A highly potent and safe pyrrolopyridine-based allosteric HIV-1 integrase inhibitor targeting host LEDGF/p75-integrase interaction site.

Allosteric integrase inhibitors (ALLINIs) are a class of experimental anti-HIV agents that target the noncatalytic sites of the viral integrase (IN) and interfere with the IN-viral RNA interaction during viral maturation. Here, we report a highly potent and safe pyrrolopyridine-based ALLINI, STP0404, displaying picomolar IC50 in human PBMCs with a >24,000 therapeutic index against HIV-1. X-ray structural and biochemical analyses revealed that STP0404 binds to the host LEDGF/p75 protein binding pocket of the IN dimer, which induces aberrant IN oligomerization and blocks the IN-RNA interaction. Consequently, STP0404 inhibits proper localization of HIV-1 RNA genomes in viral particles during viral maturation. Y99H and A128T mutations at the LEDGF/p75 binding pocket render resistance to STP0404. Extensive in vivo pharmacological and toxicity investigations demonstrate that STP0404 harbors outstanding therapeutic and safety properties. Overall, STP0404 is a potent and first-in-class ALLINI that targets LEDGF/p75 binding site and has advanced to a human trial.

Sports-Related Oral and Maxillofacial Injuries: A 5-Year Retrospective Study, Pusan National University Dental Hospital.

PURPOSE: The incidence of sports trauma is gradually increasing, and its importance is therefore also increasing. The present study was conducted to analyze the types, proportions, and tendencies of sports trauma associated with the oral and maxillofacial regions. PATIENTS AND METHODS: Patients who visited the Emergency Department of Pusan National University Dental Hospital between 2014 and 2018 for more than 5 years were surveyed through retrospective epidemiologic investigations. Type of sports was classified according to American Academy of Pediatrics classification. For statistical analysis, age and gender distribution, cause of trauma, and annual trends data were collected. RESULTS: Of the 517 patients, most of the patients were teenagers (27.9%), followed by those younger than 10 years (23.2%) (χ2 = 22.897; P = .002), and noncontact sports, which is cycling, was the most common (43.5%) cause for trauma in both adult and children groups (χ2 = 91.824; P < .001). The most common sports associated with contact sports, limited-contact sports, and noncontact sports were football (47.7%), baseball (50.0%), and cycling (74.8%), respectively. The causes for trauma injury were as follows: contact sports, other person's body (53.4%); limited-contact sports, other objects (60.9%); and noncontact sports, slip down (77.4%) (χ2 = 298.901; P < .001). The prevalence and incidence of sports injuries increase every year, and the proportion of injured patients in May was the highest (χ2 = 52.360; P = .181). The high percentage of traumatic sports demonstrated statistically significant trends (χ2 = 43.073; P = .002). Kickboard injuries showed a rapid increase recently (P for linear trend = .045), whereas other sports showed no significant trends. CONCLUSIONS: Considering the increasing incidence of sports-related injuries, oral and maxillofacial surgeons should be more concerned with maxillofacial trauma during sports.

Importance in the occurrence rate of shortest buccal bone marrow distance (<1 mm) for sagittal split ramus osteotomy.

BACKGROUND/PURPOSE: The neurosensory disturbance is a common complication following sagittal split ramus osteotomy (SSRO) whereas the shortest buccal bone marrow (SBM) is an important risk factor. The present study aimed to investigate the relationship between the occurrence rates of SBM among three skeletal patterns. METHODS: The cone-beam computed tomography (CBCT) images of 90 participants were divided into skeletal Class I, II, and III. There were six horizontal planes separated apart by a 2 mm interval; it started with plane 0 (original intact mandibular canal) to plane 5 which was 10 mm below. The data of SBM were divided into two groups (SBM ≥ 1 mm and SBM < 1 mm). With an SBM value < 1 mm, we defined a high occurrence rate of postoperative neurosensory abnormality or unfavorable split. RESULTS: The Class III patients had the smallest SBM value (1.31-1.75 mm) whereas the Class II patients had the largest SBM value (1.57-2.09 mm). For the Class III patients, the highest and lowest occurrence rates of SBM were 56.5% and 43.5% respectively. For the Class II patients, the highest and lowest occurrence rates of SBM were 37.1% and 17.7% respectively. The patients with Class III malocclusion had higher occurrence rates of SBM than the patients with Class II malocclusion. CONCLUSION: Class III had a significantly higher occurrence of probability (SBM < 1 mm) than Class II. Therefore, patients with Class III were more likely to experience postoperative neurosensory abnormalities and unfavorable split than patients with Class II.

Comparison of Antiviral Activity of Gemcitabine with 2'-Fluoro-2'-Deoxycytidine and Combination Therapy with Remdesivir against SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the coronavirus disease 2019 (COVID-19) pandemic. The virus still spreads globally through human-to-human transmission. Nevertheless, there are no specific treatments clinically approved. This study aimed to compare antiviral activity of gemcitabine and its analogue 2'-fluoro-2'-deoxycytidine (2FdC) against SARS-CoV-2 as well as cytotoxicity in vitro. Fluorescent image-based antiviral assays revealed that gemcitabine was highly potent, with a 50% effective concentration (EC50) of 1.2 µM, more active than the well-known nucleoside monophosphate remdesivir (EC50 = 35.4 µM). In contrast, 2FdC was marginally active (EC50 = 175.2 µM). For all three compounds, the 50% cytotoxic concentration (CC50) values were over 300 µM toward Vero CCL-81 cells. Western blot and quantitative reverse-transcription polymerase chain reaction analyses verified that gemcitabine blocked viral protein expression in virus-infected cells, not only Vero CCL-81 cells but also Calu-3 human lung epithelial cells in a dose-dependent manner. It was found that gemcitabine has a synergistic effect when combined with remdesivir. This report suggests that the difluoro group of gemcitabine is critical for the antiviral activity and that its combination with other evaluated antiviral drugs, such as remdesivir, could be a desirable option to treat SARS-CoV-2 infection.

Non-thermal plasma accelerates the healing process of peripheral nerve crush injury in rats.

The objective of this study was to evaluate the effect of non-thermal plasma (NTP) on the healing process of peripheral nerve crush injuries, which can occur during dental implant procedures. For this, a rat model of sciatic nerve crush injury (SNCI) was adopted. The rats were divided into three groups: non-nerve damage (non-ND), nerve damage (ND), and ND+NTP group. To evaluate the sciatic nerve (SN) function, the static sciatic index was calculated, and the muscle and SN tissues were subjected to a histologic analysis. The results showed that NTP effectively accelerated the healing process of SNCI in rats. In contrast to the ND group, which showed approximately 60% recovery in the SN function, the NTP-treated rats showed complete recovery. Histologically, the NTP treatments not only accelerated the muscle healing, but also reduced the edema-like phenotype of the damaged SN tissues. In the ND group, the SN tissues had an accumulation of CD68-positive macrophages, partially destroyed axonal fibers and myelinated Schwann cells. Conversely, in the ND+NTP group, the macrophage accumulation was reduced and an overall regeneration of the damaged axon fibers and the myelin sheath was accomplished. The results of this study indicate that NTP can be used for healing of injured peripheral nerves.

Complications Following Orthognathic Surgery for Patients With Cleft Lip/Palate.

INTRODUCTION: Cleft lip/palate is a facial anomaly caused by an abnormal developmental process. It is also the most common congenital anomaly. Orthognathic surgery is required in 25% of patients with cleft lip and palate for the correction of dentofacial deformity. There are various complications that can occur after orthognathic surgery. Complications that can occur during surgery include bleeding, improper fracture, and injuries to the inferior alveolar nerve (IAN) and lingual nerve. Meanwhile, postoperative complications include hemorrhage, edema, pain, infection, and delayed union or nonunion. This study retrospectively examines the complications that occurred after the orthognathic surgery in cleft lip/palate patients at Pusan National University Dental Hospital. PATIENTS AND METHODS: From June 1, 2008 to July 31, 2017, we selected 17 patients who underwent orthognathic surgery for cleft lip/palate at the Department of Oral and Maxillofacial Surgery, Pusan National University Dental Hospital. The patients were treated at different hospitals for all operations related to cleft lip/palate. RESULT: Intraoperative complications include hemorrhage, inadequate fracture, injury to the IAN and lingual nerve, root damage, and fistula. The patients who were evaluated included 2 patients with inadequate fracture, 3 patients with injury to the IAN, and 1 patient with fistula. Postoperative complications (e.g., as damage of the inferior alveolar nerve and velopharyngeal insufficiency) may occur, and all patients recovered during the follow-up period of 6 months or more after the surgery. The relapse rates were A-N per 14.0%, Pog-N per 15.1%, SNA 24.4%, and SNB 4.6%. There was no statistically significant difference in relapse rate. CONCLUSION: Complications that may occur after the orthognathic surgery in the patients with cleft lip/palate are similar to those without cleft lip/palate.

Fabrication of Si Micro Mold via Tribo-Nanolithography Using Micro Polycrystalline Diamond Cantilever Tool.

A direct mechanical nanomachining, tribo-nanolithography (TNL), has been developed to fabricate micro mold on Si substrate in mechanical milling mode. An atomic force microscope (AFM) tip is replaced by a lab-made Si cantilever with a polycrystalline diamond (PCD) tool. Mechanical cutting by PCD tool enables simple fabrication of micro molds with several tens of nanometers depth. Machined cavities of the Si substrate are prepared as a master mold for additional micro molding using polydimethylsiloxane (PDMS). Micro cavities are successfully duplicated in positive patterns similar to soft lithography. Normal loads of cantilever and AFM topographical images are used to discuss micro mold and micro molding characteristics.

Three-Dimensional Analysis of Perioperative Condylar Displacement After Mandibular Setback Surgery With Intended Manual Condylar Positioning.

PURPOSE: The purpose of this study is to investigate the effect of orthognathic surgery with intended manual condylar positioning on condyle by examining a change in its position. METHOD: About 18 patients with mandibular prognathism who underwent orthognathic surgery with intentional manual condyle positioning were included. Condyle displacement was analyzed through 3D cone-beam computed tomography before and after operation. The patients were categorized into 2 experimental groups: group A (1-jaw) and group B (2-jaw). The experimental groups were examined before surgery (T0), 3 days (T1), and 6 months (T2) after surgery. Condylar displacement direction was investigated in terms of bodily shift and rotational movement. RESULTS: Downward bodily shift of condyle after surgery was significantly apparent from all of the patients. Condylar bodily shift in other directions was statistically insignificant. Gross bodily shift of condyle right after surgery was anterolateral-inferior direction. In perspective of rotational movement, condyle rotated in infero-medial direction right after operation, but no significant change was presented afterwards. In addition, no significant difference in the amounts of condylar shift and pattern existed between groups A and B. CONCLUSION: Intended manual condylar positioning may minimize postoperative displacement of condyle while accomplishing skeletal stability.

Evaluation of Intersegmental Displacement After Mandibular Setback Split Ramus Osteotomy Using Modified L-Shaped Monocortical Plate: Cone-Beam Computed Tomography Superimposition.

PURPOSE: The purpose of the current study is to compare intersegmental displacements after mandibular setback sagittal split ramus osteotomy (SSRO) using 4 types of osteosynthesis methods. PATIENTS AND METHODS: This is a retrospective study of 53 subjects who presented underwent bilateral setback SSRO at Pusan National University Hospital from January 2009 to December 2013. The subjects were divided into 4 groups according to the osteosynthesis method applied: group A-modified L-type monocortical plate; B-conventional miniplate; group C-bicortical screws; group D-metal and absorbable screws. To obtain the intersegmental displacement, the mean of the differences of the 3-dimensional from T0 (2 days after surgery) to T1 (6 months after surgery) was calculated for the right and left condylar heads (condylion, Cd) and the right and left coronoid processes (Cps) using 3-dimensional imaging software (Ondemand 3D; Cybermed Co, Seoul, Korea). RESULTS: For the condylion in the x, y, z coordinate system, in group A, there were significant differences in the y-axis for the right and left Cd; in group B, significant differences in the y-axis for the right Cd and in the y- and z-axes for the left Cd; in group C, no significant differences in the axis for the Cd; and in group D, there were significant differences in the y- and z-axes for the right Cd and in the x- and y-axes for the left Cd. For the Cps, the results are not much different from the condylion movement in all group. CONCLUSION: In the current study, group C manifested the greatest displacement for the healing period. Group A did not show the significant difference to group B. In view of these results, modified L-shaped monocortical plate can be applied for osteosynthesis effectively.

The possibility of multi-layer nanofabrication via atomic force microscope-based pulse electrochemical nanopatterning.

Pulse electrochemical nanopatterning, a non-contact scanning probe lithography process using ultrashort voltage pulses, is based primarily on an electrochemical machining process using localized electrochemical oxidation between a sharp tool tip and the sample surface. In this study, nanoscale oxide patterns were formed on silicon Si (100) wafer surfaces via electrochemical surface nanopatterning, by supplying external pulsed currents through non-contact atomic force microscopy. Nanoscale oxide width and height were controlled by modulating the applied pulse duration. Additionally, protruding nanoscale oxides were removed completely by simple chemical etching, showing a depressed pattern on the sample substrate surface. Nanoscale two-dimensional oxides, prepared by a localized electrochemical reaction, can be defined easily by controlling physical and electrical variables, before proceeding further to a layer-by-layer nanofabrication process.

Selective Killing of Melanoma Cells With Non-Thermal Atmospheric Pressure Plasma and p-FAK Antibody Conjugated Gold Nanoparticles.

Melanomas are fast growing high-mortality tumors, and specific treatments for melanomas are needed. Melanoma cells overexpress focal adhesion kinase (FAK) compared to normal keratinocytes, and we sought to exploit this difference to create a selectively lethal therapy. We combined gold nanoparticles (GNP) with antibodies targeting phosphorylated FAK (p-FAK). These conjugates (p-FAK-GNP) entered G361 melanoma cells and bound p-FAK. Treatment with p-FAK-GNP decreased the viability of G361 cells in a time dependent manner by inducing apoptosis. To maximize the preferential killing of G361 cells, non-thermal atmospheric pressure plasma was used to stimulate the GNP within p-FAK-GNP. Combined treatment with plasma and p-FAK-GNP showed much higher lethality against G361 cells than HaCaT keratinocyte cells. The p-FAK-GNP induced apoptosis over 48 hours in G361 cells, whereas plasma and p-FAK-GNP killed G361 cells immediately. This study demonstrates that combining plasma with p-FAK-GNP results in selective lethality against human melanoma cells.

FOXO1 Is Involved in the Effects of Cigarette Smoke Extract on Osteoblastic Differentiation of Cultured Human Periosteum-derived Cells.

Cigarette smoke is associated with delayed fracture healing, alterations in mineral content, and osteoporosis, however, its effects on osteoblastic differentiation of osteoprogenitor cells are not fully understood. In the present study, we examined the effects of cigarette smoke extract (CSE) on osteoblastic differentiation of cultured human periosteum-derived cells. We found that CSE inhibited alkaline phosphatase (ALP) activity, mineralization and Runx2 transactivation of the periosteum-derived cells. Nucleofection of RUNX2 into the periosteum-derived cells increased expression of endogenous osteocalcin (OC) and ALP genes in osteogenic induction medium and increased OC expression in non-osteogenic medium. Treatment of the periosteum-derived cells with CSE resulted in decreased phosphorylation of AKT and forkhead box protein O1 (FOXO1). The AKT phosphorylation-resistant mutant, FOXO1-A3, inhibited transcriptional activity of RUNX2 in the periosteum-derived cells. The current study suggests one mechanism by which CSE exposure leads to inhibition of osteoblastic differentiation of cultured human periosteum-derived cells.

Stability of simultaneously placed dental implants with autologous bone grafts harvested from the iliac crest or intraoral jaw bone.

BACKGROUND: Jaw bone and iliac bone are the most frequently used autologous bone sources for dental implant placement in patients with atrophic alveolar ridges. However, the comparative long-term stability of these two autologous bone grafts have not yet been investigated. The aim of this study was to compare the stability of simultaneously placed dental implants with autologous bone grafts harvested from either the iliac crest or the intraoral jaw bone for severely atrophic alveolar ridges. METHODS: In total, 36 patients (21 men and 15 women) were selected and a retrospective medical record review was performed. We compared the residual increased bone height of the grafted bone, peri-implantitis incidence, radiological density in newly generated bones (HU values), and implant stability using resonance frequency analysis (ISQ values) between the two autologous bone graft groups. RESULTS: Both autologous bone graft groups (iliac bone and jaw bone) showed favorable clinical results, with similar long-term implant stability and overall implant survival rates. However, the grafted iliac bone exhibited more prompt vertical loss than the jaw bone, in particular, the largest vertical bone reduction was observed within 6 months after the bone graft. In contrast, the jaw bone graft group exhibited a slower vertical bone resorption rate and a lower incidence of peri-implantitis during long-term follow-up than the iliac bone graft group. CONCLUSIONS: These findings demonstrate that simultaneous dental implantation with the autologous intraoral jaw bone graft method may be reliable for the reconstruction of edentulous atrophic alveolar ridges.

Positive expression of NANOG, mutant p53, and CD44 is directly associated with clinicopathological features and poor prognosis of oral squamous cell carcinoma.

BACKGROUND: In order to predict long-term prognosis and define individual treatment modalities for patients with oral squamous cell carcinoma (OSCC), more reliable tumor biomarkers are needed during the pretreatment workup period. The present study aimed to identify more reliable immunohistochemical tumor prognostic markers in the pretreatment biopsy specimens of patients with OSCC. METHODS: We selected 57 patients who were diagnosed with primary OSCC through histopathological analysis. Pretreatment biopsy specimens were immunohistochemically analyzed for the transcription factor NANOG, cancer stem cell marker CD44, and mutant tumor protein 53 (mutant p53). The immunostaining patterns were assessed for their association with the clinicopathological features of OSCC and overall survival rates. RESULTS: Late tumor stage, positive neck node metastasis, and high-grade differentiation were associated with significantly poorer survival rates. Enhanced expression of NANOG and mutant p53 positivity were significantly associated with clinically late-stage tumors, positive neck node metastasis, histologically high-grade tumors, and poor overall survival rates. OSCCs with strong co-detection of NANOG and mutant p53 were linked to significantly lower survival rates than those with both weak NANOG expression and p53 negativity. Increased expression of CD44 had a limited correlation with unfavorable clinicopathological features. CONCLUSION: High expression of NANOG and positive expression of mutant p53 in the pretreatment biopsy specimens of patients with OSCC were associated with poor survival rates and unfavorable clinicopathological features. These results demonstrate that NANOG, mutant p53, and CD44 could be used as immunohistochemical markers in the pretreatment specimens of OSCC. In particular, analysis for co-expression of NANOG and mutant p53 should be made highly available as a tool for prognosis and selecting individual treatment modalities.

Correlation between genotype and phenotype in patients with bi-allelic SLC26A4 mutations.

Mutation of SLC26A4 is the most common cause of prelingual hearing loss in East Asia. Patients with SLC26A4 mutations have variable phenotypes ranging from non-syndromic hearing loss to Pendred syndrome. Here, we analyzed the correlation between genotype and various inner ear phenotypes and found a possible underlying mechanism. This study included 111 patients with bi-allelic SLC26A4 mutations who had bilateral enlarged vestibular aqueduct (EVA) and hearing loss. p.H723R (61%), c.919-2A>G (24%), and p.T410M (4%) were the most common mutations in Korean patients with EVAs. Residual hearing in patients with c.919-2A>G or p.T410M mutations was better than that of patients with p.H723R homozygous mutations. Interestingly, quantitative polymerase chain reaction showed normal pendrin transcript (6-17% of normal levels) was produced from patients with c.919-2A>G homozygous mutations. Surface expression ratio of pendrin and residual anion exchange activity were higher in cells transfected with p.T410M in comparison to cells transfected with p.H723R. These results suggest that there is a correlation between degree of residual hearing and the SLC26A4 genotype commonly found in the East Asian population.

Transient ipsilateral mydriasis during correction of left blowout fracture.

Mydriasis, either bilateral or unilateral, seldom occurs during reconstruction of periorbital fracture. Anisocoria, a unilateral mydriasis, requires more urgent assessment than bilateral mydriasis does. Pharmacologic agents, local anesthetic infiltration, as well as direct or indirect oculomotor nerve damage are possible causes of unilateral mydriasis. Few cases have been reported about intraoperative temporary ipsilateral mydriasis during correction of blowout fracture. We have experienced an unusual case of anisocoria and report the case with literature reviews.

Retrospective analysis on prognosis of oral cancer patients according to surgical approaches for effective cancer ablation: swing approach versus visor approach.

BACKGROUND: For the surgical treatment of oral cancer, it is sometimes necessary to expand intraoral access within the oral cavity. The "swing approach" that involves lip splitting of the mandible and temporary mandibular osteotomy and the "visor approach" that does not split the lower lip and mandible are mainly used. This study analyzed postoperative outcomes such as complications, recurrence rate, and survival rate by these two approaches. The goal of this study is to evaluate the surgical outcomes of patients using these two approaches, to propose effective perioperative management for oral cancer surgery, and to compare the prognosis of oral cancer patients. MATERIALS AND METHODS: From 2005 to 2020, 29 patients who underwent surgery at the Department of Oral and Maxillofacial Surgery of Pusan National University Dental Hospital for oral cancer lesions occurred in the mandible, floor of mouth, and tongue were selected for the study. Based on the surgical approach used, a chart review was conducted on various prognostic clinical factors such as the patients' sex and age, primary site, TNM stage, histopathologic grade, recurrence and metastasis, postoperative survival rate, adjuvant chemo-radiation therapy, satisfaction with aesthetics/function/swallowing, length of hospital stay, tracheostomy and its duration, and neck dissection and its type. Statistical analysis was conducted using SPSS 25.0 (SPSS Inc., Chicago, IL) through Fisher's exact t-test. RESULT: There was no statistically significant difference between two groups in terms of clinical and pathological findings, such as survival rate, the need for adjuvant therapies, and the local recurrence rate. Although better outcomes were observed in terms of function, aesthetics, and postoperative complications in the group with visor approach, there was still no statistically significant difference between two groups. However, the duration of hospital stay was shorter in the visor approach group. CONCLUSION: There was no statistically significant difference in clinical prognostic factors between the swing approach and the visor approach. Therefore, when choosing between the two approaches for the ablation of oral cancer, it is considered to select the surgical priority approach that can be easy access based on the size and location of the lesion. The visor approach had advantages of aesthetics and healing period.

Efficacy of tracheostomy for respiratory management in patients with advanced oral cancer.

BACKGROUND: Many studies have been reported on tracheostomy to prevent upper airway obstruction after surgery. Among these, the scoring system proposed by Cameron et al. quantifies various factors that influence postoperative respiratory failure. This system provides a basis for surgeons to decide whether to perform an elective tracheostomy. In this study, the authors applied the Cameron scoring system retrospectively to patients undergoing severe oral cancer surgery to reevaluate the indications for elective tracheostomy and to investigate its clinical efficacy in airway management. In this study, a sample of 20 patients who underwent oral cancer surgery was selected and divided into two groups: 10 underwent tracheostomy and 10 did not. The Cameron scoring scores for each patient were extracted, to verify whether elective tracheostomy was performed in accordance with the threshold scores. Differences in scores and significant clinical impact factors between the two groups were analyzed and compared. RESULT: The 10 patients who underwent tracheostomy had an average Cameron score of 6.4, all scoring above the recommended threshold of 5 for tracheostomy. For the 10 patients who did not undergo tracheostomy, the average score was 2.5, with 8 out of these 10 patients scoring below 5. Significant clinical impact factors observed included the location and size of the tumor, the performance of mandibulectomy and neck dissection, and the type of reconstruction surgery. CONCLUSION: In planning surgery for oral cancer patients, it is essential to consider the use of elective tracheostomy based on preoperative assessment of the risk of postoperative airway obstruction using tools like the Cameron scoring system, and patients' condition. Research confirms that elective tracheostomy effectively enhances airway management in patients with severe oral cancer.

JNK signaling plays an important role in the effects of TNF-α and IL-1ß on in vitro osteoblastic differentiation of cultured human periosteal-derived cells.

The purpose of this study was to examine the effects of TNF-α and IL-1ß on in vitro osteoblastic differentiation of cultured human periosteal-derived cells. To examine the effects of TNF-α and IL-1ß on in vitro osteoblastic differentiation of cultured human periosteal-derived cells, the cells cultured in the osteogenic induction medium were treated with 0.1-10 ng/ml TNF-α and 0.01-1 ng/ml IL-1ß. TNF-α and IL-1ß enhanced the alkaline phosphatase (ALP) activity and alizarin red S staining in cultured human periosteal-derived cells. However, these cytokines did not stimulate the Runt-related transcription factor (Runx) 2 activity and osteocalcin secretion. The ALP activity was decreased in the periosteal-derived cells pretreated with mitogen activated protein kinase (MAPK) inhibitors and then treated with TNF-α or IL-1ß. Among the periosteal-derived cells pretreated with MAPK inhibitors, the ALP activity was markedly decreased in the cells pretreated with SP 600125, the specific inhibitor of C-Jun N-terminal kinase (JNK). The periosteal-derived cells treated with TNF-α and IL-1ß showed an increase in extracellular signal-regulated kinase (ERK) and JNK phosphorylation. Among the ERK and JNK phosphorylation, JNK phosphorylation was strongly observed in the cells. These results suggest that TNF-α and IL-1ß increased the in vitro osteoblastic differentiation of cultured human periosteal-derived cells by enhancing the ALP activity and mineralization process, but not by Runx2 activation. The functional role of TNF-α and IL-1ß in increasing the ALP activity and mineralization of periosteal-derived cells primarily depends on the JNK signaling among the MAPK pathways.

The triazole fungicide metconazole inhibits the homodimerization of human androgen receptors to suppress androgen-induced transcriptional activation.

We assessed the mechanism of human androgen receptor-mediated endocrine-disrupting effect by a triazole fungicide, metconazole in this study. The internationally validated stably transfected transactivation (STTA) in vitro assay, which was established for determination of a human androgen receptor (AR) agonist/antagonist by using 22Rv1/MMTV_GR-KO cell line, alongside an in vitro reporter-gene assay to confirm AR homodimerization was used. The STTA in vitro assay results showed that metconazole is a true AR antagonist. Furthermore, the results from the in vitro reporter-gene assay and western blotting showed that metconazole blocks the nuclear transfer of cytoplasmic AR proteins by suppressing the homodimerization of AR. These results suggest that metconazole can be considered to have an AR-mediated endocrine-disrupting effect. Additionally, the evidence from this study might help identify the endocrine-disrupting mechanism of triazole fungicides containing a phenyl ring.