Insights into the formation and diversification of a novel chiropteran wing membrane from embryonic development.

BACKGROUND: Through the evolution of novel wing structures, bats (Order Chiroptera) became the only mammalian group to achieve powered flight. This achievement preceded the massive adaptive radiation of bats into diverse ecological niches. We investigate some of the developmental processes that underlie the origin and subsequent diversification of one of the novel membranes of the bat wing: the plagiopatagium, which connects the fore- and hind limb in all bat species. RESULTS: Our results suggest that the plagiopatagium initially arises through novel outgrowths from the body flank that subsequently merge with the limbs to generate the wing airfoil. Our findings further suggest that this merging process, which is highly conserved across bats, occurs through modulation of the programs controlling the development of the periderm of the epidermal epithelium. Finally, our results suggest that the shape of the plagiopatagium begins to diversify in bats only after this merging has occurred. CONCLUSIONS: This study demonstrates how focusing on the evolution of cellular processes can inform an understanding of the developmental factors shaping the evolution of novel, highly adaptive structures.

Physicochemical Treatments of Graphene Oxide to Improve Water Vapor/Gas Separation Performance of Supported Laminar Membranes: Sonication and H2O2 Oxidation.

We investigated the previously unexplored domain of water vapor/gas separation using graphene oxide (GO) membranes, expecting future applications, including gas dehumidifiers and superior humidity controllers. While the importance of manipulation of GO nanosheet size and surface chemistry in traditional water purification and gas separation has been acknowledged, their potential impact on water vapor/gas separation remained unexplored until now. We applied sonication and hydrogen peroxide treatments to GO water dispersions and systematically evaluated the size and surface chemistry of each GO nanosheet. Both treatments reduced the GO nanosheet size to shorten the diffusion length, which improved water permeance. In addition, hydrogen peroxide treatment improved the hydrophilicity of the nanosheet. Our novel findings demonstrate that optimization of GO nanosheet size and the increase in their hydrophilicity via hydrogen peroxide treatments for 5 h significantly enhance water permeance, leading to a remarkable water vapor permeance of 4.6 × 10-6 mol/(m2 s Pa) at 80 °C, a 3.1-fold improvement over original GO membranes, while maintaining a water vapor/nitrogen permeance ratio exceeding 10,000. These results not only provide important insights into the nature of water vapor/gas separation but also suggest innovative methods for optimizing the GO membrane structure.

Dosage-dependent effects of FGFR2W290R mutation on craniofacial shape and cellular dynamics of the basicranial synchondroses.

Craniosynostosis is a common yet complex birth defect, characterized by premature fusion of the cranial sutures that can be syndromic or nonsyndromic. With over 180 syndromic associations, reaching genetic diagnoses and understanding variations in underlying cellular mechanisms remains a challenge. Variants of FGFR2 are highly associated with craniosynostosis and warrant further investigation. Using the missense mutation FGFR2W290R , an effective mouse model of Crouzon syndrome, craniofacial features were analyzed using geometric morphometrics across developmental time (E10.5-adulthood, n = 665 total). Given the interrelationship between the cranial vault and basicranium in craniosynostosis patients, the basicranium and synchondroses were analyzed in perinates. Embryonic time points showed minimal significant shape differences. However, hetero- and homozygous mutant perinates and adults showed significant differences in shape and size of the cranial vault, face, and basicranium, which were associated with cranial doming and shortening of the basicranium and skull. Although there were also significant shape and size differences associated with the basicranial bones and clear reductions in basicranial ossification in cleared whole-mount samples, there were no significant alterations in chondrocyte cell shape, size, or orientation along the spheno-occipital synchondrosis. Finally, shape differences in the cranial vault and basicranium were interrelated at perinatal stages. These results point toward the possibility that facial shape phenotypes in craniosynostosis may result in part from pleiotropic effects of the causative mutations rather than only from the secondary consequences of the sutural defects, indicating a novel direction of research that may shed light on the etiology of the broad changes in craniofacial morphology observed in craniosynostosis syndromes.

Permeation Properties of Water Vapor through Graphene Oxide/Polymer Substrate Composite Membranes.

Graphene oxide (GO) has attracted attention as an excellent membrane material for water treatment and desalination owing to its high mechanical strength, hydrophilicity, and permeability. In this study, composite membranes were prepared by coating GO on various polymeric porous substrates (polyethersulfone, cellulose ester, and polytetrafluoroethylene) using suction filtration and casting methods. The composite membranes were used for dehumidification, that is, water vapor separation in the gas phase. GO layers were successfully prepared via filtration rather than casting, irrespective of the type of polymeric substrate used. The dehumidification composite membranes with a GO layer thickness of less than 100 nm showed a water permeance greater than 1.0 × 10-6 mol/(m2 s Pa) and a H2O/N2 separation factor higher than 104 at 25 °C and 90-100% humidity. The GO composite membranes were fabricated in a reproducible manner and showed stable performance as a function of time. Furthermore, the membranes maintained high permeance and selectivity at 80°C, indicating that it is useful as a water vapor separation membrane.

Conserved and Taxon-Specific Patterns of Phenotypic Modularity in the Mammalian Dentition.

Previous genotype:phenotype mapping of the mouse and primate dentition revealed the presence of pre- and post-canine modules in mice and anthropoid primates, as well as molar and premolar submodules in anthropoid primates. We estimated phenotypic correlation matrices for species that sample broadly across Mammalia to test the hypothesis that these modules exist across a broader range of taxa and thereby represent a conserved mammalian trait. We calculated phenotypic correlation matrices from linear dental measurements of 419 individual specimens representing 5 species from 4 mammalian orders: Artiodactyla (Odocoileus hemionus), Carnivora (Canis latrans and Ursus americanus), Didelphimorphia (Didelphis virginiana), and Primates (Colobus guereza). Our results based on hierarchical clustering indicate a generally higher correlation within incisors and among post-canine teeth. However, the post-canine phenotypic correlation matrices do not consistently exhibit the premolar and molar submodularity observed in anthropoid primates. Additionally, we find evidence of sex differences in the Odocoileus phenotypic correlation matrices: Males of this species exhibit overall higher inter-trait correlations compared to females. Our overall findings support the interpretation that incisors and post-canine dentition represent different phenotypic modules, and that this architecture may be a conserved trait for mammals.

Ecological basis and genetic architecture of crypsis polymorphism in the desert clicker grasshopper (Ligurotettix coquilletti).

Color polymorphic species can offer exceptional insight into the ecology and genetics of adaptation. Although the genetic architecture of animal coloration is diverse, many color polymorphisms are associated with large structural variants and maintained by biotic interactions. Grasshoppers are notably polymorphic in both color and karyotype, which makes them excellent models for understanding the ecological drivers and genetic underpinnings of color variation. Banded and uniform morphs of the desert clicker grasshopper (Ligurotettix coquilletti) are found across the western deserts of North America. To address the hypothesis that predation maintains local color polymorphism and shapes regional crypsis variation, we surveyed morph frequencies and tested for covariation with two predation environments. Morphs coexisted at intermediate frequencies at most sites, consistent with local balancing selection. Morph frequencies covaried with the appearance of desert substrate-an environment used only by females-suggesting that ground-foraging predators are major agents of selection on crypsis. We next addressed the hypothesized link between morph variation and genome structure. To do so, we designed an approach for detecting inversions and indels using only RADseq data. The banded morph was perfectly correlated with a large putative indel. Remarkably, indel dominance differed among populations, a rare example of dominance evolution in nature.

Simultaneous Extraction and Determination of Volatile Organic Compounds and Semi-volatile Organic Compounds in Indoor Air Using Multi-bed Solid Phase Extraction Device.

A method for the simultaneous extraction and determination of indoor volatile compounds, including volatile organic compounds (VOCs) and semi-volatile organic compounds (SVOCs), was developed using a multi-bed solid phase extraction (SPE)-type collection device. The collection device was prepared by packing styrene-divinylbenzene polymer particles and activated carbon particles. The collected analytes were completely desorbed by passing 7 mL of acetone, and the solvent was then injected into a gas chromatograph-mass spectrometry without the concentration process. Because the proposed method does not require ultrasonication and a concentration process of eluted solvent, quantitative determination of a relatively volatile compound could be achieved. The total recovery including extraction and elution recoveries for all the investigated analytes were in the range from 91.6 to 109%. The limit of quantification was less than 4.0 ng L-1 for all the investigated analytes, and relative standard deviations of the peak area of the analytes in indoor air were less than 12%. The collection device could be reused for over 50 samplings.

Emergent Coordination of the CHKB and CPT1B Genes in Eutherian Mammals: Implications for the Origin of Brown Adipose Tissue.

Mitochondrial fatty acid oxidation (FAO) contributes to the proton motive force that drives ATP synthesis in many mammalian tissues. In eutherian (placental) mammals, brown adipose tissue (BAT) can also dissipate this proton gradient through uncoupling protein 1 (UCP1) to generate heat, but the evolutionary events underlying the emergence of BAT are unknown. An essential step in FAO is the transport of cytoplasmic long chain acyl-coenzyme A (acyl-CoA) into the mitochondrial matrix, which requires the action of carnitine palmitoyltransferase 1B (CPT1B) in striated muscle and BAT. In eutherians, the CPT1B gene is closely linked to the choline kinase beta (CHKB) gene, which is transcribed from the same DNA strand and terminates just upstream of CPT1B. CHKB is a rate-limiting enzyme in the synthesis of phosphatidylcholine (PC), a predominant mitochondrial membrane phospholipid, suggesting that the coordinated expression of CHKB and CPT1B may cooperatively enhance mitochondrial FAO. The present findings show that transcription of the eutherian CHKB and CPT1B genes is linked within a unitary epigenetic domain targeted to the CHKB gene, and that that this regulatory linkage appears to have resulted from an intergenic deletion in eutherians that significantly altered the distribution of CHKB and CPT1B expression. Informed by the timing of this event relative to the emergence of BAT, the phylogeny of CHKB-CPT1B synteny, and the insufficiency of UCP1 to account for eutherian BAT, these data support a mechanism for the emergence of BAT based on the acquisition of a novel capacity for adipocyte FAO in a background of extant UCP1.

Functional Analysis of the Role of Equilibrative Nucleobase Transporter 1 (ENBT1/SLC43A3) in Adenine Transport in HepG2 Cells.

Equilibrative nucleobase transporter 1 (ENBT1/SLC43A3) has recently been identified as a purine-selective nucleobase transporter. Although it is highly expressed in the liver, its role in nucleobase transport has not been confirmed yet in hepatocytes or any relevant cell models. We, therefore, examined its role in adenine transport in the HepG2 cell line as a human hepatocyte model. The uptake of [3H]adenine in HepG2 cells was highly saturable, indicating the involvement of carrier-mediated transport. The carrier-mediated transport component, for which the Michaelis constant was estimated to be 0.268 µM, was sensitive to decynium-22, an ENBT1 inhibitor, with the half maximal inhibitory concentration of 2.59 µM, which was comparable to that of 2.30 µM for [3H]adenine uptake by ENBT1 in its transient transfectant human embryonic kidney 293 cells. Although equilibrative nucleoside transporter 1 (ENT1/SLC29A1) and ENT2/SLC29A2 are also known to be able to transport adenine, [3H]adenine uptake in HepG2 cells was not inhibited by the ENT1/2-specific inhibitor of either dipyridamole or nitrobenzylthioinosine. Finally, [3H]adenine uptake was extensively reduced by silencing of ENBT1 by RNA interference in the hepatocyte model. All these results, taken together, suggest the predominant role of ENBT1 in the uptake of adenine in HepG2 cells.

Ecological and demographic impacts of a recent volcanic eruption on two endemic patagonian rodents.

Catastrophic events can significantly impact the demographic processes that shape natural populations of organisms. However, linking the outcomes of such events to specific demographic parameters is often challenging due to a lack of detailed pre-event data. The eruption of the Puyehue-Cordon Caulle volcanic complex on 4 June 2011 had profound consequences for the biota of southwestern Argentina. Our long-term behavioral, ecological, and demographic studies of two species of tuco-tucos (Ctenomys sociabilis and C. haigi) that occur in the region most heavily impacted by ash fall from the eruption provided an unusual opportunity to assess the effects of this event on natural populations of mammals. The post-eruption density of the study population for each species was markedly reduced compared to pre-eruption values, with the relative magnitude of this reduction being greater for the group-living C. sociabilis. The more extensive data set for this species indicated that ash fall from the eruption altered the food resources available to these animals; differences in pre- and post-eruption stable isotope signatures for fur samples from C. sociabilis were consistent with observed changes in vegetation. Per capita female reproductive success was also reduced in this species during the first breeding season following the eruption. Based on our detailed demographic records for C. sociabilis, neither survival of yearling females from 2010 to 2011 nor the percentage of unmarked females in the study population in 2011 differed from pre-eruption values. Instead, the post-eruption decrease in population density for C. sociabilis appeared to reflect reduced within-population recruitment of juvenile females to the 2011 breeding population. Although the eruption did not result in the local extinction of either study population, the demographic consequences detected are likely to have impacted the effective sizes of these populations, creating important opportunities to link specific demographic parameters to previously reported decreases in genetic variability detected after this significant natural event.

Quantitative Determination of 2-Ethyl-1-hexanol, Texanol and TXIB in In-door Air Using a Solid-Phase Extraction-type Collection Device Followed by Gas Chromatography-Mass Spectrometry.

In this study, in-door air semi-volatile organic compounds (SVOCs) including 2-ethyl-1-hexanol, 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate (texanol), and 2,2,4-trimethyl-1,3-pentanediol diisobutyrate (TXIB), which are scheduled for adding as regulated compounds concerning indoor air reference values in Japan, were quantitatively extracted using a solid-phase extraction-type collection device, followed by sensitively determined by gas chromatography-mass spectrometry. The developed method has shown a good extraction recovery up to an air sampling volume of 900 L. The extracted analytes were quantitatively and rapidly eluted by 7 mL of acetone. The limit of quantification of the analytes were 0.7, 2.1 and 0.2 ng L-1 in air sample at a sampling volume of 300 mL without any concentration of a desorption solvent. The developed method was applied to simultaneous determinations of the investigated target analytes and phthalate esters in real indoor air samples.

Evidence of strong stabilizing effects on the evolution of boreoeutherian (Mammalia) dental proportions.

The dentition is an extremely important organ in mammals with variation in timing and sequence of eruption, crown morphology, and tooth size enabling a range of behavioral, dietary, and functional adaptations across the class. Within this suite of variable mammalian dental phenotypes, relative sizes of teeth reflect variation in the underlying genetic and developmental mechanisms. Two ratios of postcanine tooth lengths capture the relative size of premolars to molars (premolar-molar module, PMM), and among the three molars (molar module component, MMC), and are known to be heritable, independent of body size, and to vary significantly across primates. Here, we explore how these dental traits vary across mammals more broadly, focusing on terrestrial taxa in the clade of Boreoeutheria (Euarchontoglires and Laurasiatheria). We measured the postcanine teeth of N = 1,523 boreoeutherian mammals spanning six orders, 14 families, 36 genera, and 49 species to test hypotheses about associations between dental proportions and phylogenetic relatedness, diet, and life history in mammals. Boreoeutherian postcanine dental proportions sampled in this study carry conserved phylogenetic signal and are not associated with variation in diet. The incorporation of paleontological data provides further evidence that dental proportions may be slower to change than is dietary specialization. These results have implications for our understanding of dental variation and dietary adaptation in mammals.

Quantitative Determination of Phthalate Esters from Air Samples Using a Solid-Phase Extraction-type Collection Device.

In this study, a solid-phase extraction-type collection device, with styrene-divinylbenzene polymer particles (Sunpak-H) as the adsorbent, was used for the quantitative determination of phthalate esters in air samples. The collection and elution recoveries of eight volatile phthalate esters, i.e., dimethyl phthalate, diethyl phthalate, dipropyl phthalate, diisobutyl phthalate, dibutyl phthalate, butyl-benzyl phthalate, di(2-ethylhexyl) phthalate, and dioctyl phthalate, were quantitatively evaluated. All analytes were collected using the device up to a sampling volume of 10000 L at a sampling temperature of 35°C without breakthrough. During air collection, moisture was not trapped on the adsorbent. The collected analytes were completely eluted from the device by passing 3 mL of acetone. The eluted solvent was injected into a gas chromatography-mass spectrometry system after the eluted solvent was concentrated, if necessary. After washing the adsorbent using acetone, the device could be reused more than 50 times. The limit of quantification for the analytes was less than 1 ng L-1 in air at a sampling volume of 600 L with solvent concentration. This device was successfully applied for the quantitative determination of phthalate esters in real air samples, including indoor and in-car air.

Widespread dispersal and bio-accumulation of toxic microcystins in benthic marine ecosystems.

Freshwater cyanobacteria produce toxic microcystins (MCs), which travel from freshwater areas into the sea. The MCs produced by cyanobacteria in a freshwater reservoir were discharged frequently into the adjacent Isahaya Bay, remained in the surface sediments, and then accumulated in various macrobenthic animals on the seafloor. The MCs were transported further outside of Isahaya Bay (Ariake Bay), and the median values of the MC contents in the sediments were in the same levels in both bays, while their temporal variations were also similar during the study period. Therefore, the fluctuations of the MC contents in the surface sediments were physically controlled by the timing of the discharge from the reservoir. The MC contents in polychaetes and oysters collected in Isahaya Bay increased markedly during winter. The median values of the carbon-based MC contents in the sediments, primary consumers, and secondary consumers in the bay were 87, 160, and 250 ngMC gC-1, respectively. These results demonstrated bio-accumulation at lower trophic levels in benthic marine ecosystems. An understanding of the processes occurring between sediments and macrobenthic animals is important for clarifying MC dynamics in ecosystems.