The respiratory microbiome associated with chronic obstructive pulmonary disease comorbidity in non-small cell lung cancer.

BACKGROUND: Research has shown that some microbiomes are linked to cancer. Hence, we hypothesize that alterations in the respiratory microbiome might be associated with lung cancer. METHODS: Through droplet digital polymerase chain reaction analysis, we investigated the abundance of Acidovorax in surgically resected primary tumors and corresponding nontumor lung tissues obtained from 50 Japanese patients with non-small cell lung cancer. RESULTS: The rate of positivity for Acidovorax in tumor and nontumor tissues was 44 and 26%, respectively. The abundance of Acidovorax in tumor tissues was significantly higher in patients with nonsquamous cell carcinoma complicated by chronic obstructive pulmonary disease (COPD) and those who relapsed after surgical resection (p < 0.05). In tumor tissues, the results of the univariate and multivariate analyses revealed that only COPD exerted a direct effect on the abundance of Acidovorax (p < 0.05). Furthermore, the presence of Acidovorax was high in lung cancer patients with COPD comorbidity (65%) and TP53 gene mutation; only one of the nontumor tissues was positive for Acidovorax. In patients with lung cancer complicated by COPD, Acidovorax tended to be present in both the tumor and nontumor areas. CONCLUSIONS: This study identified novel microbiota involved in lung cancer with COPD comorbidity. The results suggested that Acidovorax may be a useful biomarker in the screening for lung cancer. Further studies are warranted to validate the clinical significance of the microbiome in a larger independent patient cohort.

Acinetobacter baumannii can be transferred from contaminated nitrile examination gloves to polypropylene plastic surfaces.

BACKGROUND: Several observational studies suggest that gloves of health care workers are major routes of multidrug-resistant Acinetobacter baumannii transmission. However, limited experimental data are available assessing Acinetobacter transmission from gloves to environmental surfaces. This study determined whether A baumannii was easily transferred from nitrile gloves to polypropylene plastic compared with other gram-negative bacteria that cause health care-associated infections in laboratory-controlled experiments. METHODS: Gloved fingerpad-to-fomite transfer efficiency was determined for drug-resistant and -sensitive strains of A baumannii, Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa. RESULTS: Only A baumannii transferred from gloves to fomites 3 minutes after the bacterial transfer event. Transfer efficiency of A baumannii was 0.1%-33% at that time point. DISCUSSION: Bacterial transfer from contaminated gloves to the hospital environment may be related to the type of contaminating bacteria, inoculated bacterial level, fomites, and glove materials. Therefore, it is important to need a comprehensive assessment of the transfer efficiency. CONCLUSIONS: A baumannii can transfer easily from nitrile gloves to fomite compared with other gram-negative bacteria that cause health care-associated infections. These findings support data from previous observational studies that gloves of health care workers can be major routes of A baumannii transmission in clinical settings.

Slightly acidic electrolyzed water disrupts biofilms and effectively disinfects Pseudomonas aeruginosa.

INTRODUCTION: Biofilm formation is an important issue in the healthcare industry, but conventional disinfectants are not effective for biofilms formed in the hospital environment and on medical instruments. In this study, aim at determine the effectiveness of slightly acidic electrolyzed water (SAEW) on biofilm removal and the disinfection of biofilm-forming Pseudomonas aeruginosa. METHODS: Mucoid and non-mucoid strains were used for biofilm formation. Biofilms were incubated with SAEW and the reduction in biofilm volume was determined based on the optical density. Furthermore, to investigate the mechanism underlying the effects of SAEW, a biofilm was produced with alginate and structural changes in response to incubation with SAEW were observed by fluorescence microscopy. The minimum bactericidal chlorine concentration of SAEW for P. aeruginosa cells was evaluated. RESULTS: The amounts of alginate and biofilm decreased by 99.9% and 56.8% immersed by 30 ppm of SAEW at 25 °C for 10 min. The effectiveness of SAEW increased as the temperature increased, and the biofilm volume was reduced by 85.4% at 45 °C. Furthermore, 30 ppm SAEW completely disinfected P. aeruginosa in the biofilm, even for immersion at 15 °C for 5 min. CONCLUSION: Our results suggest that SAEW, a low-cost and safe chlorine disinfectant, is a useful disinfectant for biofilm-forming bacteria.

Molecular Epidemiological Analysis of the Spreading Conditions of Trichophyton in Long-Term Care Facilities in Japan.

Tinea pedis and tinea unguium are common infectious diseases, and many elderly people are reported to contract these infections. In this study, to investigate whether strains of the same origin are spreading inside a long-term care facility, we analyzed Trichophyton rubrum and Trichophyton mentagrophytes, isolated from the residents and staff at the facilities located in the Kanto area, using a genomic analytical method targeting tandem repeat regions in the nontranscribed spacer (NTS) region of ribosomal DNA. Five NTS types were confirmed in T. rubrum. T. rubrum of various types (types 1 to 5) was detected at each facility, but there was no isolate specific to one facility only. Eight NTS types of T. mentagrophytes were detected, and T. mentagrophytes that carried an NTS type that was confirmed at one facility only (types C4II, F4II, and D4II) was isolated. These T. mentagrophytes sequence types were isolated from several subjects residing at the same facility. This study proved that a T. mentagrophytes strain of the same type had spread in long-term care facilities. We believe in the importance of cleaning at a long-term care facility as a countermeasure to the spread of Trichophyton species.

Discovery of Terbinafine Low Susceptibility Trichophyton rubrum strain in Japan.

This is the first confirmed report of terbinafine low susceptibility Trichophyton rubrum, BGUTR13, in Japan collected from the whole sole of the elderly over 65s with cotton swab sampling method at the special nursing care-home in 2016. We revealed BGUTR13 showed low susceptibility (MIC, >128 µg/mL) against terbinafine. But, BGUTR13 exhibited normal susceptibility to itraconazole, did not showed cross-resistance. Also, the squalene epoxidase gene of terbinafine low susceptibility strain BGUTR13 which is the target of terbinafine contained newly confirmed one mismatch. We suggested the possibility that the resistance mechanism of terbinafine low susceptibility strains is due to the loss of sensitivity of squalene epoxidase inhibitors and does not affect antifungal drugs with other different mechanisms of action.

Epidemiological Study on Trichophyton Disseminating from the Feet of the Elderly.

OBJECTIVES: To grasp the colonization status of Trichophyton in terms of spreading rate, we investigated the intergenerational epidemiological difference in the spreading rate of Trichophyton from teenagers to the elderly aged over 65. In addition, the elderly people were divided into two groups: those living at nursing homes and those living at their homes. We compared the two groups in terms of the difference in the spreading rate of Trichophyton. METHODS: Spreading rate was investigated by identifying the fungi collected by the cotton swab sampling method. The correlation between the responses to the questionnaire survey, which was conducted after the sample collection, and the spreading rate of Trichophyton was analyzed. RESULTS: The spreading rate of Trichophyton was 23.3%. It was confirmed that the spreading rate in general adults was 9.1%, whereas that in elderly people was 40.8%, which is significantly high. Also, it was confirmed that T. mentagrophytes shows a higher spreading rate among general adults, whereas T. rubrum shows a higher spreading rate among the elderly. Between the elderly living at nursing homes and those living at their homes, although no statistically significant difference was confirmed, the former tended to show a higher spreading rate than the latter. Also, the results of this study showed that spreading rate of Trichophyton and the detachment of the skin of the toes were significantly related. CONCLUSIONS: We found that the risk of spreading of Trichophyton increases with age. Particularly among elderly people aged over 65, taking some actions that prevent the spread of tinea pedis is recommended.

Oxygen-glucose deprivation increases firing of unipolar brush cells and enhances spontaneous EPSCs in Purkinje cells in the vestibulo-cerebellum.

Unipolar brush cells (UBCs) are excitatory interneurons in the granular layer of the cerebellar cortex, which are predominantly distributed in the vestibulo-cerebellar region. The unique firing properties and synaptic connections of UBCs may underlie lobular heterogeneity of excitability in the granular layer and the susceptibility to ischemia-induced excitotoxicity. In this study, we investigated the effects of oxygen-glucose deprivation (OGD) on the firing properties of UBCs and granule cells and spontaneous excitatory postsynaptic currents (sEPSCs) of Purkinje cells using whole-cell recordings. Short-term OGD induced increases in spontaneous firing of UBCs by causing membrane depolarization via the activation of NMDA receptors. UBC firing indirectly affected Purkinje cells by altering parallel fiber inputs of a subset granule cells, resulting in a marked increase in sEPSCs in Purkinje cells in vestibulo-cerebellar lobules IX-X, but not in lobules IV-VI, which have fewer UBCs. Similarly, the frequency and amplitude of sEPSCs in Purkinje cells were significantly greater in lobules IX-X than in IV-VI, even in control conditions. These results reveal that UBCs play key roles in regulating local excitability in the granular layer, resulting in lobular heterogeneity in the susceptibility to ischemic insult in the cerebellum.

Motor dysfunction in cerebellar Purkinje cell-specific vesicular GABA transporter knockout mice.

γ-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the adult mammalian central nervous system and plays modulatory roles in neural development. The vesicular GABA transporter (VGAT) is an essential molecule for GABAergic neurotransmission due to its role in vesicular GABA release. Cerebellar Purkinje cells (PCs) are GABAergic projection neurons that are indispensable for cerebellar function. To elucidate the significance of VGAT in cerebellar PCs, we generated and characterized PC-specific VGAT knockout (L7-VGAT) mice. VGAT mRNAs and proteins were specifically absent in the 40-week-old L7-VGAT PCs. The morphological characteristics, such as lamination and foliation of the cerebellar cortex, of the L7-VGAT mice were similar to those of the control littermate mice. Moreover, the protein expression levels and patterns of pre- (calbindin and parvalbumin) and postsynaptic (GABA-A receptor α1 subunit and gephyrin) molecules between the L7-VGAT and control mice were similar in the deep cerebellar nuclei that receive PC projections. However, the L7-VGAT mice performed poorly in the accelerating rotarod test and displayed ataxic gait in the footprint test. The L7-VGAT mice also exhibited severer ataxia as VGAT deficits progressed. These results suggest that VGAT in cerebellar PCs is not essential for the rough maintenance of cerebellar structure, but does play an important role in motor coordination. The L7-VGAT mice are a novel model of ataxia without PC degeneration, and would also be useful for studying the role of PCs in cognition and emotion.

[Problem and assignment for distinguishing the Usher syndrome type].

Usher syndrome is an autosomal-recessive disorder that causes bilateral sensorineural hearing loss, retinitis pigmentosa (RP), and occasionally vestibular dysfunction. Usher syndrome types 1, 2, and 3 can be distinguished by differences in audiovestibular features. The objectives of this retrospective study were to evaluate 26 patients with Usher syndrome clinically. The 26 patients (male: 12 cases, female: 14 cases) with Usher syndrome, with a clinical diagnosis based on symptoms of bilateral sensorineural hearing loss and RP, had been registered from 13 hospitals as a multicenter study. We assessed the clinical history and performed audiovestibular and ophthalmologic examinations, and genetic testing. Eleven of the patients were classified as having Usher type 1 (38.5%), 6 with Usher type 2 (23.1%), and 9 with Usher type 3 (38.5%). However, many patients with atypical Usher type 1 (70%) and type 2 (83.3%) were found compared with Usher type 3 (10%). The conductive rate of vestibular examinations including the caloric test (50%) was low. There were many variations in the clinical symptoms in Usher syndrome patients, therefore the classification of Usher types 1, 2, and 3 has been complicated. We have proposed a flowchart for the diagnosis of Usher types 1, 2, and 3.

Intraoperative molecular assessment for lymph node metastasis in head and neck squamous cell carcinoma using one-step nucleic acid amplification (OSNA) assay.

BACKGROUND: Conventional intraoperative pathological examination for Sentinel node navigation surgery (SNNS) has been controversial. We evaluated the efficacy of one-step nucleic acid amplification (OSNA) assay for intraoperative diagnosis of cervical lymph node (CLN) metastasis compared with histopathological examination in patients with head and neck squamous cell carcinoma (HNSCC). METHODS: A total of 175 CLNs dissected from 56 patients with HNSCC who underwent surgery at Aichi Cancer Center, Kyorin University, Gunma University or Fukushima Medical University, between April 2008 and December 2011 were enrolled. CLN samples were sectioned into four equal pieces, with two of each used for OSNA assay and other histopathological examinations. The diagnostic value of OSNA assay in HNSCC patients in predicting the results of histopathological diagnosis was evaluated using the area under the receiver operating characteristic (AUROC) curve. RESULTS: OSNA assay showed acceptable efficacy in the detection of pathological CLN metastasis (AUROC 0.918, 95 % confidence interval [CI] 0.852-0.984). Regarding the CK19mRNA cutoff value, the optimum cutoff point in HNSCC patients was 131 copies/µl (sensitivity: 82.4, 95 % CI 65.5-93.2; specificity: 99.3, 95 % CI 96.1-100.0; positive likelihood ratio 116.1; negative likelihood ratio 0.2]. CONCLUSIONS: We demonstrated that OSNA assay is useful in intraoperative diagnosis for CLN metastasis in patients with HNSCC. OSNA assay could be applied for SNNS in HNSCC patients.

Comparison of four reading methods of broth microdilution based on the clinical and laboratory standards institute M27-A3 method for Candida spp.

This study aimed to compare the susceptibilities of 5 reference strains and 28 isolates of Candida spp., to micafungin, amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole, and miconazole, obtained by visually determined minimum inhibitory concentration (MIC) using the agitation method (V-A), as described in the Clinical and Laboratory Standards Institute M27-A3 document; visual determinations without agitation (V-NA); and spectrophotometric determinations for the presence or absence of agitation (SP-A and SP-NA, respectively). Our results indicate that when the V-NA, SP-A, and SP-NA-the 3 alternative microdilution procedures for MIC endpoint determinations-were compared with the V-A, excellent agreements were observed between the V-NA and V-A rather than with the spectrophotometric methods (between the SP-A or SP-NA, and V-A). Furthermore, many errors occurred while using the SP-A method in the presence of agitation and some isolates showed major errors. Three of 5 isolates that showed very major errors between the spectrophotometric SP-A or SP-NA, and the reference V-A method were trailing isolates. Therefore, it was suggested that the MICs of Candida spp. obtained by the V-NA method were more precise than those by the conventional SP-A method.

Disruption of metabotropic glutamate receptor signalling is a major defect at cerebellar parallel fibre-Purkinje cell synapses in staggerer mutant mice.

Staggerer mutant mice have functional loss of a transcription factor, retinoid-related orphan receptor α (RORα), which is abundantly expressed in Purkinje cells (PCs) of the cerebellum.Homozygous staggerer (sg/sg)mice show cerebellar hypoplasia and congenital ataxia. Sg/sg mice serve as an important extreme mouse model of the hereditary spinocerebellar ataxia type 1 (SCA1), since it has been shown that RORα dysfunction is strongly correlated with SCA1 pathogenesis. However, synaptic abnormalities, especially at parallel fibre (PF)-PC synapses, in SCA1-related sg/sg mice have not been examined in detail electrophysiologically. In this study, we report that PFs can still establish functional synapses onto PCs in sg/sg mice in spite of reduction in the number of PF-PC synapses. Compared with PF-evoked EPSCs in the wild-type or heterozygotes, the success rate of the EPSC recordings in sg/sg was quite low (∼40%) and the EPSCs showed faster kinetics and slightly decreased paired pulse facilitation at short intervals. The prominent synaptic dysfunction is that sg/sg mice lack metabotropic glutamate receptor (mGluR)-mediated slow EPSCs completely. Neither intense PF stimulation nor an exogenously applied mGluR agonist, DHPG, could elicit mGluR-mediated responses.Western blot analysis in the sg/sg cerebellum revealed low-level expression of mGluR1 and TRPC3, both of which underlie mGluR-mediated slow currents in PCs. Immunohistochemical data demonstrated marked mislocalization of mGluR1 on sg/sg PCs.We found that mGluR-mediated retrograde suppression of PF-PC EPSCs by endocannabinoid is also impaired completely in sg/sg mice. These results suggest that disruption of mGluR signalling at PF-PC synapses is one of the major synaptic defects in sg/sg mice and may manifest itself in SCA1 pathology.

A new and less invasive procedure for arytenoid adduction surgery: endoscopic-assisted arytenoid adduction surgery.

OBJECTIVES/HYPOTHESIS: Arytenoid adduction (AA) is the most effective procedure for improving voice function in patients affected by unilateral vocal fold paralysis (UVFP), but it is often associated with severe complications following airway obstruction. The aim of this study is to describe a new and less invasive AA surgical procedure termed endoscopic-assisted AA surgery (EAAS) and to evaluate its voice outcomes. STUDY DESIGN: We demonstrated this method using extirpated larynges from three laryngeal cancer patients. Ten patients with severe UVFP underwent EAAS alone or combined with type I thyroplasty or lipoinjection laryngoplasty. METHODS: EAAS involves the placement of permanent adducting sutures around the muscular process (MP) of the arytenoid cartilage using two needles, a penetration needle and a loop needle, under endoscopic guidance. To define the anatomic position of the MP for safer needle insertion, the location of the MP was measured relative to three landmarks on computed tomography/X-ray images of the larynges and in resected larynges. For all patients with UVFP, the maximum phonation time, mean airflow rate, and three acoustic analysis parameters were measured before and after EAAS. RESULTS: The values of the three variables were similar in all cases. Most patients achieved a maximum phonation time of more than 10 seconds and a mean airflow rate of less than 200 mL/second. All three acoustic analysis parameters were significantly improved after surgery. CONCLUSIONS: EAAS is a simple and effective arytenoid rotation procedure.

Angiotensin II receptor blocker-induced angioedema in the oral floor and epiglottis.

We report the rare case of angioedema (also known as Quincke edema), which was induced by valsartan, an angiotensin II receptor blocker (ARB). ARBs are a new class of antihypertensive agent that is developed to exclude the adverse effects of angiotensin-converting enzyme inhibitors. In theory, ARBs do not contribute to the occurrence of angioedema because they do not increase the serum level of bradykinin, the responsible substance for angioedema. However, some reports of ARB-induced angioedema have recently been published. In this study, we present the forth case and the first Asian case of angioedema due to valsartan, which is one of the ARBs. Otolaryngologist should be wary of the prescribing ARB and discontinue ARBs treatment soon, if angioedema is recognized.

Stress-evoked tyrosine phosphorylation of signal regulatory protein α regulates behavioral immobility in the forced swim test.

Severe stress induces changes in neuronal function that are implicated in stress-related disorders such as depression. The molecular mechanisms underlying the response of the brain to stress remain primarily unknown, however. Signal regulatory protein alpha (SIRPalpha) is an Ig-superfamily protein that undergoes tyrosine phosphorylation and binds the protein tyrosine phosphatase Shp2. Here we show that mice expressing a form of SIRPalpha that lacks most of the cytoplasmic region manifest prolonged immobility (depression-like behavior) in the forced swim (FS) test. FS stress induced marked tyrosine phosphorylation of SIRPalpha in the brain of wild-type mice through activation of Src family kinases. The SIRPalpha ligand CD47 was important for such SIRPalpha phosphorylation, and CD47-deficient mice also manifested prolonged immobility in the FS test. Moreover, FS stress-induced tyrosine phosphorylation of both the NR2B subunit of the NMDA subtype of glutamate receptor and the K+-channel subunit Kvbeta2 was regulated by SIRPalpha. Thus, tyrosine phosphorylation of SIRPalpha is important for regulation of depression-like behavior in the response of the brain to stress.

Primary tumor induces sentinel lymph node lymphangiogenesis in oral squamous cell carcinoma.

The main factor that affects the prognosis of patients with oral squamous cell carcinoma (OSCC) is regional lymph node metastases, which usually spreads first to the sentinel lymph nodes (SLNs). Recent studies have demonstrated that tumor cells in several malignancies can induce lymphangiogenesis in SLNs before metastasizing. To elucidate the mechanisms of tumor dissemination of OSCC, we investigated whether primary tumors induce lymphangiogenesis within SLNs in patients with OSCC. The mRNA expression of lymphatic-specific markers, including VEGFR-3, Prox-1, and LYVE-1 in 23 metastasis-negative SLNs obtained from 10 patients with OSCC, was investigated using a quantitative real-time RT-PCR assay, and compared with control lymph nodes from patients with non-cancerous diseases. In addition, VEGF-C and VEGF-D expressions of the primary tumor were examined by immunohistochemistry. In SLNs, there were highly significant correlations between the three lymphatic markers examined. Interestingly, the level of LYVE-1 expression in SLNs, despite the absence of metastasis, was significantly higher than in control lymph nodes. Moreover, SLNs from patients with VEGF-C-positive tumor showed a significantly higher expression of VEGFR-3 than those from patients with VEGF-C-negative tumor. Our findings suggest that in OSCC, the primary tumor actively induces lymphangiogenesis in SLNs prior to the onset of metastases, and where tumor-derived VEGF-C plays an important role.

[Clinical efficacy, safety and PK-PD analysis of tebipenem pivoxil in a phase II clinical trial in otolaryngological infections].

UNLABELLED: We conducted a double-blind intergroup comparative study investigating the efficacy, safety and PK-PD analysis of the new oral carbapenem antibacterial drug tebipenem pivoxil (TBPM-PI) for the treatment of otolaryngological infections in adults to establish the recommended clinical dosage. The primary endpoint was the clinical effect of a 7-day oral administration of TBPM-PI to subjects with confirmed cases of infection by any of the 5 major bacterial species causative for otolaryngological infections (Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Moraxella catarrhalis, and Haemophilus influenzae) assigned to three groups set according to the TBPM-PI dosage, namely, a 450 mg group (150 mg t.i.d), a 500 mg group (250 mg b.i.d), and a 900 mg group (300 mg t.i.d). 1. Clinical efficacy: At the end of administration or at discontinuation, the efficacy rate for the 112 subjects in the efficacy analysis set was 72.1% (31/43 subjects) in the 450 mg group, 88.6% (31/35 subjects) in the 500 mg group, and 85.3% (29/34 subjects) in the 900 mg group. Both the 500 mg and 900 mg groups showed a high efficacy rate of over 80%. 2. Bacteriological efficacy: The disappearance rate of the pre-administration causative bacteria (5 major bacterial species) at the end of administration (at discontinuation), it was 92.2% (47/51 strains) in the 450 mg group, 94.7% (36/38 strains) in the 500 mg group, and 91.7% (33/36 strains) in the 900 mg group. All the groups showed a high disappearance rate, with no large differences among them. All strains of S. pneumoniae, including PRSP, as well as those of S. pyogenes and M. catarrhalis disappeared. The overall disappearance rate of H. influenzae was 78.6%, namely, 76.9% in the 450 mg group, 100% in the 500 mg group, and 66.7% in the 900 mg group, showing differences among the groups. 3. PK-PD: The PK-PD analysis was executed in 124 strains isolated from 111 subjects in which the plasma TBPM concentration and the MIC of causative organism were measured. The target value of the PK-PD parameter was examined from the relation between PK-PD parameter and bacteriological efficacy. The presumed target value of AUCf/MIC was 10-20, Cmaxf/MIC was 4. On the other hand, a clear relation was not found between T>MIC and the bacteriological efficacy. 4. SAFETY: The incidence of adverse reactions related to symptoms and signs was 28.8% (21/73 subjects) in the 450 mg group, 35.8% (24/67 subjects) in the 500mg group, and 30.6% (22/72 subjects) in the 900 mg group. The incidence of abnormal changes in laboratory test values was 8.2% (6/73 subjects) in the 450 mg group, 9.2% (6/65 subjects) in the 500 mg group, and 9.9% (7/71 subjects) in the 900 mg group. There were no differences in either of these categories among the groups, and the incidence was considered not to be correlated with dose. Based on the above, we considered that TBPM-PI at doses of 250 mg b.i.d (500 mg/day) promises high clinical usefulness for the treatment of otolaryngological infections in adults.

CD4+ T cell responses to HLA-DP5-restricted wild-type sequence p53 peptides in patients with head and neck cancer.

Wild-type sequence (wt) p53 peptides are attractive candidates for broadly applicable cancer vaccines. Evidence has been accumulating which indicates that CD4+ Th cells have an important role in generating and maintaining antitumor immune responses. To elucidate the nature of CD4+ Th responses to wt p53 epitopes in patients with squamous cell carcinoma of the head and neck (SCCHN), peripheral blood mononuclear cells (PBMCs) from HLA-DP5+ patients were stimulated with HLA-DP5-restricted wt p53 peptides, p53(108-122) or p53(153-166), and tested for the release of IFN-gamma and IL-5 in ELISPOT assays. Immunohistochemistry for p53 accumulation in tumors, and ELISA for serum antibodies to p53 were also performed. Eleven (57.9%) of 19 HLA-DP5+ patients but none of 5 healthy donors had detectable Th1 and/or Th2 responses to wt p53 peptides by ELISPOT assay. Among these 11 responding patients, 9 (81.8%) and all 11 (100%) patients had a tumor burden and p53 accumulation, respectively. On the other hand, two responding patients were in post-operative condition. Interestingly, among nine patients with a tumor burden, four patients with early disease showed either Th1-polarized or mixed Th1/Th2 responses, while five patients with advanced disease showed either Th2-polarized or mixed Th1/Th2 responses. Our results suggest that wt p53(108-122) and p53(153-166) peptides stimulate both Th1- and Th2-type CD4+ T cell responses in patients with SCCHN, and anti-p53 Th responses may persist even after surgical resection of the tumor; however, the presence of a tumor and its progression may affect the nature of immune responses to wt p53 peptides.

Characterization of mutant mice that express polyglutamine in cerebellar Purkinje cells.

We recently produced transgenic mice that expressed an abnormally expanded polyglutamine (polyQ) specifically in cerebellar Purkinje cells (polyQ mice). The polyQ mice showed inclusion body formation, cerebellar atrophy and severe ataxia. Here we analyzed polyQ mice using immunohistochemistry, immunoelectronmicroscopy and electrophysiology. A diffuse form of polyQ was detected in the nucleus. Interestingly, ubiquitinated large inclusions were located close to, but apparently outside of the soma of Purkinje cells. Infusion of lucifer yellow into Purkinje cells clearly indicated the traffic between the periplasmic inclusions and soma of Purkinje cells. To examine whether the formation of periplasmic inclusions was an active process or a result of cell death, the polyQ mouse cerebellum was immunolabeled for cleaved caspase-3, a marker of apoptosis. Interestingly, no Purkinje cells in P80 polyQ mice immunoreacted with the antibody. The results were substantiated by electrophysiological assay, which showed that P80 Purkinje cells with large periplasmic inclusions were functionally active: excitatory postsynaptic currents (EPSCs) were reliably evoked upon electrical stimulation of parallel fibers (PFs) or climbing fibers (CFs), and current injection into Purkinje cells generated action potentials; however, the frequency of action potentials in response to various volumes of current injection was consistently lower in polyQ mice than in wild-type animals, and aberrant innervation by multiple CFs was detected in polyQ mouse Purkinje cells. These results suggest that Purkinje cells with periplasmic inclusions were not apoptotic, but their functions were substantially impaired, which could contribute to the severe ataxic phenotype.

Changes in the characteristics of definite Meniere's disease over time in Japan: a long-term survey by the Peripheral Vestibular Disorder Research Committee of Japan, formerly the Meniere's Disease Research Committee of Japan.

CONCLUSION: The incidence of new cases of Meniere's disease (MD) in elderly patients aged 60 years or more was found to have increased over time after correction for age distribution in the overall population. Job- and care-related fatigue may be involved in the recent increase in elderly-onset cases because physical and mental fatigue can induce onset of the disease. OBJECTIVES: Changes over time in the epidemiologic characteristics of MD in Japan were analyzed. MATERIALS AND METHODS: Between 1975 and 2006, four nationwide, multi-center surveys of MD were conducted by the Meniere's Disease Research Committee of Japan (1975-1976) and the Peripheral Vestibular Disorders Research Committee of Japan (1982-1984, 1990, and 2001-2006). Information was collected by the committee members on a total of 1368 de novo cases of definite MD, 520 reported in the first survey, 290 in the second survey, 148 in the third survey, and 410 in the fourth survey. RESULTS: Clear changes were seen over time in the population-adjusted sex distribution of the disease and population-adjusted age at onset. The number of definite MD cases in females increased over time relative to the number of cases in males. The proportion of cases in which onset occurred at 60 years of age or more increased over time when the number of cases in each age group was adjusted for changes in age distribution of the population over time. From the time of the third survey, there was a slight increase in the proportion of cases with bilateral involvement.