Posterior Displacement of Meniscal Ramp Lesion Is Observed on Flexed Knee Magnetic Resonance Imaging of Anterior Cruciate Ligament-Deficient Knees.

PURPOSE: To confirm whether posterior displacement of the detached meniscocapsular structure in ramp lesions could be observed on magnetic resonance imaging (MRI) in the flexed knee position and to clarify whether this displacement on MRI is related to greater anterior knee laxity. METHODS: This retrospective comparative trial study included 50 patients with anterior cruciate ligament injuries in whom attendant ramp lesions were observed on preoperative MRI. All patients underwent 3-T MRI at 120° of knee flexion preoperatively. The gap distance of the ramp lesion on sagittal slices was measured, and a distance greater than 1 mm was classified as gap positive. Additionally, joint effusion in the posteromedial recess was quantitatively evaluated by measuring the surface area of joint fluid. Prior to surgery, the side-to-side difference in anterior tibial translation (ATT) on stress radiographs at 20° of knee flexion was measured to evaluate anterior knee laxity. During anterior cruciate ligament reconstruction, the length of the ramp lesion was measured by a scale from the posteromedial portal. RESULTS: Of the 50 knees, 29 had ramp lesion gaps greater than 1 mm on MRI at knee flexion. The gap-positive group had a significantly greater side-to-side difference in ATT (9.6 ± 3.6 mm vs 5.0 ± 2.7 mm, P < .001) and significantly larger ramp lesion length (15.7 ± 4.9 mm vs 11.7 ± 3.9 mm, P = .002) than the gap-negative group. The gap-positive group also had a significantly greater amount of joint effusion than the gap-negative group (P < .001). CONCLUSIONS: Posterior displacement of the detached meniscocapsular structure in ramp lesions can be observed on MRI at knee flexion and is related to greater anterior knee laxity, lesion size, and joint effusion. LEVEL OF EVIDENCE: Level III, retrospective comparative trial.

Magnetic Resonance Imaging Diagnosis of Medial Meniscal Ramp Lesions in Patients With Anterior Cruciate Ligament Injuries.

PURPOSE: To prospectively evaluate the sensitivity and specificity of magnetic resonance imaging (MRI) for diagnosing ramp lesions, to compare them between 1.5- and 3-T MRI, and to evaluate whether bone contusion of the posterior lip of the medial tibial plateau was associated with ramp lesions. METHODS: For 155 knees that underwent primary ACL reconstruction, we prospectively examined for ramp lesions and medial meniscal body tears on MRI. MRI diagnosis of ramp lesions required high signal irregularity of the capsular margin or separation in the meniscocapsular junction of the medial meniscus posterior horn on sagittal images. Bone contusion of the posterior lip of the medial tibial plateau was verified in 105 knees with MRI performed within 6 weeks after injury. All ramp lesions were identified by transcondylar observation during surgery. The sensitivity and specificity of MRI for ramp lesions and body tears were measured. Furthermore, we evaluated whether bone contusion of the medial tibial plateau was associated with ramp lesions. The χ-square test was used for statistical analysis. RESULTS: During surgery, ramp lesions were observed in 46 knees and medial meniscal body tears were seen in 35 knees. The sensitivity of MRI for ramp lesions was 71.7% and specificity was 90.5%. The sensitivity for ramp lesions was significantly lower than that for meniscal body tears (94.3%) (P = .01). The sensitivity of 3-T MRI (83.3%) was superior to that of 1.5-T MRI (67.6%), but not significantly different. The incidence of bone contusions was not significantly different among ramp lesions (38.5%), body tears (40.0%), or no tears (30.5%). CONCLUSIONS: The sensitivity of MRI for diagnosing ramp lesions was significantly lower than that for medial meniscal body tears. Bone contusion of the posterior lip of the medial tibial plateau on MRI was not associated with ramp lesions. LEVEL OF EVIDENCE: Level III, comparative trial.

Expression and distribution of hyaluronic acid and CD44 in unphonated human vocal fold mucosa.

OBJECTIVES: The tension caused by phonation (vocal fold vibration) is hypothesized to stimulate vocal fold stellate cells (VFSCs) in the maculae flavae (MFe) to accelerate production of extracellular matrices. The distribution of hyaluronic acid (HA) and expression of CD44 (a cell surface receptor for HA) were examined in human vocal fold mucosae (VFMe) that had remained unphonated since birth. METHODS: Five specimens of VFMe (3 adults, 2 children) that had remained unphonated since birth were investigated with Alcian blue staining, hyaluronidase digestion, and immunohistochemistry for CD44. RESULTS: The VFMe containing MFe were hypoplastic and rudimentary. The VFMe did not have a vocal ligament, Reinke's space, or a layered structure, and the lamina propria appeared as a uniform structure. In the children, HA was distributed in the VFMe containing MFe. In the adults, HA had decreased in the VFMe containing MFe. In both groups, the VFSCs in the MFe and the fibroblasts in the lamina propria expressed little CD44. CONCLUSIONS: This study supports the hypothesis that the tensions caused by vocal fold vibration stimulate the VFSCs in the MFe to accelerate production of extracellular matrices and form the layered structure. Phonation after birth is one of the important factors in the growth and development of the human VFMe.

Modifiable Lifestyle Factors and Cognitive Function in Older People: A Cross-Sectional Observational Study.

Background: The development of evidence-based interventions for delaying or preventing cognitive impairment is an important challenge. Most previous studies using self-report questionnaires face problems with reliability and consistency due to recall bias or misclassification among older people. Therefore, objective measurement of lifestyle components is needed to confirm the relationships between lifestyle factors and cognitive function. Aims: The current study examined the relationship between lifestyle factors collected with wearable sensors and cognitive function among community-dwelling older people using machine learning. Methods: In total, 855 participants (mean age: 73.8 years) wore a wristband sensor for 7.8 days on average every 3 months. Various lifestyle parameters were measured, including walking steps, conversation time, total sleep time (TST), sleep efficiency, time awake after sleep onset, awakening count, napping time, and heart rate. Random forest (RF) regression analysis was used to examine the relationships between total daily sensing data and Mini-Mental State Examination (MMSE) scores. Confounding factor analysis was conducted with models that were adjusted and unadjusted for demographic and vascular risk factors, and selected variables were assessed as risk and protective factors using partial dependence plots (PDPs). Results: Lifestyle data were collected for 31.3 ± 7.1 days per year using wristband sensors. RF regression analysis adjusted for age, gender, and education levels selected four variables, including number of walking steps, conversation time, TST, and heart rate. Moreover, walking steps, conversation time, and heart rate remained after RF regression analysis adjusted for demographic and vascular risk factors. Number of walking steps, conversation time, and heart rate were categorized as protective factors, whereas TST was categorized as a risk factor for cognitive function. Although PDPs of number of walking steps and heart rate revealed continuously increased MMSE scores, those of conversation time and TST and revealed that the tendency in the graph was reversed at the boundary of a particular threshold (321.1 min for conversation time, 434.1 min for TST). Conclusions: Lifestyle factors, such as physical activity, sleep, and social activity appear to be associated with cognitive function among older people. Physical activity and appropriate durations of sleep and conversation are important for cognitive function.

Evaluation of a deep brain stimulation electrode for laryngeal pacing.

OBJECTIVES: The purpose of the present study was to evaluate the suitability of a deep brain stimulation electrode for laryngeal pacing. Of interest was whether the smaller and more closely spaced channels could provide sufficient channel redundancy, controlled current distribution, and discrete activation of the posterior cricoarytenoid (PCA) muscle. METHODS: A study was conducted in dogs under differing states of PCA muscle innervation representing complete denervation to complete synkinetic reinnervation. In 3 animals, stimulated glottal opening was assessed in the innervated state and after chemical denervation by pancuronium bromide. In 3 additional dogs, the left side of the larynx was surgically denervated and compared to the innervated, right side to study an anatomic model of clinical paralysis. RESULTS: The thresholds were lower and the maximum level of abduction was greater for the innervated state. The stimulated glottal opening equaled that of a spontaneously breathing animal. Abductory responses were obtained across all channels in the array, demonstrating its anatomic and physiological compatibility for this application. In the denervated state, responses were only 20% of that of the innervated state with a pulse duration of 0.5 ms. The response could be enhanced to 40% and 60% by increasing the pulse duration to 1 and 2 ms. CONCLUSIONS: A deep brain stimulation electrode could effectively reanimate the PCA muscle to a normal level in a case of synkinetic reinnervation and to as much as 60% of the normal level in a case of complete denervation.

Histopathologic investigations of the unphonated human vocal fold mucosa.

CONCLUSION: Vocal fold vibration (phonation) after birth is one of the important factors in the growth and development of the human vocal fold mucosa. OBJECTIVES: Stellate cells in the maculae flavae located at both ends of the vocal fold mucosa are inferred to be involved in the metabolism of extracellular matrices. Maculae flavae are also considered to be an important structure in the growth and development of the human vocal fold mucosa. Tension caused by phonation (vocal fold vibration) is hypothesized to stimulate stellate cells to accelerate production of extracellular matrices. Vocal fold mucosae unphonated since birth were investigated histologically. SUBJECTS AND METHODS: Vocal fold mucosae, which were unphonated since birth, of three younger adults (17, 24, 28 years old) were investigated by light and electron microscopy. RESULTS: Vocal fold mucosae were hypoplastic and rudimentary and did not have a vocal ligament, Reinke's space or a layered structure. The lamina propria appeared as a uniform structure. Some stellate cells in the maculae flavae showed degeneration. Not many vesicles were present at the periphery of the cytoplasm. The stellate cells synthesized fewer extracellular matrices, such as fibrous protein and glycosaminoglycan. Cytoplasmic processes of the stellate cells were short and shrinking. The stellate cells appeared to have decreased activity.

Overexpression of small ubiquitin-related modifier-1 and sumoylated Mdm2 in oral squamous cell carcinoma: possible involvement in tumor proliferation and prognosis.

The purpose of this research was to identify a molecular clue to tumor proliferation in oral squamous cell carcinoma (SCC) and to test the value as a predictive marker for prognosis. In cDNA array analysis, small ubiquitin-related modifier-1 (SUMO-1) was expressed at much higher levels in oral SCC tissue and oral SCC cell lines than normal oral epithelium. The result was confirmed by RT-PCR analysis and Western blot analysis. Transfection of the anti-SUMO-1 antisense oligonucleotide to oral SCC cells significantly reduced proliferation of the cells. Immunoprecipitation and Western blot analyses revealed that the oncoprotein Mdm2 was present predominantly as a form of SUMO-1 congestion (sumoylation) rather than as a non-sumoylated form in both oral SCC tissues and cell lines. Immunohistological analysis revealed that patients who showed coexpression of SUMO-1 and Mdm2 experienced more frequently local recurrence after initial treatments. Multivariate analysis confirmed that the dual-high expression of SUMO-1 and Mdm2 was an independent factor for local failure. These result suggested that overexpression of Mdm2 caused by overexpression of SUMO-1 may be involved in tumor aggressiveness even in patients with early stage oral SCC. SUMO-1 may be useful as a novel target for therapy in oral SCC as well as a clinical indicator for tumor recurrence together with Mdm2.

Inducible nitric oxide synthase expression in various laryngeal lesions in relation to carcinogenesis, angiogenesis, and patients' prognosis.

CONCLUSIONS: The inducible nitric oxide synthase (iNOS) expression leading to vascular endothelial growth factor (VEGF) overexpression may be useful as a factor for predicting recurrence after initial treatment and prognosis in laryngeal squamous cell carcinoma (SCC). OBJECTIVE: We analyzed expression of iNOS, p53, and VEGF in various laryngeal lesions. MATERIALS AND METHODS: The study samples consisted of 63 SCC, 20 dysplasia, 7 polyp, and 5 normal epithelium of the larynx. The expression of iNOS, p53, and VEGF was identified by immunohistological methods. RESULTS: No positive immunostaining for iNOS, p53, and VEGF was observed in normal epithelium and polyps. In contrast, with the progression from mild/moderate dysplasia to severe dysplasia to carcinoma, their expression levels increased. In dysplasia, there was a significant positive correlation among expression of iNOS, p53, and VEGF. In SCC, iNOS expression correlated with VEGF overexpression and microvessel density, but not with p53 overexpression. In SCC, the expression of iNOS and VEGF significantly increased in patients who developed local recurrence and/or metastases after initial treatments. Kaplan-Meier analysis showed that disease-free survival was significantly shorter in patients with iNOS or VEGF expression. Multivariate analysis showed expression of iNOS and VEGF as independent indicators for poor disease-free survival.

[Therapeutic effects and prognostic factors in tonsillectomy patients with IgA nephropathy].

INTRODUCTION: Because upper respiratory tract infections, particularly tonsillitis, often precede IgA nephropathy (IgAN), IgAN is now recognized as a tonsil-related disease, and reports have shown that tonsillectomy is effective in preventing disease progression in IgAN patients. IgA may thus play an important role in IgAN pathogenesis and development. B cell activation factor belonging to the TNF family (BAFF) is expressed by monocytes, macrophages, and dendritic cells, and may be the mechanism by which macrophages and dendritic cells directly regulate human B-cell activation. BAFF acts as a potent B-cell growth factor and costimulator of immunoglobulin production, including IgA. We studied therapeutic effects and serum BAFF levels as prognostic factors. PATIENTS AND METHODS: Subjects were 41 patients undergoing tonsillectomy and followed up for at least 1 year. Serum samples were obtained from 38 with IgAN and 29 with recurrent tonsillitis as a properative control groups. The remission group consisted of patients with normal renal function without hematuria and proteinuria. Serum BAFF levels were measured by ELISA kits at our laboratory. RESULTS: Overall remission was 39.0%. The disappearance of hematuria increased for long-term follow-up groups, but proteinuria was unchanged. Serum BAFF levels among the two groups did not differ significantly. Among IgAN patients, higher BAFF levels were associated with higher serum IgA/C3 levels. The correlation was weak (p =0.026, r = 0.407). Using 3.2 ng / ml as a cut off point, we compared cumulative improvement in those with high pretreatment BAFF levels to those with low pretreatment levels. Those with high pretreatment BAFF levels had inferior cumulative improvement in hematuria. CONCLUSION: Tonsillectomy was effective in improving hematuria. BAFF appears to be a key factor in tonsillectomy among [gAN patients.

Expression of CXCR4 and its down-regulation by IFN-gamma in head and neck squamous cell carcinoma.

PURPOSE: The functional expression of CXCR4, which plays roles in cell migration and proliferation in response to its unique ligand stromal cell-derived factor-1 (SDF-1), has been reported in variety of carcinomas. However, CXCR4 expression and its functional role in head and neck squamous cell carcinomas (HNSCC) remain unclear. In this study, we investigated CXCR4 expression and analyzed its functions in HNSCC cell lines. We also attempted to regulate CXCR4 expression using cytokines, such as interleukin-1beta, tumor necrosis factor-alpha, and IFN-gamma. Finally, we investigated correlation between CXCR4 expression and clinical features in patients with HNSCC. EXPERIMENTAL DESIGN: Six HNSCC cell lines were used in this study. Reverse transcription-PCR and flow cytometry analysis were shown for CXCR4 expressions with or without stimulations of cytokines. SDF-1-mediated cell migration was assayed in Matrigel-coated chemotaxis chamber. The SDF-1-mediated cell proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The SDF-1-mediated signaling pathways were analyzed by Western blot analysis. Biopsy specimens from 56 patients with HNSCC were used for immunohistologic analysis. RESULTS: The significant CXCR4 expression was found in HSQ-89, IMC-3, and Nakamura cells. The SDF-1-mediated cell migration and proliferation were observed in CXCR4-positive cells. SDF-1 also promoted rapid phosphorylation of extracellular signal-regulated kinase 1/2 and Akt signaling pathways in CXCR4-positive cells. The SDF-1-mediated cell migration and proliferation of CXCR4-positive cells were inhibited by neutralization of CXCR4. Among three cytokines tested, IFN-gamma significantly reduced CXCR4 expression and SDF-1-induced cell migration and proliferation of CXCR4-positive cells. Immunohistologic analysis revealed that patients with advanced neck status and patients who developed distant metastases showed significantly higher CXCR4 expression, and the cause-specific survival of patients with CXCR4-expression was significantly shorter. Furthermore, multivariate analysis confirmed that CXCR4 positive was the independent factor for cause-specific death. CONCLUSION: Our results may provide an insight into future therapeutic agent that inhibits tumor metastasis and progression via down-regulating CXCR4 expression in patients with HNSCC.

Effects of injecting GABAergic agents into the medullary reticular formation upon swallowing induced by the superior laryngeal nerve stimulation in decerebrate cats.

The purpose of this study was to elucidate the role of the GABAergic system in the medullary reticular formation (MRF) in the control of swallowing. In acutely decerebrated cats (n = 12), swallowing was induced by electrical stimulation (0.3-6 V at 10-20 Hz for 10-20 s every minute) applied to the superior laryngeal nerve (SLN). The stimulus intensity was adjusted so that swallowing was induced two or four times during the period of the stimulation. Bicuculline, a GABA(A) receptor antagonist, was then injected (0.10-0.15 microl, 5 mM) into the MRF through a stereotaxically placed glass micropipette. In a total of 62 injections, 19 injections (30.6%) increased the frequency of SLN-induced swallowing when it was injected into the lateral part of the MRF corresponding to the nucleus reticularis parvocellularis (NRPv). In eight of the effective injections (42.1%) which increased the frequency of SLN-induced swallowing, SLN stimulation also induced coughing. With two injections, stimulation of the SLN-induced coughing but not facilitation of swallowing. On the other hand, an injection of 0.10-0.15 microl of 5 mM muscimol, a GABA(A) receptor agonist, into the NRPv decreased the frequency of SLN-induced swallowing. These results suggest that the NRPv neurons which are responsible for evoking swallowing are under the tonic inhibitory control of the GABAergic system.

Expressions of matrix metalloproteinases in early-stage oral squamous cell carcinoma as predictive indicators for tumor metastases and prognosis.

PURPOSE: Matrix metalloproteinase (MMP)-2 and MMP-9 are considered to play an important role in the metastasis of malignant tumors. Membrane type 1-MMP (MT1-MMP) and tissue inhibitor of metalloproteinase 2 (TIMP-2) are essential factors for the activation of pro-MMP-2. There are some reports about expressions of MMP family in relationship to clinical features of head and neck squamous cell carcinoma (SCC), but the results were not uniform and the prognostic value of their expressions remains unclear. EXPERIMENTAL DESIGN: The study group consisted of 53 Japanese patients with oral SCC of early stage (T(1-2)N(0)M(0)). Expressions of MMP-2, MMP-9, MT1-MMP, and TIMP-2 were examined using immunohistological methods on the sections of tumor biopsy samples. The intensity of MMP expression was categorized into four grades (score 0-3) by semiquantitative analysis using a computer with NIH image, and correlation between this grade and clinical aspects such as tumor recurrence, metastasis, and prognosis were examined. RESULTS: The expression score of MMP-2 correlated with that of MMP-9 (r = 0.291; P = 0.036), MT1-MMP (r = 0.286; P = 0.039), and TIMP-2 (r = 0.257; P = 0.050). Patients who developed regional lymph node and/or distant metastasis showed significantly higher scores in the expressions of MMP-9 and TIMP-2 than patients without any tumor metastases (P = 0.036 and P = 0.043, respectively). Kaplan-Meier analyses as well as univariate analyses using the Cox proportional hazards model showed that expression of MMP-9 (P = 0.0143 and P = 0.0418, respectively) and marked expression of TIMP-2 (P < 0.0001 and P = 0.0004, respectively) correlated with worse-cause-specific survival. Multivariate analysis confirmed that marked expression of TIMP-2 was the only independent factor for cause-specific death (hazard ratio, 7.543; confidence interval, 1.693-33.610; P = 0.0080). CONCLUSIONS: Expressions of MMP-9 and TIMP-2 have predictive value for tumor metastases and cause-specific survival. High expression of TIMP-2 is the most independent factor for worse prognosis in early-stage oral SCC.

Loss of p21 expression is associated with p53 mutations and increased cell proliferation and p27 expression is associated with apoptosis in maxillary sinus squamous cell carcinoma.

CONCLUSIONS: Loss of p21 expression dependent on the p53 mutation may be associated with higher tumor cell proliferation, and low p27 expression may be associated with decreased spontaneous apoptosis, resulting in poorer prognosis in patients with maxillary sinus squamous cell carcinoma (SCC). OBJECTIVE: We have previously reported that p53 mutations and decreased spontaneous apoptosis were associated with poor prognosis in maxillary sinus SCC. However, whether p21 and p27 expression and cell proliferation correlate with either p53 status, spontaneous apoptosis or prognosis in maxillary sinus SCC has not been evaluated. MATERIAL AND METHODS: Seventy patients with maxillary sinus SCC were analyzed. Tumor biopsy specimens were examined for p21 and p27 expression using an immunohistological method. The percentage of proliferating cells labeled by anti-Ki-67 mAb was expressed as the Ki-67 index (KI). RESULTS: Loss of p21 expression correlated with p53 mutations (p=0.0072). The KIs in patients without p21 expression and with p53 mutations were significantly higher than those in patients with p21 expression (p=0.0119) and those without p53 mutations (p=0.0048). Patients with p27 expression showed a significantly higher apoptotic index than those without (p=0.0012). Kaplan-Meier analysis showed that p21 expression was closely associated with prolonged disease-free survival in the group with a normal p53 status (p=0.0472). Multivariate analysis identified high KI as an independent prognostic marker (p=0.047).

[Analysis of risk factors for deficits in the quality of life in allergic rhinitis patients].

To assess the possible negative aspects in the quality of life (QOL) of patients with allergic rhinitis based on the factors such as the symptoms and examination of the nasal cavity, we examined QOL deficits using the Japanese Rhino-conjunctivitis Quality of Life Questionnaire (JRQLQ No1). Thirteen hundred seventy eight allergic rhinitis patients who visited clinics or hospitals between April and July 2003 were assessed for clinical symptoms, had the examination of their nasal cavity graded and answered QOL questionnaires. Gender, period of the disease, antigen, clinical symptoms and the results of the nasal cavity examination were evaluated and analyzed to reveal risk factors associated with negative QOL factors. These negative QOL factors tended to be more severe in women than in men, and more severe in pollen allergy than in house dust mite allergy. There was no association of the period of the disease with negative QOL factors. It was revealed that nasal discharge was the strongest risk factor for problems in usual daily activities, outdoor activities and social functioning in six QOL domains, and nasal congestion was the major factor associated with sleep problems, general physical problems and emotional function. Nasal congestion was also the strongest risk factor in the total score of the QOL questionnaire and the overall face scale of the patients. In the nasal cavity examination, the amount of aqueous secretion was the strongest risk factor for all items associated with negative QOL factors. According to the risk factors we identified, we can treat allergic rhinitis patients taking their QOL into consideration.

VEGF and bFGF expression and microvessel density of maxillary sinus squamous cell carcinoma in relation to p53 status, spontaneous apoptosis and prognosis.

We have previously reported that p53 mutations, loss of bax expression or decreased spontaneous tumor apoptosis were associated with poorer prognoses in maxillary sinus squamous cell carcinoma (SCC)(Cancer 94: 1968-1980, 2002). In the present study, we analyzed tumor angiogenesis monitored by expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and tumor microvessel density, in correlation with p53 status, spontaneous apoptosis or disease prognosis in the same group of 70 maxillary sinus SCC patients. Tumor biopsy specimens obtained prior to initiation of treatment were examined for expression of VEGF and bFGF and tumor microvessel density using immunohistological methods. Average vessel density (AVD) (range: 3-75; median: 25) and maximum vessel density (MVD) (range: 4-125; median: 53) were assessed by the number of microvessels stained with anti-CD31 mAb in tumor lesions. VEGF was expressed in 35 (50%) of 70 patients and bFGF was in 43 (61%). Patients with VEGF expression showed significantly higher levels of MVD than those without VEGF expression (57 vs. 38; P=0.019). The VEGF expression was observed more frequently in patients with p53 overexpression and/or mutation than in those with normal p53 status (P=0.048). The MVD inversely correlated with the apoptotic index (AI) defined as the number of single stranded (ss)-DNA-positive cells per 1000 tumor cells (r= -0.23; P=0.022). Patients with neck lymph node and/or distant metastases after surgery showed significantly higher levels of MVD than patients without any metastasis (64 vs. 42; P=0.048). Low histological effectiveness of radiochemotherapy correlated with bFGF expression (P=0.0059). To clarify actual prognostic factors for maxillary sinus SCC, we selected 57 patients treated uniformly with preoperative radiochemotherapy followed by maxillectomy. Kaplan-Meier analysis showed that survival was significantly worse in patients with high MVD (> or =80) than in those with low MVD (<80) (P=0.042). These data suggest that the VEGF expression in association with the p53 overexpression and/or mutations may cause increased microvascularity, decreased spontaneous apoptosis or metastases, while the bFGF expression may be associated with resistance to radiochemotherapy, thereby resulting in poorer prognoses in maxillary sinus SCC. VEGF and bFGF expression and tumor microvessel density in tumor lesions were analyzed in 70 patients with maxillary sinus squamous cell carcinoma. The VEGF expression dependent of p53 overexpression and/or mutations was associated with angiogenesis, decreased spontaneous tumor apoptosis and metastases, while the bFGF expression was associated with resistance to radiochemotherapy, resulting in poor prognosis.

P53, N- and K-Ras, and beta-catenin gene mutations and prognostic factors in nasal NK/T-cell lymphoma from Hokkaido, Japan.

We have shown previously that nasal natural killer (NK)/T-cell lymphoma was associated with Epstein-Barr virus (EBV) and had peculiar clinical features. However, little is known about its biological and genetic changes. The aim of this study is to determine the p53, N- and K-ras, and beta-catenin status in this lymphoma in relation to EBV status and clinical features. The study group consisted of 32 Japanese patients with nasal NK/T-cell lymphoma. The p53 and beta-catenin expression, phenotype, and EBV-oncogenic protein latent membrane protein type 1 (LMP-1) were determined by immunoperoxidase staining. The presence of EBV-encoded small nuclear early region (EBER) RNA was determined by in situ hybridization. The p53 mutations (exons 5 to 9), N- and K-ras mutations (exons 1 and 2), and beta-catenin mutations (exon 3) were analyzed by direct sequencing of the PCR-amplified products that were obtained from laser-microdissected tissues. CD56, CD43, and CD3 were expressed in 32 (100%), in 31 (96%), and in 18 (56%) tumors, respectively. EBER RNA was detected in 31 (96%) tumors. LMP-1 was expressed in 15 (48%) tumors, and p53 and beta-catenin protein were overexpressed in 18 (56%) and 4 (13%) tumors, respectively. Six mutations of the p53 gene, 1 mutation of each N- and K-ras gene, and 8 mutations of beta-catenin gene were detected in 6 (19%), 1 (3%), and 5 (16%) tumors, respectively. The p53 missense mutation was associated with LMP-1 expression (P = 0.038), but not with p53 overexpression. Kaplan-Meier analysis as well as univariate analysis using Cox proportional hazards model showed that high lactate dehydrogenase (LDH) level (P = 0.009, P = 0.0100, respectively), large cell, immunoblastoid polymorphous histology (P = 0.005, P = 0.0162, respectively), and p53 missense mutations (P = 0.021, P = 0.0342, respectively) were significantly related to worse cause-specific survival. Multivariate analysis showed that p53 missense mutation was the most independent among these 3 factors. Although the incidence of thep53, N- and K-ras, and beta-catenin gene mutations is not high, p53 missense mutation has a prognostic value for aggressive course in nasal NK/T-cell lymphoma.

Ambient oxygen regulates epithelial metabolism and nitric oxide production in the human nose.

The effects of ambient O(2) tension on epithelial metabolism and nitric oxide (NO) production (VNO) in the nasal airway were examined in nine healthy volunteers. Nasal VNO, O(2) consumption (VO(2)), and CO(2) production (VCO(2)) were measured during normoxia followed by gradual hypoxia from 21 to 0% O(2) concentration. Nasal VO(2), VCO(2), and respiratory quotient during normoxia were determined to be 1.19 +/- 0.04 ml/min, 1.60 +/- 0.04 ml/min, and 1.35 +/- 0.04, respectively. Hypoxia exposure to the nasal cavity significantly decreased both VCO(2) and VNO [VCO(2): 1.60 +/- 0.04 to 0.96 +/- 0.03 ml/min (P < 0.01), VNO: 530 +/- 15 to 336 +/- 9 nl/min (P < 0.01)]. VNO was reduced commensurately with gradual decline in O(2) tension, and the apparent K(m) value for O(2) was determined to be 23.0 microM. These results indicate that the nasal epithelial cells exchange O(2) and CO(2) with ambient air in the course of their metabolism and that nasal epithelial cells can synthesize NO by using ambient O(2) as a substrate. We conclude that air-borne O(2) diffuses into the epithelium where it may be utilized for either cell metabolism or NO synthesis.

Functional electrical stimulation of laryngeal adductor muscle restores mobility of vocal fold and improves voice sounds in cats with unilateral laryngeal paralysis.

Functional electrical stimulation (FES) has been proposed as a potential treatment for restoring motor functions of denervated motor systems. We investigated whether FES of paralyzed laryngeal adductor muscles could restore adduction to the vocal folds. In addition, we studied the effect of stimulated vocal fold adduction on the intensity and overall quality of voice production. We recorded movement of the vocal fold, electromyographic activity of muscles recruited for vocalization, and sound production in unanesthetized decerebrate cats during FES of the paralyzed thyroarytenoid (TA) muscle. FES of the paralyzed TA muscle induced adduction of the vocal fold. Appropriate stimulus parameters for induction was 1.5-3.0 mA intensity pulses delivered at a frequency of 30-50 pulses per second (pps). FES of the paralyzed TA muscle prolonged phonation time and increased intensity of voice sounds during vocalization induced by electrical stimulation (0.2 ms, 20-50 microA, 50 pps) of the periaqueductal gray (PAG). The quality of voice sounds evaluated by sound spectrography was shown to improve during vocalization with FES. We conclude that FES of the paralyzed laryngeal adductor muscle was effective in restoring adduction of the vocal fold and improving voice sounds impaired by unilateral laryngeal paralysis.

The neuronal circuit of augmenting effects on intrinsic laryngeal muscle activities induced by nasal air-jet stimulation in decerebrate cats.

We previously demonstrated that during nasal air-jet stimulation, both the activities of intrinsic laryngeal adductor and abductor muscles persistently increase, whereas the respiratory cycle prolongs and the activity of diaphragm decreases [Am. J. Rhinol. 9 (1995) 203-208; Neurosci. Res. 31 (1998) 137-146]. The purpose of this study was to clarify the neuronal circuit underlying the augmentation of intrinsic laryngeal muscles evoked by nasal air-jet stimulation. The immunohistologic analysis of Fos-expression was reported to determine the distribution of activated neurons in cat brainstem evoked by sneeze-inducing air puff stimulation of the nasal mucosa [Brain Res. 687 (1995) 143-154]. In sneezing cats, immunoreactivity was evoked in projection areas of the ethmoidal afferents, e.g. the subnuclei caudalis, interpolaris and in interstitial islands of the trigeminal sensory complex. Immunoreactivity was also enhanced in the solitary complex, the nucleus retroambiguus, the pontine parabrachial area and the lateral aspect of the parvocellular reticular formation [Brain Res. 687 (1995) 143-154]. In the present study, we focussed on the parvocellular reticular nucleus (PRN) as a relay of the neural circuit contributed to the augmentation of intrinsic laryngeal muscles evoked by nasal air-jet stimulation. We recorded the neuronal behavior of PRN during the nasal air-jet stimulation in precollicular-postmammillary decerebrate cats. As the results, 24% (17/71) of recorded neurons which were activated orthodromically by the electrical stimulation to anterior ethmoidal nerve, increased their firing rates in response to the nasal air-jet stimulation. Furthermore, spike-triggered averaging method revealed that four of these 17 PRN neurons activated intrinsic laryngeal muscles, suggesting that such neurons have excitatory projections to the intrinsic laryngeal muscle motoneurons in the nucleus ambiguus. These results suggest that the some of PRN neuron play a role in augmentation of the intrinsic laryngeal muscles activities during nasal air-jet stimulation.

Functional roles of nasal nitric oxide in nasal patency and mucociliary function.

The purpose of this study was twofold: (i) to clarify whether it is possible to change the amount of nasal nitric oxide (NO) artificially; and (ii) to confirm that the artificially altered nasal NO has biological properties such as vasodilatation and mucociliary function. We measured nasal NO and nasal airway resistance (NAR) in healthy subjects before and after topical administration of sodium nitroprusside (SNP), an NO donor. We also measured nasal NO and saccharin transport time after administration of SNP or L-N(G)-nitroarginine methylester (L-NAME), a NO synthase inhibitor. In addition, we examined the effect of SNP and L-NAME on nasal NO and saccharin transport time after pretreatment with flutropium bromide (FB), an anticholinergic agent. Administration of SNP elicited increases in nasal NO and NAR in a dose-dependent manner. Nasal NO decreased significantly after topical administration of L-NAME. Saccharin transport time, an indicator of mucociliary function, was shortened after topical administration of SNP and prolonged after administration of L-NAME. Moreover, these phenomena were observed even after pretreatment with FB. These results suggest that artificially altered nasal NO may affect NAR and mucociliary function.