A rumen virosphere with implications of contribution to fermentation and methane production, and endemism in cattle breeds and individuals.

Viruses have a potential to modify the ruminal digestion via infection and cell lysis of prokaryotes, suggesting that viruses are related to animal performance and methane production. This study aimed to elucidate the genome-based diversity of rumen viral communities and the differences in virus structure between individuals and cattle breeds and to understand how viruses influence on the rumen. To these ends, a metagenomic sequencing of virus-like particles in the rumen of 22 Japanese cattle, including Japanese Black (JB, n = 8), Japanese Shorthorn (n = 2), and Japanese Black sires × Holstein dams crossbred steers (F1, n = 12) was conducted. Additionally, the rumen viromes of six JB and six F1 that were fed identical diets and kept in a single barn were compared. A total of 8,232 non-redundant viral genomes (≥5-kb length and ≥50% completeness), including 982 complete genomes, were constructed, and rumen virome exhibited lysogenic signatures. Furthermore, putative hosts of 1,223 viral genomes were predicted using tRNA and clustered regularly interspaced short palindromic repeat (CRISPR)-spacer matching. The genomes included 1 and 10 putative novel complete genomes associated with Fibrobacter and Ruminococcus, respectively, which are the main rumen cellulose-degrading bacteria. Additionally, the hosts of 22 viral genomes, including 2 complete genomes, were predicted as methanogens, such as Methanobrevibacter and Methanomethylophilus. Most rumen viruses were highly rumen and individual specific and related to rumen-specific prokaryotes. Furthermore, the rumen viral community structure was significantly different between JB and F1 steers, indicating that cattle breed is one of the factors influencing the rumen virome composition.IMPORTANCEHere, we investigated the individual and breed differences of the rumen viral community in Japanese cattle. In the process, we reconstructed putative novel complete viral genomes related to rumen fiber-degrading bacteria and methanogen. The finding strongly suggests that rumen viruses contribute to cellulose and hemicellulose digestion and methanogenesis. Notably, this study also found that rumen viruses are highly rumen and individual specific, suggesting that rumen viruses may not be transmitted through environmental exposure. More importantly, we revealed differences of viral communities between JB and F1 cattle, indicating that cattle breed is a factor that influences the establishment of rumen virome. These results suggest the possibility of rumen virus transmission from mother to offspring and its potential to influence beef production traits. These rumen viral genomes and findings provide new insights into the characterizations of the rumen viruses.

Dental Arch Relationship Outcomes Following 2-Stage Palatoplasty for Japanese Patients With Complete Unilateral Cleft Lip and Palate: A 3-Center Study.

OBJECTIVE: To compare dental arch relationship outcomes following 3 different 2-stage palatal repair protocols. DESIGN: Retrospective, cross sectional. SETTING: Three cleft palate centers (A, B, C) in Japan. PATIENTS: Ninety (A: 39, B: 26, C: 25) consecutively treated Japanese patients with complete unilateral cleft lip and palate. INTERVENTIONS: In A, the soft palate and the posterior half of the hard palate were repaired at a mean age of 1 year 7 months. In B, the soft palate and hard palate were closed separately at a mean age of 1 year 6 months and 5 years 8 months, respectively. In C, the soft palate and hard palate were closed at a mean age of 1 year and 1 year 5 months, respectively. MAIN OUTCOME MEASURES: Dental arch relationships were assessed using the 5-Year-Olds' (5-Y) index by 5 raters and the Huddart/Bodenham (HB) index by 2 raters. RESULTS: Intra- and inter-rater reliabilities showed substantial or almost perfect agreement for the 5-Y and HB ratings. No significant differences in mean values and distributions of 5-Y scores were found among the 3 centers. The mean HB index scores of molars on the minor segment were significantly smaller in C than those in A and B (P < .05). CONCLUSIONS: There were no significant differences in dental arch relationships at 5 years among the times and techniques of hard palate closure. However, further analysis of the possible influence of infant cleft size as a covariable on a larger sample size is needed.

A Survey of Orthodontic Treatment in Team Care for Patients With Syndromic Craniosynostosis in Japan.

OBJECTIVE: To understand the actual condition of orthodontic treatment in team care for patients with syndromic craniosynostosis (SCS) in Japan. DESIGN: A nationwide collaborative survey. SETTING: Twenty-four orthodontic clinics in Japan. PATIENTS: A total of 246 patients with SCS. MAIN OUTCOME MEASURE: Treatment history was examined based on orthodontic records using common survey sheets. RESULTS: Most patients first visited the orthodontic clinic in the deciduous or mixed dentition phase. Midface advancement was performed without visiting the orthodontic clinic in about a quarter of the patients, and more than a half of the patients underwent "surgery-first" midface advancement. First-phase orthodontic treatment was carried out in about a half of the patients, and maxillary expansion and protraction were performed. Tooth extraction was required in about two-thirds of patients, and the extraction of maxillary teeth was required in most patients. Tooth abnormalities were found in 37.8% of patients, and abnormalities of maxillary molars were frequently (58.3%) found in patients who had undergone midface surgery below the age of 6 years. CONCLUSIONS: Many patients underwent "surgery-first" midface advancement, and visiting the orthodontic clinic at least before advancement was considered desirable. First-phase orthodontic treatment should be performed considering the burden of care. Midface advancement below the age of 6 years had a high risk of injury to the maxillary molars. This survey is considered useful for improving orthodontic treatment in team care of patients with SCS.

Evaluation of the effectiveness of intermittent mechanical pressure with short loading duration: new type of intermittent force for orthodontic treatment.

Background: Since it is difficult to precisely control the magnitude of force in orthodontic practice, controlling the duration of force is expected to prevent tissue damage. The use of an actuator as an appliance for intermittent force application is a possible solution for controlling the duration. However, effectiveness of an actuator for controlling the duration is still not clear. Aim: To determine whether a short loading duration of intermittent force has the potential to maintain a sound condition of the capillaries and induce dilation of capillaries, which is a requisite phenomenon for orthodontic tooth movement. Materials and methods: Six-week-old male hamsters with a dorsal skinfold chamber attached to an intermittent loading device were used. In three experimental groups, application of intermittent pressure that involved repetition of loading and unloading was performed. The durations of loading/unloading were 1 second/9 seconds (group T10), 1 second/19 seconds (group T20), and 1 second/29 seconds (group T30). Vessels were examined using a fluorescence microscope and a stereomicroscope for 5 days. Results: Widths of capillaries in groups T10 and T20 increased significantly (P < 0.01). In contrast, widths of capillaries in group T30 showed no significant difference from those in the control group. Only group T10 showed bleeding, obvious destruction of vessels, and a significant increase in the rate of disappearance of vessels. Conclusion: In the epidermis, although conditions are limited, a short loading duration of intermittent force maintains a sound condition of the capillaries at the tissue level and induces dilation of capillaries.

Dental arch relationship outcomes in one- and two-stage palatoplasty for Japanese patients with complete unilateral cleft lip and palate.

OBJECTIVE: To compare dental arch relationship outcomes following one- and two-stage palatal repair. DESIGN: Nonrandomized, clinical trial with concurrent control. SETTING: Hokkaido University Hospital. PATIENTS: Sixty-eight consecutively treated Japanese patients with complete unilateral cleft lip and palate. INTERVENTIONS: Thirty-one of the 68 patients underwent two-stage palatoplasty with delayed hard palate closure, and 37 patients underwent one-stage pushback palatoplasty. MAIN OUTCOME MEASURES: Dental casts were taken at 4.9 to 6.3 (mean: 5.2) years of age in the two-stage group and at 4.0 to 6.3 (mean: 5.1) years of age in the one-stage group, and dental arch relationships were assessed using the 5-Year-Olds' Index (5-Y) by four raters and the Huddart/Bodenham Index (HB) by two raters. RESULTS: Intrarater and interrater reliabilities evaluated using weighted kappa statistics were good or better for the 5-Y and HB ratings. The mean 5-Y score was 2.94 in the two-stage group and 3.13 in the one-stage group (P value was not significant). However, there was a significant difference in distributions between the groups (P < .05). The HB scores of molars were significantly greater in the two-stage group than in the one-stage group (P < .05). The rank correlation coefficients between the 5-Y and total HB score (ρ = -0.840, P < .01) and between the 5-Y and the score of the incisors in the HB (ρ = -0.814, P < .01) were significantly increased. CONCLUSIONS: These results suggest that the anteroposterior relationship was not significantly different between the groups, but the transversal relationship was better in the two-stage group than in the one-stage group.

Synthesis and evaluation of fluorescent resins with europium-ß-diketonate complex for orthodontic use.

No effective technique exists for removing adhesive remnants following bracket debonding. We propose that fluorescence imaging using europium ions (Eu3+) offers an effective solution for minimizing iatrogenic enamel damage. This study aimed to assess the impact of different mixing ratios of monomer mixtures on the photoluminescence and flexural properties of a newly developed fluorescent adhesive. Four monomer blends with varying urethane dimethacrylate (UDMA) to triethylene glycol dimethacrylate (TEGDMA) ratios were prepared and polymerized. The blends contained 0.1 wt% of tris(1,3-diphenyl-1,3-propanedionato)(1,10-phenanthroline) Eu(III), [Eu(DBM)3Phen], as the phosphor. Optical measurements and flexural tests were conducted for each resin specimen. The emission spectra exhibited narrow bands corresponding to the 4f-4f transitions of the Eu3+ ions. The photoluminescence properties remained unaffected by the mixing ratio, whereas the mechanical properties tended to improve with higher UDMA content. We conclude that the Eu(DBM)3Phen-containing resin shows promise as a fluorescent orthodontic adhesive that contributes to preserving enamel health.

Dynamic morphometric changes in the mandibular osteocytic lacunae of ovariectomized rats in response to teriparatide, as revealed by three-dimensional fluorescence analyses: Possible involvement of osteocytic perilacunar remodeling.

OBJECTIVES: Teriparatide [TPTD; human parathyroid hormone (hPTH1-34)] is an anti-osteoporotic drug with bone anabolic effects. Clinical and preclinical studies have indicated that TPTD has value in oral and maxillofacial bone therapies, including jawbone regeneration, periodontal tissue repair, and the treatment of medication-related osteonecrosis of the jaw. However, it is unclear whether the craniofacial bones respond to TPTD similarly to the axial and appendicular bones. Recent studies showed that TPTD acts on both osteocytes and osteoblasts. This study aimed to characterize distinct craniofacial bone sites, with a focus on morphometric changes in osteocytic lacunae in ovariectomized rats receiving TPTD. METHODS: Conventional bone histomorphometric analyses of mandibular and parietal bone sections were conducted. High-resolution confocal imaging-based three-dimensional fluorescence morphometric analyses of osteocytic lacunae in distinct mandibular and parietal bone sites were conducted. RESULTS: We observed dynamic changes in the morphometric characteristics of osteocytic lacunae specifically in alveolar and other mandibular bone sites upon TPTD administration. CONCLUSIONS: These findings suggest that osteocytes in mandibular bone (specifically, alveolar bone) have unique functional characteristics of osteocytic perilacunar remodeling.

Congenital and postnatal factors inducing malocclusions in Japanese unilateral cleft lip and palate patients-determination using logistic regression analysis.

Objective : To assess the congenital and postnatal factors that affect degree of malocclusion in patients with unilateral cleft lip and palate using multivariate statistical analysis. Design : Retrospective study. Patients : All information on 135 subjects with unilateral cleft lip and palate was obtained from an oral examination and radiograph at the initial examination at an orthodontic clinic and from surgical records. Plaster models were taken before orthodontic treatment. The ages of the subjects ranged from 5 to 8 years with a mean age of 6.9 years. All primary surgeries for the patients were performed at a university hospital. Main Outcome Measures : The GOSLON Yardstick was used to assess the dental arch relationships (degree of malocclusion) in patients. Family history of Class III, degree of cleft, and congenitally missing upper lateral incisor on the cleft side were chosen as congenital factors inducing malocclusion. Presurgical orthopedic treatment, cheiloplasty, and palatoplasty were chosen as postnatal factors. Associations between various factors and dental arch relationships were assessed using logistic regression analysis. Results : According to adjusted odds ratios, family history of Class III is associated with a significantly worse dental arch relationship. Palatoplasty using push-back alone correlated to a dental arch relationship that was significantly worse than palatoplasty using push-back with a buccal flap. Conclusions : Multivariate analysis shows evidence that a positive family history of Class III and palatoplasty using push-back alone are associated with worse malocclusion of unilateral cleft lip and palate patients.

Development of Autopolymerizing Resin Material with Antimicrobial Properties Using Montmorillonite and Nanoporous Silica.

Although autopolymerizing resin offers numerous applications in orthodontic treatment, plaque tends to accumulate between the appliance and the mucosa, which increases the number of microorganisms present. In this study, we added cetylpyridinium chloride (CPC) loaded montmorillonite (Mont) and nanoporous silica (NPS) to autopolymerizing resin (resin-Mont, resin-NPS) and evaluated their drug release capacity, antimicrobial capacity, drug reuptake capacity, mechanical strength, and color tone for the devolvement of autopolymerizing resin with antimicrobial properties. As observed, resin-Mont and resin-NPS were capable of the sustained release of CPC for 14 d, and a higher amount of CPC was released compared to that of resin-CPC. Additionally, resin-Mont and resin-NPS could reuptake CPC. Moreover, the antimicrobial studies demonstrated that resin-Mont and resin-NPS could release effective amounts of CPC against Streptococcus mutans for 14 d and 7 d after reuptake, respectively. Compared to resin-CPC, resin-Mont exhibited a higher sustained release of CPC in all periods, both in the initial sustained release and after reuptake. However, the mechanical strength decreased with the addition of Mont and NPS, with a 36% reduction observed in flexural strength for resin-Mont and 25% for resin-NPS. The application of these results to the resin portion of the orthodontic appliances can prevent bacterial growth on the surface, as well as on the interior, of the appliances and mitigate the inflammation of the mucosa.

Identification of 146 Metagenome-assembled Genomes from the Rumen Microbiome of Cattle in Japan.

The rumen contains a complex microbial ecosystem that degrades plant materials, such as cellulose and hemicellulose. We herein reconstructed 146 nonredundant, rumen-specific metagenome-assembled genomes (MAGs), with ≥50% completeness and <10% contamination, from cattle in Japan. The majority of MAGs were potentially novel strains, encoding various enzymes related to plant biomass degradation and volatile fatty acid production. The MAGs identified in the present study may be valuable resources to enhance the resolution of future taxonomical and functional studies based on metagenomes and metatranscriptomes.

Association between maternal feeling about pregnancy and child's lifestyle.

The number of children with undesirable lifestyles has recently increased. We tested the hypothesis that maternal feelings about pregnancy might be associated with their attitude towards promoting healthy lifestyles in their children. We used a city database collected from guardians of 204 randomly selected children aged 1 to 3 years in Nihonmatsu City, Fukushima. Maternal feeling about pregnancy was measured using a 10-point scale, and a child lifestyle score was calculated as the sum of desirable lifestyle habits (sleeping, eating, watching TV/videos, and tooth brushing). Associations between maternal feeling and her child's lifestyles were examined with multiple logistic regression analysis. For all lifestyle items, proportion of children with undesirable lifestyle habits was higher in the "unhappy group" (those who scored 1 to 9) than in the "happy group" (those who scored 10). In particular, a child's short sleeping hours (odds ratio [OR]=3.01) and lifestyle score of less than 3 ([OR] =3.60) were significantly associated with unhappy feelings. This was apparent among mothers aged 29 (median age) or younger. Our results indicate an association between a mother's unhappy feelings about pregnancy and her child's undesirable lifestyle, especially among relatively younger mothers. These findings provide public health implications important for early familial intervention to improve children's lifestyles.

A Novel Treatment with Stem Cells from Human Exfoliated Deciduous Teeth for Hypoxic-Ischemic Encephalopathy in Neonatal Rats.

Recently, cell therapy has been developed as a novel treatment for perinatal hypoxic-ischemic encephalopathy (HIE), which is an important cause of neurological disorder and death, and stem cells from human exfoliated deciduous teeth (SHED) express early markers for mesenchymal and neuroectodermal stem cells. We investigated the treatment effect of SHED for HIE in neonatal rats. Seven-day-old rats underwent ligation of the left carotid artery and were exposed to 8% hypoxic treatment. SHED (1 × 105 cells) were injected via the right external jugular vein 24 h after the insult. The effect of intravenous administration of SHED cells was evaluated neurologically and pathophysiologically. In the evaluation of engraftment using quantum dots 655, only a few SHED were detected in the injured cortex. In the immunohistological evaluation 24 h after injection, the numbers of positive cells of active caspase-3 and anti-4 hydroxynonenal antiserum were lower in the SHED group than in the vehicle group. The number of Iba-1+ cells in the cortex was higher in the SHED group. However, the proportion of M1 microglia (Iba-1+/ED-1+) was significantly decreased, whereas M2 microglia (Iba-1+/CD206+) tended to increase in the SHED group. In the behavioral tests performed 5 months after hypoxic treatment, compared to the vehicle group, the SHED group showed significant elongation of the endurance time in the rotarod treadmill test, significantly ameliorated proportion of using the impaired hand in the cylinder test, significantly lower ratio of right/left front paw area in gait analysis, and significantly higher avoidance rate in the active avoidance test. In the in vitro experiment with cultured neurons exposed to oxygen-glucose deprivation, we confirmed the neuroprotective effect of the condition medium of SHED. These results suggested that intravenous administration of SHED exerted a treatment effect both histologically and functionally, possibly via a paracrine effect.

A highly sulfated chondroitin sulfate preparation, CS-E, prevents excitatory amino acid-induced neuronal cell death.

Chondroitin sulfate (CS) is a major microenvironmental molecule in the CNS, and there have been few reports about its neuroprotective activity. As neuronal cell death by excitotoxicity is a crucial phase in many neuronal diseases, we examined the effect of various CS preparations on neuronal cell death induced by the excitotoxicity of glutamate analogs. CS preparations were added to cultured neurons before and after the administration of glutamate analogs. Then, the extents of both neuronal cell death and survival were estimated. Pre-administration of a highly sulfated CS preparation, CS-E, significantly reduced neuronal cell death induced by not only NMDA but also (S)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid or kainate. Neither CS preparations other than CS-E nor other highly sulfated polysaccharides such as heparin and dextran sulfate exerted any neuroprotective effects. NMDA-induced current in neurons was not changed by pre-administration of CS-E, but the pattern of protein-tyrosine phosphorylation was changed. In addition, the elevation of caspase 3 activity was significantly suppressed in CS-E-treated neurons. These results indicate that CS-E prevents neuronal cell death mediated by various glutamate receptors, and suggest that phosphorylation-related intracellular signals and the suppression of caspase 3 activation are implicated in neuroprotection by CS-E.

Evaluation of the maxillofacial morphological characteristics of Apert syndrome infants.

Apert syndrome is a rare craniosynostosis syndrome characterized by irregular craniosynostosis, midface hypoplasia, and syndactyly of the hands and feet. Previous studies analyzed individuals with Apert syndrome and reported some facial and intraoral features caused by severe maxillary hypoplasia. However, these studies were performed by analyzing both individuals who had and those had not received a palate repair surgery, which had a high impact on the maxillary growth and occlusion. To highlight the intrinsic facial and intraoral features of Apert syndrome, five Japanese individuals with Apert syndrome from 5 years and 2 months to 9 years and 10 months without cleft palate were analyzed in this study. A concave profile and a skeletal Class III jaw-base relationship caused by severe maxillary hypoplasia were seen in all patients. The patients exhibited anterior and posterior crossbites possibly due to a small dental arch of Maxilla.

Agenesis of third molar germs depends on sagittal maxillary jaw dimensions in orthodontic patients in Japan.

The aim of this study was to determine the correlation between congenitally missing third molar tooth germs and sagittal maxillomandibular jaw dimensions in orthodontic patients in Japan. The subjects were 391 patients from the orthodontic clinic of the Hokkaido University Dental Hospital who were less than 15 years of age. Assessments were made from panoramic radiographs and lateral cephalograms. The subjects were divided into a maxillary/mandibular third molar absent and an existent group. The ANB angle and the sagittal dimensions of the nasal floor (ANS-PNS), maxillary basal bone (Mx), mandibular corpus (Go-Pog), and mandibular basal bone (Mn) were measured. Logistic regression analysis was used to estimate associations between third molar agenesis and these measures. The following results were obtained: (1) The frequency of the maxillary third molar agenesis significantly increased with decreasing Mx (odds ratio = 0.559, 95% confidence interval = 0.377 - 0.829). The frequency of the mandibular third molar agenesis also increased with decreasing Mx (odds ratio = 0.532, 95% confidence interval = 0.330 - 0.856). (2) There were no significant correlations between Mn and mandibular third molar agenesis. These results suggest that agenesis of third molar germs does not depend on anteroposterior dimensions of the mandible but depends on anteroposterior dimensions of the maxilla in Japanese orthodontic patients.

Clinical Investigation of Congenital Factor Affecting Craniofacial Morphology of Unilateral Cleft Lip and Palate in Japanese Patients

Objective: To evaluate the craniofacial morphology of Japanese patients with unilateral cleft lip and palate and to evaluate the multiple congenital factors that affects craniofacial morphology in unilateral cleft lip and palate patients. Material and Methods: Lateral cephalograms of 140 subjects with unilateral cleft lip and palate were taken before any orthodontic treatment and alveolar bone graft. Subjects mean age was 6.85 ±1.56 years. Primary surgeries performed by surgeons from Hokkaido University Hospital. The craniofacial morphology of the 140 subjects was assessed by angular and linear cephalometric measurements. Gender, side of cleft, complete/incomplete type of cleft, presence/absence of lateral incisor in the affected side, family history of cleft and family history of skeletal Class III was chosen as congenital factors. To compare the assessments using congenital factors affecting craniofacial morphology in the unilateral cleft lip and palate subjects, angular and linear cephalometric measured values from each individual subject (control group) were converted into Z scores in relation to the means and standard deviation of the two parameters. Results: Twelve out of 13 craniofacial morphology outcomes were insignificant in 5 out of 6 dependent variables. Only 1 dependent variable showed 3/13 significant differences. Conclusion: Current study revealed the evidence that there were almost no significant differences in the craniofacial morphology outcome among various congenital factors. This will provide base line information and help determine the effectiveness of such factors.

Treatment Outcome Based on GOSLON Score of Multiple Factors in Unilateral Cleft Lip and Palate Children of Four Different Ethnic

Objective: To evaluate and compare the effect of each congenital and postnatal treatment factors in treatment outcome based on dental arch relationship (DAR) of four different populations at a time using GOSLON yardstick. Material and Methods: 432 unilateral cleft lip and palate subjects (Japanese, Malay ethnic group, Bangladeshi and Pakistani populations) age 5 to 12 years were taken before orthodontic treatment and alveolar bone grafting. The DAR was assessed by GOSLON Yardstick. Independent t-test was performed to compare the GOSLON score for each factors. A multiple comparison (Anova) was also conducted between the GOSLON score of four different populations. The significance level was set at p<0.05. Results: Statistically no significant different was found among the congenital factors. However, the family history of Class III malocclusion showed most likely to associate with, though there was no significant difference (p=0.069). Significant difference revealed between two techniques of cheiloplasty in both Malay ethnic group and Pakistani population (p=0.038 and p=0.000, respectively). Gender and Techniques of palatoplasty also showed significant difference in Pakistani population (p=0.026 and p=0.000, respectively). Japanese and Bangladeshi population showed no significant differences. Also no significant differences found between the GOSLON score and different countries. Conclusion: The treatment outcome based on DAR significantly varies in individual population [Malay ethnic group (cheiloplasty); Pakistan (gender, cheiloplasty and palatoplasty)].

Combination of Tessier clefts 3 and 4: case report of a rare anomaly with 12 years' follow-up.

Tessier cleft types 3 and 4 are both rare craniofacial anomalies. Here we present the first case of a girl born with a combined anomaly of Tessier clefts 3 and 4 with severe bilateral cleft lip, a displaced premaxilla, and three-dimensional underdevelopment of the hard and soft tissues of the maxilla and zygoma. This type of rare facial cleft poses a major operative challenge. Over a period of years, presurgical alveolar molding with an active appliance was followed by seven operations. A satisfactory esthetic outcome was obtained. A multidisciplinary approach to treatment with a plastic surgeon in charge of the operations and an orthodontist in charge of the cleft deformity is essential.