Speech evaluation and dental arch shape following pushback palatoplasty in cleft palate patients: Supraperiosteal flap technique versus mucoperiosteal flap technique.

BACKGROUND: The aim of this study was to evaluate and compare the maxillary dental arch shape and speech of cleft palate patients following pushback palatoplasty using either the supraperiosteal flap technique or the mucoperiosteal flap technique. PATIENTS: Sixty-two patients (29, cleft palate only; 33, unilateral cleft lip, alveolus and palate) operated on by the supraperiosteal technique and 47 patients (23, cleft palate only; 24 unilateral cleft lip, alveolus and palate) by the mucoperiosteal technique were reviewed in this study. STUDY DESIGN: Dental arch shape and speech proficiency at preschool and school age were evaluated in all patients. RESULTS: Dental arch shapes were classified as U type (good dental arch shape) and V type (narrow dental arch shape). In cleft palate only patients, U type was observed in 90% of the supraperiosteal group and 83% of the mucoperiosteal group. In unilateral cleft lip, alveolus and palate patients, U type was observed in 85% of the supraperiosteal group, while only in 33% of the mucoperiosteal group. In cleft palate only patients, normal speech at school age was observed 100% of the supraperiosteal group and 83% of the mucoperiosteal group. In unilateral cleft lip, alveolus and palate patients, normal speech at school age was observed in 97% of the supraperiosteal group and 75% of the mucoperiosteal group. Misarticulation was frequently found in patients with the V type of dental arch shape. CONCLUSION: It is suggested that pushback palatoplasty using the supraperiosteal technique is more advantageous for speech development compared with the mucoperiosteal technique.

Enhanced Bax in oral SCC in relation to antitumor effects of chemotherapy.

BACKGROUND: Antitumor effects of chemotherapeutic agents are commonly associated with the induction of apoptosis. Bax belongs to the Bcl-2 family and induces apoptosis. The present study was conducted to investigate the relationship between enhanced Bax expression in oral squamous cell carcinoma (SCC; cell lines and clinical cases) and the antitumor effects of chemotherapy. METHODS: In three oral SCC cell lines, Bax expression before and after treatment with chemotherapeutic agents [docetaxel (TXT), cisplatin and 5-fluorouracil] was examined by reverse transcriptase-polymerase chain reaction and immunoblotting. The effects of treatment were assessed by counting the number of viable cells and determining sub-G1 cells. Tissue samples (both biopsy specimens before chemotherapy and surgically excised specimens after chemotherapy) from nine patients with oral SCC who underwent neoadjuvant chemotherapy were immunostained for Bax. The relationship between enhancement of Bax expression and chemotherapeutic effects was established. RESULTS: Two of three cell lines did not express Bax mRNA or protein before treatment. After treatment, Bax expression was enhanced only by TXT in one cell line, but by all chemotherapeutic agents in the other two cell lines. In three of nine patients, Bax expression was not found before chemotherapy. Two of these three patients showed enhanced Bax expression after chemotherapy including TXT, but one still failed to express Bax. Both in cell lines and clinical cases, enhancement of Bax after chemotherapy was associated with antitumor effects. CONCLUSION: Certain chemotherapeutic agents enhance Bax expression in oral SCC, and it is suggested that this contributes to the antitumor effects of chemotherapy.

Comparative histology of lymph nodes from aged animals and humans with special reference to the proportional areas of the nodal cortex and sinus.

Lymph nodes are composed of a lymphocyte-rich area or cortex (subdivided into the superficial and deep cortex and the medullary cord) and another, macrophage-rich area (incorporating the subcapsular and medullary sinuses). We measured the proportional area of the cortex in lymph nodes from aged experimental mammals (rats, guinea pigs, dogs and rabbits) and elderly Japanese humans. The cervical, axillary and inguinal nodes were generally richer in cortex tissue than the pulmonary regional and mesenteric nodes. Histological heterogeneity and medullary sinus dominance were much more evident in the human nodes than in those from animals, except for the guinea pig thoracic node. Human pulmonary regional nodes were characterized by a large medullary sinus; in guinea pigs, these nodes had a similar histology but the T lymphocyte-containing areas were smaller and thinner than in humans. The paraaortic node was well developed in humans and dogs, but not in other animals tested. These species- and region-specific histological differences may influence the evaluation of experimental animal models of lymph node function, such as those recently identified for research into sentinel nodes.

Variations in the tensor veli palatini muscle with special reference to its origin and insertion.

OBJECTIVE: Previous research on the tensor veli palatini muscle (TVP) has produced conflicting descriptions of its functions and topographical relationships with other orofacial structures. The goal of this study was to describe the morphology of the TVP in a systematic and comprehensive manner. METHODS: One hundred nineteen sides of 77 human heads from donated cadavers were partially dissected under a binocular microscope. Histological examination of the hard tissue-muscle interfaces was also undertaken. RESULTS: There were two adjacent origins of the TVP: the cranial base origin (CB origin) and the auditory tube cartilage origin (AT origin). The CB origin always lay anterior to the AT origin and there was no septum or loose tissue between the two muscular laminae leading from these origins. The muscle fibers converged on a central tendinous plate in the muscle belly, which gradually became a common tendon that rounded the pterygoid hamulus before inserting into the palatine aponeurosis. Notably, secondary insertions were found on the maxillary tuber (33.6%) and/or in the submucosal tissue near the palatoglossal arch (37.8%). Maxillary insertions were almost exclusively associated with an AT origin that was wide as or wider than the CB origin. Histological observations confirmed that the hamulus acted purely as a pulley and suggested that a connecting band to the tensor tympani had no or few functions of an intermediate tendon. CONCLUSIONS: The TVP appears to act as the dilator tubae and that this function can be maintained by preserving or reconstructing the maxillary insertion during push-back surgery, even if hamulotomy is necessary.

Comparison of palatal sensitivity after treatment of cleft palate by a supraperiosteal or mucoperiosteal flap.

We compared palatal sensitivity after push-back palatoplasty in patients operated on with a conventional mucoperiosteal flap and with a supraperiosteal flap. We studied 37 patients, whose mean age was 18 years (range 11-28), with unilateral cleft lip, alveolus, and palate who required palatoplasty at our clinic; 18 patients had a supraperiosteal, and 19 a mucoperiosteal flap. Thresholds of palatal sensitivity were measured with Semmes-Weinstein monofilaments. The mean values of thresholds of palatal sensitivity in patients who had a push-back palatoplasty were higher than those in a normal control group. The mean values for patients who had a supraperiosteal flap were significantly lower than those who had a mucoperiosteal flap.

Sternocleidomastoid myocutaneous flap for intraoral reconstruction after resection of oral squamous cell carcinoma.

PURPOSE: The sternocleidomastoid (SCM) myocutaneous flap remains an important tool in head and neck reconstruction. This article retrospectively reviews 40 consecutive SCM myocutaneous flaps used for the reconstruction after resection of oral squamous cell carcinoma with respect to reliability and complications. PATIENTS: From 1987 to 1997, 40 patients underwent SCM myocutaneous flap reconstruction of the oral cavity. The age and gender of the patients, site of primary tumor TNM stage, type of associated operation, and clinical course were analyzed. RESULTS: In 8 cases, partial epithelial loss over the skin paddle occurred with survival of the muscle and at least some of the dermis. Unilateral supraomohyoid neck dissection (SND) was performed in 11 cases, and unilateral functional neck dissection, which preserves SCM and/or internal jugular vein and/or accessory nerve, in 16 cases. Pathologically positive nodes were recognized in 14 of these 27 neck dissection cases; in 11 of these 14 cases, the neck lesion was controlled. CONCLUSION: The SCM myocutaneous flap appears to be simple to use and useful for reconstruction of the defect after resection of oral carcinoma, and the indications for this flap will be extended in accordance with the recent increases in the number of supraomohyoid and functional neck dissection cases.

Afferent and efferent lymph-collecting vessels of the submandibular nodes with special reference to the lymphatic route passing through the mylohyoid muscle.

BACKGROUND: Although metastasis of cancer in the oral region to the submandibular node is well described, there has been no anatomic representation of lymph vessels penetrating the oral floor and draining into the node. MATERIALS AND METHODS: Ninety specimens were obtained from formalin-fixed, donated cadavers. Histologic observations using serial sections followed the macroscopic observations. RESULTS: In 19 of 90 specimens, we found afferent collecting lymph vessels exiting from the mylohyoid surface and draining into the preglandular submandibular node. In 3 of the 19 specimens, collecting vessels passing through the narrow muscle gap with or without arteries, veins, and nerves were identified histologically. The postglandular submandibular node was not evident in the drainage route. CONCLUSIONS: Although it carries a low incidence, because of the direct lymphatic route or pathway between the oral region and preglandular submandibular node, the pathologically positive supraomohyoid node sometimes seems to be found even in elective neck dissection. However, we speculate that sentinel node investigation would reveal the much more critical role of the jugulodigastric node not only as the actual sentinel node but also as the common terminal node along the various drainage routes from the oral region.

Enhanced antitumor effect of RGD fiber-modified adenovirus for gene therapy of oral cancer.

Current clinical success rates of adenoviral vector (Adv)-based gene therapy of squamous cell carcinoma (SCC) of the head and neck remain unsatisfactory. A major problem with this approach is thought to be related to low Adv transduction efficiency due to weak expression of the adenovirus receptor, coxsackie-adenovirus receptor (CAR), in SCC. To improve the limited infectivity of Adv in oral SCC, we constructed mutated Adv incorporating the integrin-binding motif, RGD, in the HI loop of the fiber knob. The mutated Adv infected target cells through integrins commonly expressed in oral SCC. LacZ marker gene expression after infection with this mutated Adv (Adv-F/RGD) in oral SCC cell lines that showed reduced expression of CAR was approximately 5-10 times higher than that obtained with the parental Adv containing wild-type fiber knob (Adv-F/wt). In an in vitro study, transduction of oral cancer cell lines with Adv-F/RGD expressing human IL-2 (AxCAhIL2-F/RGD) resulted in greater production of cytokine than AxCAhIL2-F/wt infection. In an in vivo therapeutic xenograft model of oral SCC in nude mice, AxCAhIL2-F/RGD demonstrated antitumor effects superior to those of AxCAhIL2-F/wt. These data suggest that exploitation of genetically altered adenovirus vectors with integrin-binding motifs may offer significant improvements in oral SCC gene therapy.

Aberrant methylation of multiple genes and clinicopathological features in oral squamous cell carcinoma.

The purpose of this study was to examine the methylation profile of various oral squamous cell carcinomas and to correlate the methylation of particular chromosomal loci with the clinicopathological features of the tumors. A semiquantitative analysis of the methylation status of 12 loci in 96 primary tumors and 13 cell lines was carried out. Methylation frequency was calculated as the percentage of methylated alleles detected by bisulfate-PCR. Of the 12 loci examined, 9 (p16INK4A, p15INK4B, p14ARF, DCC, DAP kinase, MINT1, MINT2, MINT27, and MINT31) exhibited aberrant methylation at various frequencies, whereas 3 (hMLH1, HRK, and CACNA1G) showed no methylation. Dense methylation of the 5' CpG island of DAP kinase and MINT1 was well correlated with loss of gene expression. In addition, methylation of DCC was correlated with bone invasion by gingival tumors (P = 0.036), with aggressive invasiveness of tumors of the tongue (P = 0.046), and with reduced survival (P = 0.050). Methylation of MINT1 and MINT31 also correlated with poor prognoses (P = 0.058 and 0.041), whereas methylation of p14ARF correlated with a good prognosis (P = 0.021). Cox regression analysis showed methylation of MINT31 to be an independent predictor of outcome (hazard ratio, 3.79; 95% confidence interval, 1.58-9.10) and to be associated with the T4 disease group (hazard ratio, 5.71; 95% confidence interval, 1.25-26.07). Analysis of DNA methylation is a useful approach to evaluation of the biological characteristics of oral cancers and may be a useful diagnostic indicator of patient prognosis.

Invasive front in oral squamous cell carcinoma: image and flow cytometric analysis with clinicopathologic correlation.

OBJECTIVE: Pathologists have drawn attention to the invasive tumor front (ITF) in the determination of the biologic aggressiveness of oral cancer. We have attempted to discover the prognostic significance of cancer cells with abnormal DNA content at the ITF of oral squamous cell carcinoma. STUDY DESIGN: A comparative DNA analysis by means of image cytometry and flow cytometry was conducted to confirm the usefulness of image cytometry in detecting cancer cells having abnormal DNA content at the ITF. The prognostic value of cancer cells with abnormal DNA content ws examined by a multivariate analysis for 195 patients with oral squamous cell carcinoma. RESULTS: In the comparative DNA analysis, it was suggested that image cytometry is useful for detecting cancer cells with abnormal DNA content (4c exceeding rate [4cER]), which is associated with poor prognosis of patients with oral squamous cell carcinoma. In the multivariate analysis, 3 independent factors were found to significantly influence cause-specific survival. These are, in decreasing order of influence, (1) abnormal DNA content (4cER), (2) clinical stage, and (3) growth type. CONCLUSION: The presence of cancer cells with abnormal DNA content of the ITF in conjunction with clinical findings (clinical stage and growth type) can give additional useful information when selecting treatment strategies for oral cancer patients.

Control of cell proliferation kinetics of tumor in neoadjuvant chemotherapy for advanced oral squamous cell carcinoma and its prognostic implications.

BACKGROUND: Few attempts have so far been made at studies of the cell proliferation kinetics of the tumor in neoadjuvant chemotherapy for head and neck cancer. We examined the effects of neoadjuvant chemotherapy for advanced oral squamous cell carcinoma in terms of the cell proliferation kinetics of the tumor, and attempted to correlate them with patients' survival. METHODS: Fifty-two patients with advanced oral squamous cell carcinoma who received neoadjuvant chemotherapy followed by surgery participated in this study. Cellular DNA content and mitotic index (MI) in tissue samples were measured before and after chemotherapy, using a cell image analyzer. RESULTS: A decrease in both mean DNA content (mean DNA) and MI (left-shift type of change in cell growth kinetics), indicating an accumulation of cancer cells in the G0-G1 phase, was found in 25 patients. An increase in mean DNA with decreased MI (right-shift type), which was found in 13 patients, appeared to be correlated with an accumulation of cancer cells in the S-G2 phase. Neither of these two types of change, which were considered to be a favorable effect, were found in 14 patients (ineffective type). Excellent survival rates were obtained in patients who showed favorable changes in cell growth kinetics (79% for patients with left-shift type and 92% for patients with right-shift type), whereas the survival rate for patients with the ineffective type was extremely poor (14%). The type of change in cell proliferation kinetics was a powerful independent prognostic indicator. CONCLUSION: Analysis of cell growth kinetics appears to be useful not only as a diagnostic tool to predict patient outcome but also as a means to infer the chemotherapeutic effects in oral squamous cell carcinoma.

Natural antigenic peptides from squamous cell carcinoma recognized by autologous HLA-DR8-restricted CD4+ T cells.

A large number of human tumor antigens recognized by CD8+ cytotoxic T lymphocytes (CTL) have been identified. Some of them have been employed in clinical trials and have achieved some objective responses. However, little is known about those that are recognized by CD4+ T cells, except for a very few that were identified from melanomas. Previously, we reported that an oral squamous cell carcinoma (SCC) cell line, OSC-20, was effectively lysed by HLA-DRB1*08032 (HLA-DR8)-restricted autologous CD4+ T cell line, TcOSC-20. In this study, we performed two steps of chromatographic purification of the tumor cell lysate in combination with mass spectrometry. We found one reverse-phase high-performance liquid chromatography (RP-HPLC) fraction that was effectively recognized by the T cells. We analyzed the fraction by nano-liquid chromatography/electrospray ionization ion trap mass spectrometry (LC/MS/MS) and found six representative ions. We could determine the primary amino acid sequence of each of the six ions. Three of them contained a potential HLA-DR8 binding motif, and TcOSC-20 showed a rather strong cytotoxic response to one of the synthetic peptides, namely, amino acid residues 321-336 of human alpha-enolase. Thus, several gene products of squamous cancer cells are endogenously processed and may be presented on HLA class II molecules, so that they could constitute target molecules for autologous CD4+ T cells.