Effects of Oleanolic Acid Derived from Wine Pomace on Periodontopathic Bacterial Growth in Healthy Individuals: A Randomized Placebo-Controlled Study.

Periodontal disease is caused by oral pathogenic bacteria and is associated with systemic disease and frailty. Therefore, its prevention is crucial in extending healthy life expectancy. This study aimed to evaluate the effect of orally administered oleanolic acid, extracted from wine pomace, on periodontopathic bacterial growth in healthy individuals. In this randomized, placebo-controlled, double-blind, parallel-group comparison study, 84 healthy adults were assigned to a placebo (n = 29), low-dose (n = 29, 9 mg oleanolic acid), or high-dose (n = 26, 27 mg oleanolic acid) groups. The number of oral bacteria in their saliva, collected before and 5 h after administration, was determined using the polymerase chain reaction-invader technique. The proportion of periodontopathic bacteria among the total oral bacteria in the saliva was calculated. Oleanolic acid significantly decreased the proportion of Porphyromonas gingivalis among the total oral bacteria in a dose-dependent manner (p = 0.005 (low-dose) and p = 0.003 (high-dose) vs. placebo, Williams' test). Moreover, high-dose oleanolic acid decreased the proportion of Tannerella forsythia (p = 0.064 vs. placebo, Williams' test). Periodontopathic bacteria are closely associated with the development and progression of periodontal disease; thus, the continuous daily intake of oleanolic acid derived from pomace may be helpful in maintaining a healthy oral microbiome by controlling the proportion of periodontopathic bacteria.

Dental Abnormalities in Pediatric Patients Receiving Chemotherapy.

Background: Chemotherapy is a common treatment for pediatric cancer. Although life prognosis is improving because of advances in medical science, it is important to deal with late effects such as dental abnormalities. We investigated the association between dental abnormalities and chemotherapy by age and tooth type. Methods: Among the 568 patients referred to the pediatric dentistry department of our hospital, we selected 32 patients (21 male and 11 female) who received chemotherapy between the ages of 0 and 6 and underwent panoramic examination after the age of 7. We recorded the age of chemotherapy commencement, diagnosis of systemic disease, and dental abnormalities such as congenital absence, microdonts, and short-rooted teeth. Results: Almost half of the patients had dental abnormalities such as congenital absence, microdonts, and short-rooted teeth, but there were no significant differences in the incidence of these abnormalities by age. When we analyzed the incidence of abnormal teeth by tooth type, the incidence of congenital absence was significantly higher in premolars (5.5%) and second molars (3.9%) than in incisor or canine or 1st molar (0.4%) (p < 0.01). The incidence of microdonts was significantly higher in premolars (3.9%) than in incisor or canine or 1st molar (0.2%) and second molars (0.0%) (p < 0.05). Conclusions: Patients who received chemotherapy had a high prevalence of dental abnormalities, and the incidence of abnormalities varied by tooth type. It is important to maintain long-term oral care for patients who have undergone chemotherapy even after the treatment is completed.

Investigation of periodontal disease development and Porphyromonas gulae FimA genotype distribution in small dogs.

In dogs, Porphyromonas gulae is a major periodontal pathogen with 41-kDa proteins polymerizing to form a filamentous structure called fimbriae or pili, termed FimA. FimA is classified into three genotypes: A, B, and C, and there are combinations of types A, B, C, A/B, A/C, B/C, and A/B/C. Periodontal disease is the most common oral disease in small dogs, but the periodontal disease status and P. gulae colonization at each dog age and breed remain unclear. In this study, we stratified 665 small dogs and analyzed the periodontal status and distribution of P. gulae with each FimA genotype. Dogs with periodontal disease and FimA genotype tended to increase with age. The dogs with at least one FimA genotype had significantly more severe periodontal disease compared with P. gulae-negative dogs (P < 0.01). Additionally, periodontal status was significantly associated with specific FimA genotype distribution in Toy Poodles and Chihuahuas (P < 0.05), whereas there was no such association in Dachshunds. These results suggest that the onset of periodontal disease and P. gulae colonization are related and progress with age. The relationship between periodontal disease and FimA genotype may differ depending on the dog breeds.

Inhibitory Effects of Shikonin Dispersion, an Extract of Lithospermum erythrorhizon Encapsulated in ß-1,3-1,6 Glucan, on Streptococcus mutans and Non-Mutans Streptococci.

Shikonin is extracted from the roots of Lithospermum erythrorhizon, and shikonin extracts have been shown to have inhibitory effects on several bacteria. However, shikonin extracts are difficult to formulate because of their poor water solubility. In the present study, we prepared a shikonin dispersion, which was solubilized by the inclusion of ß-1,3-1,6 glucan, and analysed the inhibitory effects of this dispersion on Streptococcus mutans and non-mutans streptococci. The shikonin dispersion showed pronounced anti-S. mutans activity, and inhibited growth of and biofilm formation by this bacterium. The shikonin dispersion also showed antimicrobial and antiproliferative effects against non-mutans streptococci. In addition, a clinical trial was conducted in which 20 subjects were asked to brush their teeth for 1 week using either shikonin dispersion-containing or non-containing toothpaste, respectively. The shikonin-containing toothpaste decreased the number of S. mutans in the oral cavity, while no such effect was observed after the use of the shikonin-free toothpaste. These results suggest that shikonin dispersion has an inhibitory effect on S. mutans and non-mutans streptococci, and toothpaste containing shikonin dispersion may be effective in preventing dental caries.

Inhibitory Effect of Adsorption of Streptococcus mutans onto Scallop-Derived Hydroxyapatite.

Hydroxyapatite adsorbs various substances, but little is known about the effects on oral bacteria of adsorption onto hydroxyapatite derived from scallop shells. In the present study, we analyzed the effects of adsorption of Streptococcus mutans onto scallop-derived hydroxyapatite. When scallop-derived hydroxyapatite was mixed with S. mutans, a high proportion of the bacterial cells adsorbed onto the hydroxyapatite in a time-dependent manner. An RNA sequencing analysis of S. mutans adsorbed onto hydroxyapatite showed that the upregulation of genes resulted in abnormalities in pathways involved in glycogen and histidine metabolism and biosynthesis compared with cells in the absence of hydroxyapatite. S. mutans adsorbed onto hydroxyapatite was not killed, but the growth of the bacteria was inhibited. Electron microscopy showed morphological changes in S. mutans cells adsorbed onto hydroxyapatite. Our results suggest that hydroxyapatite derived from scallop shells showed a high adsorption ability for S. mutans. This hydroxyapatite also caused changes in gene expression related to the metabolic and biosynthetic processes, including the glycogen and histidine of S. mutans, which may result in a morphological change in the surface layer and the inhibition of the growth of the bacteria.

Five-Year Follow-Up of a Child with Non-Syndromic Oligodontia from before the Primary Dentition Stage: A Case Report.

BACKGROUND: Congenital absence of teeth is a major dental abnormality in pediatric dentistry and the absence of six or more teeth is defined as oligodontia. Few reports of patients with non-syndromic oligodontia without systemic disease have continued dental follow-up from an early age. METHODS: We performed the five-year follow-up from before the eruption of the primary dentition of a Japanese child with non-syndromic oligodontia and analyzed changes in dental arch growth. RESULTS: At the oral examination at the age of 1 year and 2 months, eight primary incisors were congenitally absent. Therefore, we made dentures for the patient at the age of 3 years and 4 months. From the age of 5 years and 1 month, the child received articulation training for dysarthria from a speech therapist to improve the function and appearance of the oral cavity. Measurement of the patient's dental models revealed a particularly narrow dental arch, especially between the primary canines. CONCLUSIONS: Our findings highlight the importance of treatment for patients with non-syndromic oligodontia from an early age by multiple medical professionals, recognizing that the missing teeth affect the growth of the maxillofacial region.

A Third Supernumerary Tooth Occurring in the Same Region: A Case Report.

The presence of a supernumerary tooth is one of the most common dental anomalies, and surgical treatment is often required to address this anomaly. Moreover, it may lead to malocclusion, and long-term follow-up is important to monitor its status. A 4-year-and-11-month-old boy was referred to our hospital for dental caries treatment. At 5 years and 5 months of age, a radiographic examination showed a supernumerary tooth (first supernumerary tooth) near the permanent maxillary left central incisor, and it was extracted 6 months later. Eighteen months after the extraction of the first supernumerary tooth, a new supernumerary tooth (second supernumerary tooth) was detected in the same region, which was extracted when the patient was aged seven years and seven months. Seven months later, another supernumerary tooth (third supernumerary tooth) was detected and extracted immediately. However, the permanent maxillary left central incisor did not erupt spontaneously even after 6 months. Therefore, surgical exposure was performed, and the central incisor erupted into the oral cavity. This report describes our experience with this patient with three metachronous supernumerary teeth and their management until the eruption of the permanent tooth. This report highlights the importance of long-term follow-up after supernumerary tooth extraction until the permanent teeth in that region have erupted completely.

Dental radiographic information of term newborn babies within the first month: Analyzing five radiographic cases along with physical attributes in Japan.

BACKGROUND: Although dental radiography is a valuable tool for age estimation in forensic anthropology and odontology, very limited radiological data are available regarding tooth development in healthy newborn babies during the first month of life. AIM: This study aimed to describe the radiological findings of tooth development in babies aged 0 days to 1 month. DESIGN: We analyzed the postmortem findings of five newborn babies with no known natural cause of death who had undergone autopsy, computed tomography (CT), and dental radiography. We estimated the gestational age for the babies aged 0 days and analyzed the condition of mandibular symphysis, existence of tooth germs, and presence or absence of calcification of the first permanent molars of all the babies. RESULTS: The calcified form of 20 deciduous teeth, tooth germs of the permanent upper and lower first molars, and non-calcified mandibular symphysis were observed in each case. However, calcification of the first permanent molar was observed in only two 1-month-old babies. CONCLUSION: The dental radiographic findings and anthropometric measurements of non-skeletonized, non-mummified term babies confirmed calcification of all the deciduous teeth and the first permanent molar at the age of 0 days and 1 month, respectively.

Fusion of a Tooth with a Supernumerary Tooth: A Case Report and Literature Review of 35 Cases.

Tooth fusion is a dental anomaly with a prevalence of 0.1% in permanent dentition. The aim of this paper is to present the occurrence, region of fusion, and prognosis of fused tooth which is a very rare case in the literature. The authors present a very rare case with a maxillary right lateral incisor fused to a supernumerary tooth on the palatal side. The eruption was delayed, but eruption occurred naturally at 9 years and 6 months. However, cone-beam computed tomography at 11 years and 6 months revealed it was diagnosed as a fused tooth. The authors undertook a literature review over the past 10 years and found 30 articles about fused teeth between permanent teeth and supernumerary teeth. It was more common in males than females and there were no differences depending on the occurrence. Mesiodistal fusion was more common in the incisor region, while labiopalatal fusion was more common in the molar region. Most cases required dental treatment with a surgical approach, root canal treatment, or orthodontic treatment. The present study highlights the importance of long-term follow-up and suggests the region of fusion and prognosis of the fused tooth may vary depending on the occurrence.

Characterization of the unique oral microbiome of children with Down syndrome.

Down syndrome creates an abnormal oral environment, including susceptibility to periodontal disease at a young age, but there are no detailed studies of the oral microbiome in children with Down syndrome. In this study, we performed a comprehensive analysis of the oral bacteria of 40 children with Down syndrome and 40 non-Down syndrome children. Microbial DNA was extracted from dental plaque specimens and the V4 hypervariable region of the bacterial 16S rRNA gene was analyzed using the MiSeq platform. There were significant differences between the Down syndrome and non-Down syndrome groups in mean numbers of operational taxonomic units, and α- and ß-diversity (P < 0.05). Interestingly, significant differences in α- and ß-diversity between the two groups were only observed in subjects with gingival inflammation, but not in those without gingival inflammation (P < 0.05). Taxonomic analysis at the genus or species levels showed significant differences in relative abundance levels of certain bacteria between the Down syndrome and non-Down syndrome groups, including Corynebacterium, Abiotrophia and Lautropia (P < 0.05). These results suggest that children with Down syndrome may have a unique oral microbiome that could impact the development of dental diseases common in people with the syndrome.

Analysis of the feasibility of using deep learning for multiclass classification of dental anomalies on panoramic radiographs.

The aim of the feasibility study was to construct deep learning models for the classification of multiple dental anomalies in panoramic radiographs. Panoramic radiographs with single supernumerary teeth and/or odontomas were considered the "case" group; panoramic radiographs with no dental anomalies were considered the "control" group. The dataset comprised 150 panoramic radiographs: 50 each of no dental anomalies, single supernumerary teeth, and odontomas. To classify the panoramic radiographs into case and control categories, we employed AlexNet, which is a convolutional neural network model. AlexNet was able to classify whole panoramic radiographs into two or three classes, according to the presence or absence of supernumerary teeth or odontomas. The performance metrics of the three-class classification were 70%, 70.8%, 70%, and 69.7% for accuracy, precision, sensitivity, and F1 score, respectively, in the macro average. These results support the feasibility of using deep learning to detect multiple dental anomalies in panoramic radiographs.

Three-year survey of oral hygiene conditions of Cambodian public primary school children.

PURPOSE: The purpose of this repeated cross-sectional study was to investigate the prevalence and severity of oral hygiene conditions in Cambodian primary school children. METHODS: Oral examinations were conducted on 2,020 school children (1st-6th grade) at a public primary school in Siem Reap, Cambodia from 2013 to 2015, focusing on plaque adhesion, gingiva, and dental calculus deposition. Data analysis was performed on 1,998 children without any missing data, and the chi-square test was used to compare the variables. RESULTS: The prevalence of dental plaque adhesion in 2013, 2014, and 2015 was 93.6%, 93.7%, and 85.1%, respectively. The prevalence of gingivitis in 2013, 2014, and 2015 was 93.1%, 92.1%, and 88.8%, respectively. The prevalence of dental calculus deposition in 2013, 2014, and 2015 was 55.1%, 19.3%, and 34.7%, respectively. Significant differences were observed in all variables each year (P < 0.001). CONCLUSION: The findings of this study suggest that oral hygiene conditions were poor in this population.

Management of bisphosphonate preparation-treated children in the field of pediatric dentistry.

Since most of the reports of BRONJ onset are adults, in order to clarify the current situation of BRONJ onset in children, it is necessary to search for articles and report on the current status and actual conditions of surgical treatment of children with BP preparations who are being followed up in our clinic. In previous reports both inside and outside Japan, there was no mention of jaw bone necrosis during tooth extraction or surgery in children who were receiving or had a history of BP administration. There were 15 children with a history of BP administration who manage the oral cavity in our clinic. No unpleasant events in the extraction of deciduous teeth were confirmed in medical records. It is necessary to intervene early on oral management of pediatric BP-administered children, especially BP-and steroid-administered children, obtain plaque control to keep the oral cavity cleaner, respond early to infectious diseases, and manage to prevent inflammation from spreading to the jawbone. When surgical treatment is unavoidable, it is important to consider reducing the invasion as much as possible and to cooperate with the medical department such as administration of antibiotics to prevent infection.

Detecting the presence of supernumerary teeth during the early mixed dentition stage using deep learning algorithms: A pilot study.

BACKGROUND: Supernumerary teeth are a common anomaly and are frequently observed in paediatric patients. To prevent or minimize complications, early diagnosis and treatment is ideal in children with supernumerary teeth. AIM: This study aimed to apply convolutional neural network (CNN)-based deep learning to detect the presence of supernumerary teeth in children during the early mixed dentition stage. DESIGN: Three CNN models, AlexNet, VGG16-TL, and InceptionV3-TL, were employed in this study. A total of 220 panoramic radiographs (from children aged 6 years 0 months to 9 years 6 months) including supernumerary teeth (cases, n = 120) or no anomalies (controls, n = 100) were retrospectively analyzed. The CNN performances were assessed according to accuracy, sensitivity, specificity, receiver operating characteristic (ROC) curves, and area under the ROC curves for a cross-validation test dataset. RESULTS: The VGG16-TL model had the highest performance according to accuracy, sensitivity, specificity, and area under the ROC curve, but the other models had similar performance. CONCLUSION: CNN-based deep learning is a promising approach for detecting the presence of supernumerary teeth during the early mixed dentition stage.

Expression and function of variants of human catecholamine transporters lacking the fifth transmembrane region encoded by exon 6.

BACKGROUND: The transporters for dopamine (DAT) and norepinephrine (NET) are members of the Na+- and Cl--dependent neurotransmitter transporter family SLC6. There is a line of evidence that alternative splicing results in several isoforms of neurotransmitter transporters including NET. However, its relevance to the physiology and pathology of the neurotransmitter reuptake system has not been fully elucidated. METHODOLOGY/PRINCIPAL FINDINGS: We found novel isoforms of human DAT and NET produced by alternative splicing in human blood cells (DAT) and placenta (NET), both of which lacked the region encoded by exon 6. RT-PCR analyses showed a difference in expression between the full length (FL) and truncated isoforms in the brain and peripheral tissues, suggesting tissue-specific alternative splicing. Heterologous expression of the FL but not truncated isoforms of DAT and NET in COS-7 cells revealed transport activity. However, immunocytochemistry with confocal microscopy and a cell surface biotinylation assay demonstrated that the truncated as well as FL isoform was expressed at least in part in the plasma membrane at the cell surface, although the truncated DAT was distributed to the cell surface slower than FL DAT. A specific antibody to the C-terminus of DAT labeled the variant but not FL DAT, when cells were not treated with Triton for permeabilization, suggesting the C-terminus of the variant to be located extracellulary. Co-expression of the FL isoform with the truncated isoform in COS-7 cells resulted in a reduced uptake of substrates, indicating a dominant negative effect of the variant. Furthermore, an immunoprecipitation assay revealed physical interaction between the FL and truncated isoforms. CONCLUSIONS/SIGNIFICANCE: The unique expression and function and the proposed membrane topology of the variants suggest the importance of isoforms of catecholamine transporters in monoaminergic signaling in the brain and peripheral tissues.

OspF and OspC1 are Shigella flexneri type III secretion system effectors that are required for postinvasion aspects of virulence.

Shigella flexneri is the causative agent of dysentery, and its pathogenesis is mediated by a type III secretion system (T3SS). S. flexneri secretes effector proteins into the eukaryotic cell via the T3SS, and these proteins usurp host cellular functions to the benefit of the bacteria. OspF and OspC1 are known to be secreted by S. flexneri, but their functions are unknown. We transformed S. flexneri with a plasmid that expresses a two-hemagglutinin tag (2HA) in frame with OspF or OspC1 and verified that these proteins are secreted in a T3SS-dependent manner. Immunofluorescence of HeLa cells infected with S. flexneri expressing OspF-2HA or OspC1-2HA revealed that both proteins localize in the nucleus and cytoplasm of host cells. To elucidate the function of these T3SS effectors, we constructed DeltaospF and DeltaospC1 deletion mutants by allelic exchange. We found that DeltaospF and DeltaospC1 mutants invade host cells and form plaques in confluent monolayers similar to wild-type S. flexneri. However, in the polymorphonuclear (PMN) cell migration assay, a decrease in neutrophil migration was observed for both mutants in comparison to the migration of wild-type bacteria. Moreover, infection of polarized T84 intestinal cells infected with DeltaospF and DeltaospC1 mutants resulted in decreased phosphorylation of extracellular signal-regulated kinase 1/2 in comparison to that of T84 cells infected with wild-type S. flexneri. To date, these are the first examples of T3SS effectors implicated in mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway activation. Ultimately, OspF and OspC1 are essential for PMN transepithelial migration, a phenotype associated with increased inflammation and bacterial access to the submucosa, which are fundamental aspects of S. flexneri pathogenesis.

Role of C-terminal region in the functional regulation of rat serotonin transporter (SERT).

Previously, we revealed that the state of the actin cytoskeleton affects the uptake activity of the serotonin transporter (SERT). Recently, it was reported that the C-terminus of SERT interacts with MacMARCKS, a substrate of PKC that can bind to the actin cytoskeleton. To elucidate the importance of the C-terminal region in the regulation of SERT activity and the interaction with the actin cytoskeleton, we examined whether the overexpression of the C-terminus affects the transport activity of SERT. To this end, we overexpressed a GFP-fused 30-amino acid construct of the SERT C-terminus (GFP-SERT-CT) in HEK293 cells stably expressing FLAG-tagged SERT (FL-SERT-HEK293 cells). The SERT uptake activity and transporter current were attenuated in GFP-SERT-CT-expressing FL-SERT-HEK293 cells, as compared with GFP-expressing FL-SERT-HEK293 cells. Eadie-Hofstee analysis revealed that GFP-SERT-CT overexpression attenuated the SERT uptake activity by reducing the Vmax, but not changing the Km, which was consistent with the results of experiments on the cell-surface expression of SET using biotinylation/immunoblot analysis. Immunocytochemical analysis demonstrated that GFP-SERT-CT was co-localized with FLAG-SERT and cortical actin at the plasma membrane. In addition, the SERT C-terminus did not affect dopamine transporter activity. These findings showed the significance of the C-terminal region to the functional regulation of SERT, suggesting that GFP-SERT-CT acts as a molecular decoy to disrupt the interaction between SERT and the actin cytoskeleton.

Regulation of dopamine and MPP+ transport by catecholamine transporters.

Following exocytotic release of the biogenic amine neurotransmitters, norepinephrine and dopamine, are removed from the synaptic cleft by the respective transporter, norepinephrine transporter (NET) and dopamine transporter (DAT) located on the plasma membrane. The catecholamine transporters are the molecular targets for psychoactive drugs as well as drugs of abuse such as cocaine and amphetamine and the Parkinsonism-inducing neurotoxin, MPP+. Nicotine regulates the transport of catecholamines and MPP+ and may exert self-medicating effects for depression, schizophrenia and attention deficit hyperactivity disorder, and neuroprotective effects against MPP+ through the regulation of the transporters. The availability of cDNAs of these transporters has permitted detailed pharmacological studies in heterologous expression systems for determining the mechanisms of action of nicotine on transporters. Moreover, functional analysis of the effect of single amino acid substitution suggests that specific residues in DAT molecules may play a significant role in interaction with MPP+ and cocaine, and thus would promise a development of novel drugs with diverse chemical structures.