Structural and functional divergence of two fish aquaporin-1 water channels following teleost-specific gene duplication

<p>Abstract</p> <p>Background</p> <p>Teleost radiation in the oceans required specific physiological adaptations in eggs and early embryos to survive in the hyper-osmotic seawater. Investigating the evolution of aquaporins (AQPs) in these vertebrates should help to elucidate how mechanisms for water homeostasis evolved. The marine teleost gilthead sea bream (<it>Sparus aurata</it>) has a mammalian aquaporin-1 (AQP1)-related channel, termed AQP1o, with a specialized physiological role in mediating egg hydration. However, teleosts have an additional AQP isoform structurally more similar to AQP1, though its relationship with AQP1o is unclear.</p> <p>Results</p> <p>By using phylogenetic and genomic analyses we show here that teleosts, unlike tetrapods, have two closely linked AQP1 paralogous genes, termed <it>aqp1a </it>and <it>aqp1b </it>(formerly AQP1o). In marine teleosts that produce hydrated eggs, <it>aqp1b </it>is highly expressed in the ovary, whereas in freshwater species that produce non-hydrated eggs, <it>aqp1b </it>has a completely different expression pattern or is not found in the genome. Both Aqp1a and Aqp1b are functional water-selective channels when expressed in <it>Xenopus laevis </it>oocytes. However, expression of chimeric and mutated proteins in oocytes revealed that the sea bream Aqp1b C-terminus, unlike that of Aqp1a, contains specific residues involved in the control of Aqp1b intracellular trafficking through phosphorylation-independent and -dependent mechanisms.</p> <p>Conclusion</p> <p>We propose that 1) Aqp1a and Aqp1b are encoded by distinct genes that probably originated specifically in the teleost lineage by duplication of a common ancestor soon after divergence from tetrapods, 2) Aqp1b possibly represents a neofunctionalized AQP adapted to oocytes of marine and catadromous teleosts, thereby contributing to a water reservoir in eggs and early embryos that increases their survival in the ocean, and 3) Aqp1b independently acquired regulatory domains in the cytoplasmatic C-terminal tail for the specific control of Aqp1b expression in the plasma membrane.</p

Development and field evaluation of liquid inoculants with native Bradyrhizobial strains for peanut production

A critical process in the leguminous crops cycles is biological nitrogen fixation (BNF). Application of inoculants with N fixing bacteria is economically and environmentally favourable. The aim of this work was to select competitive native peanut microsymbionts, evaluate their survival in inoculant support and assess their impact on peanut (Arachis hypogaea L.) production under field conditions at Córdoba province in Argentina. The efficient N fixing Bradyrhizobium sp. J-81 and Bradyrhizobium sp. J-237, previously obtained from peanut nodules in the region of Cordoba, Argentina, were evaluated. In microcosm assays, plants inoculated with these isolates demonstrated better symbiotic parameters than those inoculated with reference strains. Different bacterial growth media and inoculant stabiliser solutions were evaluated. Balanced medium and arabic gum stabilising solution had optimal bacterial growth and the highest bacterial concentration and viability, respectively. Inoculation with either inoculants resulted in 44% greater peanut pod yield at Pizarro compared to the non-inoculated plants, although no significant differences were found with respect to commercial inoculants treatments.La fixation biologique de l’Azote (FBA) est un processus important dans le cycle de vie des légumineuses. L’application d’inoculum de bactéries fixatrices d’azote est favorable au double plan économique et environnemental. Le but de cette étude était de sélectionner des bactéries symbiotiques de l’arachide natives et compétitives, évaluer leur temps de survie dans support d’inoculum et évaluer leur impact sur la production en plein champ de l’arachide (Arachis hypogaea L.) dans la province de Córdoba en Argentine. Les bactéries fixatrices d’azote Bradyrhizobium sp. J-81 et Bradyrhizobium sp. J-237, extraites de nodules collectés sur des plants cultivés dans la région de Cordoba en Argentina, ont été évaluées. Dans des essais de microcosme, des plants inoculés avec ces isolats ont exhibés de meilleurs paramètres symbiotiques que les plants non inoculés. Différents média de culture bactérienne et supports inoculums ont été testés. Medium mixte et solution stabilisée à la gomme arabique ont respectivement exhibés la croissance optimale des bactéries et la meilleure conservation et viabilité des bactéries. L’application de n’importe quel inoculum produisit 44% plus de rendement en gousses d’arachides à Pizarro par rapport aux plantes non-inoculées, et ceci bien qu’aucune différence significative n’a été observée en comparaison avec les traitements à l’inoculum du commerce.Fil: Valetti, Lucio. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Angelini, Jorge Guillermo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Taurian, Tania. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Ibañez, Fernando Julio. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Muñoz, Vanina Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Anzuay, María Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; ArgentinaFil: Ludueña, Liliana Mercedes. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Fabra, Adriana Isidora. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentin

Isolation of a novel aquaglyceroporin from a marine teleost (Sparus auratus) Function and tissue distribution

The aquaporins (formerly called the major intrinsic protein family) are transmembrane channel proteins. The family includes the CHIP group, which are functionally characterised as water channels and the GLP group, which are specialised for glycerol transport. The present study reports the identification and characterisation of a novel GLP family member in a teleost fish, the sea bream Sparus auratus. A sea bream aquaporin (sbAQP) cDNA of 1047·bp and encoding a protein of 298·amino acids was isolated from a kidney cDNA library. Functional characterization of the sbAQP using a Xenopus oocyte assay revealed that the isolated cDNA stimulated osmotic water permeability in a mercury-sensitive manner and also stimulated urea and glycerol uptake. Northern blotting demonstrated that sbAQP was expressed at high levels in the posterior region of the gut, where two transcripts were identified (1.6·kb and 2·kb), and in kidney, where a single transcript was present (2·kb). In situ hybridisation studies with a sbAQP riboprobe revealed its presence in the lamina propria and smooth muscle layer of the posterior region of the gut and in epithelial cells of some kidney tubules. sbAQP was also present in putative chloride cells of the gill. Phylogenetic analysis of sbAQP, including putative GLP genes from Fugu rubripes, revealed that it did not group with any of the previously isolated vertebrate GLPs and instead formed a separate group, suggesting that it may be a novel GLP member.This work was supported by project PRAXIS XXI/2/2.1/BIA/211/94 from the Portuguese National Science and Technology Foundation (FCT), co-financed by EU structural funds, DG-Fisheries Project Q5RS-2002-00784 (CRYOCYTE) and an EU Biotech grant (QLRT2000-00778). C.R.A.S., J.C.R.C. and J.F. were in receipt of FCT fellowships PRAXIS XXI/BPD/22040/99, PRAXIS XXI/BD/19925/99BPD/22033/99, respectively

Role of bacterial pyrroloquinoline quinone in phosphate solubilizing ability and in plant growth promotion on strain <i>Serratia</i> sp. S119

The aim of this study was to analyze if cofactor pyrroquinoline quinone from Serratia sp. S119 is involved in the inorganic phosphate solubilization mechanism and in its ability to promote the plant growth. Site directed mutagenesis was performed to obtain a pqqE- minus mutant of strain Serratia sp. S119. The phosphate solubilization ability, gluconate and PQQ production of the mutant Serratia sp. RSL (pqqE-) was analyzed. Mutant RSL (pqqE-) showed significant decrease in P soluble and gluconic acid levels produced and undetectable levels of PQQ cofactor compared with wild-type strain. Complementation with synthetic PQQ cofactor restored P solubilization and gluconate production reaching the levels produced by wild-type strain. PqqE gene sequence indicated that it is highly conserved within Serratia strains and its product shows conserved motifs found in other PqqE proteins of several bacteria. The effect of the inoculation of the PQQ- mutant on peanut and maize plants was evaluated in pot assays. Plants growth parameters showed no differences among the different treatments indicating that PQQ from Serratia sp. S119 is not involved in the growth promotion of these plants. PQQ cofactor is essential for phosphate solubilization ability of Serratia sp. S119 but is not required for growth promotion of peanut and maize plants.Facultad de Ciencias ExactasCentro de Investigación y Desarrollo en Fermentaciones Industriale

Functional properties of sodium and calcium caseinate antimicrobial active films containing carvacrol

Active edible films were prepared by adding carvacrol into sodium caseinate (SC) and calcium caseinate (CC) matrices plasticized with two different glycerol concentrations (25 and 35 wt%) prepared by solvent casting. Functional characterisation of these bio-films was carried out by determination of some of their physico-chemical properties, such as colour, transparency, oxygen barrier, wettability, dye permeation properties and antibacterial effectiveness against Gram negative and Gram positive bacteria. All films exhibited good performance in terms of optical properties in the CIELab space showing high transparency. Carvacrol was able to reduce CC oxygen permeability and slightly increased the surface hydrophobicity. Dye diffusion experiments were performed to evaluate permeation properties. The diffusion of dye through films revealed that SC was more permeable than CC. The agar diffusion method was used for the evaluation of the films antimicrobial effectiveness against Escherichia cell and Staphylococcus aureus. Both SC and CC edible films with carvacrol showed inhibitory effects on both bacteria. (C) 2013 Elsevier Ltd. All rights reserved.This research was supported by the Ministry of Science and Innovation of Spain through the projects MAT2011-28468-C02-01, MAT2011-28468-C02-02 and HP2008-0080. M.P. Arrieta thanks Fundacion MAPFRE for "Ignacio Hernando de Larramendi 2009-Medio Ambiente" fellowship (MAPFRE-IHL-01). Authors thank Ferrer Alimentacion S.A., for providing the caseinates powders.Arrieta, MP.; Peltzer, MA.; López Martínez, J.; Garrigós Selva, MDC.; Valente, AJM.; Jimenez Migallon, A. (2014). Functional properties of sodium and calcium caseinate antimicrobial active films containing carvacrol. Journal of Food Engineering. 121:94-101. https://doi.org/10.1016/j.jfoodeng.2013.08.015S9410112

Effects of a primary care-based multifactorial intervention on physical and cognitive function in frail, elderly individuals : A randomized controlled trial

Background: Detecting and managing frailty at early stages can prevent disability and other adverse outcomes. The study aim was to evaluate whether a multifactorial intervention program could modify physical and cognitive frailty parameters in elderly individuals. Methods: We conducted a multicenter, randomized, single-blind, parallel-group trial in community-living prefrail/frail elderly individuals in Barcelona. A total of 352 patients, aged ≥65 years old with positive frailty screening, was randomized into two groups to receive a 12-week multidisciplinary intervention or usual care, with concealed allocation. The intervention consisted of: exercise training, intake of hyperproteic nutritional shakes, memory training, and medication review. Main outcome assessments with multivariate analysis were conducted at 3 and 18 months. Results: A total of 347 participants (98.6%) completed the study, mean age 77.3 years, 89 prefrail subjects (25.3%), and 75.3% female (n = 265). Eighteen-month assessments were performed in 76% of the sample. After 3 and 18 months, adjusted means difference between-groups showed significant improvements for the intervention group in all comparisons: Short Physical Performance Battery score improved 1.58 and 1.36 points (p <.001), handgrip strength 2.84 and 2.49 kg (p <.001), functional reach 4.3 and 4.52 cm (p <.001), and number of prescriptions decreased 1.39 and 1.09 (p <.001), respectively. Neurocognitive battery also showed significant improvements across all dimensions at 3 and 18 months. Conclusions: A physical, nutritional, neurocognitive, and pharmacological multifaceted intervention was effective in reversing frailty measures both at short-term and 18 months. Lasting benefits of a multi-intervention program among frail elderly individuals encourage its prioritization

Endoglin overexpression modulates cellular morphology, migration, and adhesion of mouse fibroblasts

10 p.-9 fig.-1 tab.Endoglin is the gene mutated in hereditary hemorrhagic telangiectasia type 1 (HHT1), a dominantly inherited vascular disorder. Endoglin glycoprotein is a component of the transforming growth factor type ß (TGF-ß) receptor system which is highly expressed by endothelial cells, and at lower levels on fibroblasts and smooth muscle cells, suggesting the involvement of these lineages in the HHT1 vascular dysplasia. Overexpression of endoglin in mouse NCTC929 fibroblasts led to decreased migration in chemotactic and wound healing assays, as well as changes in the cellular morphology. When plated on uncoated surfaces, endoglin transfectants formed intercellular clusters, endoglin being not specifically localized to the cell-cell junctions, but homogenously distributed on the cellular surface. Although the expression of α5ß1 integrin and of an activation epitope of ß1 integrin were unchanged, a polyclonal antibody to α5ß1 integrin was able to inhibit cluster formation, suggesting the involvement of integrin ligand/s. In fact, coating with fibronectin, laminin, or an RGD-containing 80 kDa fragment of fibronectin were able to prevent the cellular clustering. Furthermore, synthesis of plasminogen activator inhibitor 1 (PAI-1), and to a weak extent that of fibronectin, were inhibited in endoglin transfectants. Thus, the presence of endoglin in mouse NCTC929 fibroblasts is associated with reduced production of certain extracellular matrix (ECM) components, which might explain their altered morphology, migration and intercellular cluster formation.This work has been supported by grants from Camisión Interministerial de Ciencia yTecnología (CICYT-SAF97-0034 to C. Bernabéu, and CICYT-SAF97-0064-C03-02 to A. García-Pardo), Comunidad Autónoma de Madrid (CAM) and Biomed Program of the European Community (BMH4-CT95-0995 to C. Bernabéu).Peer reviewe

Structure and mechanical properties of sodium and calcium caseinate edible active films with carvacrol

Edible active films based on sodium caseinate (SC) and calcium caseinate (CC) plasticized with glycerol (G) at three different concentrations and carvacrol (CRV) as active agent were prepared by solvent casting. Transparent films were obtained and their surfaces were analysed by optical microscopy and scanning electron microscopy (SEM). The influence of the addition of three different plasticizer concentrations was studied by determining tensile properties, while Fourier transformed infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA) were used to evaluate the structural and thermal behavior of such films. The addition of glycerol resulted in a reduction in the elastic modulus and tensile strength, while some increase in the elongation at break was observed. In general terms, SC films showed flexibility higher than the corresponding CC counterparts. In addition, the presence of carvacrol caused further improvements in ductile properties suggesting the presence of stronger interactions between the protein matrix and glycerol, as it was also observed in thermal degradation studies. FTIR spectra of all films showed the characteristic bands and peaks corresponding to proteins as well as to primary and secondary alcohols. In summary, the best results regarding mechanical and structural properties for caseinates-based films containing carvacrol were found for the formulations with high glycerol concentrations.Marina Patricia Arrieta thanks Fundacion MAPFRE for "Ignacio Hernando de Larramendi 2009- Medio Ambiente" fellowship (MAPFRE-IHL-01). The Spanish Ministry of Economy and Competitiveness is acknowledged by financial support (project Ref. MAT2011-28468-C02-01). Authors thank to Ferrer Alimentacion S.A., for providing caseinates and to Prof. Juan Lopez Martinez (Polytechnic University of Valencia, Spain) for his collaboration and useful discussions.Arrieta, MP.; Peltzer, MA.; Garrigós, MDC.; Jimenez, A. (2013). Structure and mechanical properties of sodium and calcium caseinate edible active films with carvacrol. Journal of Food Engineering. 114(4):486-494. https://doi.org/10.1016/j.jfoodeng.2012.09.002S486494114

Differential localization and regulation of two aquaporin-1 homologs in the intestinal epithelia of the marine teleost Sparus aurata

11 pages, 8 figuresDifferential localization and regulation of two aquaporin-1 homologs in the intestinal epithelia of the marine teleost Sparus aurata. Am J Physiol Regul Integr Comp Physiol 294: R993–R1003, 2008. First published January 2, 2008; doi:10.1152/ajpregu.00695.2007.—Aquaporin (AQP)-mediated intestinal water absorption may play a major osmoregulatory role in euryhaline teleosts, although the molecular identity and anatomical distribution of AQPs in the fish gastrointestinal tract is poorly known. Here, we have investigated the functional properties and cellular localization in the intestine of two gilthead seabream (Sparus aurata) homologs of mammalian aquaporin-1 (AQP1), named SaAqp1a and SaAqp1b. Heterologous expression in Xenopus laevis oocytes showed that SaAqp1a and SaAqp1b were water-selective channels. Real-time quantitative RT-PCR and Western blot using specific antisera indicated that abundance of SaAqp1a mRNA and protein was higher in duodenum and hindgut than in the rectum, whereas abundance of SaAqp1b was higher in rectum. In duodenum and hindgut, SaAqp1a localized at the apical brush border and lateral membrane of columnar enterocytes, whereas SaAqp1b was detected occasionally and at very low levels at the apical membrane. In the rectum, however, SaAqp1a was mainly accumulated in the cytoplasm of a subpopulation of enterocytes spread in groups over the surface of the epithelia, including the intervillus pockets, whereas SaAqp1b was detected exclusively at the apical brush border of all rectal enterocytes. Freshwater acclimation reduced the synthesis of SaAqp1a protein in all intestinal segments, but it only reduced SaAqp1b abundance in the rectum. These results show for the first time in teleosts a differential distribution and regulation of two functional AQP1 homologs in the intestinal epithelium, which suggest that they may play specialized functions during water movement across the intestineThis work was supported by Grants from the Spanish Ministry of Education and Science (MEC; AGL2001-0364/ACU and AGL2004-00316) and the European Commission (Q5RS-2002-00784-CRYOCYTE) and by the Reference Center in Aquaculture (Generalitat de Catalunya, Spain) to J. Cerdà. Participation of M. Fabra and D. Raldúa was financed by a fellowship from the Catalan Government (DURSI, Spain) and by a “Ramón y Cajal” contract (MEC), respectively.Peer reviewe

Ovarian cysteine proteinases in the teleost Fundulus heteroclitus: Molecular cloning and gene expression during vitellogenesis and oocyte maturation

13 pages, 6 figures, 2 tablesThe cysteine proteinases cathepsins B and L are members of the multigene family of lysosomal proteases that have been implicated in the processing of yolk proteins (YPs) in teleost oocytes. However, the full identification of the type of cathepsins expressed in fish ovarian follicles and embryos, as well as their regulatory mechanisms and specific function(s), are not yet elucidated. In this study, cDNAs encoding cathepsins B, L, F, K, S, Z, C, and H have been isolated from the teleost Fundulus heteroclitus, and the analysis of their deduced amino acid sequences revealed highly similar structural features to vertebrate orthologs, and confirmed in this species the existence of cathepsin L-like, cathepsin B-like, and cathepsin F-like subfamilies of cysteine proteinases. While all identified cathepsins were expressed in ovarian follicles, the corresponding mRNAs showed different temporal expression patterns. Thus, similar mRNA levels of cathepsins L, F, S, B, C, and Z were found throughout the oocyte growth or vitellogenesis period, whereas those for cathepsin H and K appeared to decrease as vitellogenesis advanced. During oocyte maturation, a transient accumulation of cathepsins L, S, H, and F mRNAs, approximately a 3-, 1.5-, 1.6-, and 6-fold increase, respectively, was detected in ovarian follicles within the 20–25 hr after hormone stimulation, coincident with the maximum proteolysis of the oocyte major YPs. The specific temporal pattern of expression of these genes may indicate a potential role of cathepsin L-like and cathepsin F proteases in the YP processing events occurring during fish oocyte maturation and/or early embryogenesis. © 2004 Wiley-Liss, Inc.Peer reviewe