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In the tropical and subtropical coastal zone, were highly diverse fish communities occur, it is important to study the small scale fisheries exploiting these communities. For this study, 219 fishing days were carried out in Bahia de Navidad, Jalisco, Mexico. Four gillnets with different mesh sizes (76.2, 88.9, 101.6 and 114.3 mm) were used for the fishing operations, from April 1994 to March 1995 and from January 1998 to December 2000. A total of 26126 organisms weighting 11680 kg were caught. One hundred and eighty three species belonging to 57 families and 19 orders were identified. Ten species accumulated more than 60% of the total abundance and biomass, the remaining species (173) individually contributed with less than 2% of the total abundance and biomass. The most important species in the catch were Microlepidotus brevipinnis, Caranx caballus, Haemulon flaviguttatum, Scomberomorus sierra, C. caninus, C. sexfasciatus, Lutjanus guttatus and L. argentiventris. The catch per unit of effort showed a large variation during the study, both in number and biomass. On average, 110 fishes and 48.5 kg fishing day-1 were caught. The maximum values were recorded during January and October 1998 (250 fishes and 100 kg fishing day-1), and the minimum values were recorded during September and November 1994 (25 org. and 10 kg fishing day-1). The total abundance and biomass was lower during the 1994-95 period (F=6.16, 8.32, P<0.05). There were statistically significant differences among the annual catch from each gillnet. All the environmental changes caused by the ENSO event had ecological and economic effects that can be rated from moderate to severe. [ABSTRACT FROM AUTHOR]

Abstract: The effect of the water-soluble fraction of crude oil (WSF) on lipid metabolism was studied at critical metabolic points, namely fatty acid activation, enzymes of triacylglycerol and phospholipid synthesis, and membrane (lipid packing) properties in the freshwater prawn Macrobrachium borellii. To determine the effect of the contaminant, adults and embryos at different stages of development were exposed to a sublethal concentration of WSF for 7 days. After exposure, microsomal palmitoyl-CoA synthetase (ACS) showed a two-fold increase in adult midgut gland. Embryo''s ACS activity was also affected, the increment being correlated with the developing stage. Endoplasmic reticulum acylglycerol synthesis was also increased by WSF exposure in adults and stage 5 embryos, but not at earlier stages of development. Triacylglycerol synthesis was particularly increased (18.5%) in adult midgut gland. The microsomal membrane properties were studied by fluorescent steady-state anisotropy, using the rotational behavior of the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH). Microsomes from midgut gland of WSF-exposed prawn showed no differences in fluidity. Nevertheless, microsomes incubated with WSF in vitro increased their fluidity in a temperature- and WSF concentration-dependent fashion. Both, aliphatic and aromatic hydrocarbons individually tested elicited an increase in membrane fluidity at 10mg/l, but at 4mg/l only nC10–C16 aliphatics did. In vivo results indicate that WSF increased the activity of microsomal enzymes that are critical in lipid metabolism, though this change was not due to direct alterations in membrane fluidity, suggesting a synthesis induction, or an enzyme-regulatory mechanism. Nevertheless, hydrocarbons elicited membrane fluidity alterations in in vitro experiments at concentrations that could be found in the environment after an oil spill. [Copyright &y& Elsevier]

The relationship between protein stability and functional evolution is little explored in proteins purified from natural sources. Here, we investigated a novel family of egg proteins (Perivitellin-1, PV1) from Pomacea snails. Their remarkable stability and clade-related functions in most derived clades (Canaliculata and Bridgesii) make them excellent candidates for exploring this issue. To that aim, we studied PV1 (PpaPV1) from the most basal lineage, Flagellata. PpaPV1 displays unparalleled structural and kinetic stability, surpassing PV1s from derived clades, ranking among the most hyperstable proteins documented in nature. Its spectral features contribute to a pale egg coloration, exhibiting a milder glycan binding lectin activity with a narrower specificity than PV1s from the closely related Bridgesii clade. These findings provide evidence for substantial structural and functional changes throughout the genus' PV1 evolution. We observed that structural and kinetic stability decreased in a clade-related fashion and was associated with large variations in defensive traits. For instance, pale PpaPV1 lectin turns potent in the Bridgesii clade, adversely affecting gut morphology, while giving rise to brightly colored PV1s providing eggs with a conspicuous, probably warning signal in the Canaliculata clade. This work provides a comprehensive comparative analysis of PV1s from various apple snail species within a phylogenetic framework, offering insights into the interplay among their structural features, stability profiles and functional roles. More broadly, our work provides one of the first examples from natural evolution showing the crucial link among protein structure, stability and evolution of new functions., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Objective: To describe the efficacy and safety of adalimumab for the treatment of non-infectious uveitis (NIU) in four Uveitis Units from tertiary Spanish hospitals., Methods: Multicenter and retrospective clinical cohort study including all patients with NIU treated with adalimumab from January 2012 to October 2022 in four uveitis units was performed. Efficacy was measured with the number of relapses, ocular inflammation and reduction in immunosuppression and corticosteroid dosage before and after adalimumab use. We collected data regarding adverse effects and examined the immunogenicity of adalimumab., Results: One hundred and twenty-two patients (59% females), with a mean age of 48.6 years (SD = 14.8) accounting for 217 eyes were included. The majority (92.6%) were Caucasian. Uveitis analyzed were predominantly panuveitis (34.7%), bilateral (77.9%), acute (41.5%), and non-granulomatous (90%). Most of them were immune mediated (42.6%), and the main reason to initiate adalimumab was refractory disease (96.7%). The analysis was statistically significant due to the reduction in the number of immunosuppressive drugs as well as the dose of oral corticosteroids and the number of relapses during follow-up ( p  < 0.001). The decrease in ocular inflammation parameters and the improvement in visual acuity ( p < 0.05) were also significant. There were no deaths due to the drug and only one reported case of serious infection. In total, 10.9% of 73 patients tested developed anti-adalimumab antibodies and 4.1% lupus-like., Conclusions: We consider adalimumab as a leading drug in the treatment of NIU with high safety and efficacy.

Allometry refers to the relationship between the size of a trait and that of the whole body of an organism. Pioneering observations by Otto Snell and further elucidation by D'Arcy Thompson set the stage for its integration into Huxley's explanation of constant relative growth that epitomizes through the formula of simple allometry. The traditional method to identify such a model conforms to a regression protocol fitted in the direct scales of data. It involves Huxley's formula-systematic part and a lognormally distributed multiplicative error term. In many instances of allometric examination, the predictive strength of this paradigm is unsuitable. Established approaches to improve fit enhance the complexity of the systematic relationship while keeping the go-along normality-borne error. These extensions followed Huxley's idea that considering a biphasic allometric pattern could be necessary. However, for present data composing 10 410 pairs of measurements of individual eelgrass leaf dry weight and area, a fit relying on a biphasic systematic term and multiplicative lognormal errors barely improved correspondence measure values while maintaining a heavy tails problem. Moreover, the biphasic form and multiplicative-lognormal-mixture errors did not provide complete fit dependability either. However, updating the outline of such an error term to allow heteroscedasticity to occur in a piecewise-like mode finally produced overall fit consistency. Our results demonstrate that when attempting to achieve fit quality improvement in a Huxley's model-based multiplicative error scheme, allowing for a complex allometry form for the systematic part, a non-normal distribution-driven error term and a composite of uneven patterns to describe the heteroscedastic outline could be essential., (© The Author(s) 2024. Published by Oxford University Press.)

Covering: 1951 to 2022Packed with nutrients and unable to escape, eggs are the most vulnerable stage of an animal's life cycle. Consequently, many species have evolved chemical defenses and teamed up their eggs with a vast array of toxic molecules for defense against predators, parasites, or pathogens. However, studies on egg toxins are rather scarce and the available information is scattered. The aim of this review is to provide an overview of animal egg toxins and to analyze the trends and patterns with respect to the chemistry and biosynthesis of these toxins. We analyzed their ecology, distribution, sources, occurrence, structure, function, relative toxicity, and mechanistic aspects and include a brief section on the aposematic coloration of toxic eggs. We propose criteria for a multiparametric classification that accounts for the complexity of analyzing the full set of toxins of animal eggs. Around 100 properly identified egg toxins are found in 188 species, distributed in 5 phyla: cnidarians (2) platyhelminths (2), mollusks (9), arthropods (125), and chordates (50). Their scattered pattern among animals suggests that species have evolved this strategy independently on numerous occasions. Alkaloids are the most abundant and widespread, among the 13 types of egg toxins recognized. Egg toxins are derived directly from the environment or are endogenously synthesized, and most of them are transferred by females inside the eggs. Their toxicity ranges from ρmol kg -1 to mmol kg -1 , and for some species, experiments support their role in predation deterrence. There is still a huge gap in information to complete the whole picture of this field and the number of toxic eggs seems largely underestimated.

Flamingos inhabit specialized habitats and breed in large colonies, building their nests on islands that limit the access of terrestrial predators. Many aspects of their uropygial gland are still unknown. The uropygial gland, a sebaceous organ exclusive to birds, shares some histological features among species such as the presence of a capsule, adenomers with stratified epithelium and secondary and primary chambers. We found that the uropygial gland of the Chilean Flamingo (Phoenicopterus chilensis) displays most of these characteristics but lacks a primary storage chamber. This absence may be an adaptation to their aquatic environment. The uropygial secretion of this species has a variety of glycoconjugates while its lipid moiety is largely dominated by waxes and minor amounts of triacylglycerols and fatty acids. Mass spectrometry analysis of the preen wax showed branched fatty acids of varied chain length and unbranched fatty alcohols, resulting in a complex mixture of wax esters and no differences between sexes were observed. The glycoconjugates present in the preen secretion could play a role as antimicrobial molecules, as suggested for other bird species, while the absence of diester waxes in flamingos might be related with their nesting habits and limited exposure to predation. Our results were evaluated according to physiological and ecological aspects of the flamingo's biology., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

The acquisition of egg protection is vital for species survival. Poisonous eggs from Pomacea apple snails have defensive macromolecules for protection. Here we isolated and characterized a novel lectin called PdPV1 that is massively accumulated in the eggs of Pomacea diffusa and seems part of its protective cocktail. The native protein, an oligomer of ca 256 kDa, has high structural stability, withstanding 15 min boiling and denaturing by SDS. It resists in vitro proteinase digestion and displays structural stability between pH 2.0 and pH 12.0, and up to 85°C. These properties, as well as its subunit sequences, glycosylation pattern, presence of carotenoids, size and global shape resemble those of its orthologs from other Pomacea. Furthermore, like members of the canaliculata clade, PdPV1 is recovered unchanged in feces of mice ingesting it, supporting an anti-nutritive defensive function. PdPV1 also displays a strong hemagglutinating activity, specifically recognizing selected ganglioside motifs with high affinity. This activity is only shared with PsSC, a perivitelline from the same clade ( bridgesii clade). As a whole, these results indicate that species in the genus Pomacea have diversified their egg defenses: those from the bridgesii clade are protected mostly by non-digestible lectins that lower the nutritional value of eggs, in contrast with protection by neurotoxins of other Pomacea clades, indicating that apple snail egg defensive strategies are clade specific. The harsh gastrointestinal environment of predators would have favored their appearance, extending by convergent evolution the presence of plant-like highly stable lectins, a strategy not reported in other animals., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

Rationale: Proteins from the egg perivitelline fluid (PVF) are assumed to play critical roles in embryonic development, but for many groups of animals their identities remain unknown. Identifying egg PVF proteins is a critical step towards understanding their functions including their roles in evolutionary transition in habitats., Methods: We applied proteomic and transcriptomic analysis to investigate the PVF proteome of the eggs of Pomacea diffusa, an aerial ovipositing freshwater snail in the family Ampullariidae. The PVF proteins were separated with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method, and proteomic analysis was conducted using an LTQ Velos ion trap mass spectrometer coupled with liquid chromatography. Comparison of PVF proteomes and evolution analyses was performed between P. diffusa and other ampullariids., Results: In total, 32 egg PVF proteins were identified from P. diffusa. They were categorized as PV1-like subunits, immune-responsive proteins, protein degradation, signaling and binding, transcription and translation, metabolism, oxidation-reduction and proteins with unknown function. Interestingly, the proteome includes a calcium-binding protein important in forming the hard eggshell that enabled the terrestrial transition. However, it does not include PV2, a neurotoxic protein that was assumed to be present in all Pomacea species., Conclusions: The PVF proteome data from P. diffusa can help us better understand the roles that reproductive proteins played during the transition from underwater to terrestrial egg deposition. Moreover, they could be useful in comparative studies of the terrestrialization in several groups of animals that occurred independently during their evolution., (© 2019 John Wiley & Sons, Ltd.)

Gastropod Molluscs rely exclusively on the innate immune system to protect from pathogens, defending their embryos through maternally transferred effectors. In this regard, Pomacea snail eggs, in addition to immune defenses, have evolved the perivitellin-2 or PV2 combining two immune proteins into a neurotoxin: a lectin and a pore-forming protein from the Membrane Attack Complex/Perforin (MACPF) family. This binary structure resembles AB-toxins, a group of toxins otherwise restricted to bacteria and plants. Many of these are enterotoxins, leading us to explore this activity in PV2. Enterotoxins found in bacteria and plants act mainly as pore-forming toxins and toxic lectins, respectively. In animals, although both pore-forming proteins and lectins are ubiquitous, no enterotoxins have been reported. Considering that Pomacea snail eggs ingestion induce morpho-physiological changes in the intestinal mucosa of rodents and is cytotoxic to intestinal cells in culture, we seek for the factor causing these effects and identified PmPV2 from Pomacea maculata eggs. We characterized the enterotoxic activity of PmPV2 through in vitro and in vivo assays. We determined that it withstands the gastrointestinal environment and resisted a wide pH range and enzymatic proteolysis. After binding to Caco-2 cells it promoted changes in surface morphology and an increase in membrane roughness. It was also cytotoxic to both epithelial and immune cells from the digestive system of mammals. It induced enterocyte death by a lytic mechanism and disrupted enterocyte monolayers in a dose-dependent manner. Further, after oral administration to mice PmPV2 attached to enterocytes and induced large dose-dependent morphological changes on their small intestine mucosa, reducing the absorptive surface. Additionally, PmPV2 was detected in the Peyer's patches where it activated lymphoid follicles and triggered apoptosis. We also provide evidence that the toxin can traverse the intestinal barrier and induce oral adaptive immunity with evidence of circulating antibody response. As a whole, these results indicate that PmPV2 is a true enterotoxin, a role that has never been reported to lectins or perforin in animals. This extends by convergent evolution the presence of plant- and bacteria-like enterotoxins to animals, thus expanding the diversity of functions of MACPF proteins in nature., (Copyright © 2020 Giglio, Ituarte, Ibañez, Dreon, Prieto, Fernández and Heras.)

Structural knowledge of gastropod hemocyanins is scarce. To better understand their evolution and diversity we studied the hemocyanin of a caenogastropod, Pomacea canaliculata (PcH). Through a proteomic and genomic approach, we identified 4 PcH subunit isoforms, in contrast with other gastropods that usually have 2 or 3. Each isoform has the typical Keyhole limpet-type hemocyanin architecture, comprising a string of eight globular functional units (FUs). Correspondingly, genes are organized in eight FUs coding regions. All FUs in the 4 genes are encoded by more than one exon, a feature not found in non- caenogastropods. Transmission electron microscopy images of PcH showed a cylindrical structure organized in di, tri and tetra-decamers with an internal collar structure, being the di and tri-decameric cylinders the most abundant ones. PcH is N-glycosylated with high mannose and hybrid-type structures, and complex-type N-linked glycans, with absence of sialic acid. Terminal β-N-GlcNAc residues and nonreducing terminal α-GalNAc are also present. The molecule lacks O-linked glycosylation but presents the T-antigen (Gal-β1,3-GalNAc). Using an anti-PcH polyclonal antibody, no cross-immunoreactivity was observed against other gastropod hemocyanins, highlighting the presence of clade-specific structural differences among gastropod hemocyanins. This is, to the best of our knowledge, the first gene structure study of a Caenogastropoda hemocyanin., Competing Interests: The authors have declared that no competing interests exist.

Objectives: The objectives of this study were: To analyze the causes of urological consultation of the child with CP, to analyze the role of nursing in the urological care of children with CP referred to Urology, and to know the relationship between the child's urinary tract infection and CP and its degree of independence., Method: Retrospective analytical observational study based on the review of the clinical records of patients diagnosed with cerebral palsy. Forty-seven patients selected by simple randomization in an age range of 2 to 16 years were included in the study in an outpatient clinic for CP patients located in a town in northern Mexico., Results: The causes of urological consultation of the child with CP, referred by nursing were: urinary infection, phimosis, retractable testicle, cryptorchidism, hypogonadism, overactive bladder, and hypospadias. The relationship between the urinary infection of the child with CP and its degree of independence was established with the level of urinary sphincter control., Conclusions: The nursing staff is a key and decisive factor in the diagnosis, early reference for treatment and monitoring of the evolution of urinary disorders in children with CP. It is not indicated to start a toilet training program in children with CP, without having previously ruled out a urinary tract infection.

The family Ampullariidae includes both aquatic and amphibious apple snails. They are an emerging model for evolutionary studies due to the high diversity, ancient history, and wide geographical distribution. Insight into drivers of ampullariid evolution is hampered, however, by the lack of genomic resources. Here, we report the genomes of four ampullariids spanning the Old World (Lanistes nyassanus) and New World (Pomacea canaliculata, P. maculata, and Marisa cornuarietis) clades. The ampullariid genomes have conserved ancient bilaterial karyotype features and a novel Hox gene cluster rearrangement, making them valuable in comparative genomic studies. They have expanded gene families related to environmental sensing and cellulose digestion, which may have facilitated some ampullarids to become notorious invasive pests. In the amphibious Pomacea, novel acquisition of an egg neurotoxin and a protein for making the calcareous eggshell may have been key adaptations enabling their transition from underwater to terrestrial egg deposition., (© The Author(s) 2019. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Conservation of eelgrass relies on transplants and evaluation of success depends on nondestructive measurements of average leaf biomass in shoots among other variables. Allometric proxies offer a convenient way to assessments. Identifying surrogates via log transformation and linear regression can set biased results. Views conceive this approach to be meaningful, asserting that curvature in geometrical space explains bias. Inappropriateness of correction factor of retransformation bias could also explain inconsistencies. Accounting for nonlinearity of the log transformed response relied on a generalized allometric model. Scaling parameters depend continuously on the descriptor. Joining correction factor is conceived as the partial sum of series expansion of mean retransformed residuals leading to highest reproducibility strength. Fits of particular characterizations of the generalized curvature model conveyed outstanding reproducibility of average eelgrass leaf biomass in shoots. Although nonlinear heteroscedastic regression resulted also to be suitable, only log transformation approaches can unmask a size related differentiation in growth form of the leaf. Generally, whenever structure of regression error is undetermined, choosing a suitable form of retransformation correction factor becomes elusive. Compared to customary nonparametric characterizations of this correction factor, present form proved more efficient. We expect that offered generalized allometric model along with proposed correction factor form provides a suitable analytical arrangement for the general settings of allometric examination.

Unlike most of the freshwater gastropod families, the family Ampullariidae includes members that exhibit both underwater and aerial oviposition, making it an ideal model for understanding mechanisms underlying the evolutionary transition from water to land. We applied SDS-PAGE and LC-MS/MS to analyse the proteome of the egg perivitelline fluid (PVF) of Marisa cornuarietis - an aquatic ovipositing ampullariid. Comparison with the reported PVF proteomes of two aerial ovipositing ampullariids (Pomacea canaliculata and P. maculata) showed that the three species all contain several major perivitellins that nourish the embryos. However, M. cornuarietis invests more heavily on immune-related proteins, which might be due to exposure to aquatic pathogens. Interestingly, only the PVF of out-of-water egg laying species have PV2 - a neurotoxin lethal to mice, and a calcium-binding protein which might be involved in the formation of calcareous eggshell. Integrated phylogenetic, evolutionary and gene expressional analyses detected the involvement of gene duplication, positive selection and neofunctionalisation in the formation of several major PVF proteins. Overall, our study provides multiple lines of evidence of adaptive evolution in the PVF proteins, and contributes to a better understanding of how aquatic gastropod ancestors invaded terrestrial habitats. SIGNIFICANCE: Aerial egg deposition has evolved in several groups of animals, but except for Vertebrata little is known about the mechanisms underlying this critical evolution process. We compared aquatic and aerial egg laying apple snails to understand the molecular mechanisms enabling such a transition in egg laying habitat. We found that the composition of perivitelline fluid proteomes of underwater and aerial egg depositors was remarkably different, and then gene duplication and positive selection were responsible for the formation of such novel proteins than enabled the evolutionary transition., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Species beloging to the genus Pomacea (Ampullariidae), often referred as apple snails, are freshwater, amphibious snails native to South, Central and North America. Some species such as P. canaliculata have become a driver of ecosystem changes in wetlands and an important rice and taro pest after its introduction to Asia and other parts of the world. Females deposit colored egg clutches above the waterline, a reproductive strategy that exposes the eggs to harsh conditions and terrestrial predation. However, eggs have no reported predators in their native range, probably because of the acquisition of unparalleled biochemical defenses provided by a set of proteins (perivitellins) that nourish embryos and protect them from predators and abiotic factors. Notably, ingestion of egg perivitelline fluid (PVF) decreases rat growth rate and alters their gastrointestinal morphology. The aim of the study is to determine the effect of apple snail egg PVF on mice gut digestive activity, morphology and nutrient absorption., Methods: Carbohydrate digestion by intestinal disaccharidases (sucrase-isomaltase and maltase-glucoamylase) was evaluated ex vivo in mice gavaged with 1 or 4 doses of PVF. Changes in gut morphological and absorptive surface were measured. In addition, alteration on nutrient absorption rates, transport pathways and intestinal permeability was evaluated by luminal perfusions of small intestine with radiolabeled L-proline (absorbed by paracellular and transcellular pathways) and L-arabinose (absorbed exclusively by paracellular pathway)., Results: Perivitelline fluid affected mice displayed significant morphological changes in the small intestine epithelium inducing the appearance of shorter and wider villi as well as fused villi. This resulted in a diminished absorptive surface, notably in the proximal portion . Likewise, the activity of disaccharidases diminished in the proximal portion of the intestine. Total absorption of L-proline increased in treated mice in a dose-dependent manner. There were no differences neither in the ratio of paracellular-to-transcellular absorption of L-proline nor in gut permeability as revealed by the clearance of L-arabinose., Discussion: Oral administration of apple snail PVF to mice adversely alters gut morphophysiology by reducing the intestinal absorptive surface, affecting enzymes of sugar metabolism and increasing the absorption rate of nutrients without affecting the relative contribution of the absorption pathways or gut permeability. These results further support the role of PVF in passive anti-predator defenses in Pomacea snail eggs that target the digestive system., Competing Interests: The authors declare there are no competing interests.

The eggs of the freshwater Pomacea apple snails develop above the water level, exposed to varied physical and biological stressors. Their high hatching success seems to be linked to their proteins or perivitellins, which surround the developing embryo providing nutrients, sunscreens and varied defenses. The defensive mechanism has been unveiled in P. canaliculata and P. maculata eggs, where their major perivitellins are pigmented, non-digestible and provide a warning coloration while another perivitellin acts as a toxin. In P. scalaris, a species sympatric to the former, the defense strategy seems different, since no toxin was found and the major perivitellin, PsSC, while also colored and non-digestible, is a carbohydrate-binding protein. In this study we examine the structure and function of PsSC by sequencing its subunits, characterizing its carbohydrate binding profile and evaluating its effect on gut cells. Whereas cDNA sequencing and database search showed no lectin domain, glycan array carbohydrate binding profile revealed a strong specificity for glycosphingolipids and ABO group antigens. Moreover, PsSC agglutinated bacteria in a dose-dependent manner. Inspired on the defensive properties of seed lectins we evaluated the effects of PsSC on intestinal cells both in vitro (Caco-2 and IEC-6 cells) and in the gastrointestinal tract of rats. PsSC binds to Caco-2 cell membranes without reducing its viability, while a PsSC-containing diet temporarily induces large epithelium alterations and an increased absorptive surface. Based on these results, we propose that PsSC is involved in embryo defenses by altering the gut morphophysiology of potential predators, a convergent role to plant defensive lectins., Competing Interests: The authors have declared that no competing interests exist.

This article aims to present the protocol on Quality Controls in Digital Mammography published online in 2015 by the European Federation of Organisations for Medical Physics (EFOMP) which was developed by a Task Force under the Mammo Working Group. The main objective of this protocol was to define a minimum set of easily implemented quality control tests on digital mammography systems that can be used to assure the performance of a system within a set and acceptable range. Detailed step-by-step instructions have been provided, limiting as much as possible any misinterpretations or variations by the person performing. It is intended that these tests be implemented as part of the daily routine of medical physicists and system users throughout Europe in a harmonised way so allowing results to be compared. In this paper the main characteristics of the protocol are illustrated, including examples, together with a brief summary of the contents of each chapter. Finally, instructions for the download of the full protocol and of the related software tools are provided., (Copyright © 2018 Associazione Italiana di Fisica Medica. Published by Elsevier Ltd. All rights reserved.)

Background: The effects of current anthropogenic influences on eelgrass (Zostera marina) meadows are noticeable. Eelgrass ecological services grant important benefits for mankind. Preservation of eelgrass meadows include several transplantation methods. Evaluation of establishing success relies on the estimation of standing stock and productivity. Average leaf biomass in shoots is a fundamental component of standing stock. Existing methods of leaf biomass measurement are destructive and time consuming. These assessments could alter shoot density in developing transplants. Allometric methods offer convenient indirect assessments of individual leaf biomass. Aggregation of single leaf projections produce surrogates for average leaf biomass in shoots. Involved parameters are time invariant, then derived proxies yield simplified nondestructive approximations. On spite of time invariance local factors induce relative variability of parameter estimates. This influences accuracy of surrogates. And factors like analysis method, sample size and data quality also impact precision. Besides, scaling projections are sensitive to parameter fluctuation. Thus the suitability of the addressed allometric approximations requires clarification., Results: The considered proxies produced accurate indirect assessments of observed values. Only parameter estimates fitted from raw data using nonlinear regression, produced robust approximations. Data quality influenced sensitivity and sample size for an optimal precision., Conclusions: Allometric surrogates of average leaf biomass in eelgrass shoots offer convenient nondestructive assessments. But analysis method and sample size can influence accuracy in a direct manner. Standardized routines for data quality are crucial on granting cost-effectiveness of the method.

Background: Gastropoda, with approximately 80,000 living species, is the largest class of Mollusca. Among gastropods, apple snails (family Ampullariidae) are globally distributed in tropical and subtropical freshwater ecosystems and many species are ecologically and economically important. Ampullariids exhibit various morphological and physiological adaptations to their respective habitats, which make them ideal candidates for studying adaptation, population divergence, speciation, and larger-scale patterns of diversity, including the biogeography of native and invasive populations. The limited availability of genomic data, however, hinders in-depth ecological and evolutionary studies of these non-model organisms., Results: Using Illumina Hiseq platforms, we sequenced 1220 million reads for seven species of apple snails. Together with the previously published RNA-Seq data of two apple snails, we conducted de novo transcriptome assembly of eight species that belong to five genera of Ampullariidae, two of which represent Old World lineages and the other three New World lineages. There were 20,730 to 35,828 unigenes with predicted open reading frames for the eight species, with N50 (shortest sequence length at 50% of the unigenes) ranging from 1320 to 1803 bp. 69.7% to 80.2% of these unigenes were functionally annotated by searching against NCBI's non-redundant, Gene Ontology database and the Kyoto Encyclopaedia of Genes and Genomes. With these data we developed AmpuBase, a relational database that features online BLAST functionality for DNA/protein sequences, keyword searching for unigenes/functional terms, and download functions for sequences and whole transcriptomes., Conclusions: In summary, we have generated comprehensive transcriptome data for multiple ampullariid genera and species, and created a publicly accessible database with a user-friendly interface to facilitate future basic and applied studies on ampullariids, and comparative molecular studies with other invertebrates.

Plants have evolved sophisticated embryo defences by kinetically-stable non-digestible storage proteins that lower the nutritional value of seeds, a strategy that have not been reported in animals. To further understand antinutritive defences in animals, we analysed PmPV1, massively accumulated in the eggs of the gastropod Pomacea maculata, focusing on how its structure and structural stability features affected its capacity to withstand passage through predator guts. The native protein withstands >50 min boiling and resists the denaturing detergent sodium dodecyl sulphate (SDS), indicating an unusually high structural stability (i.e., kinetic stability). PmPV1 is highly resistant to in vitro proteinase digestion and displays structural stability between pH 2.0-12.0 and 25-85 °C. Furthermore, PmPV1 withstands in vitro and mice digestion and is recovered unchanged in faeces, supporting an antinutritive defensive function. Subunit sequence similarities suggest a common origin and tolerance to mutations. This is the first known animal genus that, like plant seeds, lowers the nutritional value of eggs by kinetically-stable non-digestible storage proteins that survive the gut of predators unaffected. The selective pressure of the harsh gastrointestinal environment would have favoured their appearance, extending by convergent evolution the presence of plant-like hyperstable antinutritive proteins to unattended reproductive stages in animals.

This article describes how the proteomic and transcriptomic data were produced during a study of the reproductive proteins of Pomacea maculata , an aquatic apple snail laying colorful aerial eggs, and provides public access to the data. The data are related to a research article titled 'An integrated proteomic and transcriptomic analysis of perivitelline fluid proteins in a freshwater gastropod laying aerial eggs' (Mu et al., 2017) [1]. RNA was extracted from the albumen gland and other tissues and sequenced on an Illumina Hiseq. 2000. The assembled transcriptome was translated into protein sequences and then used for protein identification. Proteins from the perivitelline fluid of P. maculata were separated in SDS-PAGE and analyzed by LTQ-Orbitrap Elite coupled to an Easy-nLC. The translated transcriptome data are provided in this article. Proteomic data (. raw file format) are available via ProteomeXchange with the identifier PXD006718.

In contrast with vitellogenin maturation, it is unknown whether gastropod perivitellin precursors are subject to large structural changes. The gastropod reproductive tract includes an accessory organ, the albumen gland (AG), that produces and secretes perivitelline fluid. In the apple snail Pomacea canaliculata, the large, reddish-pink AG provides eggs with perivitellins that are defensive against predators. Although the AG makes a considerable contribution to apple snail biomass, field observations indicate that it is rejected by avian and mammalian predators, although the underlying reason remains unknown. By analyzing the structure-function properties of P. canaliculata perivitellin precursors, we provide insight into perivitellin maturation and its relationship with apple snail predator feeding behavior. Structural analysis using small-angle X-ray scattering, absorption and fluorescence spectroscopy, circular dichroism, electrophoresis, chromatography, and partial proteolysis showed that the size, shape, and structure of perivitellin precursors resemble those of egg mature forms. Functional analysis indicates that the precursors of the defensive perivitellins ovorubin (PcOvo) and perivitellin-2 (PcPV2) are highly stable and antinutritive, withstanding proteinase digestion and displaying structural stability of their quaternary structure under a wide pH range (4.0-10.0). Furthermore, AG extracts limit a predator's ability to digest nutrients and are toxic to mice (median lethal concentration 96 h after administration: 5.9 mg/kg). Treated mice displayed neurologic signs similar to those produced by egg PcPV2. Results indicate that apple snails store active precursors of egg proteins inside the AG, providing evidence that gastropod perivitellin precursors do not experience the large structural processing of invertebrate vitellogenin maturation. These defensive proteins provide the apple snail AG with neurotoxic, antinutritive, and antidigestive activity, a likely explanation for the predators' feeding behavior.

Proteins of the egg perivitelline fluid (PVF) that surrounds the embryo are critical for embryonic development in many animals, but little is known about their identities. Using an integrated proteomic and transcriptomic approach, we identified 64 proteins from the PVF of Pomacea maculata, a freshwater snail adopting aerial oviposition. Proteins were classified into eight functional groups: major multifunctional perivitellin subunits, immune response, energy metabolism, protein degradation, oxidation-reduction, signaling and binding, transcription and translation, and others. Comparison of gene expression levels between tissues showed that 22 PVF genes were exclusively expressed in albumen gland, the female organ that secretes PVF. Base substitution analysis of PVF and housekeeping genes between P. maculata and its closely related species Pomacea canaliculata showed that the reproductive proteins had a higher mean evolutionary rate. Predicted 3D structures of selected PVF proteins showed that some nonsynonymous substitutions are located at or near the binding regions that may affect protein function. The proteome and sequence divergence analysis revealed a substantial amount of maternal investment in embryonic nutrition and defense, and higher adaptive selective pressure on PVF protein-coding genes when compared with housekeeping genes, providing insight into the adaptations associated with the unusual reproductive strategy in these mollusks., Significance: There has been great interest in studying reproduction-related proteins as such studies may not only answer fundamental questions about speciation and evolution, but also solve practical problems of animal infertility and pest outbreak. Our study has demonstrated the effectiveness of an integrated proteomic and transcriptomic approach in understanding the heavy maternal investment of proteins in the eggs of a non-model snail, and how the reproductive proteins may have evolved during the transition from laying underwater eggs to aerial eggs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Home enteral nutrition (HEN) is a well-established extra-hospital therapy that can reduce the risk of malnutrition, ensure the rapid discharge of patients from hospital and significantly reduce health care expenditure. The data reported in this study allow us to understand the relationships between mortality, the place of treatment either at patients' homes (PH) or in nursing homes (NHR) and nutritional status., Methods: Patients were analyzed according to age, gender, underlying disease, the Karnofsky Index, type of enteral access device (nasogastric tube or percutaneous endoscopic gastrostomy), weight and Body Mass Index (BMI). The duration of HEN therapy was then calculated and the outcome was established on patient mortality or survival., Results: Over an 11-year period, 3246 subjects were administered HEN therapy. The mean duration of HEN therapy was equal to 312±487 days at PH and 398±573 in NHR. The mean incidence is 406±58 patients/million inhabitants/year at PH and 319±44 in NHR (mean prevalence rate: 464±129 cases/million inhabitants at PH compared to 478±164 in NHR). Analysis of variance was used for continuous variables. The study reveals that >8% (8.6% at PH; 8.5% in NHR) of patients die within 10 days of starting HEN therapy., Conclusions: The study shows a progressive increase in HEN therapy and highlights clinical, organizational and ethical issues, which also need to be analyzed in relation to the progressively aging population.

Purpose: Demonstrate an objective procedure to quantify image quality in digital subtraction angiography (DSA) and suggest thresholds for acceptability and constancy tests., Methods: Series of images were obtained in a DSA system simulating a small (paediatric) and a large patient using the dynamic phantom described in the IEC and DIN standards for acceptance tests of DSA equipment. Image quality was quantified using measurements of contrast-to-noise ratio (CNR). Overall scores combining the CNR of 10-100 mg/ml Iodine at a vascular diameter of 1-4 mm in a homogeneous background were defined. Phantom entrance surface air kerma (Ka,e) was measured with an ionisation chamber., Results: The visibility of a low-contrast vessel in DSA images has been identified with a CNR value of 0.50 ± 0.03. Despite using 14 times more Ka,e (8.85 vs 0.63 mGy/image), the protocol for large patients showed a decrease in the overall score CNRsum of 67% (4.21 ± 0.06 vs 2.10 ± 0.05). The uncertainty in the results of the objective method was below 5%., Conclusion: Objective evaluation of DSA images using CNR is feasible with dedicated phantom measurements. An objective methodology has been suggested for acceptance tests compliant with the IEC/DIN standards. The defined overall scores can serve to fix a reproducible baseline for constancy tests, as well as to study the device stability within one acquisition series and compare different imaging protocols. This work provides aspects that have not been included in the recent European guidelines on Criteria for Acceptability of Medical Radiological Equipment., (Copyright © 2015 Associazione Italiana di Fisica Medica. Published by Elsevier Ltd. All rights reserved.)

Background: Eelgrass grants important ecological benefits including a nursery for waterfowl and fish species, shoreline stabilization, nutrient recycling and carbon sequestration. Upon the exacerbation of deleterious anthropogenic influences, re-establishment of eelgrass beds has mainly depended on transplantation. Productivity estimations provide valuable information for the appraisal of the restoration of ecological functions of natural populations. Assessments over early stages of transplants should preferably be nondestructive. Allometric scaling of eelgrass leaf biomass in terms of matching length provides a proxy that reduces leaf biomass and productivity estimations to simple measurements of leaf length and its elongation over a period. We examine how parameter variability impacts the accuracy of the considered proxy and the extent on what data quality and sample size influence the uncertainties of the involved allometric parameters., Methods: We adapted a Median Absolute Deviation data quality control procedure to remove inconsistencies in the crude data. For evaluating the effect of parametric uncertainty we performed both a formal exploration and an analysis of the sensitivity of the allometric projection method to parameter changes. We used parameter estimates obtained by means of nonlinear regression from crude as well as processed data., Results: We obtained reference leaf growth rates by allometric projection using parameter estimates produced by the crude data, and then considered changes in fitted parameters bounded by the modulus of the vector of the linked standard errors, we found absolute deviations up to 10% of reference values. After data quality control, the equivalent maximum deviation was under 7% of corresponding reference rates. Therefore, the addressed allometric method is robust. Even the smaller sized samples in the quality controlled dataset produced better accuracy levels than the whole set of crude data., Conclusions: We propose quality control of data as a highly recommended step in the overall procedure that leads to reliable allometric surrogates of eelgrass leaf growth rates. The proliferation of inconsistent replicates in the crude data points towards the importance of discarding incomplete leaves. We also recommend avoiding errors in estimating the biomass of small leaves for which precision of the used analytical scale might be an issue.

Background: Eelgrass is a cosmopolitan seagrass species that provides important ecological services in coastal and near-shore environments. Despite its relevance, loss of eelgrass habitats is noted worldwide. Restoration by replanting plays an important role, and accurate measurements of the standing crop and productivity of transplants are important for evaluating restoration of the ecological functions of natural populations. Traditional assessments are destructive, and although they do not harm natural populations, in transplants the destruction of shoots might cause undesirable alterations. Non-destructive assessments of the aforementioned variables are obtained through allometric proxies expressed in terms of measurements of the lengths or areas of leaves. Digital imagery could produce measurements of leaf attributes without the removal of shoots, but sediment attachments, damage infringed by drag forces or humidity contents induce noise-effects, reducing precision. Available techniques for dealing with noise caused by humidity contents on leaves use the concepts of adjacency, vicinity, connectivity and tolerance of similarity between pixels. Selection of an interval of tolerance of similarity for efficient measurements requires extended computational routines with tied statistical inferences making concomitant tasks complicated and time consuming. The present approach proposes a simplified and cost-effective alternative, and also a general tool aimed to deal with any sort of noise modifying eelgrass leaves images. Moreover, this selection criterion relies only on a single statistics; the calculation of the maximum value of the Concordance Correlation Coefficient for reproducibility of observed areas of leaves through proxies obtained from digital images., Results: Available data reveals that the present method delivers simplified, consistent estimations of areas of eelgrass leaves taken from noisy digital images. Moreover, the proposed procedure is robust because both the optimal interval of tolerance of similarity and the reproducibility of observed leaf areas through digital image surrogates were independent of sample size., Conclusion: The present method provides simplified, unbiased and non-destructive measurements of eelgrass leaf area. These measurements, in conjunction with allometric methods, can predict the dynamics of eelgrass biomass and leaf growth through indirect techniques, reducing the destructive effect of sampling, fundamental to the evaluation of eelgrass restoration projects thereby contributing to the conservation of this important seagrass species.

Objective: To demonstrate that problem-solving therapy is effective in reducing the burden on caregivers of children with cerebral palsy., Design: Randomized clinical trial., Location: Check primary care within a private nonprofit association., Participants: 140caregivers divided into control group (CG) and experimental group (EG)., Interventions: We performed in both groups a psychosocial intervention with a frequency of one session per week for three weeks to complete 120minutes. In the EG performed a shortened form of problem-solving therapy with a focus on caregiver burden and the CG performed an educational intervention focusing on respiratory diseases., Main Measures: The response variable corresponds to the score obtained by Zarit questionnaire. The independent variable accounted for psychosocial intervention., Results: In the EG according to Zarit questionnaire score was obtained by averaging 45.0 points pre intervention against 45.3 points in the CP after intervention Zarit was obtained by averaging 29.8 points in the EG and 44.3 points in the CG (P<.0001). The catalog groups according to their score Zarit in charge: none, mild, moderate and severe impact differences were found in the different intervention categories (Wilcoxon test Z=6.281, P<.00001)., Conclusions: Problem solving therapy is effective in reducing the burden on caregivers of children with cerebral palsy., (Copyright © 2013 Elsevier España, S.L.U. All rights reserved.)

Background: The spread of the invasive snail Pomacea canaliculata is expanding the rat lungworm disease beyond its native range. Their toxic eggs have virtually no predators and unusual defenses including a neurotoxic lectin and a proteinase inhibitor, presumably advertised by a warning coloration. We explored the effect of egg perivitellin fluid (PVF) ingestion on the rat small intestine morphology and physiology., Methodology/principal Findings: Through a combination of biochemical, histochemical, histopathological, scanning electron microscopy, cell culture and feeding experiments, we analyzed intestinal morphology, growth rate, hemaglutinating activity, cytotoxicity and cell proliferation after oral administration of PVF to rats. PVF adversely affects small intestine metabolism and morphology and consequently the standard growth rate, presumably by lectin-like proteins, as suggested by PVF hemaglutinating activity and its cytotoxic effect on Caco-2 cell culture. Short-term effects of ingested PVF were studied in growing rats. PVF-supplemented diet induced the appearance of shorter and wider villi as well as fused villi. This was associated with changes in glycoconjugate expression, increased cell proliferation at crypt base, and hypertrophic mucosal growth. This resulted in a decreased absorptive surface after 3 days of treatment and a diminished rat growth rate that reverted to normal after the fourth day of treatment. Longer exposure to PVF induced a time-dependent lengthening of the small intestine while switching to a control diet restored intestine length and morphology after 4 days., Conclusions/significance: Ingestion of PVF rapidly limits the ability of potential predators to absorb nutrients by inducing large, reversible changes in intestinal morphology and growth rate. The occurrence of toxins that affect intestinal morphology and absorption is a strategy against predation not recognized among animals before. Remarkably, this defense is rather similar to the toxic effect of plant antipredator strategies. This defense mechanism may explain the near absence of predators of apple snail eggs.

Although the process of endocytosis of the low density lipoprotein (LDL) macromolecule and its receptor have been the subject of intense experimental research and modeling, there are still conflicting hypotheses and even conflicting data regarding the way receptors are transported to coated pits, the manner by which receptors are inserted before they aggregate in coated pits, and the display of receptors on the cell surface. At first it was considered that LDL receptors in human fibroblasts are inserted at random locations and then transported by diffusion toward coated pits. But experiments have not ruled out the possibility that the true rate of accumulation of LDL receptors in coated pits might be faster than predicted on the basis of pure diffusion and uniform reinsertion over the entire cell surface. It has been claimed that recycled LDL receptors are inserted preferentially in regions where coated pits form, with display occurring predominantly as groups of loosely associated units. Another mechanism that has been proposed by experimental cell biologists which might affect the accumulation of receptors in coated pits is a retrograde membrane flow. This is essentially linked to a polarized receptor insertion mode and also to the capping phenomenon, characterized by the formation of large patches of proteins that passively flow away from the regions of membrane exocytosis. In this contribution we calculate the mean travel time of LDL receptors to coated pits as determined by the ratio of flow strength to diffusion-coefficient, as well as by polarized-receptor insertion. We also project the resulting display of unbound receptors on the cell membrane. We found forms of polarized insertion that could potentially reduce the mean capture time of LDL receptors by coated pits which is controlled by diffusion and uniform insertion. Our results show that, in spite of its efficiency as a possible device for enhancement of the rate of receptor trapping, polarized insertion nevertheless fails to induce the formation of steady-state clusters of receptor on the cell membrane. Moreover, for appropriate values of the flow strength-diffusion ratio, the predicted steady-state distribution of receptors on the surface was found to be consistent with the phenomenon of capping.

Lipid and fatty acid (FA) composition and selected oxidative stress parameters of freshwater clams (Dipolodon chilensis), from a sewage-polluted (SMA) and a clean site, were compared. Trophic markers FA were analyzed in clams and sediment. Saturated FA (SAFA), and bacteria and sewage markers were abundant in SMA sediments, while diatom markers were 50% lower. Proportions of SAFA, branched FA, 20:5n-3 (EPA) and 22:6n-3 (DHA) were higher in SMA clams. Chronic exposure of D. chilensis to increasing eutrophication affected its lipid and FA composition. The increase in EPA and DHA proportions could be an adaptive response, which increases stress resistance but could also lead to higher susceptibility to lipid peroxidation TBARS, lipofuscins (20-fold) and GSH concentrations were higher in SMA clams. FA markers indicated terrestrial plant detritus and bacteria are important items in D. chilensis diet. Anthropogenic input in their food could be traced using specific FA as trophic markers., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Despite the ecological importance of eelgrass, nowadays anthropogenic influences have produced deleterious effects in many meadows worldwide. Transplantation plots are commonly used as a feasible remediation scheme. The characterization of eelgrass biomass and its dynamics is an important input for the assessment of the overall status of both natural and transplanted populations. Particularly, in restoration plots it is desirable to obtain nondestructive assessments of these variables. Allometric models allow the expression of above ground biomass and productivity of eelgrass in terms of leaf area, which provides cost effective and nondestructive assessments. Leaf area in eelgrass can be conveniently obtained by the product of associated length and width. Although these variables can be directly measured on most sampled leaves, digital image methods could be adapted in order to simplify measurements. Nonetheless, since width to length ratios in eelgrass leaves could be even negligible, noise induced by leaf humidity content could produce misidentification of pixels along the peripheral contour of leaves images. In this paper, we present a procedure aimed to produce consistent estimations of eelgrass leaf area in the presence of the aforementioned noise effects. Our results show that digital image procedures can provide reliable, nondestructive estimations of eelgrass leaf area.

Although most eggs are intensely predated, the aerial egg clutches from the aquatic snail Pomacea canaliculata have only one reported predator due to unparalleled biochemical defenses. These include two storage-proteins: ovorubin that provides a conspicuous (presumably warning) coloration and has antinutritive and antidigestive properties, and PcPV2 a neurotoxin with lethal effect on rodents. We sequenced PcPV2 and studied whether it was able to withstand the gastrointestinal environment and reach circulation of a potential predator. Capacity to resist digestion was assayed using small-angle X-ray scattering (SAXS), fluorescence spectroscopy and simulated gastrointestinal proteolysis. PcPV2 oligomer is antinutritive, withstanding proteinase digestion and displaying structural stability between pH 4.0-10.0. cDNA sequencing and protein domain search showed that its two subunits share homology with membrane attack complex/perforin (MACPF)-like toxins and tachylectin-like lectins, a previously unknown structure that resembles plant Type-2 ribosome-inactivating proteins and bacterial botulinum toxins. The protomer has therefore a novel AB toxin combination of a MACPF-like chain linked by disulfide bonds to a lectin-like chain, indicating a delivery system for the former. This was further supported by observing PcPV2 binding to glycocalix of enterocytes in vivo and in culture, and by its hemaggutinating, but not hemolytic activity, which suggested an interaction with surface oligosaccharides. PcPV2 is able to get into predator's body as evidenced in rats and mice by the presence of circulating antibodies in response to sublethal oral doses. To our knowledge, a lectin-pore-forming toxin has not been reported before, providing the first evidence of a neurotoxic lectin in animals, and a novel function for ancient and widely distributed proteins. The acquisition of this unique neurotoxic/antinutritive/storage protein may confer the eggs a survival advantage, opening new perspectives in the study of the evolution of animal defensive strategies.

Estimation of leaf productivity in eelgrass (Zostera marina L.) is crucial for evaluating the ecological role of this important seagrass species. Although leaf marking techniques are widely used to obtain estimates of leaf productivity, the accuracy of these assessments, has been questioned mainly because these fail to account for leaf growth below the reference mark and also because they apparently disregard the contribution of mature leaf tissues to the growth rate of leaves. On the other hand, the plastochrone method is a simpler technique that has been considered to effectively capture growth in a more realistic way, thereby providing more accurate assessments of both above- and below-ground productivities. But since the actual values of eelgrass growth rates are difficult to obtain, the worth of the plastochrone method has been largely vindicated because it produces assessments that overestimate productivity as compared to estimates obtained by leaf marking. Additionally, whenever eelgrass leaf biomass can be allometrically scaled in terms of matching leaf length in a consistent way, the associated leaf growth rates can be also projected allometrically. In this contribution, we used that approach to derive an authentication of the plastochrone method and formally demonstrate that, as has been claimed to occur for leaf marking approaches, the plastochrone method itself underestimates actual values of eelgrass leaf growth rates. We also show that this unavoidable bias is mainly due to the inadequacy of single-leaf biomass assessments in providing a proxy for the growth of all leaf tissue in a shoot over a given interval. Moreover, the derived formulae give conditions under which assessments of leaf growth rates using the plastochrone method would systematically underestimate matching values obtained by leaf marking procedures. And, assessments of leaf growth rates obtained by using the present data show that plastochrone method estimations underestimated corresponding proxies obtained allometrically (27%), or through leaf marking (35%). Allometric projection is recommended as a simpler and more effective procedure to reduce the bias in eelgrass leaf productivity estimations that associates to the use of plastochrone methods.

Hydrocarbon pollution is a major environmental threat to ecosystems in marine and freshwater environments, but its toxicological effect on aquatic organisms remains little studied. A proteomic approach was used to analyze the effect of a freshwater oil spill on the prawn Macrobrachium borellii. To this aim, proteins were extracted from midgut gland (hepatopancreas) of male and female prawns exposed 7 days to a sublethal concentration (0.6 ppm) of water-soluble fraction of crude oil (WSF). Exposure to WSF induced responses at the protein expression level. Two-dimensional gel electrophoresis (2-DE) revealed 10 protein spots that were differentially expressed by WSF exposure. Seven proteins were identified using MS/MS and de novo sequencing. Nm23 oncoprotein, arginine methyltransferase, fatty aldehyde dehydrogenase and glutathione S-transferase were down-regulated, whereas two glyceraldehyde-3-phosphate dehydrogenase isoforms and a lipocalin-like crustacyanin (CTC) were up-regulated after WSF exposure. CTC mRNA levels were further analyzed by quantitative real-time PCR showing an increased expression after WSF exposure. The proteins identified are involved in carbohydrate and amino acid metabolism, detoxification, transport of hydrophobic molecules and cellular homeostasis among others. These results provide evidence for better understanding the toxic mechanisms of hydrocarbons. Moreover, some of these differentially expressed proteins would be employed as potential novel biomarkers., (Copyright © 2013 Elsevier Inc. All rights reserved.)

A phantom for image quality evaluation of digital mammography systems is presented and compared to the most widely used phantoms in Europe and the US. The phantom contains objects for subjective detection of Landolt rings (four-alternative, forced-choice task) and for objective calculation of signal-difference-to-noise ratios (SDNR), both in a titanium background within a 12-step wedge. Evaluating phantom images corresponding to exposures between 15 and 160 mAs (average glandular dose between 0.2 and 2 mGy), the resulting scores were compared to the scores obtained following the European EPQC and American College of Radiology (ACR) protocols. Scores of the Landolt test equal to 19 and 8.5 and SDNR equal to 20 and 11 were found to be equivalent to the acceptable limiting values suggested by EPQC and ACR. In addition, the Landolt and SDNR tests were shown to take into account the anatomical variations in thickness and tissue density within the breast. The simplified evaluation method presented was shown to be a sensitive, efficient and reliable alternative for image quality evaluation of mammography systems.

Pomacea canaliculata is a freshwater snail that deposits eggs on solid substrates above the water surface. Previous studies have emphasized the nutritional and protective functions of the three most abundant perivitelline fluid (PVF) protein complexes (ovorubin, PV2, and PV3) during its embryonic development, but little is known about the structure and function of other less abundant proteins. Using 2-DE, SDS-PAGE, MALDI TOF/TOF, and LC-MS/MS, we identified 59 proteins from the PVF of P. canaliculata, among which 19 are novel. KEGG analysis showed that the functions of the majority of these proteins are "unknown" (n=34), "environmental information processing" (10), 9 of which are related to innate immunity, and "metabolism" (7). Suppressive subtractive hybridization revealed 21 PVF genes to be specific to the albumen gland, indicating this organ is the origin of many of the PVF proteins. Further, the 3 ovorubin subunits were identified with 30.2-35.0% identity among them, indicating their common origin but ancient duplications. Characterization of the PVF proteome has opened the gate for further studies aiming to understand the evolution of the novel proteins and their contribution to the switch to aerial oviposition.

During reproduction vitellogenin (VTG) is transported to vitellogenic oocytes as a precursor of egg yolk lipovitellin (LV). As VTG synthesis is affected by environmental stressors, it is widely used as biomarker in endocrine disruption studies. However, it has seldom been employed to evaluate invertebrate hydrocarbon pollution. An ELISA with anti-LV antibody was developed to evaluate the impact of water-soluble fraction of crude oil (WSF) on Macrobrachium borellii vitellogenesis. Prawn VTG concentration was within the range reported for other crustaceans; LV values were positively correlated with gonadosomatic index (GSI). Females at different vitellogenic stages were exposed to a sub-lethal concentration of WSF for 7 days. Exposed animals with GSI>7 increased their VTG and LV titer as compared to control organisms (190% and 140%, respectively). VTG levels in M. borellii were upregulated and highly sensitive to WSF exposure. This assay could be employed as a biomarker for freshwater hydrocarbon pollution., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

This study presents a theoretical exploration of the effects of mechanisms that, in addition to diffusion, may influence the surface dynamics and display of unbound receptors in the low-density lipoprotein (LDL) endocytic cycle in human fibroblasts. The factors considered here are a transverse membrane flow and a generalized plaque-form insertion mode. The proposed model permits estimations of aggregation rates of unbound receptors in coated pits as well as pictorial representations of their expected steady-state display on the cell surface. Our findings show that this display is determined in a fundamental way by the ratio of the strength of the flow to the diffusion coefficient. For measured values of the diffusion coefficient and the estimated value of the flow rate strength (and independent of the receptor insertion mode), the display predicted by our model is consistent with the capping phenomenon, i.e., a gradated clustering in the direction of flow streamlines. There could be suitable characterizations of the receptor reinsertion mode that would produce a substantial reduction in the mean capture time of LDL receptors by coated pits. In any event, our results show that the existence of a transverse membrane flow precludes the display of steady-state plaque-form surface clusters.

The characterization of biomass and its dynamics provides valuable information for the assessment of natural and transplanted eelgrass populations. The need for simple, nondestructive assessments has led to the use of the leaf biomass-to-length ratio for converting leaf-length measurements, which can be easily obtained, to leaf growth rates through the plastochrone method. Using data on leaf biomass and length collected in three natural eelgrass populations and a mesocosm, we evaluated the suitability of a leaf weight-to-length ratio for nondestructive assessments. For the data sets considered, the isometric scaling that sustains the weight-to-length proxy always produced inconsistent fittings, and for leaf-lengths greater than a threshold value, the conversion of leaf length to biomass generated biased estimations. In contrast, an allometric scaling of leaf biomass and length was highly consistent in all the cases considered. And these nondestructive assessments generated reliable levels of reproducibility in leaf biomass for all the ranges of variability in leaf lengths. We argue that the use of allometric scaling for the representation of leaf biomass in terms of length provides a more reliable approach for estimating eelgrass biomass.

Apple snail perivitellins are emerging as ecologically important reproductive proteins. To elucidate if the protective functions of the egg proteins of Pomacea canaliculata (Caenogastropoda, Ampullariidae), involved in embryo defenses, are present in other Pomacea species we studied scalarin (PsSC), the major perivitellin of Pomacea scalaris. Using small angle X-ray scattering, fluorescence and absorption spectroscopy and biochemical methods, we analyzed PsSC structural stability, agglutinating activity, sugar specificity and protease resistance. PsSC aggluttinated rabbit, and, to a lesser extent, human B and A erythrocytes independently of divalent metals Ca(2+) and Mg(2+) were strongly inhibited by galactosamine and glucosamine. The protein was structurally stable between pH 2.0 to 10.0, though agglutination occurred only between pH 4.0 to 8.0 (maximum activity at pH 7.0). The agglutinating activity was conserved up to 60 °C and completely lost above 80 °C, in agreement with the structural thermal stability of the protein (up to 60 °C). PsSC was able to withstand in vitro gastrointestinal digestion, and showed no trypsin inhibition activity. The presence of lectin activity has been reported in eggs of other Pomacea snails, but here we link for the first time, this activity to an apple snail multifunctional perivitellin. This novel role for a snail egg storage protein is different from closely related P.canaliculata defensive proteins.

It has been already reported that most hemolymphatic lipids in the spider Polybetes pythagoricus are transported by HDL1 and VHDL lipoproteins. We studied in vitro the lipid transfer among midgut-diverticula (M-diverticula), and either hemolymph or purified lipoproteins as well as between hemolymphatic lipoproteins. M-diverticula and hemolymph were labeled by in vivo (14)C-palmitic acid injection. In vitro incubations were performed between M-diverticula and either hemolymph or isolated lipoproteins. Hemolymph lipid uptake was associated to HDL1 (67%) and VHDL (32%). Release from hemolymph towards M-diverticula showed the opposite trend, VHDL 75% and HDL1 45%. Isolated lipoproteins showed a similar behavior to that observed with whole hemolymph. Lipid transfer between lipoproteins showed that HDL1 transfer more (14)C-lipids to VHDL than vice versa. Only 38% FFA and 18% TAG were transferred from M-diverticula to lipoproteins, while on the contrary 75% and 73% of these lipids, respectively, were taken up from hemolymph. A similar trend was observed regarding lipoprotein phospholipids. This study supports the hypothesis that HDL1 and hemocyanin-containing VHDL are involved in the uptake and release of FFA, phospholipids and triacylglycerols in the spider P. pythagoricus. The data support a directional flow of lipids from HDL1 and VHDL suggesting a mode of lipid transport between lipoproteins and M-diverticula., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The purpose of this study is to characterize the x-ray properties of a dual-modality, anthropomorphic breast phantom whose MRI properties have been previously evaluated. The goal of this phantom is to provide a platform for optimization and standardization of two- and three-dimensional x-ray and MRI breast imaging modalities for the purpose of lesion detection and discrimination. The phantom is constructed using a mixture of lard and egg whites, resulting in a variable, tissue-mimicking structure with separate adipose- and glandular-mimicking components. The phantom can be produced with either a compressed or uncompressed shape. Mass attenuation coefficients of the phantom materials were estimated using elemental compositions from the USDA National Nutrient Database for Standard Reference and the atomic interaction models from the Monte Carlo code PENELOPE and compared with human values from the literature. The image structure was examined quantitatively by calculating and comparing spatial covariance matrices of the phantom and patient mammography images. Finally, a computerized version of the phantom was created by segmenting a computed tomography scan and used to simulate x-ray scatter of the phantom in a mammography geometry. Mass attenuation coefficients of the phantom materials were within 20% and 15% of the values for adipose and glandular tissues, respectively, which is within the estimation error of these values. Matching was improved at higher energies (>20 keV). Tissue structures in the phantom have a size similar to those in the patient data, but are slightly larger on average. Correlations in the patient data appear to be longer than those in the phantom data in the anterior-posterior direction; however, they are within the error bars of the measurement. Simulated scatter-to-primary ratio values of the phantom images were as high as 85% in some areas and were strongly affected by the heterogeneous nature of the phantom. Key physical x-ray properties of the phantom have been quantitatively evaluated and shown to be comparable to those of breast tissue. Since the MRI properties of the phantom have been previously evaluated, we believe it is a useful tool for quantitative evaluation of two- and three-dimensional x-ray and MRI breast imaging modalities for the purpose of lesion detection and characterization.