Does neutral red suit the requirements of being a reliable indicator in clearance rate measurements of suspension-feeding bivalves? An empirical refutation

One basic requirement in the measurements of feeding rates in suspension-feeding bivalves when using clearance methods is that the suspended particles that are used in the determination of these rates must reach size compatibility with full retention by the gill. In spite of this requirement, clearance of neutral red (NR) -a soluble vital dye- has long been used for this purpose and recourse to this method has even increased over the last years for determining the filtration rate as a specific physiological response to stress in ecotoxicological and biomonitoring studies. The aim of this study was to produce empirical evidence that would dismiss the suitability of the NR method for providing reliable measurements for feeding rates in suspension feeders. Experiments that were designed to this end in the blue mussel Mytilus galloprovincialis led to three main conclusions: 1) Temporal dynamics of NR retention by the mussels point to a diffusion mechanism whereby the marker would be up-taken by gill tissue rather than filtered by the muco-ciliary action of this organ. 2) The NR method largely underestimates the clearance rate (by 90% on average) when compared with conventional methods that use microscopic particles in the size-range of gill retention. 3) The close similitude between rates of retention of NR by intact mussels and mussels that had been sacrificed by sectioning apart the valves adductor muscle proves outright the lack of a quantitative relationship between this retention process and the pumping activity.This research was funded by the GIU 17/061 project from the UPV/EH

Polycomb Factor PHF19 Controls Cell Growth and Differentiation Toward Erythroid Pathway in Chronic Myeloid Leukemia Cells

Leucèmia mieloide crònica; Diferenciació eritroïdal; PolycombLeucemia mieloide crónica; Diferenciación eritroide; PolycombChronic myeloid leukemia; Erythroid differentiation; PolycombPolycomb group (PcG) of proteins are a group of highly conserved epigenetic regulators involved in many biological functions, such as embryonic development, cell proliferation, and adult stem cell determination. PHD finger protein 19 (PHF19) is an associated factor of Polycomb repressor complex 2 (PRC2), often upregulated in human cancers. In particular, myeloid leukemia cell lines show increased levels of PHF19, yet little is known about its function. Here, we have characterized the role of PHF19 in myeloid leukemia cells. We demonstrated that PHF19 depletion decreases cell proliferation and promotes chronic myeloid leukemia (CML) differentiation. Mechanistically, we have shown how PHF19 regulates the proliferation of CML through a direct regulation of the cell cycle inhibitor p21. Furthermore, we observed that MTF2, a PHF19 homolog, partially compensates for PHF19 depletion in a subset of target genes, instructing specific erythroid differentiation. Taken together, our results show that PHF19 is a key transcriptional regulator for cell fate determination and could be a potential therapeutic target for myeloid leukemia treatment.This work was supported by the Di Croce Laboratory is supported by grants from the Spanish Ministry of Science and Innovation (BFU2016-75008-P and PID2019-108322GB-100), “Fundación Vencer El Cancer” (VEC), the European Regional Development Fund (FEDER), and from AGAUR (SGR 2017-2019). We acknowledge the support of the Spanish Ministry of Science and Innovation to the EMBL partnership, the Centro de Excelencia Severo Ochoa and the CERCA Programme/Generalitat de Catalunya. PV was supported by the Fundación Científica de la Asociación Española Contra el Cáncer. SA was funded by the Ramon y Cajal program of the Ministerio de Ciencia, Innovación y Universidades, the European Social Fund under the reference number RYC-2018-025002-I, and the Instituto de Salud Carlos III-FEDER (PI19/01814)

DNA Methylation of the Gonadal Aromatase (cyp19a) Promoter Is Involved in Temperature-Dependent Sex Ratio Shifts in the European Sea Bass

15 pages, 9 figures, 1 tableTemperature changes during early embryonic and/or larval stages are able to modify sex ratios in fish and reptiles. However, the underlying mechanism by which temperature is able to modify the molecular pathways that developing gonads follow to become ovaries or testes is still unknown. One of the most interesting questions raised from previous studies with our model species, the European sea bass, was how temperature could affect the developmental fate of the gonads at a time when they were not even formed in the most rudimentary manner. This was the telltale sign of an epigenetic mechanism. In this study, DNA methylation levels of the aromatase promoter were analyzed in European sea bass exposed to different temperatures during early developmental stages. Aromatase is the enzyme that converts androgens (male hormones) into estrogens (female hormones), which are essential for ovarian development in all non-mammalian vertebrates. We show that increased temperature during a critical period in early development is able to increase DNA methylation of the aromatase promoter, preventing aromatase gene expression. We conclude that gonadal aromatase promoter methylation is most likely part of the long-sought-after mechanism connecting temperature and environmental sex determination in vertebratesLN-M and ND were supported by predoctoral scholarships and JV and LR by postdoctoral grants from the Spanish Ministry of Science and Innovation (MCINN). Research was funded by MCINN projects “Sexgene” (AGL2006-01359), “Aquagenomics” (CDS2007-0002), and “Epigen-Aqua” (AGL2010-15939) to FP and by MICINN project BFU2010-18692 and AGAUR grant to LDC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscriptPeer reviewe

Variable Capacity for Acute and Chronic Thermal Compensation of Physiological Rates Contributes to Inter-Individual Differences in Growth Rate in Mussels (Mytilus galloprovincialis)

The aim of this study was to ascertain if the capacity for acute and chronic compensation of thermal effects on physiological rates represents a trait contributing to inter-individual growth rate differences in the mussel Mytilus galloprovincialis. Juvenile mussels (10– 11 mm shell length) were collected, transported to the laboratory, and divided into two groups: one group was maintained at 20 C (warm treatment), and the other at 10 C (cold treatment). The mussels were reared at these two temperatures (continuously fed), until clear size differences allowed us to select fast- (F) and slow (S)-growing individuals from both groups (F20/S20 at 20 C and F10/S10 at 10 C). Selected F and S mussels were then exposed to three experimental temperatures (10, 15, and 20 C), and the time-course of their response, in terms of clearance rate (CR: L/h) and routine oxygen consumption (VO2: mLO2/h), was monitored. The overall growth rate of mussels in the warm treatment group was significantly higher than in the cold treatment group. For both treatments, significant differences were found in key physiological parameters between F and S mussels: F mussels had a higher CR and a larger gill surface area than their S counterparts. Although no significant differences in the thermal sensitivity of the clearance or metabolic rates were observed between F20 and S20 mussels reared at 20 C, when exposed to acute temperature changes, experiments with mussels reared at 10 C revealed a different outcome: in response to acute warming (from 10 C to 15 and 20 C), F10 were capable of compensating for the thermal effect on CR and VO2; however, no such compensatory response was observed in S10. We conclude that two significant factors contribute to endogenous differences in the growth rate of mussels: (i) the capacity to exhibit intense filtering activity, which appears to be functionally correlated with the gill surface area and (ii) the capacity to compensate for the effects of temperature on filtration and metabolic rate. The second trait does not appear to make a significant contribution to the inter-individual size-differentiation observed in mussels maintained in warm environments, but explains a significant proportion of inter-individual growth rate differences in cold environments.DP was funded by an FPI grant from the Basque Government. A part of this study was also funded by the Spanish Ministry of Economy and Competitiveness (Reference: AGL2013-49144-C3-1-R)

Methods for assessment of body tissue composition as an indication of the energetic status in bivalve populations: A comparison of biochemical and elemental analysis

Elemental (CHN) and proximate biochemical compositions of body tissues are useful tools among the ecological indices most commonly used in evaluation of the energetic status of animal populations. Gnaiger and Bitterlich (1984) supplied procedures for the interconversion between both these measurements based on stoichiometric relationships, that were further tested using gut contents and body tissue samples of freshwater fishes. Despite a lack of validation studies, the reported conversion factors have been broadly applied in the field of body composition analysis of marine invertebrates, especially bivalve mollusks. The aim of this study was to test the applicability of those conversion equivalents in analysis of the body tissues of two congeneric clam species (Ruditapes decussatus and Ruditapes philippinarum). To this end, proximate biochemical composition, as analyzed by fractionated extraction of tissues samples and quantification using spectrophotometric methods, was converted to elemental composition, and the resulting figures were compared with those of direct CHN analysis. The results of this comparison indicate good agreement within the ranges reported, provided that ninhydrin positive substances (NPS) are incorporated in the biochemical composition analyses. The magnitude of the nonprotein N component in bivalve tissues appears to complicate the reverse computation of biochemical components from elemental composition because no accurate estimation of proteins from N contents might be possible. Additionally, a specific correction of residual water in dried samples of bivalve tissues for CHN analysis should be applied. The absence of broad differences found between species reflects the morphological, evolutionary and functional proximity between them, whereas tissue differences display the differential role that each organ plays in the organism, although other sources of variability such as diet and sex should be addressed in future research.This work was supported by the Spanish Ministry of Economy and Competitiveness (AGL2013-49144-C3-1-R). K. Arranz was funded by a predoctoral research grant from Universidad del Pais Vasco/Euskal Herriko Unibertsitatea (UPV/EHU). SGIker technical and human support (UPV/EHU, MICINN, GV/EJ, ESF) is gratefully acknowledged

Parathyroid Hormone-Related Protein Promotes Inflammation in the kidney with an Obstructed ureter

Parathyroid hormone-related protein (PTHrP) promotes fibrogenesis in the acutely damaged kidney. Considering the relation between fibrosis and inflammation, we studied transgenic mice that overexpress PTHrP in the proximal tubule. When unilateral ureteric obstruction was induced in these transgenic mice, we found that they had more renal tubulointerstitial damage, leukocyte influx, and expression of proinflammatory factors than their control littermates. Reversal of PTHrP constitutive overexpression in these transgenic mice or treatment of control mice with the PTHrP antagonist (7–34) decreased this inflammatory response. Losartan, which abolished obstruction-induced endogenous PTHrP upregulation, also decreased the latter response but less effectively in transgenic mice. The PTHrP fragment (1–36) induced nuclear factor-jB (NF-jB) activation and proinflammatory cytokine overexpression in mouse cortical tubule cells in culture as well as migration of the macrophage cell line Raw 264.7. All these effects were decreased by PTHrP (7–34) and NF-jB or extracellular signal-regulated kinase (ERK) activation inhibitors. Our findings suggest a critical role of PTHrP in the renal inflammatory process that results from ureteral obstruction and indicate that ERK-mediated NF-jB activation seems to be an important mechanism whereby PTHrP triggers renal inflammatio

The use of an interstitial boost in the conservative treatment of breast cancer: how to perform it routinely in a radiotherapy department

Purpose: to demonstrate the utility of a boost with interstitial brachytherapy (BT) in breast-conserving therapy (BCT) by doing a thorough review of the literature and describing in detail our technique for delivering this boost. Material and methods: our department has been delivering the boost with interstitial BT since 1989, in most cases with rigid needles and a theoretical dosimetry. In the early years, we used low-dose-rate (LDR) with iridium-192 wires. The dose administered was 15 Gy if there were no risk factors for local relapse or 20-25 Gy in the presence of risk factors. The risk factors considered were the presence of a close margin (less than 10 mm) and an extensive intraductal component (more than 25%). After 2002, we switched to high-dose-rate (HDR); using the linear quadratic model we changed the low dose to 3 fractions of 4.5 Gy in the case of no risk factors for local relapse or to 3 fractions of 5 Gy in the presence of risk factors. Results: in 79 consecutive boost patients treated in our department between 2010 and 2011, with a median follow-up of 46 months, the local control rate was 97.47%. With respect to cosmesis, fibrosis occurred in 17 cases (21.5%) and hyperpigmentation in 26 cases (32.9%). Our hospital's results are comparable in terms of local control and cosmesis to those of other authors. Conclusions: this educational article describes our department's boost technique with rigid needles and comments briefly on our results using this technique in a group of consecutively treated patients in our department. A review of the literature and the published results on local control and cosmesis is also described

Blunted Expansion of Regulatory T Lymphocytes Is Associated With Increased Bacterial Translocation in Patients With Major Depressive Disorder

Background: Major Depressive Disorder (MDD) is associated with both proinflammatory and adaptive immune response abnormalities. Regulatory T lymphocytes (Tregs), a subtype of CD4+ T cells, are relevant for maintaining immune-inflammatory system homeostasis and control of inflammation such as the kind potentially induced by the interactions between the intestinal microbiome and gut mucosa. We investigated the Treg population and its distribution along their stages of differentiation/activation, as well as its function in MDD patients without concomitant diseases. We also studied the potential association between Treg alterations, intestinal barrier damage, and bacterial translocation. Methods: 30 MDD patients and 20 healthy controls were studied. The levels of circulating CD25FoxP3+ Tregs and their distribution on the naïve (TN), effector (TE), central (TCM), and effector memory(TEM) differentiation/activation stages were analyzed using polychromatic flow cytometry. Chemokine receptors (CCR) 2, 5, and 6, and the intracytoplasmic IL-10 expression by the Tregs were also analyzed. The serum IL-10 was measured using Luminex. The serum levels of zonulin and the intestinal fatty acid-binding protein (I-FABP), both markers of gut barrier function, and the LPS-binding protein (LBP), a marker of bacterial translocation, were measured using an enzyme-linked immunosorbent assay. Results: MDD patients had increased number of circulating Tregs cells with enhanced number of Tregs at the TN, TE, TCM, and TEM stages. The percentage of Tregs cells at TN stage was significantly higher in MDD patients. The percentage of Tregs that expressed CCR2 and CCR6 was increased as well as those expressing IL-10. MDD patients had significantly increased levels of circulating I-FABP and LBP. MDD patients with high LBP levels had a significant reduction in the number of circulating Tregs compared to normal-LBP MDD patients. Conclusions: MDD patients showed an expansion of circulating Tregs and their CD25highFoxP3+ and CD25lowFoxP3+ subsets throughout the different stages of CD4+ T lymphocyte differentiation/activation. Tregs also showed an increased frequency of cells expressing CCR6 and CCR2. IL-10 Treg production was also enhanced in MDD patients that concurrently had increased serum IL-10 levels. However, this Treg expansion was blunted in MDD patients with gut barrier damage and increased bacterial translocation.Instituto de Salud Carlos IIIComunidad de Madri

Crecimiento y supervivencia de las paralarvas del pulpo Octopus vulgaris con diferentes dietas durante el primer mes de vida

Octopus vulgaris paralarvae survival and growth during first month of live were analysed using three different enrichm ent diet types. No significant differences were observed in both paralarvae growth and survival between diets. However, significant mortality differences were observed. Diet based on Artemia salina with Ori-green supplement give the best survival p ercentage (13,11%) comparing with diets using A. salina with Isochrisys galbana (0,78%) or Multigain (1,73%) supplement

High magnetization FeCo nanoparticles for magnetorheological fluids with enhanced response

We present results concerning the fabrication of a new magnetorheological fluid with FeCo magnetic nanoparticles (NPs) as magnetic fillers. These NPs have been fabricated by using the chemical reduction technique and show a pure crystalline phase with size ranging among 30–50 nm and high magnetization, 212 ± 2 A m2 kg−1. They agglomerate due to the strong magnetic dipolar interaction among them. These FeCo nanoparticles were used to synthesize a magnetorheological fluid by using oleic acid as surfactant, mineral oil as carrier liquid and Aerosil 300 as additive to control the viscosity of the fluid. The synthesized fluid showed a strong magnetorheological response with increasing shear stress values as the magnetic field intensity increases. Thus, we have measured a superior performance up to 616.7 kA m−1, with a yield stress value of 2729 Pa, and good reversibility after demagnetization process. This value competes with the best ones reported in the most recent literature. We have compared the obtained results with our previous reported ones by using high magnetization Fe NPs fabricated by the electrical explosion of wire method (Fe-EEW).J. Berasategi, A. Gómez and M. M. Bou-Ali would like to thank the financial support provided by the Basque Government under research project PI-2017-1-0055 and MMMfavIN (KK-2020/00099, Elkartek program). V. Vadillo, J. Gutiérrez, J.M. Barandiarán, M. Insausti and I. Gil de Muro would like to thank the financial support provided also by the Basque Government under PI-2017-1-0043, the MMMfavIN (KK-2020/00099, Elkartek program) and Research Groups (IT1245-19 and IT1226-19) research projects. A. A. and A. I. gratefully acknowledge the financial support of the Basque Government (Research Groups IT-1175-19) and the Ministerio de Economía y Competitividad (PGC2018-094548-B-I00, MCIU/AEI/FEDER, UE).Peer reviewe