Molecular markers for X‐ray‐insensitive differentiated cells in the Inner and outer regions of the mesenchymal space in planarian Dugesia japonica

Planarian's strong regenerative ability is dependent on stem cells (called neoblasts) that are X‐ray‐sensitive and proliferative stem cells. In addition to neoblasts, another type of X‐ray‐sensitive cells was newly identified by recent research. Thus, planarian's X‐ray‐sensitive cells can be divided into at least two populations, Type 1 and Type 2, the latter corresponding to planarian's classically defined “neoblasts”. Here, we show that Type 1 cells were distributed in the outer region (OR) immediately underneath the muscle layer at all axial levels from head to tail, while the Type 2 cells were distributed in a more internal region (IR) of the mesenchymal space at the axial levels from neck to tail. To elucidate the biological significance of these two regions, we searched for genes expressed in differentiated cells that were locate close to these X‐ray‐sensitive cell populations in the mesenchymal space, and identified six genes mainly expressed in the OR or IR, named OR1, OR2, OR3, IR1, IR2 and IR3. The predicted amino acid sequences of these genes suggested that differentiated cells expressing OR1, OR3, IR1, or IR2 provide Type 1 and Type 2 cells with specific extracellular matrix (ECM) environments.     

Identification of the Isoform-specific Interactions between the Tail and the Head of Class V Myosin

Vertebrates have three isoforms of class V myosin (Myo5), Myo5a, Myo5b, and Myo5c, which are involved in transport of multiple cargoes. It is well established that the motor functions of Myo5a and Myo5b are regulated by a tail inhibition mechanism. Here we found that the motor function of Myo5c was also inhibited by its globular tail domain (GTD), and this inhibition was abolished by high Ca²⁺, indicating that the tail inhibition mechanism is conserved in vertebrate Myo5. Interestingly, we found that Myo5a-GTD and Myo5c-GTD were not interchangeable in terms of inhibition of motor function, indicating isoform-specific interactions between the GTD and the head of Myo5. To identify the isoform-specific interactions, we produced a number of Myo5 chimeras by swapping the corresponding regions of Myo5a and Myo5c. We found that Myo5a-GTD, with its H11-H12 loop being substituted with that of Myo5c, was able to inhibit the ATPase activity of Myo5c and that Myo5a-GTD was able to inhibit the ATPase activity of Myo5c-S1 and Myo5c-HMM only when their IQ1 motif was substituted with that of Myo5a. Those results indicate that the H11-H12 loop in the GTD and the IQ1 motif in the head dictate the isoform-specific interactions between the GTD and head of Myo5. Because the IQ1 motif is wrapped by calmodulin, whose conformation is influenced by the sequence of the IQ1 motif, we proposed that the calmodulin bound to the IQ1 motif interacts with the H11-H12 loop of the GTD in the inhibited state of Myo5.     

Effect of the presence of trophectoderm vesicles on blastocyst in relation to in vitro hatching,clinical pregnancy,and miscarriage rates

Trophectoderm vesicles (TVs) are observed in some blastocysts that penetrate cells from the zona pellucida to the outer margin. Therefore, we compared this incidence in relation to hatching, pregnancy, and miscarriage rates between conventional in vitro fertilization (c-IVF) and intracytoplasmic sperm injection (ICSI). Vitrified/warmed blastocysts (n = 112) were derived from surplus embryos. The blastocysts were then observed using time-lapse cinematography to resolve the relationship between hatching and implantation. Another study was conducted that comprised 681 embryo transfer cycles in 533 patients who received a single vitrified/warmed blastocyst from our clinic. The incidence of TV was significantly higher in embryos inseminated by ICSI compared with c-IVF [ICSI: 51/56 (91 %); c-IVF: 25/56 (45 %); P < 0.01]. The successful hatching rate was significantly lower in ICSI than in c-IVF [ICSI: 11/56 (20 %); c-IVF: 29/56 (52 %); P < 0.01]. In addition, the hatching rate was significantly lower when TVs were present (14/76; 18 %) than in non-TV embryos (26/36; 72 %) (P < 0.01). In regard to the clinical study results, no significant differences were found between the groups in the pregnancy rate (TV present group: 107/183, 58.5 %; TV absent group: 273/498, 54.8 %) and miscarriage rate (TV present group: 21/107, 19.6 %; TV absent group: 53/273, 19.4 %). In vivo, we hypothesized that hatching and hatched would occur naturally by assisting protease action in the uterus; therefore, these results suggest that the presence of TV has no effect on pregnancy rates in the clinical setting.     

The tyrosine-sorting motif of the vacuolar sorting receptor VSR4 from Arabidopsis thaliana,which is involved in the interaction between VSR4 and AP1M2, μ1-adaptin type 2 of clathrin adaptor complex 1 subunits,participates in the post-Golgi sorting of VSR4

μ1-Adaptin of adaptor protein (AP) 1 complex, AP1M, is generally accepted to load cargo proteins into clathrin-coated vesicles (CCVs) at the trans-Golgi network through its binding to cargo-recognition sequences (CRSs). Plant vacuolar-sorting receptors (VSRs) function in sorting vacuolar proteins, which are reportedly mediated by CCV. We herein investigated the involvement of CRSs of Arabidopsis thaliana VSR4 in the sorting of VSR4. The results obtained showed the increased localization of VSR4 at the plasma membrane or vacuoles by mutations in CRSs including the tyrosine-sorting motif YMPL or acidic dileucine-like motif EIRAIM, respectively. Interaction analysis using the bimolecular fluorescence complementation (BiFC) system, V10-BiFC, which we developed, indicated an interaction between VSR4 and AP1M2, AP1M type 2, which was attenuated by a YMPL mutation, but not influenced by an EIRAIM mutation. These results demonstrated the significance of the recognition of YMPL in VSR4 by AP1M2 for the post-Golgi sorting of VSR4.     

Time or retiring and sleep quality may be predictors of academic performance and psychosomatic disorder in university students

In an effort to improve school performance, we have examined the effect of sleep–wake patterns, particularly time in bed, and morning psychosomatic conditions in adolescents. A psychosomatic disorder questionnaire was administered to 135 physically healthy students, who were divided into four groups according to time they went to bed: 23–24, 24–01, 01–02 and 02–03 h. (1) The 23–24 and 24–01 h groups had significantly higher grade point averages than the 02–03 h group. (2) Partial correlations, controlling for sleep quality, revealed significant associations between time to bed and psychosomatic disorder, with regard to whole-body fatigue, lack of motivation and the desire to rest. Our results suggest that the sleep–wake pattern, especially the time of retiring, may predict academic performance and psychosomatic disorder.     

Enigmatic Diversity of the Maxillary Sinus in Macaques and Its Possible Role as a Spatial Compromise in Craniofacial Modifications

Understanding the evolutionary significance of morphological diversity is a major goal of evolutionary biology. Paranasal sinuses, which are pneumatized hollow spaces in the face, have attracted attention from researchers as one of the most intriguing traits that show unexpected variations. Macaques are one genus of primates that have accomplished adaptive radiation and therefore present an excellent opportunity to investigate the phenotypic diversification process. Using the large data set of computed tomography images of macaques (172 specimens from 17 species), we applied geometric morphometrics and multivariate analyses to quantitatively evaluate the maxillary sinus (one of the largest paranasal sinuses), the outer craniofacial shape, and nasal cavity. We then applied phylogenetic comparative methods to test their morphological interactions, phylogenetic, and ecogeographical significances. The results showed that the relative maxillary sinus size was significantly associated with the relative nasal cavity size and with the zygomaxillary surface shape. The relative nasal cavity size had ecogeographical correlations and high phylogenetic signal, whereas the zygomaxillary surface shapes were ecogeographically and phylogenetically irrelevant. The significant interactions with multiple surrounding traits that have experienced different evolutionary processes probably enable the maxillary sinus to show enigmatic diversity, which is independent of phylogeny and ecology. The pliable nature of the maxillary sinus, which is positioned between the nasal airways and the outer face, may play a role as a spatial compromise in craniofacial modifications.     

Detection of Ubiquitinated Dermcidin in Gingival Crevicular Fluid in Periodontal Disease

Periodontal disease is an inflammatory disease caused by gram-negative bacteria, such as Porphyromonas gingivalis and Actinobacllus actinomycetemcomitans. Antimicrobial peptides kill organisms, such as gram-negative and gram-positive bacteria, mycobacteria, enveloped viruses, and fungi. We previously identified the antimicrobial peptide dermcidin (DCD) in the gingival crevicular fluid (GCF) using proteomic analyses. Moreover, western blot analysis revealed that the molecular weights of GCF protein bands considerably varied (approximately 27 kDa). We attempted to explore the considerable variation in the molecular weights of protein bands using on-membrane digestion and liquid chromatography–tandem mass spectrometry (LC–MS/MS) analyses. We examined ubiquitin among the DCD-interacting proteins. In immunoprecipitation experiments, ubiquitinated DCD was detected by western blotting and by immunoprecipitation with an antibody against DCD and mono- or poly-ubiquitinated proteins. These analyses indicated the possible involvement of the ubiquitination reaction. Ubiquitinated DCD may protect against periodontal bacterial pathogen invasion in GCF.