One-hit kill: On the inactivation of RNA viruses by ultraviolet (UV)-C-induced genomic damage.

Large scale outbreaks of infectious respiratory disease have repeatedly plagued the globe over the last 100 years. The scope and strength of the outbreaks are getting worse as pathogenic RNA viruses are rapidly evolving and highly evasive to vaccines and anti-viral drugs. Germicidal UV-C is considered as a robust agent to disinfect RNA viruses regardless of their evolution. While genomic damage by UV-C has been known to be associated with viral inactivation, the precise relationship between the damage and inactivation remains unsettled as genomic damage has been analyzed in small areas, typically under 0.5 kb. In this study, we assessed genomic damage by the reduced efficiency of reverse transcription of regions of up to 7.2 kb. Our data seem to indicate that genomic damage was directly proportional to the size of the genome, and a single hit of damage was sufficient for inactivation of RNA viruses. The high efficacy of UV-C is already effectively adopted to inactivate airborne RNA viruses.

Chondrosarcoma of the jaw: a retrospective series.

OBJECTIVES: Low-grade chondrosarcoma presents with features similar to those of benign lesions, such as chondroma and synovial chondromatosis, increasing the difficulty in reaching an accurate diagnosis preoperatively. In this study, we retrospectively reviewed 10 chondrosarcoma cases and evaluated the diagnostic approaches and management modalities. STUDY DESIGN: Ten cases were included in the present study. We evaluated the clinical features, initial diagnosis, histopathology subtype, immunohistologic markers, final diagnosis, and treatment modalities. RESULTS: Most of the lesions were found in the mandible. Two cases were followed up for 1 month and 4 years, respectively as benign lesions before malignant changes were detected. With regard to chondrosarcoma histopathology subtypes, 6 cases were identified as conventional chondrosarcoma, whereas 4 cases were diagnosed as mesenchymal chondrosarcoma with aggressive behavior; of these, 3 were associated with local recurrence and metastasis. The immunohistologic markers showed no specificity for chondrosarcoma. CONCLUSIONS: Distinguishing low-grade chondrosarcoma, particularly in the temporomandibular joint, from benign lesions, such as chondroma or synovial chondromatosis, remains difficult. Currently, the correlation between clinical, radiographic, and histologic features accompanied by close follow-up is extremely important for patients diagnosed with chondrogenic lesions. Postoperative radiotherapy seems to be beneficial in patients with positive surgical margins.

Modified Fisher method for unilateral cleft lip-report of cases.

BACKGROUND: Rehabilitation of normal function and form is essential in cleft lip repair. In 2005, Dr. David M. Fisher introduced an innovative method, named "an anatomical subunit approximation technique" in unilateral cleft lip repair. According to this method, circumferential incision along the columella on cleft side of the medial flap is continued to the planned top of the Cupid's bow in straight manner, which runs parallel to the unaffected philtral ridge. Usually, small inlet incision is needed to lengthen the medial flap. On lateral flap, small triangle just above the cutaneous roll is used to prevent unesthetic shortening of upper lip. This allows better continuity of the Cupid's bow and ideal distribution of tension. CASE PRESENTATION: As a modification to original method, orbicularis oris muscle overlapping suture is applied to make the elevated philtral ridge. Concomitant primary rhinoplasty also results in good esthetic outcome with symmetric nostrils and correction of alar web. As satisfactory results were obtained in three incomplete and one complete unilateral cleft lip patients, indicating Fisher's method can be useful in cleft lip surgery with functional and esthetic outcome. CONCLUSIONS: Clinically applied Fisher's method in unilateral cleft lip patients proved the effectiveness in improving the esthetic results with good symmetry. This method also applied with primary rhinoplasty.

TSG-6 secreted by mesenchymal stem cells suppresses immune reactions influenced by BMP-2 through p38 and MEK mitogen-activated protein kinase pathway.

Bone morphogenetic protein 2 (BMP-2) has a critical function in bone and cartilage development and in repairing damaged organs and tissue. However, clinical use of BMP-2 at doses of 0.5-1 mg/ml for orthopedics has been associated with severe postoperative swelling requiring emergency surgical intervention. We determined whether a high concentration of BMP-2 induces inflammatory responses in macrophages and the suppression of osteogenesis in hMSCs. We obtained human periodontal ligament stem cells and bone marrow stem cells from the maxilla, i.e., human mesenchymal stem cells (hMSCs), from the periodontal ligament of extracted third molar teeth and from the bone marrow of the maxilla, respectively. Osteogenic differentiation was measured by alkaline phosphatase activity and alizarin red S staining. Proteins were assessed by flow cytometry, enzyme-linked immunosorbent assay, Western blot and immunocytochemistry. Changes of gene expression were measured by reverse transcription plus the polymerase chain reaction (RT-PCR) and real-time PCR. A high BMP-2 concentration inhibited the early stages of osteogenesis in hMSCs. Co-culturing THP-1 cells (human monocytic cells) with hMSCs reduced the late stages of osteogenesis compared with those seen in hMSCs alone. In addition, high-dose BMP-2 induced the expression of inflammatory cytokines in THP-1 cells and the expression of the anti-inflammatory cytokine tumor-necrosis-factor-α-inducible gene 6 protein (TSG-6) in hMSCs. Consistent with the anti-inflammatory effects of hMSCs when co-cultured with THP-1 cells, interleukin-1ß expression was downregulated by TSG-6 treatment of THP-1 cells. Our findings suggest that a high BMP-2 concentration triggers inflammation that causes inflammatory cytokine release from THP-1 cells, leading to the suppression of osteogenesis, whereas TSG-6 secreted by hMSCs suppresses inflammatory reactions through p38 and ERK in the mitogen-activated protein kinase pathway.

Extensive Surgical Procedures Result in Better Treatment Outcomes for Bisphosphonate-Related Osteonecrosis of the Jaw in Patients With Osteoporosis.

PURPOSE: To identify the risk factors associated with relapse or treatment failure after surgery for bisphosphonate-related osteonecrosis of the jaw (BRONJ) in patients with osteoporosis. PATIENTS AND METHODS: We performed a retrospective cohort study of BRONJ in patients with osteoporosis who had undergone surgical procedures from 2004 to 2016 at the Department of Oral and Maxillofacial Surgery, Seoul National University Dental Hospital. The predictor variables were a set of heterogeneous variables, including demographic (age, gender), anatomic (maxilla or mandible, or both, affected location), clinical (disease stage, etiology, comorbidities, history of intravenous bisphosphonate intake), time (conservative treatment before surgery, bisphosphonate treatment before the development of BRONJ, discontinuation of the drug before surgery, interval to final follow-up, interval to reoperation in the case of relapse or treatment failure), and perioperative variables (type of anesthesia, type of surgical procedures). The primary outcome variable was relapse after surgery that required reoperation (yes vs no). The descriptive and bivariate statistics were computed to assess the relationships between the study variables and the outcome. To determine the risk factors, we conducted a survival analysis using the Cox model. RESULTS: The final sample included 325 subjects with a median age of 75 years, and 97% were women. After surgery, 30% of patients did not completely recuperate and underwent repeat surgery. The interval from the first surgery to reoperation ranged from 10 days to 5.6 years. Relapse or treatment failure most often occurred immediately after surgery. The type of surgical procedure and mode of anesthesia were the most important factors in the treatment outcome. A drug holiday did not appear to influence the likelihood of relapse after surgery. CONCLUSIONS: Treatment of BRONJ in patients with osteoporosis might benefit from more careful and extensive surgical procedures rather than curettage performed with the patient under local anesthesia.

Valproic Acid Modulates the Multipotency in Periodontal Ligament Stem Cells via p53-Mediated Cell Cycle.

Human periodontal ligament stem cells (PDLSCs), a type of mesenchymal stem cell, are a promising source for dental regeneration and are identified in human periodontal ligaments from extracted third molars. Valproic acid (VPA) is a histone deacetylase inhibitor that has been used as a wide-spectrum antiepileptic drug and a medication for mood disorders. VPA has shown several effects on increasing the pluripotency of embryonic stem cells and controlling osteogenic differentiation, besides the prevention of seizures. However, its effect on proliferation and osteogenesis depends on the cell type and concentration. The aim of this study was to investigate the effects of cyclic and constant VPA treatment on PDLSCs. Proliferation and apoptosis of PDLSCs were determined with cyclic and constant VPA treatment. In cemento/osteogenic differentiation, osteogenic markers decreased significantly after cyclic treatment with 0.5 mM VPA. In contrast, VPA enhanced osteogenic differentiation after constant treatment. With cyclic VPA treatment, p53 levels related to apoptotic pathway decreased to induce proliferation. These findings indicated that VPA has different roles in proliferation and differentiation of PDLSCs in vitro and in vivo via p53-related pathway.

A standardized formula for aesthetic mandibular reconstruction using an osteocutaneous fibular free flap.

Ameloblastoma is the most common benign odontogenic tumor of the jaw, and expansional growth of a huge untreated ameloblastoma can result in disturbances in facial aesthetics and function, such as difficulty with mouth opening, swallowing, chewing, breathing, neurologic deficits, and pathologic fractures. Radical wide resection with safety margins and subsequent reconstruction is generally recommended. A fibular free flap (FFF) is commonly used to reconstruct the mandible in order to adequately restore both aesthetic appearance and function. The aim of this brief clinical report is to present a case of huge ameloblastoma after wide resection with free safety margins, and describe the immediate one-step mandibular reconstruction using a vascularized composite FFF. The sterolithographic( rapid prototype, RP) model, a wax pattern of the resected mandible, and a surgical fibular stent made from the wax pattern were constructed preoperatively. We suggest a standardized surgical protocol for mandibular reconstruction with FFF. FUNDING: Supported by the International Research & Development Program of the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2015K1A3A9A01028230).

Neurogenic differentiation of human dental stem cells in vitro.

OBJECTIVES: The purpose of this study was to investigate the neurogenic differentiation of human dental pulp stem cells (DPSCs), periodontal ligament stem cells (PDLSCs), and stem cells from apical papilla (SCAP). MATERIALS AND METHODS: After induction of neurogenic differentiation using commercial differentiation medium, expression levels of neural markers, microtubule-associated protein 2 (MAP2), class III ß-tubulin, and glial fibrillary acidic protein (GFAP) were identified using reverse transcriptase polymerase chain reaction (PCR), real-time PCR, and immunocytochemistry. RESULTS: The induced cells showed neuron-like morphologies, similar to axons, dendrites, and perikaryons, which are composed of neurons in DPSCs, PDLSCs, and SCAP. The mRNA levels of neuronal markers tended to increase in differentiated cells. The expression of MAP2 and ß-tubulin III also increased at the protein level in differentiation groups, even though GFAP was not detected via immunocytochemistry. CONCLUSION: Human dental stem cells including DPSCs, PDLSCs, and SCAP may have neurogenic differentiation capability in vitro. The presented data support the use of human dental stem cells as a possible alternative source of stem cells for therapeutic utility in the treatment of neurological diseases.

Effect of Curculigo orchioides on reflux esophagitis by suppressing proinflammatory cytokines.

This study was performed to investigate effects of Curculigo orchioides rhizome (curculiginis rhizome) on acute reflux esophigitis (RE) in rats that are induced by pylorus and forestomach ligation operation. Proinflammatory cytokine, as well as tumor necrosis factor (TNF)-α, interleukin (IL)-1ß and IL-6 were all assayed and the expression of TNF-α and COX2 analyzed by RT-PCR. The esophagic tissue damage of reflux esophagitis rat was increased compared to that of normal intact group. However, the esophagic damage percentage from the extract of curculiginis rhizoma (ECR) 600 mg/kg and ECR 300 mg/kg were significantly lower than that of the RE control group. Administration of α-tocopherol (30 mg/kg) and ECR (600 mg/kg, 300 mg/kg, and 150 mg/kg) had a significant effect on the gastric acid pH in rats with induced reflux esophagitis (p < 0.05). The treatment with ECR significantly reduced the production of cytokines TNF-α, IL-1ß and IL-6 levels compared to the model group (p < 0.05). The expression of TNF-α and COX2 in the intact esophageal mucosa was low while those of the RE control group were significantly higher due to an inflammatory reaction in the esophagus. Compare to the model group, treatment with α-tocopherol or ECR significantly inhibited the expression levels of COX2 and TNF-α in a dose-dependent manner. These results suggest that anti-inflammatory and protective effects of ECR could attenuate the severity of reflux esophagitis and prevent esophageal mucosal damage.

Real-time analysis of amyloid fibril formation of alpha-synuclein using a fibrillation-state-specific fluorescent probe of JC-1.

alpha-Synuclein is a pathological component of PD (Parkinson's disease) by participating in Lewy body formation. JC-1 (5,5',6,6'-tetrachloro-1,1,3,3'-tetraethylbenzimidazolyl carbocyanine iodide) has been shown to interact with alpha-synuclein at the acidic C-terminal region with a K(d) of 2.6 microM. JC-1 can discriminated between the fibrillation states of alpha-synuclein (monomeric, oligomeric intermediate and fibrillar forms) by emitting the enhanced binding fluorescence of different colours at 590, 560 and 538 nm respectively with the common excitation at 490 nm. The fibrillation-state-specific interaction of JC-1 allowed us to perform real-time analyses of the alpha-synuclein fibrillation in the presence of iron as a fibrillation inducer, rifampicin as a fibrillation inhibitor, baicalein as a defibrillation agent and dequalinium as a protofibril inducer. In addition, various alpha-synuclein fibrils with different morphologies prepared with specific ligands such as metal ions, glutathione, eosin and lipids were monitored with their characteristic JC-1-binding fluorescence spectra. FRET (fluorescence resonance energy transfer) between thioflavin-T and JC-1 was also employed to specifically identify the amyloid fibrils of alpha-synuclein. Taken together, we have introduced JC-1 as a powerful and versatile probe to explore the molecular mechanism of the fibrillation process of alpha-synuclein in vitro. It could be also useful in high-throughput drug screening. The specific alpha-synuclein interaction of JC-1 would therefore contribute to our complete understanding of the molecular aetiology of PD and eventual development of diagnostic/therapeutic strategies for various alpha-synucleinopathies.

Dequalinium-induced cell death of yeast expressing alpha-synuclein-GFP fusion protein.

Intracellular toxic effects of the dequalinium-induced protofibrils of alpha-synuclein have been investigated with the yeast system expressing alpha-synuclein-GFP fusion protein in single copy, which appears in the green halo around the plasma membrane. Intracellular responses of the green fluorescent protein were analyzed as the cells were treated with dequalinium (DQ) and lactacystin. Yeast cells expressing alpha-synuclein-GFP were susceptible to both compounds in alpha-synuclein-dependent manner. Upon DQ treatment, the green halo became smeared throughout the cytoplasm while lactacystin induced a few discrete green dots, reflecting intracellular formation of the protofibrils and the protein inclusions, respectively. The DQ-treated yeast cells were intensely stained with the nucleic acid stains of cell-permeable Hoechst 33342 and cell-impermeable propidium imidione, indicating that nucleus has been disrupted in addition to plasma membrane destabilization. Those DQ-treated yeast cells, however, still contained active mitochondria identified with MitoTracker Red. Therefore, the DQ-induced protofibrillar state of alpha-synuclein-GFP has been suggested to cause the nuclear damage either independently or in combination with the membrane destabilization without affecting mitochondria.