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Contents The CRISPR/Cas9 system has enabled the editing of mammalian genomes; however, its applicability and efficiency in the pig genome has not been studied in depth. The α-gal epitope synthesized by α-1,3-galactosyltransferase gene ( GGTA1) is known as a xenoantigen obtained upon pig-to-human xenotransplantation. We here employed the CRISPR/Cas9 system-mediated knock-in of endogenous GGTA1 via targeted homologous recombination ( HR). Linearized donors with ~800-bp homology flanking the CRISPR/Cas9 target site [exon 4 (containing ATG) of GGTA1] served as a template for gene targeting by HR. Using a targeted toxin strategy to select clones lacking α-gal epitope expression, we successfully obtained several knock-in clones within 3 weeks of initial transfection. These results suggest that the use of CRISPR/Cas9-mediated HR to knock-in a mutated fragment at defined loci represents an efficient strategy to achieve the rapid modulation of genes of interest in swine cells and is a promising tool for the creation of KO piglets. [ABSTRACT FROM AUTHOR]

The purpose of this study was to clarify the characteristics of permanent and primary tooth-crown inclinations. Landmark points from cephalograms and dental casts of two groups; 23 women (mean 20.3 +/- 3.3 years) and 11 girls (mean 5.2 +/- 0.1 years) were digitized, and the coordinates were integrated and transformed to a standardized plane. The 3-dimensional crown inclinations were projected on the sagittal plane, and the angles between the tooth vectors and the FH plane were calculated. An independent-group t-test was used to test for group differences of each tooth inclination, and correlation coefficients were generated for the inclination angles among the permanent and primary teeth. Most maxillary tooth-crown inclinations showed significant age-related differences, while only the second premolar and primary second molar differed significantly in the mandible. The maxillary molars were parallel to the corresponding mandibular molars and correlated with each other, but the primary molars were not. Significant correlations were found between inclinations of most permanent teeth, but not the primary teeth. Maxillary tooth-crown inclinations change during growth, but tooth-crown inclinations of the mandibular teeth do not. [ABSTRACT FROM AUTHOR]

The online measurement of blood gases during rewarming on hypothermic CPB was performed with sensors utilizing optical fluorescence. In 20 patients, blood samples were obtained at 25°C, 30°C, 34°C and 36°C respectively for determination of oxygen content and calculation of VO2 to determine the actual changes in VO2 which occurred and to see if the continuous online measurements of blood oxygen tensions would provide useful information. VO2 decreased to 31.28 ml/min/m2 at 25°C. close to the predicted. With rewarming, VO2 increased to 36% of predicted at 30°C. and at 36°C. was only 67% of predicted. Online measurement of PvO2 provided a reasonably accurate reflection of the changing metabolic requirements during this period, more so than PaO2. These data suggest that the continuous measurement of blood oxygen tensions, particularly PvO2 provides a simple and effective method of monitoring the adequacy of oxygen delivery. [ABSTRACT FROM PUBLISHER]

Cadherin-catenin cell-cell adhesion complexes, composed of cadherin, β-catenin or plakoglobin, and α-catenin (α-cat) molecules, are crucial for maintaining cell-cell contact and are commonly referred to as "adherens junctions (AJs)." Inactivating this system leads to loss of cell-cell contact and developmental arrest in early embryos. However, it remains unclear whether the loss of cell-cell contact affects the differentiation of embryonic cells. In this study, we explored the use of a murine embryonal carcinoma cell line, P19, as an in vitro model for early embryogenesis. P19 cells easily form embryoid bodies (EBs) and are susceptible to cellular differentiation in response to retinoic acid (RA) and teratoma formation. Using CRISPR/Cas9 technology to disrupt the endogenous α-cat gene in P19 cells, we generated α-cat knockout (KO) cells that exhibited a loss of cell-cell contact. When cultivated on non-coated dishes, these α-cat KO cells formed EBs, but their structures were labile. In the RA-containing medium, the α-cat KO EBs failed to produce differentiated cells on their outer layer and continued to express SSEA-1, an antigen specific to pluripotent cells. Teratoma formation assays revealed an absence of overt differentiated cells in tumors derived from α-cat KO P19 cells. Aggregation assays revealed the inability of the KO cells to colonize into the zona pellucida-denuded 8-cell embryos. These findings suggest that the AJs are essential for promoting the early stages of cellular differentiation and for the colonization of early-developing embryos.

Objective: A new motion capture system was developed to verify the hypothesis that "during food intake, head motion changes according to the properties of the food.", Design: Twenty healthy males and 20 healthy females with right-handed and normal occlusion participated in this study. The motion capture system used consisted of a Microsoft Xbox One Kinect Sensor® and a newly-developed program. Meatballs (solid), yogurt (paste), and water (fluid) were used as food samples. Head motion distance, head turning angle, and head forward angle were measured during food intake. Unpaired t-tests were used to analyze each head motion and compare the sexes. A one-way repeated measures analysis of variance was used to analyze each head motion for different food samples., Results: Head motion distance was significantly smaller in females for the meatball and yogurt, but not for water. There were no significant differences between the sexes for head turning angle or head forward angle. Head motion distance and head forward angle were significantly larger for water than for meatballs and yogurt. The head turning angle was significantly smaller for the meatball than for yogurt and water., Conclusions: The results indicated that females tend to consume food without moving their heads when eating solid and paste foods. As the fluidity of the food increased, the head moved in a turning motion to avoid spilling the food, and the heads tilted forward. The motion capture system used in this study was also effective in analyzing head motion during eating., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Germline manipulation at the zygote stage using the CRISPR/Cas9 system has been extensively employed for creating genetically modified animals and maintaining established lines. However, this approach requires a long and laborious task. Recently, many researchers have attempted to overcome these limitations by generating somatic mutations in the adult stage through tail vein injection or local administration of CRISPR reagents, as a new strategy called "in vivo somatic cell genome editing". This approach does not require manipulation of early embryos or strain maintenance, and it can test the results of genome editing in a short period. The newborn is an ideal stage to perform in vivo somatic cell genome editing because it is immune-privileged, easily accessible, and only a small amount of CRISPR reagents is required to achieve somatic cell genome editing throughout the entire body, owing to its small size. In this review, we summarize in vivo genome engineering strategies that have been successfully demonstrated in newborns. We also report successful in vivo genome editing through the neonatal introduction of genome editing reagents into various sites in newborns (as exemplified by intravenous injection via the facial vein), which will be helpful for creating models for genetic diseases or treating many genetic diseases.

Objective: Childhood is an important period for lip-closing strength (LCS) development, and failure to acquire LCS during childhood leads to various adverse health effects, such as mouth breathing. The purpose of this study was to examine the effectiveness of device-free lip and facial training in preschool children., Design: The participants were divided into training and control groups. Both groups comprised 123 children aged 3-4 years, and only the training group received lip and facial training (i.e., opening and closing the lips and protruding the tongue) for 1 year. A two-way repeated measures analysis of variance was applied to compare the interaction effects of LCS and facial linear distance and angle by year (initial year vs. 1 year later) and group (training vs. control group). In addition, paired t-tests were used to test the changes in LCS and facial linear distance and angle after 1 year in both groups. Furthermore, the same analysis was performed in children with weak LCS in both groups (incompetent lip seal [ILS])., Results: The LCS of children in the training group significantly increased after training compared with that in the control group, whether the analysis included all children or children with ILS alone. Lip and facial training for children with ILS reduced both the upper and lower lip protrusion; children with ILS without training had increased lip protrusion after 1 year., Conclusions: Lip and facial training for children with ILS effectively improved LCS and lip morphology, thereby preventing increased lip protrusion., Competing Interests: Declaration of Competing Interest A conflict of interest exists when an author or the author's institution has financial or personal relationships with other people or organisations that inappropriately influence (bias) his or her actions. Financial relationships are easily identifiable, but conflicts can also occur because of personal relationships, academic competition, or intellectual passion. A conflict can be actual or potential, and full disclosure to The Editor is the safest course. Nothing particular., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objectives: To investigate the effects of interventions provided by a multidisciplinary team consisting of anesthesiologists, dentists, pharmacists, and nurses at a Preoperative Clinic (POC) on postoperative outcomes., Methods: We retrospectively investigated patients who underwent preoperative evaluation at the POC at Juntendo University Hospital between May and July, 2019. Patients were divided into intervention and non-intervention groups according to whether they received intervention(s) at the POC or not. Postoperative outcomes were compared between the groups, before and after propensity score (PS) matching., Results: We investigated 909 patients who completed POC evaluation and underwent surgery. Patients in the intervention group (n = 455 [50.1%]) received at least one intervention delivered, in the order of higher delivery frequencies, by dentists, pharmacists, nurses, and anesthesiologists. Before PS matching, the intervention group was associated with older age, more frequent cardiovascular comorbidities, and higher ASA-PS grades than the non-intervention group, while neither frequencies nor severities of postoperative complications differed between the groups. These outcomes did not differ between 382 PS-matched pairs with comparable risk factors either., Conclusions: Before PS matching, postoperative outcomes did not differ between the groups, although the intervention group was associated with higher risks. These suggested that POC interventions could have improved postoperative outcomes in the higher-risk intervention group to the same level as in the non-intervention group. However, such potential beneficial effects of interventions could not be proven after PS matching. Further studies are required to elucidate effects of POC interventions on postoperative outcomes., Competing Interests: The authors declare that there are no conflicts of interest., (2023 ©The Juntendo Medical Society.)

Genome editing, as exemplified by the CRISPR/Cas9 system, has recently been employed to effectively generate genetically modified animals and cells for the purpose of gene function analysis and disease model creation. There are at least four ways to induce genome editing in individuals: the first is to perform genome editing at the early preimplantation stage, such as fertilized eggs (zygotes), for the creation of whole genetically modified animals; the second is at post-implanted stages, as exemplified by the mid-gestational stages (E9 to E15), for targeting specific cell populations through in utero injection of viral vectors carrying genome-editing components or that of nonviral vectors carrying genome-editing components and subsequent in utero electroporation; the third is at the mid-gestational stages, as exemplified by tail-vein injection of genome-editing components into the pregnant females through which the genome-editing components can be transmitted to fetal cells via a placenta-blood barrier; and the last is at the newborn or adult stage, as exemplified by facial or tail-vein injection of genome-editing components. Here, we focus on the second and third approaches and will review the latest techniques for various methods concerning gene editing in developing fetuses.

Objective: This study aims to understand the role of Cancer Philosophy Clinic activities among participants and whether participation is correlated to increase in QOL., Materials and Methods: Among the 150 Cancer Philosophy Clinics, questionnaire surveys were distributed at 28 locations that consented to participating in the study. The data was analyzed based on the respondent's situation and health related Quality of Life (QOL) prior to and after participating in Cancer Philosophy Clinic using the EQ-5D-5L questionnaire (Japanese version) regarding health related QOL prior to and after participating in Cancer Philosophy Clinic., Results: There were more female participants than male participants; 224 and 76 respectively. 46.5%, or approximately half of all participants in the Cancer Philosophy Clinic were "cancer patients," followed by 17.2% who were "family members of cancer patients," 16.6% who were "not suffering from any diseases," 11.4% who were "suffering from diseases other than cancer" and 3.2% who were classified as "other," who were bereaved family members. 51.7% were "currently receiving treatment, "32.1% were "receiving follow-up medical care, "and 15.3% were "survivors." There was 1 participant who commented, "refusing treatment." Based on an evaluation of QOL using EQ-5D-5L of 184 participants who were participating in the Cancer Philosophy Clinic, an increase in overall average index value from 0.827 to 0.867 was observed after participation compared to prior participation. In particular, there was a significant improvement in "pain/discomfort," "anxiety/depression., Conclusions: Cancer Philosophy clinic has been found important role in encouraging existing shift., Competing Interests: The authors declare that they have no conflict of interest., (© 2023 The Juntendo Medical Society.)

Objectives: A set of orofacial signs and symptoms completely or partially present in individuals who replace the correct pattern of nasal breathing with an oral or mixed pattern is defined as mouth breathing syndrome (MBS). In a previous report, it was clarified that an incompetent lip seal (ILS) affected the occurrence of MBS among primary school children. However, the factors related to MBS and the effect of ILS in preschool children remain unclear. The purpose of this study was to clarify the factors relevant to MBS in preschool children and investigate the relationship of ILS to MBS., Material and Methods: We surveyed 285 preschool children between 3 and 5 years of age. Their guardians completed the questionnaire, which consisted of 44 questions regarding the children's daily health conditions and lifestyle habits. To classify the closely related questions into their respective factors and to examine the strength of the correlation between the newly revealed factors, an exploratory factor analysis with promax rotation was performed., Results: The factor analysis identified nine items representing four factors. Factors 1-4 were defined as "diseases of the nose," "ILS," "problem with swallowing and chewing," and "eating and drinking habits," respectively. Factor 2 most strongly correlated with Factor 1, and both Factors showed a relatively strong correlation with Factor 3., Conclusions: The initial stage of MBS may be present in preschool children. ILS and diseases of the nose can cause poor development of oral functions, such as breathing and eating., (© 2022 The Authors. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)