Effects of transforming growth factor beta 1 on the plasminogen activation system, collagen and integrin synthesis, and proliferation of rabbit mandibular condylar chondrocytes.

The objective of this study was to identify the mechanism by which mandibular condyle chondrocytes regulate the extracellular matrix. Primary rabbit condylar chondrocytes were isolated, cultured, and treated with transforming growth factor beta 1 (TGF-ß1). Cells were then assayed for the following: urokinase-type plasminogen activator (uPA) and its inhibitor (PAI-1), collagen types I and II, ß1 integrin expression, and proliferative activity. TGF-ß1 induced synthesis of collagen type II, αVß1 integrin, and PAI-1. TGF-ß1 induced the growth of chondrocytes and suppressed the synthesis of uPA. Chondrocyte regulation of the extracellular matrix is mediated by TGF-ß1. Synthesis of collagen type II, αVß1 integrin, and PAI-1 is induced, while uPA is suppressed. Also, TGF-ß1 induces cellular growth.

The retromandibular transparotid approach for reduction and rigid internal fixation using two locking miniplates in mandibular condylar neck fractures.

We evaluated the safety, efficacy, and morbidity associated with the treatment of displaced mandibular condylar neck fractures using a retromandibular transparotid approach to reduce and rigidly fix using two 2.0-mm locking miniplates. Our surgical inclusion criteria were: patient selection of open reduction and fixation, displaced unilateral condylar fractures with derangement of occlusion, and bilateral condylar fractures with an anterior open bite. The study group consisted of 19 patients who underwent surgery for 19 mandibular condylar neck fractures; patients were analyzed prospectively, with more than 6 months of follow-up, and were evaluated in terms of functional results, scar formation, postoperative complications, and stability of fixation. The results showed that functional occlusion identical to the preoperative condition and correct anatomical reduction of the condylar segments in centric occlusion, followed by immediate functional recovery, was achieved in all patients. No patient suffered from any major or permanent complication postoperatively, although there were two cases (11%) of temporary facial nerve palsy, which resolved completely within 3 months. Surgical scars were barely visible. The retromandibular transparotid approach with open reduction and rigid internal fixation for displaced condylar neck fractures of the mandible is a feasible and safe, minimally invasive surgical technique that provides reliable clinical results.

Applicability of buccal fat pad grafting for oral reconstruction.

This study evaluated the applicability of pedicled buccal fat pad grafting for the reconstruction of defects surgically created during oral surgery. A buccal fat pad graft was applied in 23 patients (5 males, 18 females; mean age 68.3 years) between 2003 and 2011. The graft was used to cover surgical defects of the palate, maxilla, upper gingiva, buccal mucosa, lower gingiva, oral floor, and temporomandibular joint region. Size of the surgical defects ranged from 15mm×12mm to 30mm×40mm; size of the buccal fat pad ranged from 15mm×12mm to 43mm×38mm. A pedicled buccal fat pad was prepared by incising the maxillary vestibule following primary surgery, and the surrounding connective tissue was preserved to supply nutrition to the pedicle during surgery. The buccal fat pad was placed on the raw surface of soft tissue or bone surface and sutured to the surrounding tissue of the defect. Complete epithelialization was observed within 4 weeks postoperatively. There were no complications or functional disorders during follow-up. Buccal fat pad grafting appears to be feasible for the reconstruction of surgically induced defects, and can be extended to the palate, mandible, mouth angle, and temporomandibular joint region.

FAP-1 and NF-κB expressions in oral squamous cell carcinoma as potential markers for chemo-radio sensitivity and prognosis.

This study was designed to investigate the feasibility of using Fas-associated phosphatase-1 (FAP-1), nuclear factor kappa B (NF-κB) and p53 as markers for chemo-radio sensitivity in oral squamous cell carcinoma (OSCC). FAP-1 plays a role as an anti-apoptotic factor through Fas-dependent apoptosis after chemo-radiotherapy. NF-κB and p53 might be involved in modulation of FAP-1 expression. FAP-1, NF-κB and p53 expression were immunohistochemically examined using biopsy specimens in 50 OSCC patients treated with chemotherapy and/or radiotherapy. FAP-1 was expressed in 52%, NF-κB in 52% and p53 in 46% of patients. There was no significant difference in FAP-1, p53 or NF-κB expression according to the clinicopathological features. No correlation was found among FAP-1, p53 or NF-κB expression. FAP-1-positive cases showed a poorer survival rate than FAP-1-negative cases (P = 0.0409) and NF-κB-positive cases showed a poorer survival rate than NF-κB-negative cases (P = 0.0018). Multivariate analysis showed that FAP-1 expression, NF-κB expression, clinical stage and age were significant independent variables for survival (clinical stage: P = 0.0016; age: P = 0.0016; NF-κB: P = 0.0314; FAP-1: P = 0.0366). These results suggest that FAP-1 and NF-κB might play a role as chemo-radioresistant factor during chemo-radiotherapy, and FAP-1 and NF-κB expression in OSCC would be feasible markers for chemo-radio sensitivity and prognosis.

Treatment modalities for oral verrucous carcinomas and their outcomes: contribution of radiotherapy and chemotherapy.

BACKGROUND: This clinical study focused, firstly, on the results of treatment and, secondly, on the anaplastic transformation, of oral verrucous carcinomas (OVCs) diagnosed and treated from 1981 to 1997 at the Department of Oral and Maxillofacial Surgery at Shimane Medical University Hospital. METHODS: We analyzed the treatment modalities and outcomes for 15 patients with OVC. RESULTS: Excluding the results for 4 palliatively treated patients, the disease-free survival rates of the patients after the initial treatments, were 82% at 5 years and 66% at 10 years; for all 15 patients, these rates were 57% and 46%, respectively. Surgery alone and surgery combined with other treatments (such as radiotherapy and chemotherapy) appeared to yield disease-free survival rates to those achieved superior with other treatments whether single or combined; (78% vs 33% for 5-year disease-free survival; 52% vs 33% for 10-year disease-free survival); however, the difference was not significant (P = 0.47). Well differentiated squamous cell carcinomas (W-SCCs) (n = 5) as well as spindle cell carcinoma (n = 1) were found in subsequent operative or biopsy specimens. CONCLUSION: Surgery was the most reliable treatment method for OVC; however, radiotherapy combined with chemotherapy was the next most preferable treatment when surgery was not undertaken. We also found that highly malignant transformation (anaplastic transformation) occasionally occurred during treatments for OVC.

Nitric oxide regulates actin reorganization through cGMP and Ca(2+)/calmodulin in RAW 264.7 cells.

Nitric oxide (NO) has been reported to be involved in the regulation of pseudopodia formation, phagocytosis and adhesion in macrophages through the reorganization of actin. In the present study, we directly separated the globular (G) and filamentous (F) actin from quiescent or NO-stimulated macrophage-like cell line RAW 264.7 cells in order to investigate the dynamic redistribution of actin pools. We also focused on the regulatory mechanisms of actin assembly, induced by NO and its possible subsequent signaling pathway. We showed that predominant G-actin coexisted with Triton X-100-insoluble filamentous (TIF) and Triton X-100-soluble filamentous actin in resting RAW 264.7 cells. The exogenous NO produced by (+/-)-(E)-2-[(E)-hydroxyimino]-6-methoxy-4-methyl-5-nitro-3-hexenamide (NOR1), the endogenous NO induced by lipopolysaccharide (LPS) plus interferon-gamma (IFNgamma), and dibutyryl-cGMP increased the contents of TIF-actin in dose- and time-dependent manners and altered its morphology. The increase in the TIF-actin contents induced by NOR1 or LPS plus IFNgamma was efficiently blocked by the radical scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide and the soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one or the arginine analogue N(G)-monomethyl-L-arginine acetate, respectively. Preincubation with the calmodulin antagonist W-7 almost completely blocked the NO-induced TIF-actin increase and morphological change. On the other hand, preincubation with C3 transferase, an inhibitor of Rho protein, efficiently prevented the change in cell morphology, but had no effect on the TIF-actin increase. We postulate that cGMP and subsequent Ca(2+)/calmodulin may be key regulators of actin reorganization in NO-stimulated RAW 264.7 cells.

The effect of a traditional Chinese prescription for a case of lung carcinoma.

OBJECTIVE: To examine the effectiveness of Ninjin Yoei To (NYT; Ren-Shen-Yang-Rong-Tang in Chinese medicine; Kotaro Pharmaceutical Co., Ltd., Osaka, Japan), one of the traditional herbal medicines, against lung carcinoma. SETTING: The Nursing Center Himawari DESIGN, PATIENT, AND PREPARATION: The regular dosage of NYT (15 g/d) was prescribed for 7 weeks to one elderly patient with lung carcinoma. The daily standard dose of NYT is prepared from dried extract obtained from 12 crude natural substances, ginseng, cinnamon bark, Japanese angelica root, astragalus root, peony root, citrus unshiu peel, rehmannia root, polygala root, atractylodes rhizome, schisanda fruit, poria sclerotium, and glycyrrhiza. NYT is certified by the Japanese Ministry of Health and Welfare. RESULTS: The tumor marker levels (CEA and CA19-9) decreased and the scores of yin-yang and xu-shi inverted from negative and positive during 7 weeks. The patient's cough disappeared and her appetite recovered. CONCLUSION: NYT has a positive effect on life expectancy for patients with malignancy. The diagnostic scoring system in yin-yang and xu-shi and prescription of Chinese herb may be available to gain control over a patient's health.

Production and characterization of a monoclonal antibody specific for ubiquitin-like polypeptide responsible for nonspecific immune suppression.

Monoclonal nonspecific suppressor factor (MNSF) is a lymphokine product of a murine T cell hybridoma that inhibits the immune response in an antigen nonspecific manner. Recently, we found that a novel ubiquitin-like protein (Ubi-L), a subunit of MNSF, is responsible for its biological activity. We developed a monoclonal antibody with specific activity against Ubi-L. Inhibition experiments showed that this mAb, termed NA4, preferentially recognizes Ubi-L but not irrelevant proteins such as ubiquitin. With the use of NA4, we established an ELISA method for the quantitation of Ubi-L. By this ELISA system, approximately 40 ng/ml of MNSF was detected in the culture supernatants of concanavalin A (Con A)- or interferon gamma (IFN gamma)-activated splenocytes, whereas MNSF in the supernatant of IFN alpha- and IFN beta-stimulated splenocytes was nil. In addition, NA4 could abrogate the action of Ubi-L. Thus NA4 was confirmed to be a pertinent tool for elucidation of the underlying mechanism of action of MNSF.

Monoclonal nonspecific suppressor factor beta (MNSF beta) inhibits the production of TNF-alpha by lipopolysaccharide-activated macrophages.

The monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by a murine T cell hybridoma, shows a pleiotropic antigen-nonspecific suppressive function. Most recently, a cDNA encoding a subunit of MNSF (MNSF beta) has been isolated and characterized. Recombinant form of MNSF beta (rMNSF beta) inhibits lymphokine functions, as does native MNSF. In this study, we investigated whether rMNSF beta also affects macrophage function in terms of LPS-induced TNF-alpha production by a mouse macrophage cell line, J774. rMNSF beta suppressed the TNF-alpha production in a dose-dependent manner. This suppressive effect was remarkably reduced when rMNSF beta was added after 6 h of LPS stimulation. In addition, enhancement of TNF-alpha production by IFN-gamma was also suppressed by rMNSF beta. The suppressive effect was partly neutralized by the addition of the serine/threonine phosphatase inhibitor, okadaic acid. This finding suggests that serine/threonine protein phosphatases type 1 and/or 2A may be implicated in the mechanism of action of MNSF.