Bacterial DNA and serum IgG antibody titer assays for assessing infection of human-pathogenic and dog-pathogenic Porphyromonas species in dogs.

Periodontal disease is highly prevalent in both humans and dogs. Although there have been reports of cross-infection of periodontopathic bacteria, methods for assessing it have yet to be established. The actual status of cross-infection remains to be seen. The purpose of this study was to evaluate the utility of bacterial DNA and serum immunoglobulin G (IgG) antibody titer assays to assess infection of human-pathogenic and dog-pathogenic Porphyromonas species in dogs. Four experimental beagles were used for establishing methods. Sixty-six companion dogs at veterinary clinics visiting for treatment and prophylaxis of periodontal disease were used and divided into healthy, gingivitis, and periodontitis groups. Periodontal pathogens such as Porphyromonas gingivalis and Porphyromonas gulae were investigated as target bacteria. DNA levels of both bacteria were measured using species-specific primers designed for real-time polymerase chain reaction (PCR). Serum IgG titers of both bacteria were measured by enzyme-linked immunosorbent assay (ELISA). PCR primers were confirmed to have high sensitivity and specificity. However, there was no relationship between the amount of bacterial DNA and the severity of the periodontal disease. In addition, dogs with periodontitis had higher IgG titers against both bacteria compared to dogs in the healthy and gingivitis groups; there was cross-reactivity between the two bacteria. Receiver operating characteristic (ROC) analysis of IgG titers against both bacteria showed high sensitivity (>90 %) and specificity (>75 %). Since both bacteria were distinguished by DNA assays, the combination of these assays may be useful in the evaluation of cross-infection.

BRCA2 Frameshift Mutation in de novo Small-Cell Neuroendocrine Carcinoma of the Prostate: A Case Report.

A 66-year-old male was diagnosed with cT4N0M1b small-cell neuroendocrine carcinoma of the prostate. Four months after the administration of combined androgen blockade, multiple novel metastatic regions in the lung and liver and progression of bone metastasis were observed. The patient was referred to our hospital because of biochemical and radiographic progression after four cycles of docetaxel as a first-line therapy for castration-resistant prostate cancer. Transurethral resection of the prostate and hepatic biopsy revealed small-cell carcinoma with positive expression of neuroendocrine markers. The FoundationOne CDx next-generation sequencing test revealed several pathogenic variants, including BRCA2 (W1692fs*3), KEAP1 (R320W), and TP53 (C2385) mutation. After four cycles of chemotherapy with carboplatin plus etoposide (CE), the metastatic regions regressed markedly. The prostate-specific antigen (PSA) and neuron-specific enolase (NSE) level decreased by 96.9% and 91.6%, respectively. However, 2 months after the completion of four cycles of CE, elevation of tumor marker levels, and re-growth of the metastatic regions were observed. Although olaparib, a poly (ADP-ribose) polymerase inhibitor (PARPi), achieved a 45.2% decrease in NSE, the patient rejected to continue therapy because of G2 adverse events. After receiving an additional two cycles of CE and one cycle of cabazitaxel, the patient died because of cancer progression 24 months after the initial treatment for prostate cancer. Here, we present a case of BRCA2-altered small-cell neuroendocrine prostate cancer treated with both platinum-containing chemotherapy and PARPi. Both therapies achieved an initial response; however, durable responses were not obtained. Additional discussion regarding the optimal treatment strategy for BRCA-altered small-cell/neuroendocrine prostate cancer is required.

Development of the Japanese Parenting Style Scale and examination of its validity and reliability.

Parenting is an essential factor affecting child development. Therefore, several studies have focused on individual differences in parenting (i.e., parenting styles). However, there exist only a few useful scales in Japan, especially for parents who have preschool children. Therefore, a new scale for assessing parenting styles in Japan, based on the traditional theoretical framework, was developed, and examined for its validity and reliability. In Study 1, 82 original items were constructed and 1236 parents with preschool children completed these items. Next, 28 items for the Japanese Parenting Style Scale (JPSS) were selected based on factor analysis and the analyses of the graded response model. The JPSS included four factors: warmth, hostility, permissiveness, and harsh control. The results showed that each sub-scale had sufficient conceptual validity and internal consistency. In Study 2, the criterion-related validity of the JPSS was examined. A total of 1236 parents, non-participants in Study 1, completed the JPSS and other scales. The results showed sufficient criterion-related validity for the scale.

Analysis of subgingival microbiota in monozygotic twins with different severity and progression risk of periodontitis.

The study aims to reveal the composition of subgingival bacteria in monozygotic twins with discordant in severity and progression risk of periodontitis. Microbiome analysis indicated that most bacteria were heritable but differed in their abundance and immune response. The dysbiotic bacteria can be considered as risk markers for periodontitis progression.

Dual-Function Semaphorin 4D Released by Platelets: Suppression of Osteoblastogenesis and Promotion of Osteoclastogenesis.

Effects of the antiosteoblastogenesis factor Semaphorin 4D (Sema4D), expressed by thrombin-activated platelets (TPs), on osteoblastogenesis, as well as osteoclastogenesis, were investigated in vitro. Intact platelets released both Sema4D and IGF-1. However, in response to stimulation with thrombin, platelets upregulated the release of Sema4D, but not IGF-1. Anti-Sema4D-neutralizing monoclonal antibody (mAb) upregulated TP-mediated osteoblastogenesis in MC3T3-E1 osteoblast precursors. MC3T3-E1 cells exposed to TPs induced phosphorylation of Akt and ERK further upregulated by the addition of anti-sema4D-mAb, suggesting the suppressive effects of TP-expressing Sema4D on osteoblastogenesis. On the other hand, TPs promoted RANKL-mediated osteoclastogenesis in the primary culture of bone-marrow-derived mononuclear cells (BMMCs). Among the known three receptors of Sema4D, including Plexin B1, Plexin B2 and CD72, little Plexin B2 was detected, and no Plexin B1 was detected, but a high level of CD72 mRNA was detected in RANKL-stimulated BMMCs by qPCR. Both anti-Sema4D-mAb and anti-CD72-mAb suppressed RANKL-induced osteoclast formation and bone resorptive activity, suggesting that Sema4D released by TPs promotes osteoclastogenesis via ligation to a CD72 receptor. This study demonstrated that Sema4D released by TPs suppresses osteogenic activity and promotes osteoclastogenesis, suggesting the novel property of platelets in bone-remodeling processes.

Management of tracheostomy in COVID-19 patients: The Japanese experience.

OBJECTIVE: Involvement in the tracheostomy procedure for COVID-19 patients can lead to a feeling of fear in medical staff. To address concerns over infection, we gathered and analyzed experiences with tracheostomy in the COVID-19 patient population from all over Japan. METHODS: The data for health-care workers involved in tracheostomies for COVID-19-infected patients were gathered from academic medical centers or their affiliated hospitals from all over Japan. RESULTS: Tracheostomies have been performed in 35 COVID-19 patients with a total of 91 surgeons, 49 anesthesiologists, and 49 surgical staff members involved. Twenty-eight (80%) patients underwent surgery more than 22 days after the development of COVID-19-related symptoms (11: 22-28 days and 17: ≥29 days). Thirty (85.7%) patients underwent surgery ≥ 15 days after intubation (14: 15-21 days, 6: 22-28 days, and 10: ≥29 days). Among the total of 189 health-care workers involved in the tracheostomy procedures, 25 used a powered air-purifying respirator (PAPR) and 164 used a N95 mask and eye protection. As a result, no transmission to staff occurred during the 2 weeks of follow-up after surgery. CONCLUSION: No one involved in tracheostomy procedures were found to have been infected with COVID-19 in this Japanese study. The reason is thought to be that the timing of the surgery was quite late after the infections, and the surgery was performed using appropriate PPE and surgical procedure. The indications for and timing of tracheostomy for severe COVID-19 patients should be decided through multidisciplinary discussion.

Synthesis and Properties of a Cyclohexa-2,7-anthrylene Ethynylene Derivative.

The synthesis of a cyclohexa-2,7-(4,5-diaryl)anthrylene ethynylene (1) was achieved for the first time by using 1,8-diaryl-3,6-diborylanthracene and 1,8-diaryl-3,6-diiodoanthracene as key synthetic intermediates. Macrocycle 1 possesses a planar conformation of approximately D6h symmetry, because of the triple-bond linker between the anthracene units at the 2,7-positions. It was confirmed that macrocycle 1, bearing bulky substituents at the outer peripheral positions, behaves as a monomeric form in solution without π-stacking self-association. Macrocycle 1 has an inner-cavity size that allows specific inclusion of [9]cycloparaphenylene ([9]CPP), but not [8]CPP or [10]CPP, through an aromatic edge-to-face CH-π interaction.

Evaluation of the simulator with automatic irrigation control system designed for countermeasures of internal contamination in dental unit water lines.

The prevention of nosocomial infections is an imperative task. The dental chair unit (DCU) is an indispensable device used in dental treatment. However, it is known that the dental unit water line (DUWL) can become contaminated with biofilm, consisting mainly of heterotrophic bacteria (HB). Recently, the International Organization for Standardization specified the methods for testing DUWL contamination management. On these grounds, a simulator reproducing DUWL was prepared to standardize the examination method of the DUWL contamination. OBJECTIVES: To evaluate the reproducibility of the DUWL simulator, monitor the DUWL contamination states, and test the efficacy of a commercial decontaminant for DUWL. METHODS: The DUWL simulator was assembled by a DCU manufacturing company. The simulator's DUWL was filled with tap water (TW), and left for approximately one year. Neutral electrolyzed water (NEW) was used as a decontaminant for DUWL. Both TW and NEW were passed through DUWL in a timely manner simulating daily dental treatment. Water was sampled from the air turbine hand piece weekly for 4 weeks and used for HB culture. Contamination status was evaluated by measuring bacterial adenosine triphosphate release and by culturing on Reasoner's 2A medium. RESULTS: The DUWL released contaminated water had a bacterial count of over 6 × 104 cfu/mL. After passing NEW through DUWL for 1 week, the count drastically decreased to its basal level and remained steady for 4 weeks. However, TW showed no effect on DUWL decontamination throughout the examination periods. CONCLUSIONS: The DUWL simulator could be useful to examine the efficacy of the decontaminant for DUWL and development of new methods in DUWL contamination management.

Laryngeal suspension, combined with rehabilitation and nutritional support, improved the clinical course of a patient with sarcopenic dysphasia.

INTRODUCTION: Sarcopenic dysphasia is a relatively new disease concept describing impairments in swallowing resulting from a generalized loss of skeletal muscle mass. PRESENTATION OF CASE: In this case report, we describe the clinical history and presentation of a 76-year-old man who developed mild sarcopenic dysphasia following a period of physical inactivity after spinal stenosis surgery, which resulted in a loss of 10 kg of body weight in the 10-month period after surgery. The patient's dysphasia was managed with laryngeal suspension, performed via a minimally invasive thyromandibulopexy, in combination with rehabilitation and nutritional support. After a brief period of postoperative rehabilitation, the patient was able to eat soft meals on postoperative day 14, and a regular meal on postoperative day 18, without aspiration. We include a brief description of our surgical technique in the case report. DISCUSSION: Laryngeal suspension compensated for a decreased functional capacity of the swallowing muscles, with postoperative rehabilitation improving the strength of the swallowing muscles. Fixation of the thyroid cartilage to the mandible compensated for insufficient opening of the esophageal orifice, decreasing the pyriform sinus residue. Drawing of the thyroid cartilage in an anterosuperior position improved the anterosuperior position of the epiglottis, shortening the distance between the epiglottis and the base of the tongue, which narrowed the vallecula space and decreased vallecular residue. CONCLUSION: Based on our experience, laryngeal suspension, via minimally invasive thyromandibulopexy, could be considered to improve the outcomes of sarcopenic dysphagia, with an earlier return to eating normal meals.

Effectiveness and safety of low-concentrated ozonized water for the reduction of contamination in dental unit water lines.

Contamination of dental unit waterlines (DUWL) with heterotrophic bacteria can cause problems in immune compromised patients (aged, tumor and organ transplantation-patients). We focused on the use of low-concentrated ozonized water (OZW) as the biofilm formation restraint system for DUWL. Here, we examined the effects of low-concentrated OZW on the growth of bacteria and related biofilm formation and harmfulness to dental unit components (DUCs) in vitro. OBJECTIVES: To evaluate the bactericidal effects of OZW on biofilms in DUWL and DUC in vitro. METHODS: Low-concentrated OZW (0.4 mg/L) was generated using an OZS-PTDX generator. Heterotrophic bacterial biofilms in old DUWL tubes and Candia albicans solution (control microbe) were treated with OZW for 1 h with gentle agitation before static culturing for 96 h in Reasoner's 2A liquid media. The control solutions were 0.1% cetylpyridinium chloride (CPC), chlorinated tap water (TW), and phosphate-buffered saline (PBS). Adenosine triphosphate (ATP) amounts of the microbes were measured and the biofilms of these microbes were observed using scanning electron microscopy (SEM). Moreover, surfaces of DUC soaked in OZW and TW were observed by SEM. RESULTS: The OZW reduced ATP levels in microbes to 50% compared to TW and PBS treatment, although CPC reduced it below detection limits. SEM observation revealed deformation of microbes cultured with OZW, whereas no changes were seen on DUC surfaces. CONCLUSIONS: Low-concentrated OZW is bactericidal against heterotrophic bacteria biofilms and it is not harmful to DUC, suggesting that it might be useful in preventing DUWL contamination.

Acceleration of bone regeneration of horizontal bone defect in rats using collagen-binding basic fibroblast growth factor combined with collagen scaffolds.

BACKGROUND: Basic fibroblast growth factor (bFGF) has been applied for periodontal regeneration. However, the application depends on bone defect morphology because bFGF diffuses rapidly from defect sites. In a previous study, collagen-binding bFGF (CB-bFGF) has been shown to enhance bone formation by collagen-anchoring in the orthopedic field. The aim of this study is to demonstrate the efficacy of CB-bFGF with collagen scaffolds in bone regeneration of horizontal bone defect. METHODS: Cell proliferation activity and collagen binding activity of CB-bFGF was confirmed by WST-8 assay and collagen binding assay, respectively. The retention of CB-bFGF in the collagen sheet (CS) was measured by fluorescence imaging. The rat horizontal alveolar bone defect model was employed to investigate the efficacy of CB-bFGF with collagen powder (CP). After 4 and 8 weeks, the regenerative efficacy was evaluated by microcomputed tomography, histological, and immunohistochemical analyses. RESULTS: CB-bFGF had a comparable proliferation activity to bFGF and a collagen binding activity. CB-bFGF was retained in CS longer than bFGF. At 8 weeks postoperation, bone volume, bone mineral content, and new bone area in CB-bFGF/CP group were significantly increased compared with those in other groups. Furthermore, epithelial downgrowth was significantly suppressed in CB-bFGF/CP group. At 4 weeks, the numbers of osteocalcin, proliferating cell nuclear antigen, and osteopontin-positive cells at the regeneration site in CB-bFGF/CP group were greater than those in other groups. CONCLUSIONS: CB-bFGF/CP effectively promoted bone regeneration of horizontal bone defect possibly by sustained release of bFGF. The potential of CB-bFGF composite material for improved periodontal regeneration in vertical axis was shown.

Ultrasound-Guided Fine-Needle Aspiration in the Neck Region Using an Optical See-Through Head-Mounted Display: A Randomized Controlled Trial.

OBJECTIVES: The purpose of this study was to examine the feasibility of an optical see-through head-mounted display (OST-HMD) to improve ergonomics during ultrasound-guided fine-needle aspiration (FNA) in the neck region. METHODS: This randomized controlled study compared an OST-HMD with a normal ultrasound monitor during an ultrasound-guided FNA in the neck region. Patients with a neck tumor were recruited and randomized into one of two groups. Two practitioners performed ultrasound-guided FNA with or without the HMD, as indicated. An independent researcher measured the procedure time, the number and time of head movements, as well as the number of needle redirections. In addition, practitioners completed questionnaires after performing the FNA on each patient. RESULTS: In 93% of the sessions with the OST-HMD, practitioners performed ultrasound-guided FNA without turning the patients' heads. There was no difference in procedural time and number of needle redirections between the two groups. Results from the questionnaire revealed not only good wearability and low fatigue, but also the practitioners' preference for the HMD. CONCLUSIONS: The OST-HMD improved the practitioners' ergonomics and can be adopted for performing ultrasound-guided interventional procedures in clinical settings.

Polymorphisms in the UGT1A1 gene predict adverse effects of irinotecan in the treatment of gynecologic cancer in Japanese patients.

Irinotecan is a key chemotherapeutic drug used to treat many tumors, including cervical and ovarian cancers; however, irinotecan can cause toxicity, particularly in the presence of uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1) gene polymorphisms, which are associated with reduced enzyme activity. Here, we investigated the prevalence of three different variants of UGT1A1 (UGT1A1*6, UGT1A1*27 and UGT1A1*28) and their relationships with irinotecan-induced adverse events in patients with gynecologic cancer, who are treated with lower doses of irinotecan than patients with other types of solid tumors. Fifty-three female patients treated with irinotecan and 362 female patients not treated with irinotecan were screened for UGT1A1*6, UGT1A1*27 and UGT1A1*28. Homozygosity for UGT1A1*6 or heterozygosity for UGT1A1*6/*28 was associated with a high risk of severe absolute neutrophil count decrease or diarrhea (odds ratios: 16.03 and 31.33, respectively). In contrast, serum bilirubin levels were not associated with irinotecan toxicity. Homozygosity for UGT1A1*6/*6 and heterozygosity for UGT1A1*6/*28 were associated with an increased risk of absolute neutrophil count and/or diarrhea in Japanese gynecologic cancer patients, despite the lower doses of irinotecan used in these patients. UGT1A1*6 and UGT1A1*28 are potential predictors of severe absolute neutrophil decrease and diarrhea caused by low-dose irinotecan in gynecologic cancer patients.

MicroRNA-196a is a putative diagnostic biomarker and therapeutic target for laryngeal cancer.

BACKGROUND: MicroRNA (miRNA) is an emerging subclass of small non-coding RNAs that regulates gene expression and has a pivotal role for many physiological processes including cancer development. Recent reports revealed the role of miRNAs as ideal biomarkers and therapeutic targets due to their tissue- or disease-specific nature. Head and neck cancer (HNC) is a major cause of cancer-related mortality and morbidity, and laryngeal cancer has the highest incidence in it. However, the molecular mechanisms involved in laryngeal cancer development remain to be known and highly sensitive biomarkers and novel promising therapy is necessary. METHODOLOGY/PRINCIPAL FINDINGS: To explore laryngeal cancer-specific miRNAs, RNA from 5 laryngeal surgical specimens including cancer and non-cancer tissues were hybridized to microarray carrying 723 human miRNAs. The resultant differentially expressed miRNAs were further tested by using quantitative real time PCR (qRT-PCR) on 43 laryngeal tissue samples including cancers, noncancerous counterparts, benign diseases and precancerous dysplasias. Significant expressional differences between matched pairs were reproduced in miR-133b, miR-455-5p, and miR-196a, among which miR-196a being the most promising cancer biomarker as validated by qRT-PCR analyses on additional 84 tissue samples. Deep sequencing analysis revealed both quantitative and qualitative deviation of miR-196a isomiR expression in laryngeal cancer. In situ hybridization confirmed laryngeal cancer-specific expression of miR-196a in both cancer and cancer stroma cells. Finally, inhibition of miR-196a counteracted cancer cell proliferation in both laryngeal cancer-derived cells and mouse xenograft model. CONCLUSIONS/SIGNIFICANCE: Our study provided the possibilities that miR-196a might be very useful in diagnosing and treating laryngeal cancer.

Traditional Chinese Medicine for treatment of laryngeal papillomatosis.

OBJECTIVE: The objective of this study was to assess the clinical effectiveness of Traditional Chinese Medicine (TCM) as an adjuvant therapy for recurrent respiratory papillomatosis. DESIGN: The design of this study was a case series. LOCATION AND SUBJECTS: Fifty-one (51) patients with laryngeal papillomatosis who were treated at Keio University Hospital between May 1981 and April 2008 were incorporated in this study. INTERVENTIONS: Individually formulated TCM was orally administered postsurgically to 20 patients with aggressive laryngeal papillomatosis requiring multiple laser ablations (at least biannually) because of frequent recurrence. Eight (8) patients were excluded because of discontinued visits or medication (6 patients), or malignant transformation of the lesion (2 patients). The remaining 12 patients were enrolled in this study. OUTCOME MEASURES: Retrospective chart review and review of the patients' recorded laryngeal images were performed. Clinical response to TCM was measured by the surgical necessity and Derkay's severity score for each patient, followed by statistical analyses. RESULTS: Surgeries were required statistically less often (p = 0.0029) after TCM administration compared with the pre-TCM period. Furthermore, Derkay's severity score was significantly lower (p = 0.022) at the patients' last visit compared with the score before TCM administration. CONCLUSIONS: TCM may be a useful adjuvant therapy to treat aggressive laryngeal papillomatosis. Further studies are necessary to clarify the pharmacological mechanism of TCM in the treatment of laryngeal papillomatosis.

Injection laryngoplasty with calcium phosphate cement.

OBJECTIVE: To assess the feasibility of injection laryngoplasty with calcium phosphate cement (CPC), which is an injectable paste, self-hardening, and which recrystallizes to calcium hydroxylapatite after injection. STUDY DESIGN: A case series with planned data collection. METHODS: Fifty-six patients with unilateral vocal fold paralysis, who received injection laryngoplasty with CPC between August 2003 and August 2007 with a minimum follow-up period of six months, were enrolled in this study. Volumetric and migration analysis for injected CPC were performed using CT after surgery. Vocal function was also assessed by GRBAS subjective voice assessment scale and maximum phonation time (MPT), acoustic analysis including period perturbation quotient (PPQ), amplitude perturbation period (APQ), and noise-to-harmonics ratio (NHR). RESULTS: No remarkable migration or absorption of injected CPC was observed on CT up to two years after surgery. The average remaining volume of CPC was 87.8% +/- 5.3% two years after injection compared to immediately after injection. Significant improvements in GRBAS scales, MPT, PPQ, APQ, and NHR were observed postoperatively. No adverse effects were observed. CONCLUSIONS: Our clinical experience revealed that CPC was safe, nonabsorbable, and effective. Injection laryngoplasty with CPC may be a useful option in the treatment of glottic insufficiency.

[CHEP with the total removal of the arytenoid on the tumor-bearing side].

A supracricoid laryngectomy with cricohyoidoepiglottopexy (CHEP) consists of the resection of the whole thyroid cartilage and paraglottic space, while preserving the cricoid cartilage, the hyoid bone, most of the epiglottis and the arytenoids. Laryngeal reconstruction is achieved be suturing the cricoid cartilage and the hyoid bone. This procedure is mainly indicated for large T2 glottic carcinomas and provides a complete resection and laryngeal preservation without requiring a permanent tracheostomy. Although bilateral arytenoids are usually preserved to ensure better laryngeal function after CHEP, we unavoidably had to remove the arytenoid on the tumor-bearing side during a complete resection performed in a 56-year-old male with a rT2 tumor who had undergone radiation and demonstrated impaired vocal fold motion. Despite the resection of one arytenoid, the final laryngeal function proved to be satisfactory. CHEP should be utilized as an alternative surgical modality for conventional vertical partial laryngectomies or total laryngectomies. CHEP with the total removal of the arytenoid on the tumor-bearing side may be a useful laryngeal preservation procedure for the treatment of patients with glottic carcinoma associated with an impaired vocal fold motion or a fixed vocal fold.