COVID-19 transmission in dental and oral/maxillofacial surgical practice during pandemic: questionnaire survey in 51 university hospitals in Japan.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic has become a major public health problem. Dental procedures that generate aerosols are considered to impose a high risk of infection; therefore, dental professionals, such as dentists and dental hygienists, may be at high risk of viral transmission. However, few studies have reported COVID-19 clusters in dental care settings. AIM: To investigate whether dental and oral/maxillofacial procedures are associated with the occurrence of COVID-19 clusters and measures taken to prevent nosocomial infection in dental clinics. METHODS: An online questionnaire survey on clinical activities (administrative control), infection control measures (environmental/engineering control, personal protective equipment, etc.), and confirmed or probable COVID-19 cases among patients and clinical staff was administered to the faculties of the dental and oral/maxillofacial surgical departments of university hospitals. FINDINGS: Fifty-one faculty members completed the questionnaire. All members were engaged in the treatment of dental and oral surgical outpatients and actively implemented standard precautions. Fourteen faculty members treated patients with COVID-19, but no infections transmitted from the patients to the medical staff were observed. In seven facilities, patients were found to have the infection after treatment (medical staff came in close contact), but there was no transmission from patients to medical staff. Four facilities had medical staff with infections, but none of them exhibited disease transmission from staff to patients. CONCLUSION: COVID-19 clusters are unlikely to occur in dental and oral surgical care settings if appropriate protective measures are implemented.

Efficacy and Safety of Biosimilar Filgrastim for Cord Blood Transplantation: A Single-Institution Retrospective Analysis.

BACKGROUND: Filgrastim has been widely used for hematopoietic recovery after hematopoietic stem cell transplantation. Recently, biosimilar filgrastim (BF) has been approved for the same indications as for the originator filgrastim (OF). However, evidence of the efficacy and safety of BF for unrelated cord blood transplant (CBT) is unreported. Therefore, we evaluated the efficacy and safety of BF and OF (historical control) for CBT. METHODS: Twenty-two consecutive patients with hematologic malignant tumors were assessed. Patients received BF (n = 12) or OF (n = 10) from day 1 after CBT for hematopoietic recovery. The time to hematopoietic recovery, total filgrastim dose, duration of filgrastim administration, total transfusion units, incidences of engraftment, documented infection, febrile neutropenia, acute and chronic graft-vs-host disease, incidence and severity of adverse events, hospitalization duration, and 100-day and 1-year overall survival were evaluated. RESULTS: The median total dose of BF and OF used for hematopoietic recovery were 9.68 and 10.80 mg, respectively. There were no significant between-group differences in time to hematopoietic recovery, total filgrastim dose, duration of filgrastim administration, total transfusion units, incidences of engraftment, documented infection, febrile neutropenia, acute and chronic graft-vs-host disease, incidence and severity of adverse events, hospitalization duration, and overall survival. Multivariate analysis demonstrated that filgrastim type was not a significant factor for neutrophil recovery. Median total filgrastim costs per patient were 446,405 and 910,320 yen for BF and OF, respectively. CONCLUSIONS: BF is as safe and effective as OF for hematopoietic recovery after CBT. BF is a useful option for CBT owing to its economic benefits.

Effect of pre-reacted glass-ionomer filler extraction solution on demineralization of bovine enamel.

OBJECTIVE: To determine the effect of pre-reacted glass-ionomer (PRG) filler extraction solution on the demineralization of bovine enamel by measuring changes in the ultrasound transmission velocity. METHODS: The specimens were prepared by cutting bovine teeth into enamel blocks. The specimens were immersed in buffered lactic acid solution for 10 minutes twice a day, and then stored in artificial saliva. Other specimens were stored in PRG filler extraction solution for 10 minutes, followed by 10-minute immersion in the buffered lactic acid solution twice a day. The propagation time of longitudinal ultrasonic waves was measured by a pulser receiver. Six specimens were used for each condition, and analyses of variance followed by Tukey tests (α=0.05) were done. RESULTS: No changes in sonic velocity were found for specimens stored in the PRG filler extraction solution, indicating that the PRG extraction solution had an effect on inhibiting the demineralization of bovine enamel. CONCLUSIONS: The results obtained with the use of an ultrasound measurement technique suggested that PRG filler extraction solution has the ability to prevent demineralization of enamel.

Cyclooxygenase-2 activity is important in craniofacial fracture repair.

The aim of this study was to examine the effect of cyclooxygenase (COX)-2 on bone repair after craniofacial fracture in mice. A 4-mm fracture was created in the parietal bone of 8-week-old male COX-2 wild-type (COX-2(+/+)) and knockout (COX-2(-/-)) mice. Ribonucleic acid was extracted from the fractured bone and analysed. For morphological and histological analysis, the mice were killed 8 and 12 weeks after treatment, and sections were prepared. Three-dimensional computed tomography was performed, and the sections were stained with hematoxylin-eosin for histological examination. Expression of COX-2 messenger ribonucleic acid was induced in COX-2(+/+) mice, but not in COX-2(-/-) mice. Ossification at the fracture site was almost complete 12 weeks after fracture in COX-2(+/+) mice. In COX-2(-/-) mice, incomplete union had occurred at the fracture site. In both types of mice, the fracture site contained no cartilaginous tissue, and the callus formed from the periosteal side. These results suggest that COX-2 plays an important role in craniofacial fracture repair and that COX-2-selective non-steroidal anti-inflammatory drugs might interfere with fracture repair of the membranous viscerocranium in the clinical setting.

Contribution of postmortem multidetector CT scanning to identification of the deceased in a mass disaster: Experience gained from the 2009 Victorian bushfires.

CT scanning of the deceased is an established technique performed on all individuals admitted to VIFM over the last 5 years. It is used primarily to assist pathologists in determining cause and manner of death but is also invaluable for identification of unknown deceased individuals where traditional methods are not possible. Based on this experience, CT scanning was incorporated into phase 2 of the Institute's DVI process for the 2009 Victorian bushfires. All deceased individuals and fragmented remains admitted to the mortuary were CT scanned in their body bags using established protocols. Images were reviewed by 2 teams of 2 radiologists experienced in forensic imaging and the findings transcribed onto a data sheet constructed specifically for the DVI exercise. The contents of 255 body bags were examined in the 28 days following the fires. 164 missing persons were included in the DVI process with 163 deceased individuals eventually identified. CT contributed to this identification in 161 persons. In 2 cases, radiologists were unable to recognize commingled remains. CT was utilized in the initial triage of each bag's contents. If radiological evaluation determined that bodies were incomplete then this information was provided to search teams who revisited the scenes of death. CT was helpful in differentiation of human from non-human remains in 8 bags, recognition of human/animal commingling in 10 bags and human commingling in 6 bags. In 61% of cases gender was able to be determined on CT using a novel technique of genitalia detection and in all but 2 cases this was correct. Age range was able to be determined on CT in 94% with an accuracy of 76%. Specific identification features detected on CT included the presence of disease (14 disease entities in 13 cases), medical devices (26 devices in 19 cases) and 274 everyday metallic items associated with the remains of 135 individuals. CT scanning provided useful information prior to autopsy by flagging likely findings including the presence of non-human remains, at the time of autopsy by assisting in the localization of identifying features in heavily disfigured bodies, and after autopsy by retrospective review of images for clarification of issues that arose at the time of pathologist case review. In view of the success of CT scanning in this mass disaster, DVI administrators should explore the incorporation of CT services into their disaster plans.

Functions of glutamate transporters in cerebellar Purkinje cell synapses.

Glutamate transporters play a critical role in the maintenance of low extracellular concentrations of glutamate, which prevents the overactivation of post-synaptic glutamate receptors. Four distinct glutamate transporters, GLAST/EAAT1, GLT-1/EAAT2, EAAC1/EAAT3 and EAAT4, are distributed in the molecular layer of the cerebellum, especially near glutamatergic synapses in Purkinje cells (PCs). This review summarizes the current knowledge about the differential roles of these transporters at excitatory synapses of PCs. Data come predominantly from electrophysiological experiments in mutant mice that are deficient in each of these transporter genes. GLAST expressed in Bergmann glia contributes to the clearing of the majority of glutamate that floods out of the synaptic cleft immediately after transmitter release from the climbing fibre (CF) and parallel fibre (PF) terminals. It is indispensable to maintain a one-to-one relationship in synaptic transmission at the CF synapses by preventing transcellular glutamate spillover. GLT-1 plays a similar but minor role in the uptake of glutamate as GLAST. Although the loss of neither GLAST nor GLT-1 affects cerebellar morphology, the deletion of both GLAST and GLT-1 genes causes the death of the mutant animal and hinders the folium formation of the cerebellum. EAAT4 removes the low concentrations of glutamate that escape from uptake by glial transporters, preventing the transmitter from spilling over into neighbouring synapses. It also regulates the activation of metabotropic glutamate receptor 1 (mGluR1) in perisynaptic regions at PF synapses, which in turn affects mGluR1-mediated events including slow EPSCs and long-term depression. No change in synaptic function is detected in mice that are deficient in EAAC1.

Unilateral expansion of a narrow mandibular dental arch combined with bimaxillary osteotomies in a patient with hypoglossia.

A 23-year-old female with hypoglossia, who had a narrow mandibular dental arch, was treated using the gradual expansion technique. Three lower incisors were missing and the right molar occlusion showed a scissor bite. Her speech was acceptable. Gradual unilateral expansion of the mandibular alveolar bone was performed. Orthodontic tooth alignment was performed prior to surgical treatment. A tooth-borne expander was devised using a hyrax-type screw to move the inclined right alveolar bone into an upright position. Alveolar bone osteotomies were performed under general anesthesia and the expander was placed in the mandibular dental arch. After a 5-day latency period, the screw was activated for 21 days. After expansion, the width of the mandibular dental arch increased by 10mm at the first molar region and the right molars were moved to an upright position. After a consolidation period of 7 days, simultaneous two-jaw surgery that combined Le Fort I osteotomy and intraoral vertical ramus osteotomies was performed to obtain a stable occlusion. After post-surgical orthodontic and prosthodontic treatment, her occlusion improved without deterioration of her speech. The results indicate that this technique is useful for unilateral expansion of distorted mandibular alveolar process.

Influence of arthroscopically observed fibrous adhesions before and after joint irrigation on clinical outcome in patients with chronic closed lock of the temporomandibular joint.

The aim of this study was to investigate the changes in the state of arthroscopically observed fibrous adhesions (FA) after visually guided irrigation (VGIR) and the influence of FA on clinical outcome in patients with chronic closed lock of the temporomandibular joint (TMJ). Forty-eight TMJs of 48 patients with unilateral chronic closed lock were enrolled in this study. All 48 joints underwent VGIR twice. After the first VGIR (immediately before the second VGIR), clinical outcome was assessed as regards maximal interincisal opening (MIO) and self-evaluated TMJ pain (VAS). Thirty patients were symptom-free (good outcome group) and the remaining 18 patients had symptoms (poor outcome group). In each group, the changes of the MIO, VAS and severity of FA (FA score) after the first VGIR were studied. The influence of FA score in the first and second VGIR on clinical outcome was analyzed by logistic regression analysis. There was no joint with disappearance or reduction of FA after the first VGIR. In both groups, MIO and VAS were significantly improved after the first VGIR even though the state of FA became significantly worse. The multivariate logistic regression analysis showed that the risk of poor outcome for FA scores in the first and second VGIR were 0.89-times (95% CI: 0.33-2.40, P=0.82) and 1.76-times (95% CI: 0.54-5.73, P=0.35), respectively. The dose-response relationships between FA scores in the first or second VGIR were not significant. In conclusion, our results indicate that the presence of FA or a postoperative worsening of FA (including postoperative new FA formation) seems not to affect the clinical outcome as regards MIO and VAS in patients with chronic closed lock of the TMJ.

Immunohistochemical localization of cyclooxygenase-1 and -2 in synovial tissues from patients with internal derangement or osteoarthritis of the temporomandibular joint.

This study examined the immunohistochemical expression and localization of cyclooxygenase-1 and -2 (COX-1 and COX-2) in synovial tissues from patients with internal derangement (ID) or osteoarthritis (OA) of the temporomandibular joint (TMJ). Synovial tissues from patients with condylar fractures of the mandible were studied as control. Synovial tissues from 13 TMJs of 10 patients with ID or OA and from 5 TMJs of 4 patients with fractures were examined for COX-1 and COX-2 expression by immunohistochemical staining using two monoclonal antibodies. In addition, whether the COX-2 expression grade correlated with the synovitis score and clinical findings was assessed. COX-2 was expressed in the synovial lining, infiltrating mononuclear cells, fibroblast-like cells, and blood vessels, including CD31-positive endothelial cells, in the synovium of patients with ID or OA. Expression levels of COX-1 in synovial lining cells and endothelial cells were similar in the specimens obtained from the patients with ID or OA and those obtained from the controls. The expression of COX-2 positively correlated with arthroscopic findings of synovitis (p = 0.55, P = 0.023) and with joint pain (p = 0.56, P = 0.021). These results suggest that up-regulation of COX-2 in synovium may play a part in the pathogenesis of synovitis in patients with ID or OA of the TMJ.

Suppression of the morphine-induced rewarding effect and G-protein activation in the lower midbrain following nerve injury in the mouse: involvement of G-protein-coupled receptor kinase 2.

The present study was designed to investigate whether a state of neuropathic pain induced by sciatic nerve ligation could alter the rewarding effect, antinociception, and G-protein activation induced by a prototype of mu-opioid receptor agonist morphine in the mouse. The sciatic nerve ligation caused a long-lasting and profound thermal hyperalgesia. Under this neuropathic pain-like state, an i.c.v. morphine-induced place preference was observed in sham-operated mice but not in sciatic nerve-ligated mice. However, no differences in the antinociceptive effect of i.c.v.-administered morphine were noted between the groups. The increases in the binding of guanosine-5'-o-(3-[(35)S]thio)triphosphate induced by morphine in lower midbrain membranes including the ventral tegmental area, which contributes to the expression of the rewarding effect of opioid, were significantly attenuated in sciatic nerve-ligated mice. On the other hand, there were no differences in the stimulation of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding to pons/medulla membranes, which plays an important role in the antinociception of mu-opioid receptor agonists, between the groups. In addition, no changes in levels of guanosine-5'-o-(3-[(35)S]thio)triphosphate binding by either the selective delta- or kappa-opioid receptor agonists were noted in membrane of the lower midbrain and limbic forebrain membranes obtained from sciatic nerve-ligated mice. Reverse transcription-polymerase chain reaction analysis showed that sciatic nerve ligation did not alter the mRNA product of mu-opioid receptors in the lower midbrain, indicating that a decrease in some mu-opioid receptor functions may result from the uncoupling of mu-opioid receptors from G-proteins. We found a significant increase in protein levels of G-protein-coupled receptor kinase 2, which causes receptor phosphorylation in membranes of the lower midbrain but not in the pons/medulla, obtained from mice with nerve injury, whereas there were no changes in the protein level of phosphorylated-protein kinase C in the lower midbrain. These results suggest that the uncoupling of mu-opioid receptors from G-proteins by G-protein-coupled receptor kinase 2 in the lower midbrain may, at least in part, contribute to the suppression of the rewarding effect of morphine under neuropathic pain.

Dentoalveolar reconstruction of a missing premaxilla using bone graft and endosteal implants.

We report here on a patient with bilateral cleft lip and palate (BCLP) and a missing premaxilla, who underwent dentoalveolar reconstruction of the cleft and premaxillary alveolus using endosteal implants after bone grafting. The patient, whose maxillary incisors and premaxilla were missing, had corticocancellous bone grafting from the iliac crest, followed by excellent bone formation at the anterior alveolus. After the placement of the endosteal implants and the completion of the pre-surgical orthodontic alignment, orthognathic surgery was performed for the restoration of a Class III open bite. After post-operative orthodontic preparation, the final fixed prostheses were completed. This treatment procedure offers an option of dentoalveolar reconstruction for BCLP patients with an excised premaxilla.

Partial inferior turbinectomy during secondary alveolar bone grafting.

This study examined the characteristics and outcome of patients undergoing partial inferior turbinectomy during secondary alveolar bone grafting. Thirty-three of 55 patients with cleft lip and palate or cleft lip and alveolus who underwent secondary alveolar bone grafting concurrently received partial inferior turbinectomy to ensure that the height of the nasal floor was similar on the cleft side and non-affected side. At the time of surgery, patients who underwent turbinectomy were significantly older than those who did not undergo the procedure. The proportion of patients who underwent turbinectomy was significantly higher among patients with cleft lip and palate than among those with cleft lip and alveolus. These differences apparently reflected the developmental stage of the inferior turbinate and the relative severity of alveolar and palatal defects. In most patients who underwent partial inferior turbinectomy, postoperative X-ray films revealed excellent bone formation at the graft site. Our findings suggest that partial inferior turbinectomy during secondary alveolar bone grafting is a very useful procedure that facilitates dissection to the height of the nasal floor, reconstruction of the mucosal nasal floor, and formation of a sufficient bone bridge. It also promotes alveolar cleft closure, especially in patients with wide bone defects.

Auxin- and abscisic acid-dependent osmoregulation in protoplasts of Phaseolus vulgaris pulvini.

Protoplasts isolated from the laminar pulvinus of Phaseolus vulgaris and bathed in a medium containing KCl as the major salt were found to swell in response to IAA and to shrink in response to ABA. The protoplasts of flexor cells and those of extensor cells responded similarly. The results indicate that the cellular content of osmotic solutes is enhanced by IAA and reduced by ABA. The IAA-induced swelling was abolished when either the K(+) or the Cl(-) of the bathing medium was replaced by an impermeant ion or when the medium was adjusted to neutral pH (instead of pH 6). The response was inhibited by vanadate. It is concluded that the swelling is caused by enhanced influxes of K(+) and Cl(-), which probably occur through K(+) channels and Cl(-)/H(+) symporters, respectively. The ABA-induced shrinking was inhibited by 5-nitro-2-(3-phenylpropylamino)-benzoic acid, an anion-channel inhibitor, suggesting that it is caused by Cl(-) efflux through anion channels and charge-balancing K(+) efflux through outward-rectifying K(+) channels. It appears that the two plant hormones act on pulvinar motor cells to regulate their turgor pressure, as they do in stomatal guard cells. The findings are discussed in relation to the pulvinar movements induced by environmental stimuli.

Dendritic spine geometry is critical for AMPA receptor expression in hippocampal CA1 pyramidal neurons.

Dendritic spines serve as preferential sites of excitatory synaptic connections and are pleomorphic. To address the structure-function relationship of the dendritic spines, we used two-photon uncaging of glutamate to allow mapping of functional glutamate receptors at the level of the single synapse. Our analyses of the spines of CA1 pyramidal neurons reveal that AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid)-type glutamate receptors are abundant (up to 150/spine) in mushroom spines but sparsely distributed in thin spines and filopodia. The latter may be serving as the structural substrates of the silent synapses that have been proposed to play roles in development and plasticity of synaptic transmission. Our data indicate that distribution of functional AMPA receptors is tightly correlated with spine geometry and that receptor activity is independently regulated at the level of single spines.

Mapping of heme-binding domains in soluble guanylyl cyclase beta1 subunit.

Soluble guanylyl cyclase (sGC) is activated upon the interaction of NO with heme in the sGC beta1 subunit. To identify the domains contributing to heme-binding, we constructed a series of deletion mutants of the beta1 subunit, and evaluated their heme-binding capability. Deletion mutants consisting of residues 1-120 [beta1(1-120)] and 80-385 [beta1(80-385)] were the shortest mutants exhibiting heme binding among the C-terminal and N-terminal-truncated mutants, respectively. The region common to both beta1(1-120) and beta1(80-385), i.e., residues 80-120, is therefore essential for heme binding, although the residues 341-385 play an auxiliary role in heme binding. Two deletion mutants, beta1(80-195) and beta1(60-195), which include only the essential region, exhibited strong heme binding and spectral properties similar to those of the nitrosyl complex of native sGC. Thus, these heme-binding core proteins may serve as model proteins for future studies on the tertiary structure of the nitrosyl complex of sGC.

Vestibuloplasty after secondary alveolar bone grafting.

OBJECTIVE: This paper introduces a surgical technique for vestibuloplasty after secondary alveolar bone grafting of patients with cleft lip and palate (CLP). This paper also reports on the patients who underwent this modified vestibuloplasty. SURGICAL PROCEDURE: The vestibuloplasty technique described in this paper consists of: (1) reduction of submucosal scar tissue of the upper lip, (2) V-Y plasty of the superficial mucosa, (3) placement of horizontal mattress sutures between nostril floor skin and freed marginal mucosa, (4) application of artificial skin to cover the exposed periosteal surface, and (5) use of a removable retention splint. CONCLUSION: This surgical procedure appears to be very useful for patients with CLP. The technique enables the surgeon to obtain an adequate sulcus depth around the graft area. In addition, this technique releases the mucosal scar contraction and improves the shape and mobility of the upper lip.

Visualization of IP(3) dynamics reveals a novel AMPA receptor-triggered IP(3) production pathway mediated by voltage-dependent Ca(2+) influx in Purkinje cells.

IP(3) signaling in Purkinje cells is involved in the regulation of cell functions including LTD. We have used a GFP-tagged pleckstrin homology domain to visualize IP(3) dynamics in Purkinje cells. Surprisingly, IP(3) production was observed in response not only to mGluR activation, but also to AMPA receptor activation in Purkinje cells in culture. AMPA-induced IP(3) production was mediated by depolarization-induced Ca(2+) influx because it was mimicked by depolarization and was blocked by inhibition of the P-type Ca(2+) channel. Furthermore, trains of complex spikes, elicited by climbing fiber stimulation (1 Hz), induced IP(3) production in Purkinje cells in cerebellar slices. These results revealed a novel IP(3) signaling pathway in Purkinje cells that can be elicited by synaptic inputs from climbing fibers.

IGF-I-induced apoptosis in LM2d6 cultured at a low concentration of fetal bovine serum.

We examined the effects of IGF-I (1-1000 ng/ml) on cell proliferation in LM2d6 mouse fibroblast cells at 0.1, 1.0 and 5.0% fetal bovine serum (FBS). In medium containing 0.1% FBS, treatment of LM2d6 cells with IGF-I significantly reduced the cell number in a dose- and time-dependent manner, whereas no effects were seen at 1 or 5% FBS. Treatment of the cells with 0.1% FBS for 72 h caused DNA laddering and nuclear condensation. However, Scatchard analysis for IGF-I binding sites on the cells revealed that both the number and the affinity of IGF-I receptors were not greater than that of Balb/3T3 cells. Furthermore, the apoptotic action of Long (R(3))-IGF-I, an analogue of IGF-I that has a reduced affinity for IGF binding proteins, was not greater than that of IGF-I. Taken together, we conclude that IGF-I reduces cell proliferation at low levels of FBS due to the induction of apoptosis. This effect is probably not caused by an excess production of IGF binding proteins in LM2d6 cells.

Reduced IP3 sensitivity of IP3 receptor in Purkinje neurons.

The inositol 1,4,5-trisphosphate receptor (IP3R) is highly expressed in Purkinje neurons (PNs) and is thought to be essential for the induction of long-term depression at parallel-fiber-PN synapses. Here, by imaging the fluorescence intensity of the low-affinity Ca2+ indicator inside the Ca2+ stores in the permeabilized single PNs, we analyzed the kinetics of Ca2+ release via the IP3R in controlled cytoplasmic environments. The rate of Ca2+ release is dependent on the IP3 concentration with an EC50 of 25.8 microM, which is > 20-fold greater than that of the IP3R in the isolated preparations or in peripheral cells. This property would be advantageous in inducing the release of Ca2+ in a localized space adjacent to the site of synaptic inputs.