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Biology subjects

Ohara, H.

Publications and source records attributed to Ohara, H..

3 recordsLinked to original sources

Retinoid signaling promotes frontonasal identity while repressing maxillary identity during craniofacial development

Vertebrate facial development depends on the correct specification of embryonic facial prominences, a process known to be governed by region-specific reciprocal signaling pathways between the craniofacial ectoderm, endoderm and mesenchyme. This process is further modulated by transcriptional and epigenetic mechanisms that regulate the expression of essential genes in mesenchymal progenitors derived from cranial neural crest cells. Retinoid signaling plays a critical role in facial development, and both gain-and loss-of-function results in a wide spectrum of facial defects, including orofacial clefts. In this study, we identified retinoid signaling as a critical regulator of cranial neural crest cell specification toward a frontonasal process identity in mice. Rdh10 oxidizes vitamin A (all-trans retinol) to retinal, which is a rate limiting step in the synthesis of retinoic acid. Rdh10 loss-of-function resulted in ectopic formation of whisker pads - a derivative of the maxillary process - within the frontonasal process region. This transformation was evidenced by the mis-expression of maxillary specific transcription factors including Meis2 and Lhx6 in the frontonasal mesenchyme. Furthermore, these transcription factors exhibited increased chromatin accessibility at their consensus binding sites following the loss of retinoid signaling in frontonasal cranial neural crest cells. These results indicate that retinoid signaling acts as a critical regulator specifying frontonasal identity and fate of cranial neural crest cells as they migrate into the frontonasal process, while concomitantly repressing maxillary process fate. These results not only advance our understanding of frontonasal prominence specification and the evolutionary development of craniofacial structures but also offers valuable insights into the etiology and pathogenesis of craniofacial malformations such as orofacial clefts.

developmental biology↗

Antibacterial Effect of Oral Care Gel-Containing Hinokitiol and 4-isopropyl-3-methylphenol Against Intraoral Pathogenic Microorganisms

ObjectiveDeterioration of oral hygiene is closely related to increase severity and mortality of corona virus disease-19 (COVID-19), and also contribute to the development of various diseases such as aspiration pneumonia or Alzheimers. Oral care is attracting high interest in Japan, which has entered a super-aging society. In this study, we aimed to investigate whether commercially available Hinora(R) (HO) that oral care gels-containing hinokitiol and 4-isopropyl-3-methylphenol (IPMP) have biofilm formation inhibitory and antibacterial activities against various intraoral pathogen microorganisms. MethodCandida spp., Aggregatibacter actinomycetemcomitans, Staphylococcus aureus, and Pseudomonas aeruginosa were selected during the study period, all which were analyzed using antimicrobial disc, microorganism turbidity, and crystal violet assays. In addition, the germ tube test using C. albicans was performed with a modification of Mackenzies method. Images for morphological observation of the germ tubes were acquired with an inverted microscope. For comparison between products, we used Refrecare(R) (RC), which contains only hinokitiol (not containing IPMP). ResultsAll the intraoral pathogenic microorganisms showed drug susceptibility against undiluted form HO and/or RC. In particular, HO was more effective at lower concentrations than RC. In the HO-added group, inhibition circles were observed in all bacteria except P. aeruginosa when added at a concentration of 0.5 g/mL or more. The optical density values at 590 nm (crystal violet) and/or 600 nm (microorganism turbidity) of all the fungi and bacteria were significantly lower when cultured in medium with HO. Inhibition of growth or biofilm formation was observed when HO was added at a concentration of 0.05 g/mL or higher. To investigate the action mechanism of HO, germ tube tests were performed in C. albicans. The results show that culturing C. albicans in soyabean-casein digest broth with HO (0.05 g/mL) significantly suppressed germ tube formation. ConclusionsThese data suggest that oral care gel-containing hinokitiol and IPMP has strong biofilm formation inhibitory, antifungal and antibacterial effects against Candida fungi and multiple intraoral pathogenic microorganisms. Therefore, it may be a promising treatment option for oral infections.

microbiology↗

Self-organization, quality control, and preclinical studies of human iPSC-derived retinal sheets for tissue-transplantation therapy

Three-dimensional retinal organoids (3D-retinas) are a promising graft source for transplantation therapy. We previously developed self-organizing culture for 3D-retina generation from human pluripotent stem cells (hPSCs). Here we present a quality control method and preclinical studies for tissue-sheet transplantation. Self-organizing hPSCs differentiated into both retinal and off-target tissues. Gene expression analyses identified the major off-target tissues as eye-related, cortex-like, and spinal cord-like tissues. For quality control, we developed a qPCR-based test in which each hPSC-derived neuroepithelium was dissected into two tissue-sheets: inner-central sheet for transplantation and outer-peripheral sheet for qPCR to ensure retinal tissue selection. During qPCR, tissue-sheets were stored for 3-4 days using a newly developed preservation method. In a rat tumorigenicity study, no transplant-related adverse events were observed. In retinal degeneration model rats, retinal transplants differentiated into mature photoreceptors and exhibited light responses in electrophysiology assays. These results demonstrate our rationale toward self-organizing retinal sheet transplantation therapy.

neuroscience↗