Binding of APC and dishevelled mediates Wnt5a‐regulated focal adhesion dynamics in migrating cells

Wnt5a is a representative ligand that activates the Wnt/β‐catenin‐independent pathway, resulting in the regulation of cell adhesion, migration, and polarity, but its molecular mechanism is poorly understood. This report shows that Dishevelled (Dvl) binds to adenomatous polyposis coli (APC) gene product, and this binding is enhanced by Wnt5a. Dvl was involved in the stabilization of the plus end dynamics of microtubules as well as APC. Frizzled2 (Fz2) was present with Wnt5a at the leading edge of migrating cells and formed a complex with APC through Dvl. Fz2 also interacted with integrins at the leading edge, and Dvl and APC associated with and activated focal adhesion kinase and paxillin. The binding of APC to Dvl enhanced the localization of paxillin to the leading edge and was involved in Wnt5a‐dependent focal adhesion turnover. Furthermore, this new Wnt5a signalling pathway was important for the epithelial morphogenesis in the three‐dimensional culture. These results suggest that the functional and physical interaction of Dvl and APC is involved in Wnt5a/Fz2‐dependent focal adhesion dynamics during cell migration and epithelial morphogenesis.     

Applying Data Envelopment Analysis to Preventive Medicine: A Novel Method for Constructing a Personalized Risk Model of Obesity

Data envelopment analysis (DEA) is a method of operations research that has not yet been applied in the field of obesity research. However, DEA might be used to evaluate individuals’ susceptibility to obesity, which could help establish effective risk models for the onset of obesity. Therefore, we conducted this study to evaluate the feasibility of applying DEA to predict obesity, by calculating efficiency scores and evaluating the usefulness of risk models. In this study, we evaluated data from the Takahata study, which was a population-based cohort study (with a follow-up study) of Japanese people who are >40 years old. For our analysis, we used the input-oriented Charnes-Cooper-Rhodes model of DEA, and defined the decision-making units (DMUs) as individual subjects. The inputs were defined as (1) exercise (measured as calories expended) and (2) the inverse of food intake (measured as calories ingested). The output was defined as the inverse of body mass index (BMI). Using the β coefficients for the participants’ single nucleotide polymorphisms, we then calculated their genetic predisposition score (GPS). Both efficiency scores and GPS were available for 1,620 participants from the baseline survey, and for 708 participants from the follow-up survey. To compare the strengths of the associations, we used models of multiple linear regressions. To evaluate the effects of genetic factors and efficiency score on body mass index (BMI), we used multiple linear regression analysis, with BMI as the dependent variable, GPS and efficiency scores as the explanatory variables, and several demographic controls, including age and sex. Our results indicated that all factors were statistically significant (p < 0.05), with an adjusted R² value of 0.66. Therefore, it is possible to use DEA to predict environmentally driven obesity, and thus to establish a well-fitted model for risk of obesity.     

Interaction between FliJ and FlhA,Components of the Bacterial Flagellar Type III Export Apparatus

A soluble protein, FliJ, along with a membrane protein, FlhA, plays a role in the energy coupling mechanism for bacterial flagellar protein export. The water-soluble FliH_X-FliI₆ ATPase ring complex allows FliJ to efficiently interact with FlhA. However, the FlhA binding site of FliJ remains unknown. Here, we carried out genetic analysis of a region formed by well-conserved residues—Gln38, Leu42, Tyr45, Tyr49, Phe72, Leu76, Ala79, and His83—of FliJ. A structural model of the FliI₆-FliJ ring complex suggests that they extend out of the FliI₆ ring. Glutathione S-transferase (GST)-FliJ inhibited the motility of and flagellar protein export by both wild-type cells and a fliH-fliI flhB(P28T) bypass mutant. Pulldown assays revealed that the reduced export activity of the export apparatus results from the binding of GST-FliJ to FlhA. The F72A and L76A mutations of FliJ significantly reduced the binding affinity of FliJ for FlhA, thereby suppressing the inhibitory effect of GST-FliJ on the protein export. The F72A and L76A mutations were tolerated in the presence of FliH and FliI but considerably reduced motility in their absence. These two mutations affected neither the interaction with FliI nor the FliI ATPase activity. These results suggest that FliJ(F72A) and FliJ(L76A) require the support of FliH and FliI to exert their export function. Therefore, we propose that the well-conserved surface of FliJ is involved in the interaction with FlhA.     

Canopy-related adult density and sex-related flight activity of Monochamus alternatus (Coleoptera: Cerambycidae) in pine stands

Monochamus alternatus Hope is an important vector of the pinewood nematode Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle, which causes pine wilt disease. To determine canopy-related differences in the activity density of M. alternatus adults in pine stands, sticky screen traps with no lure were suspended under and in the canopy at 3.8 and 10 m above the ground of a Pinus densiflora Siebold et Zuccarini stand during a flight season. The five upper and five lower traps caught 338 and 54 adults, respectively, during a flight season from 9 June to 12 October 2011, indicating that activity density was six times as high in the canopy as under it. Male-biased sex ratio was observed for trap catches during a season: The sex ratio was male-biased in the early half of the flight season and 1:1 in the late half. There was no difference in sex ratio between the inside and outside of the canopy. To determine the sex-related difference in flight activity, the adults were captured by two different measures in two other mixed stands of P. densiflora and P. thunbergii Parlatore. Kicking tree trunks to cause beetles to drop revealed a 1:1 sex ratio of catches, whereas sticky screen traps tended to show a male-biased sex ratio in a 4-year study, suggesting that males were more active fliers than females.     

Observations of the migration of adult Dasylepida ishigakiensis (Coleoptera: Scarabaeidae) from belowground to the soil surface and their reproductive status by MRI

Field observations indicate that white grub beetles Dasylepida ishigakiensis Niijima et Kinoshita (Coleoptera: Scarabaeidae) pupate approximately 40 cm underground and then emerge from the soil surface as adults. Although the factors influencing adult emergence from the soil have been studied, the migration of adults from belowground to the surface has never been characterized. In this study, the vertical movement of virgin adults from natural eclosion depths to their emergence from the soil surface was investigated. Virgin adults were introduced at a depth of 40 cm from the soil surface in wooden-framed glass cases, and the vertical position of each individual was checked every day. Both females and males emerged near the soil surface at approximately day 50 after they were transferred from 24 °C to 18 °C conditions. The maturity of the ovaries in unemerged and emerged females was also observed by magnetic resonance imaging. All females that had emerged from the soil possessed developed oocytes, and most possessed mature eggs. The cross-sectional area of eggs was determined in magnetic resonance images and compared between unemerged and emerged females, and the egg areas of emerged females were significantly larger than those of unemerged females. Our results indicate that the timing of female emergence from soil corresponded to ovary maturity in this species.     

Serratia Protease

A strain of Serratia, isolated from an intestinal canal of a silkworm, produced a large quantity of protease. The enzyme was extracellular and was named Serratiopeptidase, tentatively. Protease production of this strain was over 3 times as much as that of Serratia marcescens which was known as a protease-producing organism. The highly purified enzyme was prepared from the culture supernatant through ammonium sulfate precipitation, acetone fractionation, DEAE-cellulose column chromatography and gel filtration on Sephadex G-75.

The purified enzyme moved homogeneously with a sedimentation constant, s_20,w of 3.8 S in ultracentrifugation and the molecular weight was determined to be 6.0 × 10₄ by the Archibald method. Determination of the ultraviolet absorption spectrum indicated the E^1%_{280 mμ,1 cm} was 13.0. Neither carbohydrate nor sulfur-containing amino acid was detected in the purified enzyme preparation. The enzyme showed maximal activity at pH 9.0 and at 40°C, and was stable under lower temperatures over the pH range from 5 to 10, whereas it was unstable at 37°C in alkaline conditions. The enzyme was completely inactivated by heating at 55°C for 15 min.     

2- or 4-Thiazolylmethyl Cyclopropanecarboxylates: Their Synthesis and Insecticidal Activities

From a consideration of the structure-activity relationships, nineteen 2- or 4-thiazolylmethyl cyclopropanecarboxylates, including three benzyl-thiazolylmethyl chrysanthemates (IVf, Xb and Xc), were prepared from thioamides and α-haloketones or α-haloaldehydes and examined for insecticidal activity against houseflies. All those with benzyl substitutions on the thiazole ring were ineffective, as were the other substituents. Only 4,5,6,7-tetrahydrobenzothiazolylmethyl chrysanthemate and 2,2,3,3-tetramethylcyclopropanecarboxylate (Xf and Xlf) possessed about the same order of activity as allethrin, and their activities were 0.61 and 1.56 μg/fly respectively.     

Metabolism of l-Tyrosine in Aspergillus sojae

Circadian rhythmicity was investigated in isolated small intestine and mucosal epithelial cells from rats on restricted-feeding regimen (food available from 17:00 to 23:00 every day). In the isolated intestine, daily rhythms synchronized to meal-timing were found in the activity patterns of l-leucine, l-lysine and d-glucose transport, and mucosal γ-glutamyltransferase and sucrase, and in the rates of lactate formation from glucose; the nadirs occurred at 12:00 and the peaks at 23:00. These same patterns were also noted with the mucosal epithelial cells prepared at distinct times of day from rats on meal-feeding regimen. The fasted rat intestine responded to refeeding with prompt increase in transport activity, i.e., out of phase with the original rhythm. Intraperitoneal administration of cyloheximide suppressed the daily rise in leucine transport activity, indicating that the transport rhythm was entrained by or closely associated with the rhythmic fluctuation in protein synthesis in the epithelial cells, which in turns is cued by the feeding schedule. The kinetic parameters estimated for leucine transport, the apparent affinity constant for transport and the maximal transport rate, were significantly higher at high activity periods. It is suggested that the rhythmic increase in transport activity is not only associated with membrane hyperpolarization but may be mediated by the emergence of a high capacity transport system.     

Influence of Saturated and Polyunsaturated Egg Yolk Phospholipids on Hyperlipidemia in Rats

The supplementation of egg yolk phospholipid (PL) containing phosphatidylcholine (PC) and phosphatidylethanolamine (PE) to a cholesterol-free purified diet causes a reduction in the serum cholesterol level in rats [J. Nutr., 112,1805 (1982)]. The present study was carried out to determine if dietary egg yolk PL also exerts this hypocholesterolemic action in rats given a high cholesterol diet and if this action is influenced by the constituent fatty acids. Egg yolk PL suppressed the elevation of serum cholesterol irrespective of its fatty acid composition, while purified PC had no effect, suggesting that the ethanolamine portion is responsible for this hypocholesterolemic effect. Egg yolk PL and PC containing longer-chain polyunsaturated fatty acids (arachidonic and docosahexaenoic acids) lowered the serum triglyceride level, while their hydrogenated forms did not. The present results, therefore, indicate that the hypolidemic effect of dietary egg yolk PL can be modulated by the combination of the constituent fatty acids as well as the base moieties. This hypolipidemic effect, however, appeared not to be related to the activities of adipocyte lipoprotein lipase and serum lecithin: cholesterol acyltransferase.