. In this study, we tested whether geographic location or genetic structure of conspecific sponges influences their microbial assembly. For that, we used three sponge species with different rates of gene flow, and collected samples along their entire distribution range (two from the Mediterranean and one from the Southern Ocean) yielding a total of 393 samples. These three sponge species have been previously analysed by microsatellites or single nucleotide polymorphisms, and here we investigate their microbiomes by amplicon sequencing of the microbial 16S rRNA gene. The sponge Petrosia ficiformis, with highly isolated populations (low gene flow), showed a stronger influence of the host genetic distance on the microbial composition than the spatial distance. Host‐specificity was therefore detected at the genotypic level, with individuals belonging to the same host genetic cluster harbouring more similar microbiomes than distant ones. On the contrary, the microbiome of Ircinia fasciculata and Dendrilla antarctica ‐ both with weak population structure (high gene flow) ‐ seemed influenced by location rather than by host genetic distance. Our results suggest that in sponge species with high population structure, the host genetic cluster influence the microbial community more than the geographic location.
Summary
The diversity and function of sponge‐associated symbionts is now increasingly understood; however, we lack an understanding of how they dynamically behave to ensure holobiont stability in the ...face of environmental variation. Here, we performed a metatransciptomic analysis on three microbial symbionts of the sponge Cymbastela concentrica in situ over 14 months and through differential gene expression and correlation analysis to environmental variables uncovered differences that speak to their metabolic activities and level of symbiotic and environmental interactions. The nitrite‐oxidizing Ca. Porinitrospira cymbastela maintained a seemingly stable metabolism, with the few differentially expressed genes related only to stress responses. The heterotrophic Ca. Porivivens multivorans displayed differential use of holobiont‐derived compounds and respiration modes, while the ammonium‐oxidizing archaeon Ca. Nitrosopumilus cymbastelus differentially expressed genes related to phosphate metabolism and symbiosis effectors. One striking similarity between the symbionts was their similar variation in expression of stress‐related genes. Our time‐series study showed that the microbial community of C. concentrica undertakes dynamic gene expression adjustments in response to the surroundings, tuned to deal with general stress and metabolic interactions between holobiont members. The success of these dynamic adjustments likely underpins the stability of the sponge holobiont and may provide resilience against environmental change.
The free‐living (FL) and particle‐attached (PA) marine microbial communities have repeatedly been proved to differ in their diversity and composition in the photic ocean and also recently in the ...bathypelagic ocean at a global scale. However, although high taxonomic ranks exhibit preferences for a PA or FL mode of life, it remains poorly understood whether two clear lifestyles do exist and how these are distributed across the prokaryotic phylogeny. We studied the FL (<0.8 μm) and PA (0.8–20 μm) prokaryotes at 30 stations distributed worldwide within the bathypelagic oceanic realm (2150–4000 m depth) using high‐throughput sequencing of the small subunit ribosomal RNA gene (16S rRNA). A high proportion of the bathypelagic prokaryotes were mostly found either attached to particles or freely in the surrounding water but rarely in both types of environments. In particular, this trait was deeply conserved through their phylogeny, suggesting that the deep‐ocean particles and the surrounding water constitute two highly distinct niches and that transitions from one to the other have been rare at an evolutionary timescale. As a consequence, PA and FL communities had clear alpha‐ and beta‐diversity differences that exceeded the global‐scale geographical variation. Our study organizes the bathypelagic prokaryotic diversity into a reasonable number of ecologically coherent taxa regarding their association with particles, a first step for understanding which are the microbes responsible for the processing of the dissolved and particulate pools of organic matter that have a very different biogeochemical role in the deep ocean.
Bacteroidetes is one of the dominant phyla of ocean bacterioplankton, yet its diversity and population structure is poorly understood. To advance in the delineation of ecologically meaningful units ...within this group, we constructed near full‐length 16S rRNA gene clone libraries from contrasting marine environments in the NW Mediterranean. Based on phylogeny and the associated ecological variables (depth and season), 24 different Bacteroidetes clades were delineated. By considering their relative abundance (from iTag amplicon sequencing studies), we described the distribution patterns of each of these clades, delimiting them as Ecologically Significant Taxonomic Units (ESTUs). Spatially, there was almost no overlap among ESTUs at different depths. In deep waters there was predominance of Owenweeksia, Leeuwenhoekiella, Muricauda‐related genera, and some depth‐associated ESTUs within the NS5 and NS2b marine clades. Seasonally, multi‐annual dynamics of recurring ESTUs were present with dominance of some ESTUs within the NS4, NS5 and NS2b marine clades along most of the year, but with variable relative frequencies between months. A drastic change towards the predominance of Formosa‐related ESTUs and one ESTU from the NS5 marine clade was typically present after the spring bloom. Even though there are no isolates available for these ESTUs to determine their physiology, correlation analyses identified the environmental preference of some of them. Overall, our results suggest that there is a high degree of niche specialisation within these closely related clades. This work constitutes a step forward in disentangling the ecology of marine Bacteroidetes, which are essential players in organic matter processing in the oceans.
Bisphenol A (BPA) is an endocrine disruptor used in manufacturing of plastic devices, resulting in an ubiquitous presence in the environment linked to human infertility, obesity or cardiovascular ...diseases. Both transcriptome and epigenome modifications lie behind these disorders that might be inherited transgenerationally when affecting germline. To assess potential effects of paternal exposure on offspring development, adult zebrafish males were exposed to BPA during spermatogenesis and mated with non-treated females. Results showed an increase in the rate of heart failures of progeny up to the F2, as well as downregulation of 5 genes involved in cardiac development in F1 embryos. Moreover, BPA causes a decrease in F0 and F1 sperm remnant mRNAs related to early development. Results reveal a paternal inheritance of changes in the insulin signaling pathway due to downregulation of insulin receptor β mRNAs, suggesting a link between BPA male exposure and disruption of cardiogenesis in forthcoming generations.
•We examine the effects of adult male exposure to BPA on the progeny (F1 and F2).•Paternal exposure promotes similar cardiac malformations to those caused by direct exposure.•BPA applied during spermatogenesis decrease the insra and insrb transcripts in spermatozoa.•Sperm insrb transcript controls embryonic expression being the downregulation inherited by F1.•Paternal BPA exposure impairs heart development in F1 and F2 disrupting insulin signaling pathway.
Paternal bisphenol A exposure impairs cardiac development throughout generations.
Uncontrolled extracellular matrix (ECM) production by fibroblasts in response to injury contributes to fibrotic diseases, including idiopathic pulmonary fibrosis (IPF). Reactive oxygen species (ROS) ...generation is involved in the pathogenesis of IPF. Transforming growth factor‐β1 (TGF‐β1) stimulates the production of NADPH oxidase 4 (NOX4)‐dependent ROS, promoting lung fibrosis (LF). Dysregulation of microRNAs (miRNAs) has been shown to contribute to LF. To identify miRNAs involved in redox regulation relevant for IPF, we performed arrays in human lung fibroblasts exposed to ROS. miR‐9‐5p was selected as the best candidate and we demonstrate its inhibitory effect on TGF‐β receptor type II (TGFBR2) and NOX4 expression. Increased expression of miR‐9‐5p abrogates TGF‐β1‐dependent myofibroblast phenotypic transformation. In the mouse model of bleomycin‐induced LF, miR‐9‐5p dramatically reduces fibrogenesis and inhibition of miR‐9‐5p and prevents its anti‐fibrotic effect both in vitro and in vivo. In lung specimens from patients with IPF, high levels of miR‐9‐5p are found. In omentum‐derived mesothelial cells (MCs) from patients subjected to peritoneal dialysis (PD), miR‐9‐5p also inhibits mesothelial to myofibroblast transformation. We propose that TGF‐β1 induces miR‐9‐5p expression as a self‐limiting homeostatic response.
Synopsis
miR‐9‐5p is discovered as an anti‐fibrotic miRNA that targets TGBR2 and NOX4 to inhibit the transformation of fibroblasts into myofibroblasts. TGF‐β1 itself is pro‐fibrogenic, but promotes miR‐9‐5p expression, thus inducing inhibition of its own pro‐fibrogenic role.
Reactive oxygen species and TGF‐β1 induce miR‐9‐5p expression.
miR‐9‐5p inhibits TGFBR2 and NOX4 expression by binding to their 3′‐UTRs.
miR‐9‐5p inhibits TGF‐β‐mediated pro‐fibrogenic signaling in human lung fibroblasts and attenuates the development of experimental pulmonary fibrosis.
Peritoneal mesothelial fibrogenesis is also significantly reduced by miR‐9‐5p.
miR‐9‐5p is discovered as an anti‐fibrotic miRNA that targets TGBR2 and NOX4 to inhibit the transformation of fibroblasts into myofibroblasts. TGF‐β1 itself is pro‐fibrogenic, but promotes miR‐9‐5p expression, thus inducing inhibition of its own pro‐fibrogenic role.
Medical education should enhance empathy. We examined, using self-assessment instruments and standardized patients (SPs), the impact on empathy, of a multi-year intervention (years 4–6 of medical ...training) that uses reflective learning approaches.
241 final-year medical students participated; 110 from the 2018 graduation class (non-intervention group) and 131 from the 2019 graduation class (intervention group). Participants completed two self-reported empathy questionnaires – the Jefferson Scale of Empathy-Students (JSE-S) and the Interpersonal Reactivity Index (IRI) – and a personality questionnaire, the NEO Five-Factor Inventory. Additionally, SPs in a simulated station assessed participants’ empathy with two patient-reported instruments: the Consultation and Relational Empathy (CARE) scale and the Jefferson Scale of Patient Perceptions of Physician Empathy (JSPPPE).
Empathy scores were significantly higher in the intervention group compared to the non-intervention group when assessed by the SP (p < 0.001). No differences were found in self-reported questionnaires between the two groups.
A longitudinal, multi-year reflection-based intervention enhanced empathy amongst medical students as assessed by SPs, but not when assessed by student self-reported measures.
Multi-year reflective learning interventions during clinical training nurture empathy in medical students. Assessments completed by SPs or patients may enhance the evaluation of empathy.
•Workshops in the last three years of medical training enhanced students’ empathy.•Differences in empathy were found by standardized patient evaluations.•No differences were found by student self-reported instruments.•Increased empathy was found in female students, those with volunteers and those preferring family medicine.•Different empathy evaluation tools are closely related.
Display omitted
•Analysis of 14 aminoglycosides in milk, liver and honey samples was achieved.•Separation of aminoglycosides was obtained with zwitterionic stationary phases.•Formic acid mobile phase ...provided an excellent sensitivity.•Cation-exchange cartridges provided the best recoveries in all matrices.
In this study, fourteen highly polar aminoglycoside (AGs) antibiotics were selected. Various stationary phases were tested, including Obelisc R, ZIC-HILIC, BEH amide and aminopropyl. The nature of the stationary phase, mobile phase (water or buffer solutions and acetonitrile), pH (percentage of formic acid), gradient conditions and injection solvents were systematically studied as relevant parameters for tuning retention selectivity and detectability of AGs in liquid chromatography electrospray tandem mass spectrometry (LC–(ESI)–MS/MS). Only the two zwitterionic phases (Obelisc R and ZIC-HILIC) achieved a proper chromatographic separation considering interferences due to the crosstalk effect in low resolution mass spectrometers. The water/acetonitrile mobile phase containing 1% formic acid used with Obelisc R provided more sensitivity than the highly concentrated buffered mobile phases required for ZIC-HILIC. A solid phase extraction (SPE) clean-up procedure with polymeric weak cation exchange (WCX) cartridges was optimized for honey, milk and liver samples. Different brands of cartridges and elution solvents were tested, and the Taurus WCX offered the best recovery rate with a buffer elution at pH 3. The final optimized method was validated in these matrices according to Decision 2002/657/EC. A monitoring campaign for sixty honey, milk and liver samples was carried out at the Food Authority Control in Geneva. The concentration of dihydrostreptomycin (DSTP) found in one ovine liver exceeded the established maximum residue levels (MRLs) within the European and Swiss legislations but it was compliant taking into account the validation data.
In this review we provide an overview of the components of the spermatozoa playing an important role in reproductive success beyond fertilization, showing the relationship between the integrity of ...the diverse elements and the development of a healthy offspring. The present knowledge about fish sperm chromatin organization, epigenetic modifications of DNA and histones and sperm-borne RNAs, essential in controlling embryo development, is summarized, pointing out the possibility of using specific genes or transcripts as biomarkers of sperm quality. Data about commercial species are reported when available and more detailed information about zebrafish sperm is presented.
Considering the implications that the integrity of sperm genome and epigenome has on the preservation of a proper genotype and phenotype in the progeny, the methods applied for the study of chromatin damage and for the study of transcriptome are described. Moreover we discuss some injuring agents affecting paternal information, from the presence of contaminants in the aquatic environment, to the reproductive practices applied in fish farming. The consequences of fertilizing with damaged spermatozoa, as well as the zygotic ability to repair damage are also reviewed.
Role of the sperm on achieving high rates of healthy fries.
•Knowledge on sperm chromatin organization, epigenetic patterning and cytoplasmic RNAs is reviewed.•The effects of environmental and husbandry factors on the sperm genome, epigenome and transcriptome are reported.•The link between paternal information stability and offspring outcome highlights the need for new markers of sperm quality.
Empathy is an essential competence in the medical field. There are no validated patient-rated empathy measures in Spanish (Spain). The Consultation and Relational Empathy (CARE) measure is a widely ...used patient-rated measure of physician empathy.
To analyse the reliability and validity of the Spanish (Spain) version of the CARE measure in primary care.
After translation, back translation and pilot testing, a convenient sample of 369 patients recruited through 21 primary care physicians in five primary care centres in Pamplona (Navarre, Spain) completed the Spanish (Spain) CARE (Sp-CARE) measure. The number of 'does not apply' or blank responses was calculated to assess acceptability. We analysed internal reliability by means of Cronbach's alpha and ordinal alpha and homogeneity with corrected item-total correlations. The construct validity was examined by confirmatory factor analysis (CFA) and concurrent validity by Spearman's correlation.
We observed high acceptability; only 37 (1%) responses were marked 'does not apply' and only 3 (0.08%) were left blank. Cronbach's alpha and ordinal alpha for the Sp-CARE measure were 0.953 and 0.970, respectively, and all corrected item-total correlations exceeded the accepted cut of 0.30, demonstrating high internal reliability and homogeneity. CFA corroborated the one-factor structure proposed in the original version. The Sp-CARE measure total score was significantly correlated with overall patient satisfaction (Spearman's rho 0.45, P < 0.001).
The results support the reliability and validity of the Sp-CARE measure as a patient-rated empathy measure in the primary care setting.
PDF
