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Sexual conflict is common in animals, and female sexual cannibalism represents an extreme form of sexual conflict. Males in many species have evolved a variety of strategies to circumvent or decrease the risk of female sexual cannibalism. Opportunistic mating, by which a male mates with a female when she is disturbed or when she is feeding or undertaking moulting, is one of such kinds of strategies, and widely occurs in many animals, especially in spiders. However, whether the occurrence of male opportunistic mating depends on the intensity of female sexual cannibalism remains largely unexplored. We predicted a positive correlation between them. In this study, we tested this prediction by performing a series of mating trials in the laboratory using 3 species of web-building spiders with different intensities of female sexual cannibalism: Nephila pilipes, Nephilengys malabarensis, and Parasteatoda tepidariorum. We found that the occurrence of male opportunistic mating was positively, though not statistically significantly, correlated with the intensity of female sexual cannibalism, thus supporting our hypothesis. All together, we provide evidence that male opportunistic mating may have evolved to respond to the selection pressure posed by female sexual cannibalism.

Decorating behavior is common in various animal taxa and serves a variety of functions from camouflage to communication. One predominant function cited for decoration is to avoid predators. Conspicuous, disc-like (discoid) silk decorations spun by orb-web Argiope juvenile spiders are hypothesized, among others, to defend spiders against visual predators by concealing spider outlines on the web, deflecting attacks, shielding them from view, or masquerading as bird-droppings. However, the direct evidence is limited for a specific mechanism by which discoid decorations may deter predators. Here we evaluate the mechanisms by which discoid decorations may defend Argiope juveniles against naïve chicks. Using visual modeling, we show that avian predators are able to distinguish spiders from discoid decorations. Using chick predation experiments, we found that the naïve chicks readily pecked any objects, ruling out the possibility of their neophobia. Significantly more chicks attacked spiders when they were exposed to chicks, regardless of whether their webs had discoid decorations, but few chicks attacked spiders when they were behind the decorations. We also found that significantly few chicks attacked decorations when spiders were absent or behind the decorations. We thus conclude that discoid decorations function to deter avian predators by shielding the spider from view or distracting, not by deflecting attacks, concealing the spider’s outline, or masquerading as bird-droppings. This study sheds light on the study of other similar anti-predator strategies, in a wide range of spider species and other animals that use decorating strategies.

Insects' intestinal microbes have profound effects on the host's physiological traits, which can impact their physiology at both the local (gut) and systemic (body) levels. Nevertheless, the molecular mechanisms underlying host-microbiota interactions, especially in non-model insects, remain elusive. Recently, tissue-specific transcriptomic analysis has been highlighted as a robust tool in studying host-microbe interactions. Plagiodera versicolora is a worldwide leaf-eating pest that primarily feeds on willows and poplar. The interplay between gut microflora and this host beetle has yet to be studied. Herein, we investigate the effects of the gut microbiota on the body mass of P. versicolora larvae, compare the nutrition status of larvae in absence and presence of gut microbiota, and profile gut bacterial loads throughout its developmental larval stages. We then perform comparative transcriptomic analysis of gut and body tissues in axenic and non-axenic larvae. Finally, we confirm the expression patterns of representative genes in nutritional metabolism and immunity. Results show that weight growth is retarded in conventional larvae, with a concomitant increase of total bacterial load by the 5th development day, and germ-free larvae have a higher glucose content than conventional-reared larvae. Both nutritional and immunological analyses indicate that gut bacteria are a burden in the beetle's larval development. These findings elucidate the impacts of gut microbiota on P. versicolora, and provide insight into tissue-specific responses to gut microflora in this pest at the genetic level, boosting our understanding of the molecular mechanisms underlying host-microbe interactions in leaf beetles and beyond.

NJ001 is a monoclonal antibody that can specifically recognize the SP70 antigen on lung adenocarcinoma cells. The goal of this study was to explore its utility in targeted imaging. Subcutaneous xenograft and orthotopic lung tumor implantation BALB/c mouse models were established. Near-infrared fluorescent CF750-labeled NJ001 was injected into two tumor mouse models. Mice that received orthotopic lung tumor implantation were also injected with NJ001-conjugated nanomagnetic beads intravenously, and then underwent micro-CT scanning. Meanwhile, mice with lung tumor were intravenously injected with normal saline and bare nanomagnetic beads as a control. Fluorescence could be monitored in the mice detected by anti-SP70 fluorescence imaging, which was consistent with tumor burden. Signal intensities detected with SP70-targeted micro-CT scans were greater than those in control mice. More importantly, orthotopic tumor lesions could be found on the fourth week with SP70-targeted imaging, which was 2 weeks earlier than detection in the control. Our results suggest that SP70 is a promising target for molecular imaging, and molecularly targeted imaging with an NJ001-labeled probe could be applied for the early detection of lung adenocarcinoma.

Intestinal organoids are three-dimensional (3D) cultures with a long history of being studied in vivo. They constitute a model that has been widely applied to many aspects of bioengineering and regenerative medicine. Recently, increasing demand has created an urgent need for more efficient approaches to organoid culture. Here, we present an overview of some vital factors that may affect organoid formation and maturation and explain some currently available advanced methods using pluripotent stem cells (PSCs) and scaffolding to produce intestinal organoids that best resemble the organ of origin. These methods are compared with traditional two-dimensional (2D) culturing methods. In addition, we note some remaining challenges and explore other potential factors that may help improve 3D culture technology in the future.

Mating may change a male’s behaviour by increasing its motivation to engage in a contest, and enabling it to win in subsequent male–male contests. To test this hypothesis, we recorded male contests in the wolf spider, Venonia coruscans (Araneae: Lycosidae), testing a male’s motivation to fight under three different resource value conditions. First, we staged contests between two males in two different resource value conditions, on an egg-produced female’s web and then on a virgin female’s web, to test a male’s fighting ability. After determining each male’s fighting ability, we allowed each loser that lost its contests under both resource value conditions to mate with a virgin female and then introduced the previous contest winner to the web where the loser had mated. We found that without mating, the losers always lost their contests, regardless of the resource value conditions. However, once they had mated, the losers fiercely attacked the previous winners, and most won the contests back. Our study therefore provides evidence that a male’s motivation to fight can be changed under certain circumstances (e.g. mating) and can greatly influence contest outcomes in male–male competition in a mating context.

Prey vary dramatically in quality, and maternal diet is generally assumed to substantially influence offspring survivorship, growth, and reproduction in spiders. Numerous studies that have tested this hypothesis have focused exclusively on parental generation or have considered relatively few fitness components of juvenile offspring. However, maternal diet may have a substantial effect on fitness performance beyond juvenile offspring. Here, we investigated the influence of one-time maternal feeding on multiple offspring fitness components, including the survival rate and growth of juvenile offspring as well as the mating and reproductive success of adult offspring in Hylyphantes graminicola, a sheetweb spider with an extremely short lifespan (∼1 month). We fed field-collected adult female spiders two different diets only once immediately before oviposition: midges (Tendipes sp.) only (MO) or flies (Drosophila melanogaster) only (FO). Juvenile offspring of MO females had significantly higher survival rate, faster growth, and larger male size at maturity than FO offspring. Although maternal diet did not significantly influence mating behavior or fecundity of female offspring overall, those of MO females laid eggs earlier and their eggs also hatched earlier and had a higher hatching rate than those of FO females. Intriguingly, one-time maternal feeding was sufficient to have such an influence on offspring fitness even beyond juvenile offspring in H. graminicola. This one-time maternal effect may be widespread in other spiders and other invertebrates with a short lifespan. This article has an associated First Person interview with the first author of the paper., Summary: A single meal immediately before oviposition is sufficient to influence offspring survival, growth, and daughters' reproduction in the sheetweb spider Hylyphantes graminicola.

Background Mortality due to septic shock is relatively high. The dynamic monitoring of plasma cell-free DNA (cfDNA) can guide the treatment of septic shock. Case summary Herein, we present a typical case of septic shock syndrome caused by the bacilli Acinetobacter baumannii and Pantoea. The patient complained of abdominal pain, fever and chills upon admission to the Emergency Department. Marked decreases in white blood cells and procalcitonin (PCT) were observed after the patient received continuous renal replacement and extracorporeal membrane oxygenation. Plasma cfDNA levels were consistently high, peaking at 1366.40 ng/mL, as measured by a duplex real-time PCR assay with an internal control, which was developed as a novel method for the accurate quantification of cfDNA. The patient died of septic shock on HD 8, suggesting that cfDNA could be used to monitor disease progression more effectively than PCT and the other inflammatory factors measured in this case. Conclusion CfDNA may be a promising marker that complements other inflammatory factors to monitor disease progression in patients with septic shock.

Prey proteins and lipids greatly impact predator life-history traits. However, life-history plasticity offers predators the opportunity to tune the life-history traits in response to the limited macronutrients to allocate among traits. A fast-growing predator species with a strict maturation time may be more likely to consume nutritionally imbalanced prey. Here, we tested this hypothesis by examining the effect of the protein-to-lipid ratio in prey on a small sheet web-building spider, Hylyphantes graminicola, with a short life span, using adult Drosophila melanogaster as the prey. By manipulating the macronutrient content of the prey to generate three prey types with different protein-to-lipid ratios (i.e. high, intermediate and low), we demonstrated that the majority of the spiders that consumed only these flies could reach full maturity. However, juvenile spiders that consumed high-lipid (low protein-to-lipid ratio) flies had a higher rate of mortality than those consuming medium-protein and high-protein flies. The prey protein-to-lipid ratio had no significant effects on the developmental duration and size at maturity. Although the prey protein-to-lipid ratio had no significant influence on mating behaviour and female fecundity, females reared on high-lipid flies exhibited a significant delay in oviposition compared with those reared on high-protein flies. We conclude that high-lipid prey has negative effects on the survival and reproductive function of H. graminicola Our study thus provides clear evidence that low plasticity with fast development to a certain size means a high nutritional requirement for protein at a cost of lower survival and prolonged time to egg laying when prey have low protein-to-lipid content in H. graminicola.

Anticardiolipin antibody (ACA) is a kind of autoantibody and is one of the antiphospholipid antibodies (aPLs). Phospholipids with a negative charge on platelets and endothelial cell membranes are ACA target antigens. ACA is common in systemic lupus erythematosus and other autoimmune diseases and is closely associated with thrombosis, thrombocytopenia, and spontaneous abortion. In 1983, Harris established a method for detecting ACA, and research on the antibody has gained worldwide attention and has developed rapidly. For this review, we browsed articles that cover most of the ACA-related studies in the last 25 years and extracted influential ideas and conclusions in this field.

Visual signals are widely used by animals in foraging, mating and predator avoidance. Body coloration plays an important role in the reproductive behavioural interactions of many diurnal animal species; however, its role in the courtship of nocturnal species has received little attention. We investigated the role of conspicuous body coloration, a white stripe on the forehead region, in the reproductive interactions of the nocturnal brown huntsman spider, Heteropoda venatoria. We performed a series of mating trials in which we manipulated the body coloration of males and the vision of females and determined the probabilities of mating success of males in different treatments. Results showed that males that courted females with intact vision had a significantly lower probability of mating success than males that courted females whose vision was blocked. Females with intact vision were also more likely to choose males with a white stripe than those whose stripe was removed. Therefore, we have unambiguously demonstrated for the first time that body coloration plays an important role as a visual signal in the mating of nocturnal spiders.

SP70 is a novel tumor biomarker in patients with nonsmall cell lung cancer (NSCLC). However, its role as a marker for predicting the response to chemotherapy for patients with advanced NSCLC has not been investigated. A total of 152 patients were enrolled. Serum SP70, carcinoembryonic antigen (CEA), cytokeratin 19 fragment (CYFRA21‐1), and neuron‐specific enolase (NSE) were detected before and after 2 cycles of chemotherapy. The correlation between serum tumor biomarker levels and chemotherapy responses and their association with epidermal growth factor receptor (EGFR) mutation status and progression‐free survival (PFS) were analyzed. Serum SP70 levels were significantly decreased after chemotherapy in the partial remission (PR) group (P

Elevated CO2 concentrations can change the multi-level nutritional relationship of the ecosystem through the cascading effect of the food chain. To date, few studies have investigated the effects of elevated CO2 concentration on the Araneae species through the tritrophic system. Hylyphantes graminicola (Araneae: Linyphiidae) is distributed widely in Asia and is a dominant predator in cotton fields. This study investigated chemical components in the food chain of cotton (Gossypium hirsutum)—cotton aphid (Aphis gossypii)—predator (H. graminicola) and compared the development, reproduction, and predation of H. graminicola under ambient (400 ppm) and elevated concentration of CO2 (800 ppm). The results showed that the elevated CO2 concentration increased the chemicals of cotton and cotton aphid, but it did not affect the nutrients, development, reproduction, and predation of the spider. However, the survival rate of the spider was significantly decreased in elevated CO2. The results will further our understanding of the role of natural enemies in an environment with elevated CO2 concentration.

Animals that use deceptive visual signals to attract prey often employ colour mimicry or bioluminescence but less commonly self-excreted lures. The conspicuous web decorations and silks of some web-building spiders have been shown to visually lure prey in the daytime. However, it remains unknown whether spider webs can also lure prey at night with these self-produced silks. In this study, we tested this hypothesis by manipulating the coloration of the webs of Psechrus clavis and the presence of the web dwellers. We monitored the foraging performance of four treatment groups (spider present, normal web; spider present, blackened web; spider absent, normal web; spider absent, blackened web) and measured the reflectance spectra of spider body parts and web silks. We found that both spider body colour and web silk can lure insect prey at night. Lepidopteran insects comprised the majority of attracted prey. This study is the first to empirically demonstrate that animals can use self-produced substances to visually lure prey in a nocturnal environment and improve foraging success.

Spider silks are protein-based fibers that are incorporated into webs with the unique combination of high mechanical toughness and resistance to microbial degradation. While spiders are undoubtedly exposed to saprophytic microorganisms in their native habitats, such as the forest understory and bush, their silks have rarely been observed to decompose in either field or laboratory studies. We performed cross-streaking assays using silk from three spider species and four bacterial strains and found no inhibition zones, indicating the absence of antibacterial properties. We also cultured all bacteria directly upon silk in Luria-Bertani (LB) broth (full nutrients), phosphate-buffered saline (PBS; no nutrients) and nitrogen-free glucose broth (NFG; full nutrients, no nitrogen), and found that bacteria grew readily on silk in LB broth but not in PBS or NFG buffer. Our results indicate that spider silk's resistance to bacterial degradation is likely due to bacteriostatic rather than antibacterial mechanisms when nitrogen is inaccessible.

Body coloration serves a variety of purposes in animals. Diurnal and nocturnal predators such as spiders may use their body coloration to lure prey. We predicted here that the white patches on the forelegs on females of the nocturnal semi-aquatic spider Dolomedes raptor lure prey, explaining why they are primarily displayed when the spider forages along the water edge. To test our prediction, we developed a color vision model assessing whether the patches are visible to pygmy grasshoppers, the spider's primary prey. We conducted a field experiment using cardboard dummies that resemble D. raptor in size, shape, and color, but with half of them lacking leg patches, and we staged interactions between pygmy grasshoppers and D. raptor with and without leg patches in a greenhouse. We found the white patches to be visible to grasshoppers. The dummies with white patches attracted more grasshopper prey than the dummies without the patches. Moreover, grasshoppers were more attracted to spiders when their white patches were present. Our results supported the hypothesis that the white patches of D. raptor lure prey. Our findings, nevertheless, could not be explained as the spider's body coloration acting as a sensory trap but it should not be ruled out. More studies on a wider range of predators and prey will give more meaningful insights into the co-evolution of predatory lures and prey sensory modalities.

The slow progress in clinical applications of stem cells and the bewildering mechanisms involved have puzzled many researchers. Recently, the increasing evidences have indicated that cells have superior plasticity in vivo or in vitro, spontaneously or under extrinsic specific inducers. The concept of stem cells may be challenged, or even replaced by the concept of cell plasticity when cell reprogramming technology is progressing rapidly. The characteristics of stem cells are manifestations of cellular plasticity. Incorrect understanding of the concept of stem cells hinders the clinical application of so-called stem cells. Understanding cellular plasticity is important for understanding and treating disease. The above issues will be discussed in detail to prove the reconceptualization of stem cells from cellular plasticity.

Lactation is a mammalian attribute, and the few known nonmammal examples have distinctly different modalities. We document here milk provisioning in a jumping spider, which compares functionally and behaviorally to lactation in mammals. The spiderlings ingest nutritious milk droplets secreted from the mother’s epigastric furrow until the subadult stage. Milk is indispensable for offspring survival in the early stages and complements their foraging in later stages. Maternal care, as for some long-lived vertebrates, continues after the offspring reach maturity. Furthermore, a female-biased adult sex ratio is acquired only when the mother is present. These findings demonstrate that mammal-like milk provisioning and parental care for sexually mature offspring have also evolved in invertebrates, encouraging a reevaluation of their occurrence across the animal kingdom, especially in invertebrates.

Nocturnal animals frequently possess seemingly conspicuous colour patterns that can function in a variety of ways (e.g. prey attraction, camouflage, predator avoidance, etc.). The use of colour patterns in intraspecific signalling, especially reproductive activities, in nocturnal animals has received relatively little attention. This study tested for a dual function of colour in the nocturnal fishing spider, Dolomedes raptor (Araneae: Pisauridae), whose males develop dimorphic white stripes at sexual maturation. We tested for a role in foraging as well as mate assessment. First, quantifications of the natural variation of male stripes indicated a correlation between stripe area and male body size and weight. Subsequent diet experiments confirmed that the area of a male's white stripes are body size-dependent and thus could potentially convey information to choosey females about male quality. Field experiments used dummies resembling D. raptor in appearance to test a prey attraction function of the white stripes. We found that dummies with the white stripes present attracted significantly more prey than those without stripes. Finally, we used males with manipulated phenotypes in laboratory mating trials and found that males with intact white stripes were significantly more likely to be accepted by females than those with the white stripes eliminated. Together, our results document a nutrient-dependent trait that functions not only in strengthening foraging success, but also in a mating context, increasing male mating success. We suggest that the role of these male white stripes in reproduction has been facilitated by their function in foraging.

Body coloration has been known to play an important role in animal visual communication. Attraction of prey by visual signals, such as bright body coloration, has been mainly observed in diurnal predators. For nocturnal cursorial predators, however, this foraging tactic has been largely ignored, because one would not expect it to occur under low ambient illumination at night and because of the technical constraints of conducting research in dim light conditions. Metabolic constraints prevent most cursorial spiders moving continuously for a long time to improve prey encounter rate. Any adaptation that induces prey to orient towards these cursorial predators would greatly enhance their foraging success. Bright body coloration of certain nocturnal invertebrates may play such a role. In this study, we investigated colour-mediated prey attraction in a nocturnal cursorial predator, the brown huntsman spider, Heteropoda venatoria , which has a conspicuous moustache-like white stripe on the forehead. We conducted field experiments using dummy and real spiders and monitored with infrared video cameras the responses of nocturnal prey to treatment groups with the white stripe intact or removed. The results showed that in both dummy and real spiders the presence of the white stripe can significantly increase prey attraction rate. Nocturnal flying insects such as moths comprised the majority of attracted prey. We conclude that cursorial invertebrate predators can visually lure prey, especially flying insects such as moths, by their conspicuous body coloration at night.

Sexually selected ornaments and signals are costly to maintain if they are maladaptive in nonreproductive contexts. The jumping spider Cosmophasis umbratica exhibits distinct sexual dichromatism with males displaying elaborate UV body markings that signal male quality. Female C. umbratica respond favorably to UV-reflecting males and ignore males that have their UV masked. However, Portia labiata, a UV-sensitive spider-eating specialist and a natural predator of C. umbratica, is known to use UV reflectance as a cue when hunting prey. We investigated the cost of these UV signals in C. umbratica in terms of their predation risk. Under experimental conditions, three choice scenarios were presented to P. labiata individuals. Choices by P. labiata were made between male C. umbratica with and without the UV signal; a UV-reflecting male and non-UV-reflecting female; and a UV-masked male and female. The presence and absence of UV signals was manipulated using an optical filter. Portia labiata exhibited a strong bias toward UV+ individuals. These results suggest the sexually selected trait of UV reflectance increases the visibility of males to UV-sensitive predators. The extent of this male-specific UV signal then is potentially moderated by predation pressure. Interestingly though, P. labiata still preferred males to females irrespective of whether UV reflectance was present or not. This suggests P. labiata can switch cues when conditions to detect UV reflectance are not optimal.

Web building has been such a highly successful foraging innovation among spiders that the vast majority of extant spiders are web builders. The structure of spider webs varies substantially between species, and web building has even been lost completely in some clades. Examples of different web forms include the classic orb webs, which may be orientated vertical to the ground or horizontal, sheet webs, and cobwebs, which consist of three-dimensional meshwork and ascending sticky threads for support and capture of prey. The architecture of webs may also vary within clades and even within species. This may be a consequence of: (i) individuals adapting their web structures to the environment; e.g., larger webs are built in areas where more space is available, (ii) spiders varying their webs to tune its performance, e.g., when spiders are exposed to different prey, or (iii) silk expression constraints, e.g., when on diets lacking certain nutrients. We review the literature, focusing on contributions from the Neotropical region, showing that spider webs vary in structure and function at multiple levels and so must be considered a dynamic, variable, extended phenotype of its builder. Webs accordingly depict the foraging, mating, and defensive strategies, and physiological status, of the spider.

While molting occurs in the development of many animals especially in arthropods, post-maturity molting (PMM, organisms continue to molt after sexual maturity) has received little attention. Mechanism of molting has been studied intensively, however, the mechanism of PMM remains unknown although it is suggested to be crucial for the development of body size. In this study, we investigated factors that potentially induce PMM in the golden orb-web spider Nephila pilipes, which has the greatest degree of sexual dimorphism among terrestrial animals. We manipulated the mating history and the nutrient consumption of the females to examine whether they can affect PMM. The results showed that female spiders under low nutrition were more likely to molt as adults, and mating had no significant influence to the occurrence rate of PMM. Moreover, spiders that experienced PMM lived longer than those without and their body sizes were significantly increased. Therefore, we concluded that it is the nutritional condition rather than the mating history that has affected PMM.

Emasculation—males becoming effectively sterile by self-removing their genitals—has long been considered a peculiar evolutionary phenomenon with unknown function, taxonomically restricted to few spiders and flies. In spiders, emasculation results in half or full eunuchs when males sever one or both sperm transferring organs, palps. Three types of emasculation, pre-maturation, mating, and post-mating are known in spiders, all having evolved multiple times. Males practicing pre-maturation emasculation sever one of their palps while still immature, then engage in strict monogyny via genital plugging and spontaneous death. Emasculation during mating also results in genital plugs, but half eunuchs have another chance to mate. So far, the behavior of those males that become eunuchs post-mating by self-removing disfigured palps has not been investigated empirically. We test the mechanism and adaptive significance of post-mating emasculation in coin spiders (Herennia multipuncta) and use phylogenetic reconstruction to understand its evolutionary history. Our laboratory assays corroborate three hypotheses related to mate monopolization: (1) The plugging hypothesis—predicting genital plugs to prevent female remating; (2) The better-fighter hypothesis—predicting enhanced eunuch aggressiveness toward rivals; and (3) The gloves-off hypothesis—predicting increased eunuch endurance. The support for these hypotheses in spiders practicing emasculation during and after mating reinforces recent phylogenetic interpretations of these two emasculation types being evolutionarily linked in the family Nephilidae. We weigh the evidence in support of three different, but equally parsimonious scenarios of nephilid emasculation evolution. We conclude that emasculation is an adaptive, sexually selected trait that calls for further comparative and experimental research.

Tissue-engineered liver using primary hepatocytes has been considered a valuable new therapeutic modality as an alternative to whole organ liver transplantation for different liver diseases. The development of clinically feasible liver tissue engineering approaches, however, has been hampered by the poor engraftment efficiency of hepatocytes. We developed a three-dimensional (3D) culture system using a microgravity bioreactor (MB), biodegradable scaffolds and growth-factor-reduced Matrigel to construct a tissue-engineered liver for transplantation into the peritoneal cavity of non-obese diabetic severe combined immunodeficient mice. The number of viable cells in the hepatic tissue constructs was stably maintained in the 3D MB culture system. Hematoxylin-eosin staining and zonula occludens-1 expression revealed that neonatal mouse liver cells were reorganized to form tissue-like structures during MB culture. Significantly upregulated hepatic functions (albumin secretion, urea production and cytochrome P450 activity) were observed in the MB culture group. Post-transplantation analysis indicated that the engraftment efficiency of the hepatic tissue constructs prepared in MB cultures was higher than that of those prepared in the static cultures. Higher level of hepatic function in the implants was confirmed by the expression of albumin. These findings suggest that 3D MB culture systems may offer an improved method for creating tissue-engineered liver because of the higher engraftment efficiency and the reduction of the initial cell function loss.

In a mating system in which females are monandrous and males are polygynous, females may incur a risk by mating with males with possible sperm depletion following consecutive matings. Here, we examined the effects of male mating history on male mating success and female reproductive fitness in the wolf spider Pardosa pseudoannulata by performing mating trials and sperm counting experiments. Results showed that male mating history had a significant negative impact on subsequent copulation success but had little effect on courtship duration and courtship intensity. In addition, neither male courtship intensity nor morphological measurements of males and females had significant effects on male mating success. Furthermore, male mating history had no obvious impact on the fecundity of inseminated females, with no significant differences observed in the oviposition rate of females, the numbers and the carapace width of the second-instar spiderlings between treatments. Results showed that the number of sperm decreased significantly after mating but could be replenished, with no significant differences observed between groups in which males had rested for 7 d. These findings suggest that polygynous male spiders may recharge their sperm during the mating season, but how females differentiate the mating status of males remains unknown.

Hepatocytes can be generated from embryonic stem cells (ESCs) using inducers such as chemical compounds and cytokines, but issues related to low differentiation efficiencies remain to be resolved. Recent work has shown that overexpression of lineage-specific transcription factors can directly cause cells phenotypic changes, including differentiation, trans-differentiation, and de-differentiation. We hypothesized that lentivirus-mediated constitutive expression of forkhead box A2 (Foxa2) and hepatocyte nuclear factor 4 alpha (Hnf4a) could promote inducing mouse ESCs to hepatocyte-likes cells. First, ESC lines that stably expressed Foxa2, Hnf4a, or Foxa2/Hnf4a were constructed via lentiviral expression vectors. Second, observations of cell morphology changes were made during the cell culture process, followed by experiments examining teratoma formation. Then, the effects of constitutive expression of Foxa2 and Hnf4a on hepatic differentiation and maturation were determined by measuring the marker gene expression levels of Albumin, α-fetoprotein, Cytokeratin18, and α1-antitrypsin. The results indicate that constitutive expression of Foxa2 and Hnf4a does not affect ESCs culture, teratoma formation, or the expression levels of the specific hepatocyte genes under autonomous differentiation. However, with some assistance from inducing factors, Foxa2 significantly increased the hepatic differentiation of ESCs, whereas the expression of Hnf4a alone or Foxa2/Hnf4a could not. Differentiated CCE-Foxa2 cells were more superior in expressing several liver-specific markers and protein, storing glycogen than differentiated CCE cells. Therefore, our method employing the transduction of Foxa2 would be a valuable tool for the efficient generation of functional hepatocytes derived from ESCs.

The ideal cells for tissue engineering should have the following characteristics: easy obtainment, safety, immune privilege, the capability of self-renewal, and multipotency. Adipose-derived stem cells (ADSCs) are a promising candidate. However, the immunogenicity of allogeneic mesenchymal stem cells limits their long-term benefits. In this study, we introduced human cytomegalovirus US2/US3 gene into the ADSCs to decrease the expression of MHC I protein of ADSCs and reduce the activation of T-cells of the recipient animals. Moreover, the biosafety and biological characteristics of ADSCs transfected with the US2/US3 genes (ADSCs-US2/US3) were similar to normal ADSCs. Then we took ADSCs-US2/US3 to construct a tissue-engineered bone for repairing bone defects in pigs and found that there were no great differences in repair effects or healing time between the allogeneic ADSCs-US2/US3 group and the autologous ADSC group. These results suggest that allogeneic ADSCs-US2/US3 have the advantages of biological safety, low immunogenicity, and effective osteogenesis. Such barely immunogenic ADSCs will be crucial for the success of future tissue-regenerative approaches.

Embryonic stem (ES) cells are pluripotent cells that are capable of differentiating all the somatic cell lineages, including those in the liver tissue. We describe the generation of functional hepatic-like cells from mouse ES (mES) cells using a biodegradable polymer scaffold and a rotating bioreactor that allows simulated microgravity. Cells derived from ES cells cultured in the three-dimensional (3D) culture system with exogenous growth factors and hormones can differentiate into hepatic-like cells with morphologic characteristics of typical mature hepatocytes. Reverse-transcription polymerase chain-reaction testing, Western blot testing, immunostaining, and flow cytometric analysis show that these cells express hepatic-specific genes and proteins during differentiation. Differentiated cells on scaffolds further exhibit morphologic traits and biomarkers characteristic of liver cells, including albumin production, cytochrome P450 activity, and low-density lipoprotein uptake. When these stem cell-bearing scaffolds are transplanted into severe combined immunodeficient mice, the 3D constructs remained viable, undergoing further differentiation and maturation of hepatic-like cells in vivo. In conclusion, the growth and differentiation of ES cells in a biodegradable polymer scaffold and a rotating microgravity bioreactor can yield functional and organizational hepatocytes useful for research involving bioartificial liver and engineered liver tissue.

Developmental characteristics of the predatory bug Orius similis fed on Tetranychus cinnabarinus were investigated at three constant temperatures (25, 28, and 31°C) under laboratory conditions (75 ± 5% relative humidity and a 14L : 10D photo- period). The survival of nymphs was highest at 28°C (75.57%) and at this temperature female adults O. similis had the longest ovi- position period (21.1 d), the greatest fecundity (40.3 eggs) and the highest potential intrinsic rate of increase (rm: 0.108 d -1 ). These results suggest that O. similis can maintain greater population densities at 28°C than at the other temperatures tested. In addition, the functional response indicates that the attack rate of O. similis (1.04) and the maximum prey capacity (30.7 spider mites per bug in 24 h) are greatest at 28°C. The results of this study provide useful information on the biology and time when to release O. similis in order to reduce the abundance of T. cinnabarinus in cotton fields.

Adipose tissue-derived stem cells (ADSC) have been shown to possess stem cell properties such as transdifferentiation, self-renewal and therapeutic potential. However, the property of ADSC to accommodate immune system is still unknown. In this study, ADSC were cocultured with allogenetic dendritic cells (DC), and then treated DC were mixed with allogenetic CD4+ T cells. The results demonstrated that ADSC could downregulate costimulatory molecules, including CD80, CD83, CD86, and cytokine secretion such as interleukin (IL)-12 and tumor necrosis factor (TNF)-α, while upregulate indoleamine-2,3-dioxygenase (IDO) of allogenetic DC. In addition, treated DC could inhibit CD4+ Tcell activation and naive T cells toward Th1 polarization. The results suggest that ADSC could negatively modulate immunity and induce immune tolerance, which provide a promising strategy in transplantation or autoimmune disease.

To counter female resistance to mating and cannibalism, males of many animal species have evolved a variety of behavioural adaptations. Here we investigated a novel copulatory courtship behaviour, mate binding, in which the male deposits fine silk onto the female’s body in between copulation bouts, in an orb-web nephilid spider, Nephila pilipes. We hypothesized that mate binding might reduce female aggressiveness and sexual cannibalism and that both tactile and chemical cues play a role. We performed a series of mating trials, in which we blocked (1) the females’ tactile perception, (2) the females’ chemoreceptors, and (3) both types of communication. We also manipulated male spinnerets and thus male silk production. As predicted, mate binding reduced both female resistance to repeated mating and levels of sexual cannibalism. Our results suggest that both tactile and chemical cues are crucial for mate binding to succeed in rendering females less aggressive, but that tactile cues are more important. We conclude that mate binding prolongs total copulation duration, whereby the male maximizes his paternity. Therefore, mate binding may serve as a mechanism countering sexual conflict over repeated mating and sexual cannibalism.

Spider webs are made of silk, the properties of which ensure remarkable efficiency at capturing prey. However, remaining on, or near, the web exposes the resident spiders to many potential predators, such as ants. Surprisingly, ants are rarely reported foraging on the webs of orb-weaving spiders, despite the formidable capacity of ants to subdue prey and repel enemies, the diversity and abundance of orb-web spiders, and the nutritional value of the web and resident spider. We explain this paradox by reporting a novel property of the silk produced by the orb-web spider Nephila antipodiana (Walckenaer). These spiders deposit on the silk a pyrrolidine alkaloid (2-pyrrolidinone) that provides protection from ant invasion. Furthermore, the ontogenetic change in the production of 2-pyrrolidinone suggests that this compound represents an adaptive response to the threat of natural enemies, rather than a simple by-product of silk synthesis: while 2-pyrrolidinone occurs on the silk threads produced by adult and large juvenile spiders, it is absent on threads produced by small juvenile spiders, whose threads are sufficiently thin to be inaccessible to ants.

Male–male competition for females can significantly affect a male’s reproductive success and hence his fitness. Game theory predicts that an individual should avoid fighting when its future reproductive potential is high, but should fight forcefully when its future reproductive potential is insignificant. When mates are scarce, extreme competition and fatal fighting is expected. We recently showed that Nephilengys malabarensis eunuchs, i.e. sterile spider males that lost their genitals during copulation, become more aggressive during male–male contests. Here, we add crucial comparative data by exploring eunuch fighting behaviour in Nephilengys livida from Madagascar, specifically by testing the ‘better fighter hypotheses’ in a laboratory setting. Similar to N. malabarensis, N. livida copulations resulted in total male castration with the severed palp plugging the female genitals in 70.83% cases, which mostly (63.63%) prevented subsequent copulations. Unexpectedly, however, N. livida eunuchs exhibited lower aggressiveness than virgin males. We interpret these results in the light of different mating biology between the so far studied species known for the eunuch phenomenon, which might reflect differing plug effectiveness due to variation in genital anatomy in N. livida, N. malabarensis and Herennia multipuncta. However, detected differences in aggressive behaviour of N. livida versus N. malabarensis eunuchs might also be explained by the species’ ecology, with lower population densities resulting in a relaxed male–male competition making excessive aggression and mate guarding redundant. This study thus questions the generality of overt aggressiveness in mated males with no reproductive value, and highlights the importance of understanding the natural history of species in the question.

The larval behaviors probably associated with respiration are reported in different larval instars of Luciola substriata Gorham (Coleoptera: Lampyridae). The behavior involved exposing eighth abdominal segment with spiracles to the atmosphere when swim behavior occurred to 1st–2nd and 3rd–6th instar larvae. In addition, 3rd–6th instar larvae exposed second to seventh abdominal segments with spiracles to air when swimming. Swimming and the behavior that uses the force of body curving travelling in a head-to-tail direction produces a sinusoidal wave. However, probably 1st–2nd instar larvae used both tracheal gills and the exposed posterior spiracle for respiration. Whereas 3rd–6th instar larvae lack tracheal gills, they probably rely only on tracheal respiration. The survival times of 1st–2nd instar larvae were nine times to 3rd–6th instar larvae when the water in the beakers was isolated from atmosphere by a thin plastic membrane. The potential for different selective pressures acting on these ear...

The role of background matching in camouflage has been extensively studied. However, contour modification has received far less attention, especially in twig-mimicking species. Here, we studied this deceptive strategy by revealing a special masquerade tactic, in which the animals protract and cluster their legs linearly in the same axis with their bodies when resting, using the spider Ariamnes cylindrogaster as a model. We used cardboard papers to construct dummies resembling spiders in appearance and colour. To differentiate the most important factors in the concealment effect, we manipulated body size (long or short abdomen) and resting postures (leg clustered or spread) of the dummies and recorded the responses of predators to different dummy types in the field. The results showed that dummies with clustered legs received significantly less attention from predators, regardless of the body length. Thus, we conclude that A. cylindrogaster relies on the resting posture rather than body size for predator avoidance. This study provides, to the best of our knowledge, empirical evidence for the first time that twig-mimicking species can achieve effective camouflage by contour modification.

Nephila are known for the greatest degrees of sexual size dimorphism among orb weaving spiders (Araneoidea) and thus among terrestrial animals. However, a meaningful quantification of the dimorphism is lacking and the proximate developmental mechanisms of female gigantism are poorly understood, being attributed solely to female delayed maturation. Here we show that females in the giant wood spider Nephila pilipes (Fabricius 1793) become giants through facultative post-maturity molting, a phenomenon resulting in female carapaces on average 4.27 times longer than males' (ranging from 3 to 6.4 times), and female mass averaging 125 times the male's (ranging from 28 to 502 times). Although the small males follow a typical developmental pathway and reach maturity with their final molt, the females mature at varying sizes and instars and then continue to grow by molting the entire exoskeleton except their genitals. The newly discovered phenomenon of additional, single-sex, adult, non-genital molting may...

Adipose-derived stem cells (ADSCs) represent a valuable source of stem cells for regenerative medicine, but the loss of their stemness during in vitro expansion remains a major roadblock. We employed a microgravity bioreactor (MB) to develop a method for biomaterial-free-mediated spheroid formation to maintain the stemness properties of ADSCs. ADSCs spontaneously formed three-dimensional spheroids in the MB. Compared with monolayer culture, the expression levels of E-cadherin and pluripotent markers were significantly upregulated in ADSC spheroids. Spheroid-derived ADSCs exhibited increased proliferative ability and colony-forming efficiency. By culturing the spheroid-derived ADSCs in an appropriate induction medium, we found that the multipotency differentiation capacities of ADSCs were significantly improved by spheroid culture in the MB. Furthermore, when ADSCs were administered to mice with carbon tetrachloride-induced acute liver failure, spheroid-derived ADSCs showed more effective potentials to rescue liver failure than ADSCs derived from constant monolayer culture. Our results suggest that spheroid formation of ADSCs in an MB enhances their stemness properties and increases their therapeutic potential. Therefore, spheroid culture in an MB can be an efficient method to maintain stemness properties, without the involvement of any biomaterials for clinical applications of in vitro cultured ADSCs.

Apert syndrome (AS) is a type of autosomal dominant disease characterized by premature fusion of the cranial sutures, severe syndactyly, and other abnormalities in internal organs. Approximately 70% of AS cases are caused by a single mutation, S252W, in fibroblast growth factor receptor 2 (FGFR2). Two groups have generated FGFR2 knock-in mice Fgfr2 S252W/+ that exhibit features of AS. During the present study of AS using the Fgfr2 S252W/+ mouse model, an age-related phenotype of bone homeostasis was discovered. The long bone mass was lower in 2 month old mutant mice than in age-matched controls but higher in 5 month old mutant mice. This unusual phenotype suggested that bone marrow-derived mesenchymal stem cells (BMSCs), which are vital to maintain bone homeostasis, might be involved. BMSCs were isolated from Fgfr2 S252W/+ mice and found that S252W mutation could impair osteogenic differentiation BMSCs but enhance mineralization of more mature osteoblasts. A microarray analysis revealed that Wnt pathway inhibitors SRFP1/2/4 were up-regulated in mutant BMSCs. This work provides evidence to show that the Wnt/β-catenin pathway is inhibited in both mutant BMSCs and osteoblasts, and differentiation defects of these cells can be ameliorated by Wnt3a treatment. The present study suggested that the bone abnormalities caused by deregulation of Wnt pathway may underlie the symptoms of AS.

Ethnopharmacological relevance Salvia miltiorrhiza Bunge (Labiatae sp. plant, Chinese name Danshen) and Panax notoginseng (Burk.) F. H. Chen (Araliaceae plant, Chinese name Sanqi) have a long history in treating coronary heart disease, cerebrovascular disease and inner ear disorders in traditional Chinese medicine. To provide a rational basis for the use of these herbs in clinical practice, we investigated the in vivo distribution and pharmacokinetics of marker agents in Danshen and Sanqi via intravenous and inner ear administration and explored the potential interactions of these agents in compound prescription. Materials and methods Guinea pigs were given Danshen extracts (salvianolic acid B, tanshinone IIA), Sanqi extracts (Panax notoginseng saponins) and combination of the two extracts via intravenous and intratympanic administration (IT). Samples from the brain, inner ear perilymph (PL), cerebrospinal fluid (CSF) and plasma were collected at different time points. The concentration of salvianolic acid B (Sal B), tanshinone IIA (Ts IIA), notoginsenoside R1 (R1), ginsenoside Rg1 (Rg1) and ginsenoside Rb1 (Rb1) was determined by high-performance liquid chromatography coupled with a diode array detector (DAD). Pharmacokinetic parameters were estimated using non-compartmental methods. Results Local drug application via inner ear greatly improved drug distribution within the PL, CSF and brain tissues compared with intravenous administration (IV). The values of Cmax and AUC(0–t) after IT were significantly higher than IV. In comparison with IT of Danshen and Sanqi alone, the pharmacokinetic parameters for R1, Rg1, Rb1, Sal B and Ts IIA were markedly different in the compound prescription. The compound compatibility enhanced the transport of Danshen components into the brain through the inner ear and apparently prolonged the retention time in CSF while decreasing the distribution of Sanqi components in the inner ear and brain. Conclusions The results indicated that local drug application to the inner ear was a more effective delivery route than systemic administration. Co-administration of Danshen and Sanqi could cause significant pharmacokinetic herb–herb interactions in guinea pigs. The multiple active components via inner ear administration might be promising candidates for the treatment of inner ear and brain diseases.

A S252W mutation of fibroblast growth factor receptor 2 (FGFR2), which is responsible for nearly two-thirds of Apert syndrome (AS) cases, causes retarded development of the skeleton and skull malformation resulting from premature fusion of the craniofacial sutures. We utilized a Fgfr2(+/S252W) mouse (a knock-in mouse model mimicking human AS) to demonstrate decreased bone mass due to reduced trabecular bone volume, reduced bone mineral density, and shortened growth plates in the long bones. In vitro bone mesenchymal stem cells (BMSCs) culture studies revealed that the mutant mice showed reduced BMSC proliferation, a reduction in chondrogenic differentiation, and reduced mineralization. Our results suggest that these phenomena are caused by up-regulation of p38 and Erk1/2 phosphorylation. Treatment of cultured mutant bone rudiments with SB203580 or PD98059 resulted in partial rescue of the bone growth retardation. The p38 signaling pathway especially was found to be responsible for the retarded long bone development. Our data indicate that the S252W mutation in FGFR2 directly affects endochondral ossification, resulting in growth retardation of the long bone. We also show that the p38 and Erk1/2 signaling pathways partially mediate the effects of the S252W mutation of FGFR2 on long bone development.

There is increasing evidence that mesenchymal stem cells (MSCs) derived from different tissues could act as an alternative source of mature hepatocytes for treatment of acute liver failure (ALF). Human umbilical cord matrix stem cells (hUCMSCs) represent a novel source of MSCs. We examined the therapeutic potential and the different mechanisms of hUCMSCs by their transplantation into nonobese diabetic severe combined-immunodeficient (NOD-SCID) mice with carbon tetrachloride (CCl(4))-induced ALF in comparison to adult human hepatocytes (AHHs). The characteristics of isolated hUCMSCs were determined from MSCs and hepatocyte marker expression, hepatic function, and differentiation. Native hUCMSCs constitutively expressed some hepatic markers, though weaker hepatocyte-specific functions were observed when compared to AHHs. When native hUCMSCs or AHHs were transplanted into livers of NOD-SCID mice with ALF induced by CCl(4), both hUCMSCs and AHHs provided a significant survival benefit and prevented the release of liver injury biomarkers. hUCMSCs were found to engraft within the recipient liver and differentiated into functional hepatocytes, whereas the HepPar1-/albumin (ALB)-positive cells of the hUCMSC group were less than the AHH group in the recipient liver. Higher values of human ALB in the serum of mice-transplanted AHHs were determined in comparison with levels in mice-transplanted hUCMSCs. The analysis of mouse serum cytokine levels showed that hUCMSC transplantation was even more effective than treatment with AHHs and successfully downregulated the systemic inflammatory cytokines such as interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6, IL-10, and IL-1 receptor antagonist (IL-1RA). Furthermore, paracrine effects produced by hUCMSCs were identified by indirect coculture with damaged mouse hepatocytes (MHs) induced by CCl(4). Coculture with hUCMSCs significantly increased the viability, ALB secretion of damaged MHs, and greatly enhanced the regeneration of MHs in vitro when compared with AHHs. These data suggest that direct transplantation of native hUCMSCs can rescue ALF and repopulate livers of mice through paracrine effects to stimulate endogenous liver regeneration rather than hepatic differentiation for compensated liver function, which is the primary effect of AHHs. Thus, hUCMSCs can be a potential alternative source of AHHs for cell therapy of ALF and eliminate the shortage of hepatocytes.

Preservation of hepatocyte functions in vitro will undoubtedly help the management of acute liver failure. The coculture system may be able to prevent functional decline of hepatocytes. It has already been shown that hepatocytes, when cocultured with bone marrow mesenchymal stem cells, could undergo long-term culture in vitro without loss of functions. In this study, human orbital fat-derived stem cells were isolated and cocultured with rat hepatocytes. When treated with serum from an acute liver failure patient, rat hepatocyte monoculture showed reduction of cell viability and loss of liverspecific functions. However, rat hepatocytes in the coculture system were still able to secret albumin and synthesize urea. IL-6 was significantly elevated in the coculture of rat hepatocyte with orbital fat-derived stem cells, and it might be the key immunoregulator which protects rat hepatocytes against inflammation. Our data confirmed that orbital fat-derived stem cells, or other adipose tissue-derived stem cells, are an ideal candidate to support rat hepatocyte functions in vitro.

Embryonic stem cells (ESCs) can have unlimited proliferation potential and the capability to differentiate into almost all types of tissues. The use of ESCs as a novel source of hepatocytes has therefore recently received much attention. To date, however, the efficient and mass differentiation of ESCs into hepatocytes in vitro has been proven to be difficult. In this study, we investigated a three-dimensional (3D) mouse ESC hepatic differentiation system. Cells derived from 5-day-old embryoid bodies were mixed with Matrigel and immediately seeded in a biodegradable polymer scaffold composed of poly-l-lactic acid and polyglycolic acid. Exogenous growth factors and hormones were added sequentially for 20 days to promote hepatic histogenesis. The system enabled spontaneous spheroid formation and mass cultivation of embryoid body-derived cells. The 3D differentiated hepatocyte-like cells were able to express several liver-specific markers and proteins, secrete albumin, store glycogen, and allow the uptake of low-density lipoproteins. These findings suggest that this supportive 3D environment with appropriate inducing factors could provide mouse ESC-derived cells with an environment closely mimicking that of hepatogenesis in vivo.

Orthotopic liver transplantation is, to date, the only proven effective treatment for end-stage liver disease. However, it suffers from lack of donors and immunorejection. Here, we speculate that co-transplantation of induced pluripotent stem (iPS) cells derived hepatocytes and mesenchymal stem cells (MSCs) may offer an alternative way to treat patients with end-stage liver disease. Recently, progress on iPS cells, homogeneous differentiation of hepatocyte-like cells from embryonic stem cells (ESCs), and paracrine effects by MSCs highlight the possibility. Safe, efficient and rapid generation of iPS cells has been reliably produced by several experimental laboratories. Methods for highly efficient and homogeneous differentiation of ESCs into functional hepatocytes have been established as well. Moreover, paracrine effects by MSCs have been proven to play an important role in liver regeneration and repair, and the effects can be used as an enhancer for engraftment. All of these remarkable developments lead to this hypothesis which may offer a novel therapeutic strategy for treatment of patients with end-stage liver disease, though some issues about safety and efficacy need to be further guaranteed.

Chemoreception is a key feature in selection of host plant by phytophagous insects, and odorant-binding proteins (OBPs) are involved in chemical communication of both insects and vertebrates. The legume pod borer, Maruca vitrata Fabricius (Lepidoptera: Crambidae) is one of the key pest species of cowpea and widely distributed throughout tropical and subtropical regions, causing up to 80% of yield loss. In this study, we investigated the electrophysiological responses of female M. vitrata to floral volatiles from V. unguiculata. Seventeen electroantennogram-active compounds were identified from floral volatiles of V. unguiculata by coupled gas chromatography-electroantennography (GC-EAD) and gas chromatography-mass spectrometry (GC-MS). Then, we cloned two novel full-length GOBP genes (MvitGOBP1 and MvitGOBP2) from the antennae of M. vitrata using reverse transcription PCR. Protein sequence analysis indicated that they shared high sequence similarity with other Pyralididae insect GOBPs and had the typical six-cysteine signature. Real-time PCR analysis indicated that MvitGOBP1-2 mRNA was highly expressed in the antennae of female adult with several thousands-fold difference compare to other tissue. Next, the recombinant MvitGOBP1-2 was expressed in Escherichia coli and purified using Ni ion affinity chromatography. Fluorescence binding assays demonstrated that MvitGOBP1-2 had different binding affinities with 17 volatile odorant molecules including butanoic acid butyl ester, limonene, 4-ethylpropiophenone, 1H-indol-4-ol, butanoic acid octyl ester and 2-methyl-3-phenylpropanal. In the field trapping experiment, these six floral volatiles could effectively attract female moths and showed significant difference compared with the blank lure. These results suggested that MvitGOBPs and the seventeen floral volatiles are likely to function in the olfactory behavior response of female moths, which may have played crucial roles in the selection of oviposition sites. The six compounds that we have identified from the volatiles of V. unguiculata may provide useful information for exploring efficiency monitoring and integrated pest management strategies of this legume pod borer in the field.

Introduction Embryonic stem (ES) cells are considered a potentially advantageous source of hepatocytes for both transplantation and the development of bioartificial livers. However, the efficient large-scale generation of functional hepatocytes from ES cells remains a major challenge, especially for those methods compatible with clinical applications. Methods In this study, we investigated whether a large number of functional hepatocytes can be differentiated from mouse ES (mES) cells using a simulated microgravity bioreactor. mES cells were cultured in a rotating bioreactor in the presence of exogenous growth factors and hormones to form embryoid bodies (EBs), which then differentiated into hepatocytes. Results During the rotating culture, most of the EB-derived cells gradually showed the histologic characteristics of normal hepatocytes. More specifically, the expression of hepatic genes and proteins was detected at a higher level in the differentiated cells from the bioreactor culture than in cells from a static culture. On further growing, the EBs on tissue-culture plates, most of the EB-derived cells were found to display the morphologic features of hepatocytes, as well as albumin synthesis. In addition, the EB-derived cells grown in the rotating bioreactor exhibited higher levels of liver-specific functions, such as glycogen storage, cytochrome P450 activity, low-density lipoprotein, and indocyanine green uptake, than did differentiated cells grown in static culture. When the EB-derived cells from day-14 EBs and the cells’ culture supernatant were injected into nude mice, the transplanted cells were engrafted into the recipient livers. Conclusions Large quantities of high-quality hepatocytes can be generated from mES cells in a rotating bioreactor via EB formation. This system may be useful in the large-scale generation of hepatocytes for both cell transplantation and the development of bioartificial livers.