Smart Pneumatic Artificial Muscle Using a Bend Sensor like a Human Muscle with a Muscle Spindle.

Shortage of labor and increased work of young people are causing problems in terms of care and welfare of a growing proportion of elderly people. This is a looming social problem because people of advanced ages are increasing. Necessary in the fields of care and welfare, pneumatic artificial muscles in actuators of robots are being examined. Pneumatic artificial muscles have a high output per unit of weight, and they are soft, similarly to human muscles. However, in previous research of robots using pneumatic artificial muscles, rigid sensors were often installed at joints and other locations due to the robots' structures. Therefore, we developed a smart actuator that integrates a bending sensor that functions as a human muscle spindle; it can be externally attached to the pneumatic artificial muscle. This paper reports a smart artificial muscle actuator that can sense contraction, which can be applied to developed self-monitoring and robot posture control.

Sialic acid-binding lectin from bullfrog eggs inhibits human malignant mesothelioma cell growth in vitro and in vivo.

Malignant mesothelioma is an aggressive cancer that results from exposure to asbestos. The therapeutic options for this type of cancer are limited; therefore, the development of novel therapeutic agents is urgently required. Sialic acid-binding lectin isolated from Rana catesbeiana oocytes (cSBL) is a novel therapeutic candidate for cancer, which exhibits antitumor activity mediated through RNA degradation. In the present study, we evaluated the effect of cSBL in vitro and in vivo. Xenograft-competent H2452 and MSTO human mesothelioma cell lines were treated with cSBL, and the pathway by which cSBL induces apoptosis was analyzed. In vivo studies were performed using nude mice inoculated with one of the two cell lines, and the effects of cSBL and pemetrexed were monitored simultaneously. Furthermore, the pharmacological interactions between the three agents (pemetrexed, cisplatin and cSBL) were statistically assessed. It was demonstrated that cSBL treatments caused morphological and biochemical apoptotic changes in both cell lines. Caspase cascade analysis revealed that an intrinsic pathway mediated cSBL-induced apoptosis. The administration of cSBL significantly inhibited tumor growth in two xenograft models, without any adverse effects. Furthermore, the combination index and dose reduction index values indicated that the cSBL + pemetrexed combination showed the highest synergism, and thus potential for reducing dosage of each drug, compared with the other combinations, including the existing pemetrexed + cisplatin regimen. cSBL exerted prominent antitumor effects on malignant mesothelioma cells in vitro and in vivo, and showed favorable effects when combined with pemetrexed. These results suggest that cSBL has potential as a novel drug for the treatment of malignant mesothelioma.

Synergistic anti-tumor effect of bullfrog sialic acid-binding lectin and pemetrexed in malignant mesothelioma.

Malignant mesothelioma is an aggressive cancer with limited therapeutic options. Sialic acid-binding lectin isolated from Rana catesbeiana oocytes (cSBL) is a multifunctional protein with anti-cancer activity. The effects of pemetrexed, cisplatin, and cSBL were evaluated in mesothelioma and normal mesothelial cell lines. We evaluated cytotoxicity, apoptosis, caspase-3 cleavage and activation, cell proliferation, cell cycle arrest, and levels of cell cycle proteins in H28 cells treated with pemetrexed, cisplatin, and cSBL alone or in combination. Treatment with cSBL alone was cytotoxic to mesothelioma cells. The anti-cancer effect of cSBL was observed in a broader range of cell lines and exhibited greater cancer cell selectivity than pemetrexed or cisplatin. Combination treatment with pemetrexed + cSBL resulted in greater dose-dependent cytotoxicity than pemetrexed + cisplatin, the standard of care in mesothelioma. The synergistic effect of pemetrexed + cSBL was mediated by the cytostatic effect of pemetrexed and the cytotoxic effect of cSBL. It thus appears that cSBL has therapeutic potential for the treatment of mesothelioma.

Octopamine and cooperation: octopamine regulates the disappearance of cooperative behaviours between genetically unrelated founding queens in the ant.

We investigated whether octopamine (OA) is associated with the disappearance of cooperation in Polyrhachis moesta ant queens. Queens of P. moesta facultatively found the colony with genetically unrelated queens. The founding queens perform frequent food exchange with these non-related queens and partake in cooperative brood rearing, whereas single colony queens exclude non-related queens via aggressive behaviour. Thus, aggression is a factor that reduces cooperation. Given that aggression is generally associated with brain OA in insects, we hypothesized that OA controls the behavioural change in cooperation in the ant queen, via an increase in aggression. To test this hypothesis, we compared the amounts of OA and related substances in the brain between founding and colony queens, and observed the interaction of founding queens following oral OA administration. The brain OA levels in colony queens were significantly higher than those in founding queens. Oral administration of OA to founding queens caused significantly less trophallaxis and allogrooming behaviour than in the control founding queens, but with no significant increase in aggression. These results suggest that OA promotes the disappearance of cooperation in founding queens of P. moesta. This is the first study to reveal the neuroendocrine mechanism of cooperation in ant queens.

Identification, characterization and full-length sequence analysis of a novel dsRNA virus isolated from the arboreal ant Camponotus yamaokai.

A novel dsRNA virus was identified from the arboreal ant Camponotus yamaokai. The complete nucleotide sequence analysis of the virus revealed that the virus consisted of 5704 bp with two ORFs. ORF1 (3084 nt) encoded a putative capsid protein. ORF2 (1977 nt) encoded a viral RNA-dependent RNA polymerase (RdRp). ORF2 could be translated as a fusion with the ORF1 product by a - 1 frameshift in the overlapping ORF1. Phylogenetic analyses based on the RdRp revealed that the virus from C. yamaokai was most likely a novel totivirus, but it was not closely related to the previously known totiviruses in arthropods. Transmission electron microscopy revealed isometric virus particles of ~30 nm diameter in the cytoplasm, which was consistent with the characteristics of the family Totiviridae. The virus was detected by reverse transcription-PCR in all caste members and developmental stages of ants, including eggs, larvae, pupae, adult workers, alates (male and female) and queens. To our knowledge, this is the first report of a member of the family Totiviridae in a hymenopteran; the virus was designated Camponotus yamaokai virus.

Colony fusion in a parthenogenetic ant, Pristomyrmex punctatus.

In the ant Pristomyrmex punctatus Smith (Hymenoptera: Formicidae), all young workers lay a small number of eggs parthenogenetically. Some colonies consist of monoclonal individuals that provide high inclusive fitness, according to the kin selection theory. However, in some populations, a majority of the colonies contain multiple lineages. Intracolonial genetic variation of parthenogenetic ants cannot be explained by the multiple mating of single founderesses or by the foundation of a colony by multiple foundresses, which are the usual causes of genetically diverse colonies in social insects. Here, we hypothesized that the fusion of established colonies might facilitate the formation of multiclonal colonies. Colony fusion decreases indirect benefits because of the reduction in intracolonial relatedness. However, when suitable nesting places for overwintering are scarce, colony fusion provides a strategy for the survival of colonies. Here, ants derived from different colonies were allowed to encounter one another in a container with just one nesting place. Initially, high aggression was observed; however, after several days, no aggression was observed and the ants shared the nest. When the fused colonies were allowed to transfer to two alternative nests, ants from different colonies occupied the same nest. This study highlights the importance of limiting the number of nesting places in order to understand the genetic diversity of parthenogenetic ant colonies.

Polymorphisms of the Dopamine D4 Receptor Gene in Stabled Horses are Related to Differences in Behavioral Response to Frustration.

In stabled horses, behavioral responses to frustration are often observed, especially around feeding time. These behavioral responses are a useful indicator of their welfare. In this study, we investigated the association between this behavioral indicator and DRD4 gene polymorphisms in stabled horses. Twenty one horses housed in two stables were used. The horses were observed for approximately 4 h around feeding over three or more days using focal-sampling and instantaneous-sampling. Horses were genotyped for the A-G substitution in the DRD4 gene. The effects of the A-G substitution (with or without the A allele in the DRD4 gene), the stables, and their interaction on the frequency of behavioral responses to frustration were analyzed using general linear models. The total time budget of behavioral responses to frustration was higher in horses without the A allele than in those with the A allele (P = 0.007). These results indicate that the A-G substitution of the DRD4 gene is related to frustration-related behavioral responses in stabled horses. Appropriate consideration should be made for the DRD4 gene polymorphism when the welfare of stabled horses is assessed, based on this behavioral indicator.

Inhibitory effects of daiokanzoto (da-huang-gan-cao-tang) on p-glycoprotein.

We have studied the effects of various Kampo medicines on P-glycoprotein (P-gp), a drug transporter, in vitro. The present study focused on Daiokanzoto (Da-Huang-Gan-Cao-Tang), which shows the most potent inhibitory effects on P-gp among the 50 Kampo medicines studied, and investigated the P-gp inhibitory effects of Daiokanzoto herbal ingredients (rhubarb and licorice root) and their components by an ATPase assay using human P-gp membrane. Both rhubarb and licorice root significantly inhibited ATPase activity, and the effects of rhubarb were more potent than those of licorice root. The content of rhubarb in Daiokanzoto is double that in licorice root, and the inhibition patterns of Daiokanzoto and rhubarb involve both competitive and noncompetitive inhibition, suggesting that the inhibitory effects of Daiokanzoto are mainly due to rhubarb. Concerning the components of rhubarb, concentration-dependent inhibitory effects were observed for (-)-catechin gallate, (-)-epicatechin gallate, and (-)-epigallocatechin gallate. In conclusion, rhubarb may cause changes in the drug dispositions of P-gp substrates through the inhibition of P-gp. It appears that attention should be given to the interactions between these drugs and Kampo medicines containing rhubarb as an herbal ingredient.

The importance of workers for queen hibernation survival in Camponotus ants.

The higher proportion of polygynous ant species in northern areas indicates that cold climates influence queen number per colony. It is unclear, however, what ecological and physiological factors facilitate the dominance of polygynous species in cold climates. This is the case in two common arboreal ants in Japan-Camponotus yamaokai and C. nawai-which are quite similar in morphology, but different in social structure and geographical distribution. Polygynous C. yamaokai inhabits colder areas, whereas monogynyous C. nawai inhabits warmer climates. We compared queen survival in both ants at low temperature to evaluate whether interspecific difference in cold tolerance can explain the geographical distribution. We examined the influence of cohabitation with other individuals, as well as individual cold tolerance. Experimental groups with different caste compositions were prepared and maintained under conditions simulating in the laboratory climates of the northern limit of C. nawai. Wintering experiments revealed that C. yamaokai queens survived longer than C. nawai queens under solitary conditions, although half of the queens died in less than a month, even in C. yamaokai. Queens hibernating with workers survived longer than solitary queens, but queen number did not affect queen survival. Cohabitation with workers allowed 80% of C. yamaokai queens to survive more than two months. Under field conditions, monogynous C. nawai foundresses overwinter without workers, whereas new queens of polygynous C. yamaokai always overwinter with many workers. Thus, the geographical distribution of these ants appears to depend on the overwintering behavior of new queens.

Effects of Kampo medicines on P-glycoprotein.

The Kampo medicines are more and more often used in recent years, usually together with the western drugs. The need for the investigation of drug interactions between Kampo medicines and western drugs are, therefore, widely recognized. Among the various possible causes for the drug-drug interactions, those related to pharmacokinetics such as drug metabolism and transport are regarded as most frequent and clinically important. In the present study, the effects of Kampo medicines on the P-glycoprotein (P-gp), one of the major drug transporters, were investigated in in vitro studies using human P-gp membranes. The P-gp activity in the presence and absence of commonly used 50 Kampo medicines was evaluated by the ATPase assay detecting the inorganic phosphate produced by the ATP hydrolysis. The ATPase activity was inhibited by most of the Kampo medicines studied, indicating the possibility of their inhibiting the P-gp. The degree of inhibition in the presence of verapamil, a P-gp substrate, showed a significant correlation with that in the absence of verapamil. Furthermore, the inhibitory effect of the Kampo medicines on the ATPase activity correlated with their licorice root (kanzo) content, suggesting the contribution of licorice root in the P-gp inhibition. Because licorice root is one of the most common ingredients in the Kampo medicines and is also often used in the food as a sweetener, it might be necessary to pay attention on the interaction between the licorice root-containing drug/food and the number of drugs transported by P-gp.

Effects of Kampo medicines on CYP and P-gp activity in vitro.

The Kampo medicines are more and more often used in recent years, usually together with the Western drugs. The need for the investigation of drug interactions between Kampo medicines and Western drugs are, therefore, widely recognized. In the present study, the effects of 3 Kampo medicines (Rikkunshito, Yokukansan and Boiogito) on the activity of cytochrome P450 (CYP), a superfamily of drug-metabolizing enzymes, were investigated in an in vitro study using human CYP recombinants. Their effects on the P-glycoprotein (P-gp), one of the major drug transporters, were also evaluated by the ATPase assay using human P-gp membranes and verapamil as a substrate. The inhibition rate of Rikkunshito, Yokukansan and Boiogito on human CYP3A4, 2C9, 2C19, 2D6 and 2E1 was less than 50% at the concentrations below 0.1 mg/ml except for the inhibition of CYP2D6 by Boiogito. Furthermore, none of the Kampo medicines affected the ATPase activity at the concentrations lower than 0.1 mg/ml, either in the absence or presence of verapamil, indicating their low inhibitory potency against P-gp. These findings indicate that Rikkunshito, Yokukansan and Boiogito are unlikely to cause clinically relevant drug interactions involving the inhibition of major CYP isozymes and P-gp.

Mate preference in males of the cabbage butterfly, Pieris rapae crucivora, changes seasonally with the change in female UV color.

We initially investigated whether females of the cabbage butterfly, Pieris rapae crucivora, exhibit a seasonal change in ultraviolet wing color, which is a key stimulus for mate recognition by conspecific males, and whether and how a seasonal change affects the mating behavior of the males. We found that female UV wing color changes seasonally, the color being more pronounced in summer than in spring or autumn. We also demonstrated that male mate preference changes seasonally, concomitantly with the change in female UV color. Specifically, males appearing in summer exhibit a mating preference for summer-form females over spring- or autumn-form females, while those appearing in spring or autumn exhibit no seasonal preference, thereby facilitating more effective mate location. Our results suggest that this field of study will require more strictly controlled experimental investigation in which the seasonal change in UV color is considered when UV-influenced mating behaviors such as mate choice are investigated.

Clonal structure affects the assembling behavior in the Japanese queenless ant Pristomyrmex punctatus.

The queenless ant Pristomyrmex punctatus (Hymenoptera: Myrmicinae) has a unique society that differs from those of other typical ants. This species does not have a queen, and the workers lay eggs and produce their clones parthenogenetically. However, a colony of these ants does not always comprise members derived from a single clonal line. In this study, we examined whether P. punctatus changes its "assembling behavior" based on colony genetic structure. We prepared two subcolonies--a larger one comprising 200 individuals and a smaller one comprising 100 individuals; these subcolonies were established from a single stock colony. We investigated whether these subcolonies assemble into a single nest. The genetically monomorphic subcolonies (single clonal line) always fused into a single nest; however, the genetically polymorphic subcolonies (multiple clonal lines) did not tend to form a single colony. The present study is the first to demonstrate that the colony genetic structure significantly affects social viscosity in social insects.

Population structure of the large Japanese field mouse, Apodemus speciosus (Rodentia: Muridae), in suburban landscape, based on mitochondrial D-loop sequences.

Genetic structure of the large Japanese field mouse populations in suburban landscape of West Tokyo, Japan was determined using mitochondrial DNA control region sequence. Samples were collected from six habitats linked by forests and green tract along the Tama River, and from two forests segregated by urban areas from those continuous habitats. Thirty-five haplotypes were detected in 221 animals. Four to eight haplotypes were found within each local population belonging to the continuous landscape. Some haplotypes were shared by two or three adjacent local populations. On the other hand, two isolated habitats were occupied by one or two indigenous haplotypes. Significant genetic differentiation between all pairs of local populations, except for one pair in the continuous habitats, was found by analysis of molecular variance (amova). The geographical distance between habitats did not explain the large variance of pairwise F(ST)-values among local populations. F(ST)-values between local populations segregated by urban areas were higher than those between local populations in the continuous habitat, regardless of geographical distance. The results of this study demonstrated quantitatively that urban areas inhibit the migration of Apodemus speciosus, whereas a linear green tract along a river functions as a corridor. Moreover, it preserves the metapopulation structure of A. speciosus as well as the corridors in suburban landscape.