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Ants exhibit cooperative behaviors and advanced forms of sociality that depend on pheromone-mediated communication. Odorant receptor neurons (ORNs) express specific odorant receptors (ORs) encoded by a dramatically expanded gene family in ants. In most eusocial insects, only the queen can transmit genetic information, restricting genetic studies. In contrast, workers in Harpegnathos saltator ants can be converted into gamergates (pseudoqueens) that can found entire colonies. This feature facilitated CRISPR-Cas9 generation of germline mutations in orco, the gene that encodes the obligate co-receptor of all ORs. orco mutations should significantly impact olfaction. We demonstrate striking functions of Orco in odorant perception, reproductive physiology, and social behavior plasticity. Surprisingly, unlike in other insects, loss of OR functionality also dramatically impairs development of the antennal lobe to which ORNs project. Therefore, the development of genetics in Harpegnathos establishes this ant species as a model organism to study the complexity of eusociality.

Insects, such as the malaria vector mosquito, Anopheles gambiae, depend upon chemoreceptors to respond to volatiles emitted from a range of environmental sources, most notably blood meal hosts and oviposition sites. A subset of peripheral signaling pathways involved in these insect chemosensory-dependent behaviors requires the activity of heteromeric odorant receptor (OR) ion channel complexes and ligands for numerous A. gambiae ORs (AgOrs) have been identified. Although AgOrs are expressed in nonhead appendages, studies characterizing potential AgOr function in nonolfactory tissues have not been conducted. In the present study, we explore the possibility that AgOrs mediate responses of spermatozoa to endogenous signaling molecules in A. gambiae. In addition to finding AgOr transcript expression in testes, we show that the OR coreceptor, AgOrco, is localized to the flagella of A. gambiae spermatozoa where Orco-specific agonists, antagonists, and other odorant ligands robustly activate flagella beating in an Orco-dependent process. We also demonstrate Orco expression and Orco-mediated activation of spermatozoa in the yellow fever mosquito, Aedes aegypti. Moreover, we find Orco localization in testes across distinct insect taxa and posit that OR-mediated responses in spermatozoa may represent a general characteristic of insect reproduction and an example of convergent evolution.

Background. Recent studies have suggested inferior outcome of patients treated with rabbit ATG (Thymoglobulin) as compared to horse ATG (ATGAM , Pfizer or Lymphoglobulin, Genzyme- the latter no longer available); other studies have shown comparable responses and survival. However these studies are based on a relatively small number of patients and a short follow up. Aim of the study. The aim of this study was to assess real life outcome of a large number of AA patients , treated in Europe and Asia with rabbit ATG (Thymoglobulin, SANOFI) and cyclosporin, as first line treatment . Methods and patients Eligible for this study were patients with AA treated with Thymoglobulin between 2001 and 2008 (n=501) and 2009-2012 (n=473) in Europe (n=519) or Asia (n=457). Median year of treatment was 2008 : median age (20 and 21 years), interval diagnosis treatment (23 and 25 days) and severity of the disease (46% and 48% with vSAA) were comparable in the 2 time periods. Early mortality. Mortality Response. Overall response was recorded in 799 patients. At 6 months , responses were comparable in the 2 time periods: 56% vs 57%, and at 1 year, 75% vs 73%. Response rates at 6 months were age dependent: 60%, 58%, 52%, 40% respectively in patients aged 0-20, 21-40, 41-60, >60. When non responders at 3 months were re-evaluated at 1 year, 59% had responded , 26% were non responders , 5% had died, and 10% had received other treatment. Survival: The actuarial 10 year survival for the entire population was 73%, and 72%, when patients were censored as surviving at transplant. The actuarial survival has significantly improved after year 2008, from 71% to 81% , not because of more transplants, which have remained stable (29% vs 30% before or after 2008). Other predictors of 10 year survival in univariate analysis were the following : 89%, 86%, 59% for complete, partial responders and non responders (p0.5x10^9/L (p 60 days (p=0.002); 82%, 72%, 66%, 27% in patients aged 0-20, 21-40, 41-60, > 60 years (p Conclusions. With a current overall early mortality ( Disclosures Bacigalupo: PIERRE FABRE: Speakers Bureau; SANOFI: Speakers Bureau. Carlo:Pfizer, Novartis: Consultancy. Kojima:SANOFI: Honoraria, Research Funding.

The surface-modification of paper substrates with functional layers is gaining increasing interest, both from academic and industrial research. In this case, the cellulosic paper (CP) surface was functionalized with zwitterionic poly-(3-dimethyl(methacryloyloxyethyl) ammoniumpropane sulfonate) (CP-g-P(DMAPS) via surface-initiated atom transfer radical polymerization (ATRP) technique for enhancing blood compatibility. An obvious increase in graft yield of the functional P(DMAPS) with polymerization time was observed. The new CP-g-P(DMAPS) produced was investigated for its hemocompatibility. The hemocompatibility studied including platelet and whole blood ceels adhesion tests, hemolysis assay, morphological changes of red blood cells (RBCs), coagulation time tests, and complement activation, platelet activation at the molecular level. Most assays had remarkable differences in the presence of the new zwitterionic CP, indicated the importance of the zwitterion for hemocompatibility of CP.

Background Two sibling members of the Anopheles gambiae species complex display notable differences in female blood meal preferences. An. gambiae s.s. has a well-documented preference for feeding upon human hosts, whereas An. quadriannulatus feeds on vertebrate/mammalian hosts, with only opportunistic feeding upon humans. Because mosquito host-seeking behaviors are largely driven by the sensory modality of olfaction, we hypothesized that hallmarks of these divergent host seeking phenotypes will be in evidence within the transcriptome profiles of the antennae, the mosquito’s principal chemosensory appendage. Results To test this hypothesis, we have sequenced antennal mRNA of non-bloodfed females from each species and observed a number of distinct quantitative and qualitative differences in their chemosensory gene repertoires. In both species, these gene families show higher rates of sequence polymorphisms than the overall rates in their respective transcriptomes, with potentially important divergences between the two species. Moreover, quantitative differences in odorant receptor transcript abundances have been used to model potential distinctions in volatile odor receptivity between the two sibling species of anophelines. Conclusion This analysis suggests that the anthropophagic behavior of An. gambiae s.s. reflects the differential distribution of olfactory receptors in the antenna, likely resulting from a co-option and refinement of molecular components common to both species. This study improves our understanding of the molecular evolution of chemoreceptors in closely related anophelines and suggests possible mechanisms that underlie the behavioral distinctions in host seeking that, in part, account for the differential vectorial capacity of these mosquitoes.

Olfactory-driven behaviors are central to the lifecycle of the malaria vector mosquito Anopheles gambiae and are initiated by peripheral signaling in the antenna and other olfactory tissues. To continue gaining insight into the relationship between gene expression and olfaction, we have performed cohort comparisons of antennal transcript abundances at five time points after a blood meal, a key event in both reproduction and disease transmission cycles. We found that more than 5,000 transcripts displayed significant abundance differences, many of which were correlated by cluster analysis. Within the chemosensory gene families, we observed a general reduction in the level of chemosensory gene transcripts, although a subset of odorant receptors (AgOrs) was modestly enhanced in post–blood-fed samples. Integration of AgOr transcript abundance data with previously characterized AgOr excitatory odorant response profiles revealed potential changes in antennal odorant receptivity that coincided with the shift from host-seeking to oviposition behaviors in blood-fed female mosquitoes. Behavioral testing of ovipositing females to odorants highlighted by this synthetic analysis identified two unique, unitary oviposition cues for An. gambiae, 2-propylphenol and 4-methylcyclohexanol. We posit that modest, yet cumulative, alterations of AgOr transcript levels modulate peripheral odor coding resulting in biologically relevant behavioral effects. Moreover, these results demonstrate that highly quantitative, RNAseq transcript abundance data can be successfully integrated with functional data to generate testable hypotheses.

Background The recA/RAD51 gene family encodes a diverse set of recombinase proteins that affect homologous recombination, DNA-repair, and genome stability. The recA gene family is expressed across all three domains of life - Eubacteria, Archaea, and Eukaryotes - and even in some viruses. To date, efforts to resolve the deep evolutionary origins of this ancient protein family have been hindered by the high sequence divergence between paralogous groups (i.e. ~30% average pairwise identity). Results Through large taxon sampling and the use of a phylogenetic algorithm designed for inferring evolutionary events in highly divergent paralogs, we obtained a robust, parsimonious and more refined phylogenetic history of the recA/RAD51 superfamily. Conclusions In summary, our model for the evolution of recA/RAD51 family provides a better understanding of the ancient origin of recA proteins and the multiple events that lead to the diversification of recA homologs in eukaryotes, including the discovery of additional RAD51 sub-families.

“Water-in-water” emulsions of cross-linked and hydrophobically associating cationic polyacrylamide (CHCPAM) with cationic groups (methacrylatoethyl trimethyl ammonium chloride, DMC), hydrophobic groups (octadecyl methacrylate, OA), and cross-linked groups (N, N’-methylene bisacrylamide, MBA) were prepared by dispersion polymerization. The structure of the copolymer was confirmed by FTIR and 1HNMR analyses. The prepared “water-in-water” emulsions possessed high solids content, low viscosity, good stability, and water solubility. Optical microscope images showed that the diameters of most emulsion particles were several microns. Appropriate concentrations of OA and MBA for a stable dispersion system were in the range of 0 to 0.1 wt% and 0 to 50 ppm. “Water-in-water” emulsions of CHCPAM can be directly used as a retention aid without further processing. They have a comparative retention rate, better anti-shearing ability, and better salt resistance, compared to commercial CPAM. The cross-linked structure of CHCPAM contributed to the anti-shearing ability. Intermolecular and intramolecular hydrophobic association of CHCPAM was the most important factor in improving resistance to salt.

Ants are a highly successful family of insects that thrive in a variety of habitats across the world. Perhaps their best-known features are complex social organization and strict division of labor, separating reproduction from the day-to-day maintenance and care of the colony, as well as strict discrimination against foreign individuals. Since these social characteristics in ants are thought to be mediated by semiochemicals, a thorough analysis of these signals, and the receptors that detect them, is critical in revealing mechanisms that lead to stereotypic behaviors. To address these questions, we have defined and characterized the major chemoreceptor families in a pair of behaviorally and evolutionarily distinct ant species, Camponotus floridanus and Harpegnathos saltator. Through comprehensive re-annotation, we show that these ant species harbor some of the largest yet known repertoires of odorant receptors (Ors) among insects, as well as a more modest number of gustatory receptors (Grs) and variant ionotropic glutamate receptors (Irs). Our phylogenetic analyses further demonstrate remarkably rapid gains and losses of ant Ors, while Grs and Irs have also experienced birth-and-death evolution to different degrees. In addition, comparisons of antennal transcriptomes between sexes identify many chemoreceptors that are differentially expressed between males and females and between species. We have also revealed an agonist for a worker-enriched OR from C. floridanus, representing the first case of a heterologously characterized ant tuning Or. Collectively, our analysis reveals a large number of ant chemoreceptors exhibiting patterns of differential expression and evolution consistent with sex/species-specific functions. These differentially expressed genes are likely associated with sex-based differences, as well as the radically different social lifestyles observed between C. floridanus and H. saltator, and thus are targets for further functional characterization. Our findings represent an important advance toward understanding the molecular basis of social interactions and the differential chemical ecologies among ant species., Author Summary Chemical communication is an important factor in the regulation of social interaction in animals. The family of eusocial insects commonly known as ants offers an almost unique opportunity for examining the genetic basis for the chemosensory pathways that underlie ant sociality. In order to address this issue, we have manually and comprehensively reannotated the chemoreceptor repertoire in a pair of evolutionarily and behaviorally divergent ant species, Camponotus floridanus and Harpegnathos saltator. In addition, we have used next-generation RNA sequencing to examine the chemosensory receptor transcriptome between males and females within these species. Our analysis demonstrates rapid gene birth-and-death for the ant odorant and gustatory receptor gene families, as well as clear differences in the expression of particular subsets of chemoreceptor genes between males and females. Finally, we have begun to examine the odor space within these discrete social units by heterologous characterization of the first C. floridanus odorant receptor that also exhibits sex-specific differential expression. Taken together, our results provide a foundation for future studies of the genetic basis for the chemical signaling and chemical ecology underlying the dramatically different social lifestyles exhibited by these and other species of ants.

A facile process to prepare self-reinforced antibacterial paper (SRAP) was developed by in situ synthesis of zinc oxide (ZnO) in partially dissolved cellulose. The SRAP was fabricated by impregnating filter paper in zinc chloride (ZnCl 2 ) solution and then reacting with sodium hydroxide (NaOH). Filter paper was firstly impregnated with ZnCl 2 solution of 65 wt% concentration for 5 seconds at 80 °C, and then pressed at 3.85 kPa for 5 seconds to remove excess liquid. Subsequently, the paper was soaked in a 0.8 wt% NaOH solution for 1 hour, and then washed with deionized water, and dried finally to yield SRAP. Energy dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS) and scanning electron microscope (SEM) were used to characterize the SRAP. The results revealed that the SRAP contained intact cellulose fibers as the skeleton, gelled cellulose as the matrix, and clusters of nano ZnO particles as the filler. The SRAP had a much higher density, tensile and burst strength, compared with the untreated cellulose paper, and the folding strength was enhanced by more than fifteen times. In addition, the SRAP had outstanding antibacterial properties due to the presence of nano ZnO particles.

A phylogentic analysis reveals that many gene duplications occurred prior to the split in animals, fungi and plants., Background Gene duplication is considered a major driving force for evolution of genetic novelty, thereby facilitating functional divergence and organismal diversity, including the process of speciation. Animals, fungi and plants are major eukaryotic kingdoms and the divergences between them are some of the most significant evolutionary events. Although gene duplications in each lineage have been studied extensively in various contexts, the extent of gene duplication prior to the split of plants and animals/fungi is not clear. Results Here, we have studied gene duplications in early eukaryotes by phylogenetic relative dating. We have reconstructed gene families (with one or more orthogroups) with members from both animals/fungi and plants by using two different clustering strategies. Extensive phylogenetic analyses of the gene families show that, among nearly 2,600 orthogroups identified, at least 300 of them still retain duplication that occurred before the divergence of the three kingdoms. We further found evidence that such duplications were also detected in some highly divergent protists, suggesting that these duplication events occurred in the ancestors of most major extant eukaryotic groups. Conclusions Our phylogenetic analyses show that numerous gene duplications happened at the early stage of eukaryotic evolution, probably before the separation of known major eukaryotic lineages. We discuss the implication of our results in the contexts of different models of eukaryotic phylogeny. One possible explanation for the large number of gene duplication events is one or more large-scale duplications, possibly whole genome or segmental duplication(s), which provides a genomic basis for the successful radiation of early eukaryotes.

Background Histone methylation can dramatically affect chromatin structure and gene expression and was considered irreversible until recent discoveries of two families of histone demethylases, the KDM1 (previously LSD1) and JmjC domain-containing proteins. These two types of proteins have different functional domains and distinct substrate specificities. Although more and more KDM1 and JmjC proteins have been shown to have histone demethylase activity, our knowledge about their evolution history is limited. Results We performed systematic phylogenetic analysis of these histone demethylase families and uncovered different evolutionary patterns. The KDM1 genes have been maintained with a stable low copy number in most organisms except for a few duplication events in flowering plants. In contrast, multiple genes for JmjC proteins with distinct domain architectures were present before the split of major eukaryotic groups, and experienced subsequent birth-and-death evolution. In addition, distinct evolutionary patterns can also be observed between animal and plant histone demethylases in both families. Furthermore, our results showed that some JmjC subfamilies contain only animal genes with specific demethylase activities, but do not have plant members. Conclusion Our study improves the understanding about the evolutionary history of KDM1 and JmjC genes and provides valuable insights into their functions. Based on the phylogenetic relationship, we discussed possible histone demethylase activities for several plant JmjC proteins. Finally, we proposed that the observed differences in evolutionary pattern imply functional divergence between animal and plant histone demethylases.

Figure S7. Gene body transcriptional changes between biological replicates after normalization. gb: gene body regions: gbd: read density in gene body regions; WT1: wildtype liver replicate 1; WT2; wildtype liver replicate 2; KO1: Hdac3-deleted liver replicate 1; KO2: Hdac3-deleted liver replicate 2. (PPTX 47Â kb)

Figure S2. Transcriptional levels of common and unique enhancers identified in K562 GRO/PRO-seq data. (PPTX 88Â kb)

Figure S2. Transcriptional levels of common and unique enhancers identified in K562 GRO/PRO-seq data. (PPTX 88Â kb)

Figure S4. The relationship of transcriptional changes between promoter proximal levels (x-axis) and gene body levels (y-axis) in mice-liver Hdac3 knockout (green), triptolide (purple) and flavopiridol (blue) treatment. (PPTX 711Â kb)

Figure S7. Gene body transcriptional changes between biological replicates after normalization. gb: gene body regions: gbd: read density in gene body regions; WT1: wildtype liver replicate 1; WT2; wildtype liver replicate 2; KO1: Hdac3-deleted liver replicate 1; KO2: Hdac3-deleted liver replicate 2. (PPTX 47Â kb)