Multiple N-cadherin enhancers identified by systematic functional screening indicate its Group B1 SOX-dependent regulation in neural and placodal development

AbstractNeural plate and sensory placodes share the expression of N-cadherin and Group B1 Sox genes, represented by Sox2. A 219-kb region of the chicken genome centered by the N-cadherin gene was scanned for neural and placodal enhancers. Random subfragments of 4.5 kb average length were prepared and inserted into tkEGFP reporter vector to construct a library with threefold coverage of the region. Each clone was then transfected into N-cadherin-positive (lens, retina and forebrain) or -negative embryonic cells, or electroporated into early chicken embryos to examine enhancer activity. Enhancers 1–4 active in the CNS/placode derivatives and non-specific Enhancer 5 were identified by transfection, while electroporation of early embryos confirmed enhancers 2–4 as having activity in the early CNS and/or sensory placodes but with unique spatiotemporal specificities. Enhancers 2–4 are dependent on SOX-binding sites, and misexpression of Group B1 Sox genes in the head ectoderm caused ectopic development of placodes expressing N-cadherin, indicating the involvement of Group B1 Sox functions in N-cadherin regulation. Enhancers 1, 2 and 4 correspond to sequence blocks conserved between the chicken and mammalian genomes, but enhancers 3 and 5 are unique to the chicken

Utilization of online real–time feedback application service “Mentimeter” in the lecture of Operative Dentistry for steps in order to pass the National Board Dental Examination at Matsumoto Dental University

Summary Many attempts to utilize ICT (Information and Communication Technology) tools have been made to promote active and interactive learning between teachers and students. A major area has been the introduction of education using clickers for dental students, and some reports indicated their effects. However, the introduction of mobile devices and software and their maintenance and management very costly. On the other hand, almost allundergraduate students of Matsumoto Dental University have a smartphone, and can connect to the Internet in the lecture room. Therefore, we conducted online exercises to pre- pare for the National Board Dental Examination using students’ smartphones and the online real–time voting service“Mentimeter” at the lecture of Operative Dentistry for 6th–year dental students of Matsumoto Dental University, in the school year of 2019. We also conducted some anonymous online questionnaires using Mentimeter. Thirty percent of the participating students answered that they were not good at conservation and restoration studies. Although 97% of students answered that the test questions conducted in the exercise using Mentimeter were difficult, 92% of the students answered that the exercise was effective in deepening their knowledge of Operative Dentistry. The results of the questionnaires suggested the effectiveness of the interactive lecture using Mentimeter in order to deepen and retain knowledge of Operative Dentistry

Attempt to introduce a new educational program in the clinical clerkship of operative dentistry for 5th grade students, School of Dentistry,Matsumoto Dental University Part2.Simulation training of direct composite veneer restoration

Summary A simulation practice of direct composite veneer restoration was newly started during the clinical training period of the 5th year conservatory course of Matsumoto Dental University since the AY 2020. This manuscript presents an overview of the program and examines its educational effects based on the evaluation of an anonymous questionnaire by students who participated in the program in 2020. 72 students in total participated in the 5th year clinical internship in AY 2020, and the program was conducted 9 times for 8 students each in the Department of Operative Dentistry. After a lecture and demonstration by the instructor, four pairs of two students each were assigned to practice direct composite veneer restoration without tooth reduction by applying the layering technique. After the practice, a lecture on the technique,materials and equipment used was given, followed by exercises on the relevant questions of the National Dental Examination that were asked in the past. After the program, an anonymous questionnaire was administered to the participants regarding their level of knowledge of direct composite veneer restoration before and after the program and their self–evaluation of the completion of their practice. The response rate of the questionnaires was 100%. After the program, 68% of the respondents answered that they “understood the direct composite veneer restoration very well” and 31% answered that they “understood it somewhat well. None of the respondents answered that they “did not understand much” or “did not understand at all. When asked whether the contents of the program deepened their knowledge of operative dentistry, 83% of the respondents answered “Very effective” and 15% answered “Somewhat effective”, and none of the respondents answered “Not very effective” or “Not at all effective”. These results suggested that the program was effective in deepening the knowledge level of operative dentistry

Down-regulation of circadian clock gene period 2 in uterine endometrial stromal cells of pregnant rats during decidualization

Circadian rhythms are modulated in a variety of peripheral tissues, including in the uterus where endometrial stromal cells (UESCs) undergo proliferation and differentiation (decidualization) during gestation. Here the authors focused on circadian rhythms in UESCs during implantation and decidualization in rodents. As revealed by analyses of cultured UESCs from pregnant Per2 promoter-dLuc transgenic rats, Per2 oscillation of ～24h was observed in response to dexamethasone. Per2 oscillation was enhanced in UESCs during implantation, whereas they were attenuated during decidualization. In vivo studies showed that PER2 protein in the uteri displayed a peak at zeitberger time 4 (ZT 4) (day 4.50 of gestation) and a trough at ZT 12 (day 4.83), indicating its circadian rhythmicity. Conversely, no significant circadian rhythm of the PER2 protein was observed during decidualization. Fluorescent immunohistochemical studies also supported circadian rhythmicity of the PER2 protein in its intracellular distribution. In accordance with Per2 mRNA expression, a circadian rhythm of vascular endothelial growth factor (Vegf) gene expression, having several E-box or E-boxlike sites at the upstream of the transcription start site, was observed during implantation, showing a peak at ZT 0 and a trough at ZT 12. In contrast, Vegf mRNA expression displayed no circadian rhythm during decidualization. Collectively, the present results prove that Per2 oscillation is down-regulated in UESCs during decidualization. It is strongly suggested that cellular differentiation in UESCs interferes with circadian clockwork

Down-regulation of circadian clock gene period 2 in uterine endometrial stromal cells of pregnant rats during decidualization

Circadian rhythms are modulated in a variety of peripheral tissues, including in the uterus where endometrial stromal cells (UESCs) undergo proliferation and differentiation (decidualization) during gestation. Here the authors focused on circadian rhythms in UESCs during implantation and decidualization in rodents. As revealed by analyses of cultured UESCs from pregnant Per2 promoter-dLuc transgenic rats, Per2 oscillation of ～24h was observed in response to dexamethasone. Per2 oscillation was enhanced in UESCs during implantation, whereas they were attenuated during decidualization. In vivo studies showed that PER2 protein in the uteri displayed a peak at zeitberger time 4 (ZT 4) (day 4.50 of gestation) and a trough at ZT 12 (day 4.83), indicating its circadian rhythmicity. Conversely, no significant circadian rhythm of the PER2 protein was observed during decidualization. Fluorescent immunohistochemical studies also supported circadian rhythmicity of the PER2 protein in its intracellular distribution. In accordance with Per2 mRNA expression, a circadian rhythm of vascular endothelial growth factor (Vegf) gene expression, having several E-box or E-boxlike sites at the upstream of the transcription start site, was observed during implantation, showing a peak at ZT 0 and a trough at ZT 12. In contrast, Vegf mRNA expression displayed no circadian rhythm during decidualization. Collectively, the present results prove that Per2 oscillation is down-regulated in UESCs during decidualization. It is strongly suggested that cellular differentiation in UESCs interferes with circadian clockwork