Deformation Matching: Force Computation Based on Deformation Optimization.

The tactile information to be presented to a user during interaction with a virtual object is calculated by simulating the contact between the object model and user model. In the simulation, a distributed force is applied to the contact area on the skin tissue of users' hands and results in deformation of the skin tissue. The skin deformation caused by the distributed force is the target contact state that should be presented by the device. However, most multipoint haptic displays do not have sufficient degrees of freedom (DoF) to represent the target contact state. This paper presents the concept and formulation of "deformation matching," whereby the output force is calculated to minimize the error between the target skin deformation and skin deformation that can be realized by the limited DoF device's output force. For comparison, the conventional concept of "force matching" was also formulated. The difference in human perception between these two concepts in the expression of friction was investigated through experiments using a pin-array tactile display capable of stimulating 128 points. It was demonstrated that the perception of the friction coefficient was more sensitive and the perception of the friction direction was more accurate in deformation matching than in force matching.

Impact Vibration Source Localization in Two-Dimensional Space Around Hand.

This article investigated the localization ability of an impulse vibration source outside the body in two-dimensional space. We tested whether humans can recognize the direction or distance of an impulse vibration source when using their hand to detect spatiotemporal vibrotactile information provided by the propagated vibrational wave from the source. Specifically, we had users put their hands on a silicone rubber sheet in several postures. We asked users to indicate the position of the vibration source when a location on the sheet was indented. Experimental results suggested that the direction of the impact vibration source can be recognized to some extent, although recognition accuracy depends on hand posture and the position of the vibration source. The best results were achieved when the fingers and palm were grounded and a vibration source was presented around the middle fingertip, and the directional recognition error in this case was 6 °. In contrast, results suggest it is difficult to accurately recognize the distance of the vibration. The results of this study suggest a new possibility for directional display where vibrotactile actuators are embedded at a distance from the user's hand.

Influence of Sparse Contact Point and Finger Penetration in Object on Shape Recognition.

Making a virtual object shape recognizable using a haptic display is one of the major themes of haptic research. In previous works, multipoint haptic displays have been developed that had a high contact point density between the users' finger skin and the virtual object. However, the ideal contact point density that enables intuitive shape recognition has not been determined yet. Meanwhile, there is also a fundamental problem; that is, real fingers and virtual objects do penetrate, which cannot be solved with such wearable displays. This article investigated the influence of both contact point density and penetration on the shape recognition performance. We prepared a real testing environment where the user touched the real object, and where we could simulate both the sparse contact point and the penetration. Specifically, users' fingers wore thin film coated with glass particles and they touched the urethane foams that deformed flexibly. The result of experiments showed a broad trend where the sparseness of the contact and the softness of the object influenced the exploration time required to achieve recognition. In addition, the result suggested that the larger contact density could make up for the problem of penetration. We confirmed it by conducting two different tasks: (1) exploring the object surface with the index finger and (2) grasping the object surface with the thumb and the index finger.

C-Mannosyl tryptophan increases in the plasma of patients with ovarian cancer.

Ovarian cancer survival is poor, in part, because there are no specific biomarkers for early diagnosis. C-Mannosyl tryptophan (CMW) is a structurally unique glycosylated amino acid recently identified as a novel biomarker of renal dysfunction. The present study investigated whether blood CMW is altered in patients with ovarian cancer and whether differences in blood CMW can distinguish benign from malignant ovarian tumors. Plasma samples were obtained from 49 patients with malignant, borderline or benign ovarian tumors as well as from seven age-matched healthy women. CMW was identified and quantified in these samples using ultra-performance liquid chromatography with fluorometry. Plasma CMW was significantly higher in the malignant tumor group than in the borderline and benign tumor groups, and higher in the combined tumor group (malignant, borderline or benign) compared with healthy controls. Receiver operating characteristic curve analysis of plasma CMW distinguished malignant tumors from borderline/benign tumors [area under the curve (AUC)=0.905]. Discrimination performance was greater than that of cancer antigen (CA) 125 (AUC=0.835), and CMW + CA125 combined achieved even greater discrimination (AUC=0.913, 81.8% sensitivity, 87.5% specificity, 93.1% positive predictive value and 70.0% negative predictive value). Plasma CMW differentiates malignant ovarian cancer from borderline or benign ovarian tumors with high accuracy, and performance is further improved by combined CMW and CA125 measurement.

A novel assay for detection and quantification of C-mannosyl tryptophan in normal or diabetic mice.

C-Mannosyl tryptophan (C-Man-Trp) is a unique molecule in that an α-mannose is connected to the indole C2 carbon atom of a Trp residue via C-glycosidic linkage. Although serum C-Man-Trp may be a novel biomarker of renal function in humans, the biological significance of C-Man-Trp has yet to be fully investigated. In this study, a novel assay system for C-Man-Trp was established using hydrophilic-interaction liquid chromatography, followed by detecting the fluorescence intensity or mass abundance of C-Man-Trp. Using this system, we systematically assessed the amount of free monomeric C-Man-Trp in different tissues of mice. The tissue level of C-Man-Trp was high, especially in the ovaries and uterus. Other organs with high levels of C-Man-Trp included the brain, spleen, lungs, bladder, and testes. The level was low in skeletal muscle. We also investigated whether the tissue level of C-Man-Trp is affected in diabetes. In KK-Ay diabetic mice, the level of urinary C-Man-Trp excretion was increased, and the tissue levels of C-Man-Trp were decreased in the liver but increased in the kidney. These results demonstrate that C-Man-Trp is differentially distributed in numerous tissues and organs in mice, and the levels are altered by disordered carbohydrate metabolism such as diabetes.

Water temperature-dependent degradation of environmental DNA and its relation to bacterial abundance.

Environmental DNA (eDNA) is DNA shed by organisms into surrounding environments such as soil and water. The new methods using eDNA as a marker for species detection are being rapidly developed. Here we explore basic knowledge regarding the dependence of the eDNA degradation rate on time and water temperature, and the relationship between eDNA degradation and bacterial abundance. This subject has not been well clarified, even though it is essential for improving the reliability of eDNA analysis. To determine the time- and water temperature-dependent degradation of eDNA, river water was sampled and eDNA concentrations were determined for ayu sweetfish (Plecoglossus altivelis altivelis) and common carp (Cyprinus carpio) at seven time points, over a 48-h period, and at three different water temperatures. The degradation of eDNA was modeled for each species using an existing exponential decay model with an extension to include water temperature effects. The degradation models were constructed for ayu sweetfish as Nt = 229,901.2 × exp [- (0.01062 × k - 0.07081) × t] and for common carp as Nt = 2,558.0 × exp [- (0.01075 × k - 0.07372) × t]. Nt is the DNA concentration at time t (elapsed time in hours) and k is the water temperature (°C). We also measured the concentration of eDNA derived from purified genomic DNA of the common carp, which was spiked into aquarium water without the target species, and we measured the bacterial abundance in the sample water after 12 and 24 h of incubation. Environmental DNA degradation was accelerated at higher water temperatures (generalized linear model, GLM; p < 0.001), but bacterial abundance did not have a significant effect on eDNA degradation (GLM, p = 0.097). These results suggest that the proper treatment of this temperature effect in data interpretations and adjustments would increase the reliability of eDNA analysis in future studies.

Expression of Heat Shock Protein Genes in Different Developmental Stages and After Temperature Stress in the Maize Weevil (Coleoptera: Curculionidae).

The maize weevil, Sitophilus zeamais Motschulsky, is a major pest of rice and other postharvest grain stocks in tropical countries. Heating and cooling treatments have been adopted to control this pest. Because heat shock protein (hsp) genes respond to temperature stress, we examined the association of hsp genes with development and thermal stress in S. zeamais. The temperature response of the insect to heat and cold treatments was assessed at four developmental stages: egg, larva, pupa, and adult. LT50 values at high temperatures were similar among the four developmental stages, while adults were the most tolerant to low temperatures, and eggs, larvae, and pupae exhibited similar LT50 values. Expression levels of three hsps--Szhsp70, Szhsc70, and Szhsp90--fluctuated substantially throughout the four stages at a rearing temperature of 28°C. Heat shock and cold shock increased the expression of all three hsps, and the highest upregulation was observed at 40°C, although the intensity of upregulation varied among the three genes: strongly in Szhsp70, moderately in Szhsp90, and slightly in Szhsc70. Basal expression of the three hsps at 28°C and gene responses to heat and cold shock also varied significantly at the tissue level.

Trehalase activity in fungus-growing termite, Odontotermes feae (Isoptera: Termitideae) and inhibitory effect of validamycin.

Trehalase is the hydrolytic enzyme that catalyzed the hydrolysis of trehalose to glucose. In this study, trehalase activity in the fungus-growing termite, Odontotermes feae Wasmann had been examined. Trehalase activity in digestive tract and carcass of O. feae was higher than that in wood-feeding termite, Coptotermes gestroi Wasmann. The intestinal tract of worker caste of O. feae was the main source of trehalase compared with that in salivary, fat body, and carcass. In particular, the highest activity was found in the midgut and hindgut parts. More specifically, the contents of midgut and hindgut had higher enzyme activity compared with that trehalase prepared from their epithelial tissue. The enzyme activity of gut trehalase in three different termite castes, worker, soldier, and reproductive, had been determined. The result showed that female alate had the highest activity, followed by worker, male alate, and soldier castes. Trehalose concentration in the reproductive caste was at lowest level, while soldier and worker contained the high trehalose concentration. This study indicates that high trehalase activity locates in the midgut and hindgut contents and change in trehalase activity in fungus-growing termite is caste-specific. Validamycin inhibited trehalase activity of O. feae in vivo and caused high mortality, indicating that this trehalase inhibitor is valuable tools for termite control.

Hormonal regulation of the death commitment in programmed cell death of the silkworm anterior silk glands.

During larval-pupal transformation, the anterior silk glands (ASGs) of the silkworm Bombyx mori undergo programmed cell death (PCD) triggered by 20-hydroxyecdysone (20E). Under standard in vitro culture conditions (0.3 ml of medium with 1 µM 20E), ASGs of the fourth-instar larvae do not undergo PCD in response to 20E. Similarly, larvae of the fifth instar do not respond to 20E through day 5 of the instar (V5). However, ASGs of V6 die when challenged by 20E, indicating that the glands might be destined to die before V6 but that a death commitment is not yet present. When we increased the volume of culture medium for one gland from 0.3 to 9 ml, V5 ASGs underwent PCD. We examined the response of ASGs to 20E every day by culturing them in 9 ml of medium and found that ASGs on and after V2 were fully responsive to 20E. Because pupal commitment is associated with juvenile hormone (JH), the corpora allata (a JH secretory organ) were removed on day 3 of the fourth larval instar (IV3), and their ASGs on V0 were cultured with 20E. Removal of the corpora allata allowed the V0 larval ASGs to respond to 20E with PCD. In contrast, topical application of a JH analogue inhibited the response to 20E when applied on or before V5. We conclude that the acquisition of responsiveness to 20E precedes the loss of JH sensitivity, and that the death commitment in ASGs occurs between V5 and 6.

Programmed cell death of larval tissues induced by juvenile hormone in the bamboo borer, Omphisa fuscidentalis.

Programmed cell death (PCD) plays a critical role during animal development through the destruction of unneeded cells and tissues. In some insects, the prothoracic glands (PGs) and anterior silk glands (ASGs) are larval-specific tissues that are normally eliminated by PCD after pupation. Previous studies report that juvenile hormone analog (JHA) terminates the larval diapause of Omphisa fuscidentalis by increasing the hemolymph ecdysteroids that trigger PCD. Because JHA may indirectly induce the PCD of the PGs and ASGs of Omphisa diapausing larvae, the effects of JHA on the induction of PCD were determined. The application of 1µg JHA induced PCD in the PGs and ASGs of larvae identified as stage G0 (prior to pupation). The injection of 1µg 20E triggered the PCD of the ASGs when the larvae expressed a G0-G1 morphology, whereas PCD occurred in the PGs on day 1 post-injection. Histological studies revealed similar patterns of morphological changes during the PG and ASG PCD in the JHA- and 20E-treated larvae. Furthermore, to confirm that PCD was induced by a high ecdysteroid level that increases after JHA application, the expression profiles of EcR-A and EcR-B1 in the PGs and ASGs from the JHA-treated larvae were examined, and the results showed that the expression levels of EcR-A and EcR-B1 mRNA increased during the G0 stage. These results suggest that JHA may be involved in PCD by increasing the ecdysteroid titer, leading to termination of the larval diapause period in Omphisa fuscidentalis.

Improved photocurrent of a poly (3,4-ethylenedioxythiophene)-ClO4⁻/TiO2 thin film-modified counter electrode for dye-sensitized solar cells.

We prepared a poly(3,4-ethylenedioxythiophene) (PEDOT)-ClO4⁻-supported TiO2 thin-film electrode as a counter electrode on a transparent conductive oxide glass electrode for a dye-sensitized solar cell (DSSC) using a combination of sol-gel and electropolymerization methods. The photocurrent-voltage characteristics indicate that DSSCs with PEDOT-ClO4⁻/TiO2 thin-film counter electrodes had a high photovoltaic conversion efficiency similar to that of PEDOT-ClO4⁻/TiO2 particle composite-film electrodes. Furthermore, it was found that the photocurrent was increased by attaching a reflector to the opposite side of the transparent counter electrode.

Larval fat body cells die during the early pupal stage in the frame of metamorphosis remodelation in Bombyx mori.

In holometabolus insects, morphology of the larval fat body is remodeled during metamorphosis. In higher Diptera, remodeling of the fat body is achieved by cell death of larval fat body cells and differentiation of the adult fat body from primordial cells. However, little is known about remodeling of the fat body at pupal metamorphosis in Lepidoptera. In this study, we found that cell death of the larval fat body in Bombyx mori occurs at shortly after pupation. About 30% of the fat body cells underwent cell death on days 1 and 2 after pupation. The cell death involved genomic DNA fragmentation, a characteristic of apoptosis. Surgical manipulation and in vitro culture of fat body cells revealed that 20-hydroxyecdysone and juvenile hormone had no effect on either initiation or progression of cell death. During cell death, a large increase in activity of caspase-3, a key enzyme of cell death, was observed. Western blot analysis of the active form of caspase-3-like protein revealed that the length of caspase-3 of B. mori was much larger than that of caspase-3 in other species. The results suggest that larval fat body cells of B. mori are removed through cell death, which is mediated by a caspase probably categorized in a novel family.

Steroidal regulation of hydrolyzing activity of the dietary carbohydrates in the silkworm, Bombyx mori.

Blood sugar is an essential energy source for growth and development and is maintained at a constant level through precise regulation of formation and utilization. Sugars are produced from dietary carbohydrates by enzymatic hydrolysis in the digestive tract, which are under the homeostatic control of paracrine and prandial mechanisms in mammals. Here, we show that dietary carbohydrates hydrolyzing activity of the digestive tract is developmentally regulated by the steroid hormone ecdysone in the silkworm, Bombyx mori. The dietary carbohydrates hydrolyzing activity remained high throughout the last larval period and then decreased to negligible levels until the pupal period. However, dietary carbohydrates digestive activities were constitutively high when the steroidogenic organ, prothoracic glands were ablated. The prothoracic glands produced and released a large amount of ecdysone at the end of the larval period, suggesting that ecdysone is responsible for the decrease in dietary carbohydrates hydrolyzing activity. In fact, ecdysone decreased the activity to negligible levels in silkworms lacking the prothoracic glands. The present results indicate that the dietary carbohydrates hydrolyzing activity is regulated by ecdysone and that an increase in ecdysone titer decreases that activity at the end of the larval period, suggesting that ecdysone is essential for metabolic coordination during development.

Glucose oxidase prevents programmed cell death of the silkworm anterior silk gland through hydrogen peroxide production.

During pupal metamorphosis, the anterior silk glands (ASGs) of the silkworm Bombyx mori degenerate through programmed cell death (PCD), which is triggered by 20-hydroxyecdysone (20E). 20E triggers the PCD of the ASGs of day 7 fifth instar (V7) larvae but not that of V5 larvae. When V7 ASGs were cocultured with V5 ASGs in the presence of 20E, neither culture of ASGs underwent PCD. The 20E-induced PCD of V7 ASGs was also inhibited when they were incubated in conditioned medium that was prepared by incubating V5 ASGs for 48 h, an indication that V5 ASGs released an inhibitor of 20E-induced PCD during incubation. The inhibitor was purified from conditioned medium and identified as glucose oxidase (GOD). GOD catalyzes the oxidation of glucose to gluconolactone, and generates hydrogen peroxide as a byproduct. We found that hydrogen peroxide is the molecule that directly inhibits the action of 20E and may act to protect the ASGs from early execution of PCD during the feeding stage. GOD was localized in the inner cavity of the gland, and was discharged to the outside of the ASGs with the silk thread at the onset of spinning. Thus, the spinning behavior, occurring at the beginning of the prepupal period, plays an important role in controlling the time at which ASGs undergo PCD in response to 20E.

Physiological requirements for 20-hydroxyecdysone-induced rectal sac distention in the pupa of the silkworm, Bombyx mori.

Successful insect development is achieved via appropriate fluctuation of ecdysteroid levels. When an insect's ecdysteroid level is disrupted, physiological and developmental defects occur. In the pupa of the silkworm, Bombyx mori, the rectal sac is an essential organ that operates as a repository for degraded ecdysteroids, and it can be distended by administration of 20-hydroxyecdysone (20E). Our previous study showed that rectal sac distention appears 4 days after 20E administration. Hemolymph ecdysteroid levels, however, decrease to lower level during this period. Thus, the timing of the rectal sac distention does not match with that of ecdysteroid elevation. Here, we examine how 20E induces rectal sac distention. A ligature experiment and ecdysteroid quantification showed that continuous 20E stimulation induces rectal sac distention. Thorax tissue contributed to the continuous 20E stimulation needed to induce distention. Ecdysteroid released from the thorax tissue may be converted to 20E by ecdysone 20-hydroxylase to produce continuous 20E stimulation. Thus, the ecdysone metabolic pathway plays a critical role in rectal sac distention.

Ca2+-PKC-caspase 3-like protease pathway mediates DNA and nuclear fragmentation in ecdysteroid-induced programmed cell death.

20-Hydroxyecdysone (20E) induces programmed cell death in the anterior silk gland of the silkworm. Here, we report the direct interaction between Ca(2+) and protein kinase C (PKC)-caspase 3-like protease pathway in the 20E-induced cell death. The calcium ionophore can mimic 20E effects in inducing DNA and nuclear fragmentation, but such mimicry is only possible in the glands precultured for 18 h with 20E. The simultaneous presence of translation inhibitor with 20E in the preculture showed that de novo protein synthesis was needed to mimic 20E effects by the calcium ionophore. Both a PKC inhibitor and a caspase 3 inhibitor inhibited the mimicking effects. After substitution of the calcium ionophore for 20E, caspase 3-like protease was fully activated 12h later, and DNA and nuclear fragmentation occurred faster than continuous 20E stimuli. The results show the presence of a Ca(2+)-PKC-caspase 3-like protease pathway in 20E signaling, and possible involvement of the pathway up to the mobilization of Ca(2+) in regulating the timing of cell death in vivo.

Manduca sexta prothoracicotropic hormone: evidence for a role beyond steroidogenesis.

Prothoracicotropic hormone (PTTH) is a homodimeric brain peptide hormone that positively regulates the production of ecdysteroids by the prothoracic gland of Lepidoptera and probably other insects. PTTH was first purified from heads of adult domestic silkworms, Bombyx mori. Prothoracic glands of Bombyx and Manduca sexta undergo apoptosis well before the adult stage is reached, raising the recurring question of PTTH function at these later stages. Because Bombyx has been domesticated for thousands of years, the possibility exists that the presence of PTTH in adult animals is an accidental result of domestication for silk production. In contrast, Manduca has been raised in the laboratory for only five or six decades. The present study found that Manduca brains contain PTTH at all stages examined post-prothoracic gland apoptosis, i.e., pharate adult and adult life, and that PTTH-dependent changes in protein phosphorylation and protein synthesis were observed in several reproductive and reproduction-associated organs. The data indicate that PTTH indeed plays a role in non-steroidogenic tissues and suggest possible future avenues for determining which cellular processes are being so regulated.

Rectal sac distention is induced by 20-hydroxyecdysone in the pupa of Bombyx mori.

Holometabolous insects do not excrete but store metabolic wastes during the pupal period. The waste is called meconium and is purged after adult emergence. Although the contents of meconium are well-studied, the developmental and physiological regulation of meconium accumulation is poorly understood. In Bombyx mori, meconium is accumulated in the rectal sac; thereby, the rectal sac distends at the late pupal stage. Here, we show that rectal sac distention occurs between 4 and 5 days after pupation. The distention is halted by brain-removal just after larval-pupal ecdysis but not by brain-removal 1 day after pupation. In the pupae, brain-removal just after ecdysis kept the hemolymph ecdysteroid titer low during early and mid-pupal stages. An injection of 20-hydroxyecdysone (20E) evoked the distention that was halted by brain-removal in a dose-dependent manner. Therefore, brain-removal caused the lack of ecdysteroid, and rectal sac distention did not appear in the brain-removed pupae because of the lack of ecdysteroid. We conclude that rectal sac distention is one of the developmental events regulated by 20E during the pupal period in B. mori.

Intracellular mobilization of Ca2+ by the insect steroid hormone 20-hydroxyecdysone during programmed cell death in silkworm anterior silk glands.

20-Hydroxyecdysone (20E) triggers programmed cell death (PCD) and regulates de novo gene expression in the anterior silk glands (ASGs) of the silkworm Bombyx mori. PCD is mediated via a nongenomic pathway that includes Ca2+ as a second messenger and the activation of protein kinase C/caspase-3-like protease; however, the steps leading to a concomitant buildup of intracellular Ca2+ are unknown. We employed pharmacological tools to identify the components of this pathway. ASGs were cultured in the presence of 1 microM 20E and one of the following inhibitors: a G-protein-coupled receptor (GPCR) inhibitor, a phospholipase C (PLC) inhibitor, an inositol 1,4,5-trisphosphate receptor (IP3R) antagonist, and an L- or T-type Ca2+ channel blocker. The T-type Ca2+ channel blocker inhibited 20E-induced nuclear and DNA fragmentation; in contrast, PCD was induced by 20E in Ca2+-free medium, indicating that the source of Ca2+ is an intracellular reservoir. The IP3R antagonist inhibited nuclear and DNA fragmentation, suggesting that the endoplasmic reticulum may be the Ca2+ source. Finally, the GPCR and PLC inhibitors effectively blocked nuclear and DNA fragmentation. Our results indicate that 20E increases the intracellular level of Ca2+ by activating IP3R, and that this effect may be brought about by the serial activation of GPCR, PLC, and IP3.

Expression of 20-hydroxyecdysone-induced genes in the silkworm brain and their functional analysis in post-embryonic development.

The insect brain is the center of developmental control, from which ecdysone governs brain morphogenesis and regulates gene expression cascades associated with molting and metamorphosis. In order to identify the 20-hydroxyecdysone (20E)-inducible genes responsible for molting and metamorphosis, we constructed a 20E-induced subtraction complementary DNA library from the fifth instar larval brain of the silkworm Bombyx mori. We isolated 10 genes, designated as bombeil-1 to bombeil-10, three of which did not show any sequence similarity to previously identified Bombyx genes. Whole-mount in situ hybridization revealed that all of these bombeil messenger RNAs were exclusively located in two pairs of lateral neurosecretory cells in the larval brain, known as prothoracicotropic hormone (PTTH)-producing cells. RNA-interference knockdown targeting bombeil-2 resulted in larval-pupal molt defects, and adult wing and leg malformations. These results, together with the cell-specific co-localization of bombeil transcripts with PTTH, suggest that bombeil genes play important roles during larval-pupal-adult development.