Organization of pERK-immunoreactive cells in trigeminal spinal nucleus caudalis and upper cervical cord following capsaicin injection into oral and craniofacial regions in rats.

To define the somatotopic arrangement of neurons in the trigeminal spinal subnucleus caudalis and upper cervical cord activated by acute noxious stimulation of various orofacial sites, pERK expression was analyzed in these neurons. After capsaicin injection into the tongue, lower gum, upper and lower lips, or mental region, pERK-like immunoreactive (pERK-LI) cells were distributed mainly in the dorsal half of the trigeminal spinal nucleus interporalis (Vi) and caudalis (Vc) transition zone (Vi/Vc zone), middle Vc, and Vc and upper cervical cord transition zone (Vc/C2 zone). pERK-LI cells were distributed throughout the dorsal to ventral portion of the Vi/Vc zone, middle Vc, and Vc/C2 zone following capsaicin injection into the anterior hard palate, upper gum, buccal mucosa, or vibrissal pad and in the ventral portion of the Vi/Vc zone, middle Vc, and Vc/C2 zone following snout, ophthalmic, or ocular injection of capsaicin. The rostrocaudal distribution area of pERK-LI cells was more extensive from the Vi/Vc zone to the Vc/C2 zone after intraoral injection than that after facial injection, and the rostrocaudal distribution of pERK-LI cells from the Vi/Vc zone to the Vc/C2 zone had a somatotopic arrangement, with the snout being represented most rostrally and ophthalmic, ocular, or mental regions represented most caudally. These findings suggest that the pERK-LI cells expressed from the Vi/Vc zone to the Vc/C2 zone following injection of capsaicin in facial and intraoral structures may be differentially involved in pain perception in facial and intraoral sites.

Effects of IL-6 and soluble IL-6 receptor on the expression of cartilage matrix proteins in human chondrocytes.

Elevated concentrations of interleukin (IL)-6 and soluble IL-6 receptor (sIL-6Ralpha) in synovial fluid have been implicated in joint cartilage destruction. We examined the effect of IL-6 and sIL-6Ralpha on cell growth, alkaline phosphatase (ALPase) activity, and the expression of Sox-9, type II collagen, aggrecan core, link protein, BMP-7, and BMP receptors in human chondrocytes. Cell proliferation increased slightly in the presence of both IL-6 and sIL-6Ralpha, whereas ALPase activity decreased markedly. The expression of Sox-9 and aggrecan core did not change in the presence or absence of IL-6 and sIL-6Ralpha, whereas the expression of type II collagen, link protein, BMP-7, and BMP receptors increased in the presence of both IL-6 and sIL-6Ralpha. These results suggest that IL-6 and sIL-6Ralpha suppress the differentiation of chondrocytes and induce the repair of arthrodial cartilage through an increase in the expression of cartilage matrix proteins, BMP-7, and BMP receptors in the cells.

Butyrate inhibits oral cancer cell proliferation and regulates expression of secretory phospholipase A2-X and COX-2.

Although surgical resection is the first choice for oral cancer, the development of new anti-cancer drugs is of great interest. The effect of the histone deacetylation inhibitor, sodium butyrate (NaBu) on oral cancer cell (OCC) HSC-3 and HSC-4 proliferation in vitro was investigated. The synthesis of rate-limiting enzymes such as sPLA2 (-IIA, -V, -X) and COX-2 was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, as well as PGE2 by ELISA. NaBu acted in a concentration-dependent manner. Over 3 mM, it inhibited OCC proliferation, due to increased p21 expression and cell cycle arrest in the G2/M-phase. At low concentration (< or = 1 mM), NaBu showed no effects or enhanced cell proliferation. NaBu also regulated COX-2 and sPLA2-X expression, and augmented PGE2 synthesis in OCC. These results indicate that NaBu is a novel candidate agent for the treatment of oral cancer. The treatment efficacy must be investigated in additional experiments considering NaBu concentration and tumor cell heterogeneity.

Changes in extracellular activin A:follistatin ratio during differentiation of a mesenchymal progenitor cell line, ROB-C26 into osteoblasts and adipocytes.

We investigated the effects of BMP-2 and dexamethasone (Dex) on follistatin (FS) and activin A expressions in a mesenchymal progenitor cell line, ROB-C26 (C26). C26 cells stimulated to differentiate into osteoblastic cells by blocking myogenic differentiation after BMP-2 treatment and into adipocytes with Dex treatment. Alkaline phosphatase (ALP) mRNA expression and its activity in the confluent C26 cells were dose- and time-dependently stimulated by BMP-2, but inhibited by Dex. The stimulatory effect on FS and activin A mRNA expressions by BMP-2 and Dex were dose-dependent. Cycloheximide pre-treatment indicated that FS and activin A expressions appear to be the direct target of BMP-2 and Dex signaling. BMP-2 time-dependently increased FS and activin A levels. Dex also increased FS level, but induced a time-dependent biphasic effect on activin A level, a decrease (2-6 h) followed by an increase (12-72 h). The data of the ratio of activin A concentration in the culture media to that of FS (activin A:FS ratio) measured by ELISA showed that BMP-2-induced osteoblastic differentiation involved an activin-dominant microenvironment, whereas Dex-induced adipocyte differentiation involved a FS-dominant microenvironment. Excess FS suppressed the stimulatory ALP activity of BMP-2, whereas activin A prevented not only Dex-induced inhibitory ALP activity, but also adipogenesis via suppression of the adipocyte transcriptional factor cascade. These results indicate that BMP-2-induced activin-dominant microenvironment may be critical for osteoblastic differentiation by restricting the antagonistic effects of FS on BMP activity, while Dex-induced FS-dominant microenvironment may be critical for adipocyte differentiation by restricting the inhibitory action of activin A on adipocyte differentiation.

Augmenting effect of serotonin on the voltage-dependent Ca2+ current and subsequently activated K+ current in Aplysia neurons.

Receptor-induced activation of protein kinase C (PKC) plays an important role in modulation of various types of ionic channels in neurons. For example, PKC causes facilitation or long-lasting activation of certain ionic channels involved in spike firing after the receptor stimulation. We investigated the effect of serotonin (5-HT) on the voltage-dependent Ca(2+) channels in RB and RC neurons of Aplysia ganglia under voltage clamp. An outward current response was induced by voltage change of the cell membrane from -60 mV to +10 mV. Application of 5-HT significantly augmented the outward current response to the voltage change. Both the outward current and the augmenting effect of 5-HT markedly decreased when examined in either Ca(2+)-free, 10 mM tetraethylammonium, or 0.3 mM Cd(2+)-solution, indicating the current to be Ca(2+)-activated K(+) current produced by Ca(2+) entry. Intracellular application of either guanosine 5'-O-(2-thiodiphosphate) or cholera toxin (CTX), reagents for G-proteins, irreversibly blocked the augmenting effect of 5-HT. Application of phorbol dibutylate (PDBu), an activator of PKC, augmented the outward current and the effect of 5-HT was occluded after PDBu application. Staurosporine, a specific inhibitor of PKC, markedly suppressed the augmenting effects of both 5-HT and PDBu on the outward current. However, either 5-HT or PDBu did not augment the Ca(2+)-activated K(+) current induced by intracellular injection of Ca(2+) but rather depressed it. These results suggest that stimulation of 5-HT receptor may activate a novel type of CTX-sensitive G-protein and subsequent PKC, and that phosphorylation of voltage-dependent Ca(2+) channels may result in the increase in Ca(2+) entry and subsequent Ca(2+)-activated K(+) current. The mechanism may contribute to retain the long-lasting activation without broadening of the spike width during the excitatory response to 5-HT in these neurons.

Differential involvement of cyclase- versus non-cyclase-coupled D1-like dopamine receptors in orofacial movement topography in mice: studies with SKF 83822.

Though orofacial movements are fundamental motor patterns that are known to be regulated critically by D1-like dopamine receptors, these processes remain poorly understood. This uncertainty is heightened by evidence for putative D1-like receptors that are linked not only to adenylyl cyclase (AC) but also to phospholipase C (PLC). Using a new method, we have characterised four topographies of orofacial movement in the mouse using the novel D1-like agonist SKF 83822, which stimulates AC but not PLC. These were compared with responses to SKF 83959, which stimulates PLC but not AC. Also, effects were characterised using the D1-like antagonist SCH 23390 and the D2-like antagonist YM 09151-2. SKF 83822 induced vertical jaw movements with incisor chattering but inhibited horizontal jaw movements; there was little effect on tongue protrusions. Vertical jaw movements induced by SKF 83822 were inhibited by SCH 23390 but uninfluenced by YM 09151-2, while YM 09151-2 released horizontal jaw movements; thus, D1-like agonist-induced, AC-mediated vertical jaw movements constitute a 'pure' D1-like-dependent process that does not involve D1-like:D2-like interactions, while horizontal jaw movements involve oppositional interactions. Orofacial movements in mice appear to consist of at least four phenomenologically dissociable topographies that are mechanistically distinct. They are regulated differentially by AC- and/or PLC-dependent processes and these processes involve distinct D1-like:D2-like interactions.

Electrophysiological and pharmacological characterization of the K(ATP) channel involved in the K+-current responses to FSH and adenosine in the follicular cells of Xenopus oocyte.

The follicular cells surrounding Xenopus oocyte under voltage clamp produce K(+)-current responses to follicle-stimulating hormone (FSH), adenosine (Ade), and intracellularly applied cAMP. We previously reported that these responses are suppressed by the stimulation of P2Y receptor through phosphorylation by PKC presumably of the ATP-sensitive K(+) (K(ATP)) channel. This channel comprises sulfonylurea receptors (SURs) and K(+) ionophores (Kirs) having differential sensitivities to K(+) channel openers (KCOs) depending on the SURs. To characterize the K(+) channels involved in the FSH- and Ade-induced responses, we investigated the effects of various KCOs and SUR blockers on the agonist-induced responses. The applications of PCO400, cromakalim (Cro), and pinacidil, but not diazoxide, produced K(+)-current responses similar to the FSH- and Ade-induced responses in the magnitude order of PCO400 > Cro >> pinacidil in favor of SUR2A. The application of glibenclamide, phentolamine, and tolbutamide suppressed all the K(+)-current responses to FSH, Ade, cAMP, and KCOs. Furthermore, both the FSH- and Ade-induced responses were markedly augmented during the KCO-induced responses, or vice versa. The I-V curves for the K(+)-current responses induced by Cro, Ade, and FSH showed outward rectification in normal [K(+)](o), but weak inward rectification in 122 mM [K(+)](o). Also, stimulations of P2Y receptor by UTP or PKC by PDBu markedly depressed the K(+)-current response to KCOs in favor of Kir6.1, as previously observed with the responses to FSH and Ade. These results suggest that the K(+)-current responses to FSH and Ade may be produced by the opening of a novel type of K(ATP) channel comprising SUR2A and Kir6.1.

The effect of IL-1beta on the expression of inflammatory cytokines and their receptors in human chondrocytes.

Cytokines released at sites of inflammation and infection can alter the normal processes of cartilage turnover, resulting in pathologic destruction or formation. Interleukin (IL)-1beta plays a central role in the pathophysiology of cartilage damage and degradation in arthritis. In the present study, we examined the effect of IL-1beta on the expression of IL-1beta, IL-6, IL-8, IL-11, tumor necrosis factor-alpha (TNF-alpha), and their receptors in human chondrocytes. The cells were cultured either with or without 100 U/ml of IL-1beta for up to 28 days. The level of expression of the cytokines and their receptors was estimated by determining mRNA levels using real-time PCR or by determining protein levels using ELISA. The expression of IL-1beta, IL-8, and TNF-alpha markedly increased in the presence of IL-1beta after day 14 of culture. The expression of IL-6 and IL-11 increased greatly in the presence of IL-1beta on day 1 and after day 14 of culture. The expression of IL-1beta, IL-8, IL-11, and TNF-alpha receptors significantly decreased in the presence of IL-1beta after day 14 of culture, whereas the expression of IL-6 receptor significantly increased. The expression of these cytokines, except for IL-6, decreased with the addition of human IL-1 receptor antagonist. These results suggest that IL-1beta promotes the resolution system of cartilage matrix turnover through an increase in inflammatory cytokine production by chondrocytes and that it also may promote the autocrine action of IL-6 through an increase in IL-6 receptor expression in the cells.

Phosphorylation of Extracellular Signal-Regulated Kinase in medullary and upper cervical cord neurons following noxious tooth pulp stimulation.

The phosphorylated Extracellular Signal-regulated Kinase (pERK) and Fos expression and masticatory muscle activity were analyzed in rats with capsaicin-induced acute inflammation of the tooth pulp in order to clarify the role of the spinal trigeminal nucleus and upper cervical spinal cord in tooth pulp pain. Digastric and masseteric muscle activities were significantly increased following capsaicin injection into the molar tooth pulp but not after vehicle treatment. The pERK-like immunoreactive (LI) neurons were observed in the subnuclei interpolaris-caudalis transition (Vi/Vc) zone, the paratrigeminal nucleus (Pa5) and the superficial laminae of the caudal Vc/C2 zone. The pERK expression was detected as early as 2 min and peaked at 5 min after capsaicin or vehicle injection. The pERK expression in the Vi/Vc zone and Pa5 was bilateral, whereas it was predominantly ipsilateral in the caudal Vc/C2 zone. The capsaicin treatment of the whisker pad produced pERK expression in the rostro-caudal middle portion of the ipsilateral Vc, but small number of pERK-LI cells were observed after vehicle treatment. The pERK expression was similar in the Vi/Vc zone following capsaicin injection into the upper or lower molar tooth pulp, whereas the pERK expression was in the lateral portion of the caudal Vc/C2 zone after upper molar injection and restricted to the medial portion of the Vc/C2 zone after the lower molar capsaicin. These data were confirmed with Western blots. There were differences in the distribution of Fos protein-like immunoreactive (LI) cells and pERK-LI cells following tooth pulp stimulation. After capsaicin application into the upper molar tooth pulp, no pERK-LI cells were observed in the ventral part of the Vi/Vc zone, whereas many Fos protein-LI cells were expressed in this region. The difference in the distribution pattern of pERK- and Fos protein-LI cells in the Vi/Vc zone suggests their differential temporal expression profiles after capsaicin. The present findings suggest that tooth-pulp-driven neurons in the spinal trigeminal nucleus are involved in tooth pulp pain through activation of the intracellular signal transduction pathway that involves earlier ERK phosphorylation and subsequent Fos expression.

Unusual cyst-like lesions in the parapharyngeal space associated with recurrence of tongue carcinoma.

A 54-year-old male presented with the complaint of a painful sore on the left side of his tongue. Our examination found an ulcer 15 x 20 mm in size on the left edge of the tongue, with peripheral indurations. The lesion was diagnosed histopathologically as squamous cell carcinoma (T2N0M0). Consequently, the lesion was surgical removed and radical neck dissection was performed. Four months after the operation, two unusual cyst-like lesions were identified in the parapharyngeal space by CT and MRI. A biopsy specimen revealed recurrent carcinoma with a cyst-like structure. The route of the tumor metastasis into the parapharyngeal space was obscured, but it was speculated that the excessive lymph accumulation was due to a lymphatic occlusion caused by the surgical procedure, proliferation of the metastatic carcinoma, or stagnation and accumulation of tissue fluid caused by parapharyngeal invasion by the recurrent lesion.

Postoperative mental disorder following prolonged oral surgery.

A prolonged period of oral surgery is a potential risk factor of postoperative mental disorders although no such report has been published to date. We retrospectively studied perioperative features in 36 patients who underwent prolonged (10 hours or more) of oral surgery. Patients were categorized as pre-delirium (Pre-D) when they manifested 1 or 2 symptoms and delirium (D) when they showed more than 2 symptoms, according to the modified International Classification of Diseases, 10th edition. Of the 36 patients who returned to a normal mental state without drug therapy, 13 were classified as D and 14 were Pre-D. A number of patients had moderate complications preoperatively, and massive hemorrhaging occurred during the operation in some Pre-D and D patients. Age was greater in D (62.0 +/- 9.9 years) than in Pre-D (56.0 +/- 13.8 years) patients. Propofol-based general anesthesia was most commonly employed. The time prior to appearance of pre-delirium was significantly shorter in D (30.0 +/- 16.7 hours) than in Pre-D (55.0 +/- 35.0 hours) group patients. Our results indicate that, in general, patients predisposed to postoperative mental disorders have moderate complications preoperatively, are generally older than 50-years-old, receive propofol-based general anesthesia and/or experience a massive hemorrhage during the operation.

Masticatory efficiency before and after surgery in oral cancer patients: comparative study of glossectomy, marginal mandibulectomy and segmental mandibulectomy.

This study evaluated the effect of oral cancer surgery on masticatory efficiency. Masticatory efficiency was measured using the ATP absorption method. Eating ability was measured using a questionnaire. Two groups were employed as controls: The "normal occlusion group" consisted of subjects who had a complete set of natural maxillary teeth opposed to mandibular teeth, and the "unilateral occlusion group" consisted of subjects who had lost their molar and premolar teeth on one side of the mandible as a result of caries or periodontal diseases. Three treatment groups, each of 6 patients, were studied: a glossectomy group, a marginal mandibulectomy group and a segmental mandibulectomy group. There were no differences in masticatory efficiency between two control groups. Masticatory efficiencies of the three oral cancer treatment groups were lower than in the unilateral occlusion group, even 12 months after surgery. Masticatory efficiency of the glossectomy group was significantly higher 12 months after surgery compared with pre-surgery. Masticatory and eating abilities of the marginal mandibulectomy group and the segmental mandibulectomy were reduced at 3 and 6 months after surgery. The masticatory efficiency 12 months after surgery was higher in the marginal mandibulectomy group than the segmental mandibulectomy group, although the difference was not statistically significant. The self assessed eating ability 12 months after surgery was significantly higher in the marginal mandibulectomy group than the segmental mandibulectomy group. These results suggest that discontinuation of the mandible may lead patients to eat only foods that do not require a substantial amount of chewing. Hence, the quality of life of patients in the marginal mandibulectomy group was considered to be better than that in the segmental mandibulectomy group.