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Gut dysbiosis induces 'leaky gut,' a condition associated with diabetes, NASH, and various auto-immune diseases. Porphyromonas gingivalis is a periodontopathic bacterium which causes periodontal tissue breakdown, and often enters the systemic blood flow. Oral administration of P. gingivalis induced gut dysbiosis in mice model, but no systemic administration of P. gingivalis has been reported thus far. In the present study, we investigated the effect of P. gingivalis-derived lipopolysaccharide (Pg-LPS) on the intestinal flora of our established mouse model. Eight-week-old C57BL/6J mice were intraperitoneally administered Pg-LPS. Three months later, DNA was extracted from stool, and RNA from the small and large intestines. After euthanizing the mice, pathological sections of the intestinal tract were prepared and stained with hematoxylin and eosin (HE). Tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, and IL-6 expression levels were evaluated using quantitative PCR. 16S rRNA gene PCR amplicon analysis data were acquired using NGS. Microbial diversity and composition were analyzed using Quantitative Insights into Microbial Ecology 2. Furthermore, alterations in microbial function were performed by PICRUSt2. No significant inflammatory changes were observed in the HE. No significant differences in the mRNA levels of IL-1β, IL-6, and TNF-α were observed between the groups. Pg-LPS administration decreased the abundance of Allobacterium in the gut. A predictive metagenomic analysis by PICRUSt2 and STAMP showed that 47 pathways increased and 17 pathways decreased after Pg-LPS administration. Systemic application of periodontal pathogens may cause changes in the intestinal flora which may affect the physiological functions of the intestinal tract.

Background/Aim: Alzheimer’s disease is the most common type of neurodegenerative disorder in elderly individuals worldwide. Increasing evidence suggests that periodontal diseases are involved in the pathogenesis of Alzheimer’s disease, and an association between periodontitis and amyloid-β deposition in elderly individuals has been demonstrated. The aim of the present study was to examine the effects of systemic administration of Porphyromonas gingivalis-derived lipopolysaccharide (PG-LPS) on neprilysin expression in the hippocampus of adult and senescence-accelerated mice. Materials and Methods: PG-LPS diluted in saline was intraperitoneally administered to male C57BL/6J and senescence-accelerated mouse prone 8 (SAMP8) mice at a dose of 5 mg/kg every 3 days for 3 months. Both C57BL/6J and SAMP8 mice administered saline without PG-LPS comprised the control group. The mRNA expression levels of neprilysin and interleukin (IL)-10 were evaluated using the quantitative reverse transcriptase-polymerase chain reaction. The protein levels of neprilysin were assessed using western blotting. Sections of the brain tissues were immunohistochemically stained. Results: The serum IL-10 concentration significantly increased in both mouse strains after stimulation with PG-LPS. Neprilysin expression at both mRNA and protein levels was significantly lower in the SAMP8 PG-LPS group than those in the SAMP8 control group; however, they did not differ in PG-LPS-treated or non-treated C57BL/6J mice. Additionally, the immunofluorescence intensity of neprilysin in the CA3 region of the hippocampus in PG-LPS-treated SAMP8 mice was significantly lower than that in control SAMP8 mice. Conclusion: Porphyromonas gingivalis may reduce the expression of neprilysin in elderly individuals and thus increase amyloid-β deposition.

Highlights • MMTAH is a new clinical entity, misdiagnosed as temporomandibular joint disorder. • MMTAH is diagnosed based on a square mandible, cord-like masseter muscle aponeurosis, and limited mouth-opening. • Aponeurectomy combined with coronoidectomy is the treatment of choice for a better prognosis. • It is important to evaluate the patient’s compliance with mouth-opening training before surgery., Introduction Masticatory muscle tendon-aponeurosis hyperplasia (MMTAH) is a new clinical entity that presents mainly with trismus due to hyperplasia of the masseter aponeurosis and temporalis muscle tendon. However, the etiological factors of this disease are unknown; it is often mistreated as temporomandibular joint disorder (TMD). Presentation of case We report a 32-year-old female patient complaining of bilateral pain in her jaw and difficulty opening her mouth. She was first diagnosed as TMD and treated with a splint; however, her symptoms did not improve. Clinical examination revealed a square mandible, tenderness in the left and right temporalis muscles and masseter muscles, and tenderness along the anterior border of the masseter muscle. Her maximum mouth-opening was 30 mm. Short TI inversion recovery magnetic resonance imaging showed areas of low intensity at the anterior border of the masseter muscle and around the coronoid process where the temporalis muscle tendon attaches. Consequently, the diagnosis made based on the clinical and radiographic findings was MMTAH. Bilateral coronoidectomy was performed, followed by a rehabilitation program for six months. The maximum opening was maintained at 48 mm two years after the operation. Discussion MMTAH was treated as type 1 TMD until it was recognized as a new disease at the conference for the Japanese Society for Oral and Maxillofacial Surgeons. Since then, many clinicians have become aware of this particular condition, and different treatment modalities have been proposed. Conclusion Clinicians should consider MMTAH as a differential diagnosis when the patient’s chief complaint is gradually decreasing mouth-opening.

Introduction Vertical maxillary excess, a common orthodontic problem that leads to long faces and open bites, can be repositioned with a Le Fort I osteotomy. However, the Le Fort I osteotomy poses the risk of a variety of complications including descending palatine artery (DPA) injury. Although several Le Fort I osteotomy modifications were reported to avoid complications associated with this osteotomy, only a few of such studies were conducted in Japan, and details remain scarce. Patients and methods We performed a literature review regarding modifications of Le Fort I osteotomies, including Le Fort I with a horseshoe osteotomy, modified horseshoe osteotomy, unilateral horseshoe osteotomy, pyramidal osteotomy, and U-shaped osteotomy. We identified eight relevant studies conducted in Japan; one study did not provide the number of patients examined. The 77 patients (seven studies) with vertical maxillary excess who underwent orthognathic surgery were ≥17 years old. Discussion There were no severe complications after the modified Le Fort I osteotomies. The postoperative maxillary changes obtained by the conventional horseshoe, modified horseshoe, unilateral type of horseshoe, pyramidal, and U-shaped osteotomies were nearly repositioned to the planned position and remained stable for ≥12 months post-surgery. Conclusion Our review indicates that preserving the DPA can lower the incidence of intra- and post-operative complications. Each modification of the Le Fort I osteotomy (i.e., conventional horseshoe, modified horseshoe, unilateral horseshoe, pyramidal, and U-shaped osteotomy) has its respective advantages and indications., Highlights • Vertical maxillary excess (excessive maxilla growth) impairs functions/aesthetics • Vertical maxillary excess is treated with a conventional Le Fort I osteotomy. • Extensive bone removal for the descending palatine artery with impaction is needed. • Various modifications of Le Fort I provide better maxilla repositioning. • Modifications can preserve the DP artery and minimize complications.

Curcumin, a yellow phytochemical found in the rhizomes of Curcuma longa, has various biological effects, including anti-oxidant and anti-inflammatory activities. In the present study, we examined the effect of curcumin on the expression of inflammatory cytokines in human gingival epithelial progenitor cells (HGEPs) stimulated for a prolonged period with lipopolysaccharide (LPS) derived from Porphyromonas gingivalis. The cells were alternately cultured with LPS and/or curcumin every 3 days for 18 days. The expression levels of TNF-α, IL-1β, IL-6, TIMP-1, and MMP-9 in the HGEPs were evaluated by quantitative real-time polymerase chain reaction. Enzyme-linked immunosorbent assay was used to measure the concentrations of these five proteins in the supernatant and nuclear factor (NF)-κB in the nuclear extracts. Curcumin inhibited the mRNA expression levels of TNF-α, IL-1β, IL-6, and MMP-9 in HGEPs treated with curcumin over a prolonged period. Similarly, the expression levels of IL-1β, IL-6, and MMP-9 were decreased in the culture supernatants. NF-κB activity was also inhibited in the cells cultured with curcumin. In conclusion, these findings indicate that curcumin inhibits the expression of inflammatory cytokines and MMP-9 in primary gingival epithelial cells stimulated with P. gingivalis-derived LPS via NF-κB activation.

RNase 7 is a skin-derived antimicrobial peptide expressed in various epithelial tissues. It is upregulated by stimulation with microbes and pro-inflammatory cytokines. Herein, we examined the expression levels of RNase 7 in tissues from normal and inflamed oral epithelia and salivary glands via immunohistochemistry. RNase 7 was expressed mainly in the surface layers of the parakeratinized and orthokeratinized oral epithelium. In addition, it was strongly and weakly expressed in oral lichen planus and radicular cysts, respectively. RNase 7 was constitutively expressed in salivary glands, particularly in the serous and duct cells. In the case of Sjogren's syndrome, RNase 7 was strongly expressed in serous, ductal, and mucous cells in areas with lymphocytic infiltration. The localization patterns of RNase 7 were similar to those of other epithelial antimicrobial peptides, including beta-defensins. Thus, epithelial antimicrobial peptides may act against microbial infections in a coordinated manner in oral epithelia and salivary glands.

Although epidemiological studies have shown a relationship between periodontal disease and pancreatic cancer, the molecular mechanisms involved remain unclear. In this study, the effects of systemic administration of Porphyromonas gingivalis lipopolysaccharide (PG-LPS) on gene expression were comprehensively explored in mouse pancreas that did not demonstrate any signs of inflammation. PG-LPS was prepared in physiological saline and intraperitoneally administered to male mice at a concentration of 5 mg/kg every 3 days for 1 month. After extracting total RNA from the excised mice pancreas, a comprehensive DNA microarray analysis of gene expression was performed. Tissue specimens were also subjected to hematoxylin&ndash, eosin staining and immunohistochemistry using anti-regenerating islet-derived 3A and G (Reg3A/G) antibody. ImageJ software was used to quantify the area of Reg3A/G positive cells in pancreatic islets by binarizing image date followed by area extraction. The results were compared using Mann&ndash, Whitney U test. Data are presented as mean &plusmn, standard deviation (SD) with p &lt, 0.05 considered as significant. Reg3G, a gene related to pancreatic cancer, was one of the 10 genes with the highest levels of expression in the pancreas stimulated with PG-LPS. The comprehensive analysis revealed a 73-fold increase in Reg3G expression level in the PG-LPS group when compared with the control group, in addition, the expression level of Reg3A was increased by 11-fold in the PG-LPS group. Image analysis showed that the ratio of Reg3A/G positive cells was higher in the PG-LPS group than the control. Immunostaining showed the presence of Reg3A/G-positive cells in the alpha-cell equivalent areas around the islets of Langerhans in the PG-LPS group. These results support the notion that periodontal disease may be a risk factor for pancreatic cancer.

Psychological stress is involved in the development of various oral diseases. Alterations in the levels of cytokines in the saliva of patients with stress-related oral diseases have been reported. However, the inconsistencies in the results of these studies might be attributed to differences in the local and systemic factors in the oral cavities of the patients. We examined the effect of chronic stress on three major inflammatory cytokines Interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α in the saliva and salivary glands of chronically stressed mice. Six-week-old C57BL/6 J mice were randomly divided into a control and a stress group. The mice in stress group were exposed to 4 h of stress daily for 10 days and subsequently saliva, as well as the submandibular glands, were collected from both groups. The expression levels of cytokines in the saliva were examined by enzyme-linked immunosorbent assay. The submandibular glands were subjected to histopathological and mRNA expression analyses. IL-1β was significantly elevated in saliva of the chronic stressed mice. Furthermore, the mRNA expression levels of both IL-1β and IL-6 were significantly elevated in the submandibular gland of chronic stressed mice. IL-1β may be a potential salivary biomarker in response to chronic stress in mice.

BackgroundDNA hypermethylation of tumor suppressor genes is observed in precancerous lesions and oral cancer of individuals with the habits of betel quid (BQ) chewing.SIRT1has been identified as playing a role in the maintenance of epithelial integrity, and its alteration is often related to carcinogenesis. However, the methylation and transcription status ofSIRT1in patients with BQ chewing-related oral cancer has not been investigated. We examined the methylation status ofSIRT1in paraffin-embedded tissue samples of oral squamous cell carcinoma (OSCC) obtained from BQ chewing and non-chewing patients and in tissue samples from healthy control subjects. In addition, we examined whether the hypermethylation ofSIRT1followed by its transcriptional downregulation in the human gingival epithelial cells could be caused by arecoline, a major component of BQ. Furthermore, we investigated the methylation status ofSIRT1in smear samples of macroscopically healthy buccal mucosa from subjects with a habit of BQ chewing.ResultsSIRT1was significantly hypermethylated in tissue samples of OSCC from BQ chewers and non-chewers than in oral mucosa from healthy control subjects. Results also showed that the hypermethylation level ofSIRT1was significantly higher in OSCC of patients with BQ chewing habits than in those of non-chewing habits (p< 0.05). Our in vitro model showed that hypermethylation is followed by downregulation of the transcriptional level ofSIRT1(p< 0.05). The methylation levels ofSIRT1in the smear samples obtained from BQ chewing individuals were significantly higher than those in the samples obtained from individuals that did not chew BQ. The duration of BQ chewing habits was correlated positively to the frequency ofSIRT1hypermethylation (p< 0.05).ConclusionsOur results suggest that DNA hypermethylation ofSIRT1is involved in the occurrence of oral cancer in BQ chewing patients and that hypermethylation in the oral mucosa of BQ chewers could be a predictive marker for the occurrence of malignant transformation. This is the first report that showed DNA hypermethylation in clinically healthy oral epithelium of BQ chewers. Our study shows evidence that DNA hypermethylation may be an early event of oral carcinogenesis prior to observable clinical changes.

Background: Betel quid chewing is known as a crucial risk factor for oral diseases such as periodontal diseases, oral cancer, and precancerous lesions in Southeast Asian countries. Although abnormal oral bacterial flora may be linked to betel quid related-oral diseases such as oral cancer, precancerous lesions, and periodontal diseases, little information is available on alterations of their oral flora thus far. To identify these alterations, we analyzed the oral flora in betel quid chewers (BQC) and non-chewers (NC) in Sri Lanka. Methods: Samples obtained from buccal swabs of BQC and NC were analyzed with a next generation sequencer. Data were processed and analyzed using the QIIME software package. Mann–Whitney U test and Permutational multivariate analysis of variance were used for statistical analyses. P values

Adenomatous ductal proliferation/hyperplasia (ADP/H) is a rare hyperplastic condition of the salivary gland. It is mostly associated with other salivary gland pathologies such as tumors and inflammations, and is incidentally found in tissue sections during histopathological examinations of those diseases. Herein, we report a case of ADP/H in the parotid gland not associated with any other pathological lesions, and present a review of the literature on this condition. A 60-year-old Japanese female complained of swelling on the left side of parotid region. Clinical examination revealed a swelling on the lower lobe of the left parotid gland. The lesion was firm but non-tender and was not attached to adjacent structures. A clinical diagnosis of benign salivary gland tumor was reached, and surgical excision was performed under general anesthesia. Histopathological examination revealed an intact parotid gland capsule with isomorphic and basaloid cells within scanty cytoplasm. In addition, an admixture of hyperplasia and proliferation of the intercalated ducts, the presence of zymogen granules, the absence of solid nests, and a peripheral palisaded arrangement of the cells were observed. Based on these findings, a diagnosis of ADP/H was confirmed. ADP/H is a non-tumorous lesion; therefore, tumor involvement should be ruled out before the diagnosis is reached.

Introduction Mucoceles are mucous extravasation phenomena resulting from spontaneous ductal rupture or, less commonly, a traumatic cutting of a salivary excretory duct. Occasionally, the appearance of a mucocele closely resembles a neoplastic lesion, and it can be difficult to diagnose. Presentation of case A 74-year-old Japanese male patient was referred to our clinic with painless swelling related to the upper right canine-premolar area. Preoperative imaging of the lesion revealed that it was fluid-filled and the histopathological biopsy suggested a myxoid tumor. We excised the lesion with the patient under general anesthesia, using safety margins based on the histology of the biopsy specimen to reach the definitive diagnosis and treatment. The final pathological diagnosis was a mucocele in the vicinity with an aberrant small salivary gland. Follow-up visits showed complete healing of the epithelium, and no recurrence has been observed during the subsequent 30 months. Discussion In cases with a difficult definitive diagnosis based on medical history, diagnostic imaging and pathological examination, it is important to proceed carefully with the possibility of a tumor in mind. Conclusion This case emphasizes that some mucous cysts are challenging for clinicians in terms of diagnosis and treatment due to an uncommon presentation., Highlights • Mucoceles are relatively easily diagnosed with a clear clinical picture/history. • Mucocele often occurs on the lower lip due to frequent trauma from biting. • At an unusual site such as gingiva, it is likely to be diagnosed as a neoplasm. • We treated a maxillary gingiva mucocele initially diagnosed as a myxoid tumor.

Secretory carcinoma (SC) of the salivary gland was recently added to the fourth edition of the World Health Organization classification of head and neck tumors. Some salivary tumors, including acinic cell carcinoma, have been reclassified as SC. Most of these tumors are located on the parotid gland with very few cases reported in the minor salivary glands of the buccal mucosa. Herein, we present a case of SC of buccal mucosa, which appeared clinically as a benign lesion in a 54-year-old Japanese female patient. Histopathologically, the tumor cells presented with an eosinophilic cytoplasm with microcytic structure along with eosinophilic secretory material and hemosiderin deposit. Immunohistochemical staining revealed strongly positive staining for S100, vimentin, and mammaglobin and negative staining for DOG-1. The tumor was finally diagnosed as secretory carcinoma of the buccal mucosa. We present a review of the medical literature of SC arising from minor salivary glands. We found only 15 cases of SC of buccal mucosa out of 63 cases of SC in the minor salivary glands. They showed good prognoses and only one case of SC in the buccal mucosa exhibited local recurrence and lymph node metastases.