Search

ETS homologous factor (EHF) belongs to the epithelium-specific subfamily of the E26 transformation-specific (ETS) transcription factor family. Currently, little is known about EHF’s function in cancer. We previously reported that ETS1 induces expression of the ZEB family proteins ZEB1/δEF1 and ZEB2/SIP1, which are key regulators of the epithelial–mesenchymal transition (EMT), by activating the ZEB1 promoters. We have found that EHF gene produces two transcript variants, namely a long form variant that includes exon 1 (EHF-LF) and a short form variant that excludes exon 1 (EHF-SF). Only EHF-SF abrogates ETS1-mediated activation of the ZEB1 promoter by promoting degradation of ETS1 proteins, thereby inhibiting the EMT phenotypes of cancer cells. Most importantly, we identified a novel point mutation within the conserved ETS domain of EHF, and found that EHF mutations abolish its original function while causing the EHF protein to act as a potential dominant negative, thereby enhancing metastasis in vivo. Therefore, we suggest that EHF acts as an anti-EMT factor by inhibiting the expression of ZEBs, and that EHF mutations exacerbate cancer progression.

Background The efficacy and safety of 300 index of reactivity (IR) tablets of house dust mite (HDM) allergen extracts in Japanese pediatric (5‐16 years old) patients with allergic rhinitis (AR) were assessed in a double‐blind, randomized, placebo‐controlled study (JAPIC CTI‐152981). Methods Patients were randomized 1:1 to HDM sublingual tablets or placebo once daily for 52 weeks. The primary end‐point was average adjusted symptom score (AASS; average of daily Rhinitis Total Symptom Scores, comprising sneezing, rhinorrhea, nasal congestion, and nasal pruritus, adjusted for rescue medication use), analyzed during Weeks 48‐52 (mixed‐effects model for repeated measures). Results Of 438 patients randomized, 403 (92%; 193 active, 210 placebo) completed the study. AASS (least‐squares [LS] mean [SE]) during Weeks 48‐52 was significantly (P = 0.0005) lower in the active group compared with placebo (6.32 [0.20] vs 7.27 [0.19]; relative LS mean difference, −13%). Immunological responses (IgE and IgG4 antibodies specific to antigens of two HDM species) were significantly greater in the active group compared with placebo (P

Background: To evaluate the long-term safety of subcutaneous immunotherapy with TO-204, a standardized house dust mite (HDM) allergen extracts, we conducted a multicenter, open label clinical trial. Methods: Japanese patients aged 5–65 years were eligible for the study, if they had HDM-induced allergic rhinitis (AR), allergic bronchial asthma (BA), or both. TO-204 was administered in a dose titration scheme, and the maintenance dose was determined according to the predefined criteria. The treatment period was 52 weeks, and patients who were willing to continue the treatment received TO-204 beyond 52 weeks. This clinical trial is registered at the Japan Pharmaceutical Information Center (Japic CTI-121900). Results: Between July 2012 and May 2015, 44 patients (28 with AR and 16 with allergic BA) were enrolled into the study. All patients were included in the analysis. The duration of treatment ranged from 23 to 142 weeks and the median maintenance dose was 200 Japanese allergy units (JAU). Adverse events occurred in 22 patients (50%). The most common adverse event was local reactions related to the injection sites. Four patients experienced anaphylactic reactions when they were treated with the dose of 500 JAU. Two patients experienced anaphylactic shock with the doses of 1000 JAU at onset. These 6 patients could continue the study with dose reduction. Conclusions: Safety profile of TO-204 was acceptable in Japanese patients with HDM-induced AR or allergic BA. Higher doses should be administered carefully, because the risk of anaphylaxis increased at doses of 500 or 1000 JAU. Keywords: Allergen immunotherapy, Allergic bronchial asthma, Allergic rhinitis, Clinical trial, House dust mite

The lack of available tumor antigens with strong immunogenicity, human leukocyte antigen restriction, and immunosuppression via regulatory T-cells (Tregs) and myeloid-derived suppressor cells are limitations for dendritic cell (DC)–based immunotherapy in patients with advanced head and neck cancer (HNC). We sought to overcome these limitations and induce effective antitumor immunity in the host. The effect of low-dose docetaxel (DTX) treatment on DC maturation was examined in an ex vivo study, and a phase I clinical trial of combination therapy with direct peritumoral immature DC (iDC) injection with OK-432 and low-dose cyclophosphamide (CTX) plus DTX was designed. Low-dose DTX did not negatively affect iDC viability and instead promoted maturation and IL-12 production. Five patients with metastatic or recurrent HNC were enrolled for the trial. All patients experienced grade 1 to 3 fevers. Intriguingly, elevated CD8+ effector T-cells and reduced Tregs were observed in four patients who completed two treatment cycles. All patients were judged to have progressive disease, but tumor regressions were observed in a subset of targeted metastatic lesions in two of five patients. Our results show that the combination of direct peritumoral iDC injection with OK-432 and low-dose CTX plus DTX is well tolerated and should give rise to changing the immune profile of T-cell subsets and improvement of immunosuppression in advanced HNC patients. Additionally, our ex vivo data on the effect of low-dose DTX treatment on DC maturation may contribute to developing new combination therapies with low-dose chemotherapy and immunotherapy.

Hypereosinophilic syndrome (HES) is a systemic disease characterized by an increased peripheral blood eosinophil count accompanied by systemic organ dysfunction. HES is classified into idiopathic HES, primary (neoplastic) HES (HESN), and secondary (reactive) HES (HESR). In this case report, a patient who developed peripheral blood eosinophilia and granulation tissue in the pharynx and paranasal sinus, which was initially diagnosed as chronic eosinophilic leukemia (CEL), categorized as HESN, but was eventually identified after the patient had died as natural killer/T-cell (NK/T) lymphoma, nasal type (ENKL), categorized as HESR, is presented. ENKL-induced HES is very rare but must be considered.

Objectives Intraoperative identification of tumor margins is essential to achieving complete tumor resection. However, the process of intraoperative pathological diagnosis involves cumbersome procedures, such as preparation of cryosections and microscopic examination, thus requiring more than 30 min. Moreover, intraoperative diagnoses made by examining cryosections are occasionally inconsistent with postoperative diagnoses made by examining paraffin-embedded sections because the former are of poorer quality. We sought to establish a more rapid accurate method of intraoperative assessment. Materials and methods A diagnostic algorithm of head and neck squamous cell carcinoma (HNSCC) using machine learning was constructed by mass spectra obtained from 15 non-cancerous and 19 HNSCC specimens by probe electrospray ionization mass spectrometry (PESI-MS). The clinical validity of this system was evaluated using intraoperative specimens of HNSCC and normal mucosa. Results A total of 114 and 141 mass spectra were acquired from non-cancerous and cancerous specimens, respectively, using both positive- and negative-ion modes of PESI-MS. These data were fed into partial least squares-logistic regression (PLS-LR) to discriminate tumor-specific spectral patterns. Leave-one-patient-out cross validation of this algorithm in positive- and negative-ion modes showed accuracies in HNSCC diagnosis of 90.48% and 95.35%, respectively. In intraoperative specimens of HNSCC, this algorithm precisely defined the borders of the cancerous regions; these corresponded with those determined by examining histologic sections. The procedure took approximately 5 min. Conclusion This diagnostic system, based on machine learning, enables accurate discrimination of cancerous regions and has the potential to provide rapid intraoperative assessment of HNSCC margins.

Background House dust mite (HDM) respiratory allergy is a common and burdensome disease in children and adolescents. There are few HDM allergy immunotherapy trials in children with perennial allergic rhinitis. This post hoc analysis used pooled data to evaluate efficacy and safety of the SQ HDM sublingual immunotherapy (SLIT) tablet in adolescents (12-17 years). Methods In 2 double-blind, placebo-controlled trials conducted in North America and Japan, respectively, subjects aged 12+ years with HDM allergic rhinitis were randomized to up to one year of treatment. The primary endpoint in both trials was the average total combined rhinitis score (TCRS) during the last 8 weeks of treatment in the active group compared to placebo. Data from subjects aged 12–17 years were pooled (N=395). Results In the pooled adolescent subpopulation, average TCRS improved 22% with 12 SQ-HDM versus placebo (absolute treatment difference of 1.04; p

Background: Complementary and alternative medicine (CAM) is extensively used in patients with allergic diseases worldwide. The purpose of this study was to investigate the actual situation of CAM practice in the treatment of allergic rhinitis. Methods: We distributed questionnaires to otolaryngologists at 114 facilities in Japan. The subjects who participated in this study included children

Bothersome symptoms of hay fever impair not only patients' quality of life but also their labor productivity and learning efficiency. Excessive daytime sleepiness (EDS) caused by hay fever is thought to be one of the reasons for these impairments. The purpose of this study was to investigate the relationship between the severity of springtime hay fever and EDS by using a questionnaire. The questionnaire included information about age, sex, height, weight, severity of hay fever, treatment for hay fever, smoking and alcohol consumption habit, history of drug use for sleeping, existence of snoring, and Japanese version of the Epworth Sleepiness Scale. After excluding responses containing insufficient data, responses from 1,734 patients were considered as eligible. By performing logistic regression analysis, we analyzed the effect of the aforementioned parameters on the comorbidity of EDS and snoring. The odds ratio (OR) to comorbid EDS was significantly higher in the moderate and severe hay fever groups than in the asymptomatic hay fever group (moderate: OR=1.76, p=0.014, severe: OR=2.53, p

Pneumonia is the third leading cause of death in Japan, several cases of which are associated with aspiration pneumonia. However, no quantitative indicator exists for swallowing function evaluation. Therefore, we decided to use pharyngeal ultrasound video to establish the indicator noninvasively. We herein propose a method to obtain the movement speed and opening condition of the esophagus as well as the flow speed of the bolus, which are considered effective indicators of food-correspondent aspiration. Additionally, using this method, the flow characteristics of the esophageal wall and the bolus can be visualized and compared by extracting the elements at the time of swallowing three liquids of different viscosities.

In Japan, a country with an unparalleled super-aged society, the seventh leading cause of death is aspiration pneumonia at 2.7%, with the overall death rate due to pneumonia being 9.9%. The probability of aspiration of food related to the degree of swallowing function is unknown. In this study, multichannel analysis was used to measure the time gap between signals from multiple microphones when recording the sound of swallowing. In addition, the change in timing of sounds I, II, and III was confirmed depending on the amount and viscosity of swallowed food. In the multichannel analysis, a slight deviation of the signal occurs for each microphone, suggesting that the index can be extracted from the multichannel analysis. It was determined that when the amount of material to be swallowed and the viscosity change, the time of the entire set of sounds and that of each section changes accordingly.

Thickening agents are commonly used to prevent aspiration, a condition that can prove fatal in the elderly. However, no established indicators exist that show the extent to which sticky food reduces the risk of aspiration. VideoEndoscopy (VE) and VideoFluorography (VF) are the classic inspection methods used for evaluating the function of swallowing, but they are both have limited utility in that they are invasive. In this study, we propose a non-invasive method that exploits esophagus ultrasound videos to estimate the internal flow characteristics of foods, and facilitates quantitative evaluation of the swallowing function. The method combines optical flow with Maximally Stable Extremal Regions (MSER) to extract the movement velocity and position of the esophagus and bolus. The results suggest that movement velocity could be used as an indicator to quantify the internal flow characteristics of foods. The displacement of the esophagus indicates the esophageal opening and could be used as an indicator to evaluate swallowing.

Sclerosing polycystic adenosis (SPA) is a rare salivary gland disease. Histologically it resembles a low-grade ductal carcinoma in situ or sclerosing adenosis of the breast, characterized by lobular proliferation of ducts with apocrine cellular features surrounded by fibrosclerotic stroma. Although SPA is typically benign, recurrence is not uncommon, and cases with a malignant component have been documented. Thus, complete excision is desirable but preoperative diagnosis is challenging. A 12-year-old boy presented with a painless mass in the right neck. We identified a well-demarcated mass in the right parotid region measuring approximately 2 cm using cervical echography and magnetic resonance (MR) imaging. Fine-needle aspiration (FNA) revealed two cell types. There were loosely cohesive clusters of polymorphic epithelioid cells with irregular nuclei and abundant vacuolated cytoplasm containing zymogen granules. Some of these cells were binuclear. The other cell types represented normal ductal cells. The original cytological diagnosis was Warthin tumor. Right parotidectomy was performed. Histologically, we observed proliferation of ducts with granular, vacuolated, zymogen granules, and apocrine-like features in the cytoplasm with hyalinizing sclerotic stroma and some binuclear cells. Four years after parotidectomy, there has been no recurrence or malignant transformation.Cytological diagnosis of SPA is challenging on FNA specimens since SPA is a very rare entity of the salivary gland that can mimic other salivary gland neoplasms. A mixture of apocrine-like cells and sebaceous-like cells, nuclear pleomorphism, and zymogen granules can help to diagnose this rare lesion during the initial cytological diagnosis.

The Eustachian tube insufflation method is widely used for diagnosis and medical treatment. However, because the Eustachian tube insufflation method is auscultated by only the doctor who enforces it, the Eustachian tube insufflation sound is not left behind as data, and diagnosis may change depending on the doctors because the sound on which the diagnosis is based is subjective. Therefore, this research considered using feature extraction for classifying the difference in the Eustachian tube insufflation sound according to the disease condition using a self-organizing map (SOM) for the purpose of improving the objectivity and reliability of diagnosis by quantification of the Eustachian tube insufflation method. In this study, various feature vectors to be input to the SOM were prepared and examined, and the result of refinement concerning the optimal condition is reported.

Background House dust mite (HDM) is the major indoor allergen for allergic diseases such as allergic rhinitis (AR) and asthma. Although sublingual immunotherapy is a curative treatment for HDM-induced AR, data from large-scale studies are limited. We evaluated the efficacy and safety of HDM tablets in adolescent and adult patients (aged 12–64 years) with HDM-induced AR with or without intermittent asthma. Methods In a double-blind trial in Japan, 968 subjects were randomized 1 : 1 : 1 to 300 index of reactivity (IR), 500 IR, or placebo groups. The primary endpoint was the Average Adjusted Symptom Score (AASS) in the last eight weeks of the 52-week treatment. Secondary endpoints included individual nasal and ocular symptom scores, rescue medication use, and the Japanese Rhinoconjunctivitis Quality of Life Questionnaire (JRQLQ) scores. Results The AASS in the last eight weeks of treatment significantly improved in both the 300 IR and the 500 IR groups compared to that in the placebo group (P < 0.001). In the 300 IR group, the onset of action occurred at week 8–10. All four nasal symptoms significantly improved in both active treatment groups; rescue medication use and JRQLQ outcome improved in the 300 IR group. Most adverse events (AEs) were mild, and 16 serious AEs (SAEs) were reported; however, none of them were drug-related. Conclusions One-year treatment with 300 IR and 500 IR HDM tablets was effective without major safety concerns. The recommended therapeutic dose for AR is 300 IR.

Objective Sublingual immunotherapy (SLIT) appears to offer practical advantages for the treatment of allergic rhinitis (AR). Based on a review of the scientific literature, we present recommendations as guiding principles to administer SLIT safely. Methods Clinical questions concerning SLIT were prepared. Literature published between January 2003 and December 2012 was searched from PubMed, the Cochrane Library, and Japana Centra Revuo Medicina. Qualified studies were analyzed and the results were evaluated, consolidated, and codified. We answered 17 clinical questions and, based on this, presented evidence-based recommendations. Results Sublingual immunotherapy improved symptoms (e.g., quality of life [QOL]) and reduced medication scores in seasonal AR and perennial AR. Most SLIT-induced adverse effects were local oral reactions, although systemic adverse effects such as gastrointestinal symptoms, urticaria, and asthma are occasionally reported. There have been no reports of lethal anaphylactic reactions by SLIT. When SLIT is continued for 3–4 years, its effect persists long after discontinuation. Conclusion A correct diagnosis of AR and sufficient informed consent from patients are required before initiating SLIT. Sublingual immunotherapy should be continued for 3 years or longer. The initial administration of SLIT during the uptitration of an allergen vaccine and the general condition of patients are critical for the safe performance of SLIT.

Background: Sublingual immunotherapy (SLIT) is a safe and effective treatment for allergic rhinitis (AR). However, many issues regarding SLIT remain to be resolved, including the optimal timing of administration. This study investigated the effect of time of day on SLIT efficacy with the goal of optimizing the therapeutic outcome. Methods: We performed prophylactic SLIT at different times of day (10 a.m. or 10 p.m.) in 2 mouse models of AR: an ovalbumin (OVA)-induced AR model and Cry j 1-induced AR model, and compared the effects. Results: In the OVA-induced AR model, mice sublingually receiving OVA at 10 a.m. exhibited a greater decrease in total and OVA-specific IgE levels than mice treated at 10 p.m. In addition, mice treated at 10 a.m. exhibited reductions in OVA-specific IL-4, IL-10, and IL-13 production by splenocytes relative to mice treated at 10 p.m. Furthermore, we observed a more efficient capture of sublingually administered OVA in submandibular lymph nodes at 10 a.m. than at 10 p.m. in mice. Similar results were observed in the Cry j 1-induced AR model using Japanese cedar pollen extract for SLIT. Conclusions: Given the allergen-specific antibody and T cell responses, we suggest that SLIT may be more effective in the resting phase than in the active phase (note that mice are nocturnal animals). Thus, we propose that a chronotherapeutic approach should be considered for SLIT to maximize its effectiveness.

Japanese cedar pollen (JCP) and house dust mite (HDM) are two major allergens that cause allergic rhinitis (AR) in Japan and the prevalence of AR is increasing. Pharmacothearpy is a commonly used treatment, but the level of patient satisfaction is very low. Allergen immunotherapy (AIT) is the only therapeutic modality that provides not only symptom relief but also quality of life improvement that leads to a high rate of satisfaction. In particular, sublingual immunotherapy (SLIT) is a safe and effective treatment for AR. Here we introduce a large-scale double-blind, placebo-controlled trial of SLIT in Japanese patients using JCP droplets or HDM tablets conducted in Japan. The immediate future of SLIT in Japan is also discussed. Keywords: Allergen immunothearpy, Allergic rhinitis, HDM allergen, Japanese cedar pollinosis, Sublingual immunotherapy

Background The SQ house dust mite (HDM) sublingual immunotherapy (SLIT)-tablet (TO-203, Torii, Japan/ALK, Denmark) treatment has been effective against respiratory allergic diseases in patients aged ≥12 years during European, Japanese, and North American trials. This trial was conducted to investigate the efficacy and safety of this treatment in Japanese children (5-17 years) with moderate-to-severe HDM allergic rhinitis (AR). Methods In this randomized, double-blind, placebo-controlled trial, 458 Japanese children were randomly assigned to a daily SQ HDM SLIT-tablet [10 000 Japanese Allergy Unit (JAU), equivalent to 6 SQ-HDM in Europe and the US] or placebo (1:1) treatment for 1 year. Inclusion required an AR symptom score of ≥7 on at least 7 days during a 14-day run-in period while symptomatic treatment was withdrawn. The primary endpoint was the total combined rhinitis score (TCRS) comprising AR symptom and medication scores during the last 8 weeks of the treatment period. Results The analysis of primary endpoint demonstrated statistically significant absolute reduction in TCRS of 1.22 with a relative difference of 23% (95% confidence interval, 14% to 31%) in the 10 000 JAU compared with placebo. Predefined stratified analyses revealed the same degree of efficacy of 1.11 (P = 0.002), 21% (8% to 32%) and 1.36 (P = 0.001), 26% (11% to 38%), respectively, in pediatric (5-11 years) and adolescent subjects (12-17 years). The treatment was well tolerated by both pediatric and adolescent subjects. Conclusion This trial, for the first time, demonstrated the efficacy and safety of the HDM SLIT-tablet in pediatric patients with moderate-to-severe HDM AR (JapicCTI-152953).

Thyroid-like low-grade nasopharyngeal papillary adenocarcinoma (TL-LGNPPA) is an extremely rare neoplasm characterized by morphological analogy to papillary thyroid carcinoma and abnormal expression of thyroid transcription factor-1 (TTF-1). Here we report a novel case of TL-LGNPPA with literature review. The patient was a 43-year-old woman complaining of nasal obstruction. Laryngoscopic study and computed tomography identified a pedunculated mass located on the posterior edge of the left nasal septum. Histologically, the tumor consisted of papillary growth of cuboidal or columnar epithelium. Tubular architecture and a spindle cell component were also observed focally. Some tumor cells exhibited intra-nuclear cytoplasmic inclusions. Immunohistochemically, the neoplastic cells were positive for pancytokeratin (AE1/AE3), CK7, CK19, TTF-1, vimentin and HBME1, but negative for thyroglobulin, Pax8 and CK5/6. Ki67-labeling index reached 5% in the most concentrated spot. Despite the morphological and immunohistochemical similarity to papillary thyroid carcinoma, no BRAF V600E mutation was detected by mutation-specific immunohistochemistry. The patient had neither local recurrence nor distant metastasis 19 months after removal of the tumor.

We experienced two cases of esophageal web accompanying severe stricture that were treated by endoscopic incisions with an insulated-tip knife (IT-knife). With attention paid to the mucosa at the stricture, the lesion was incised with an IT-knife without complications. Sato's curved laryngoscope was used even in cervical esophageal lesions and an excellent field was secured.

Background Allergic rhinitis (AR) is a very common disorder peaking in the teenage years that is mediated by hypersensitivity responses to environmental allergens. Although it is well established that the ORMDL3 locus at chromosome 17q21 is associated with susceptibility to bronchial asthma, the genetic influences of the polymorphisms of the locus in allergic rhinitis are unclear. Objective To examine whether the polymorphisms in the 17q21 asthma susceptibility locus are associated with allergic rhinitis in the Japanese population. Methods We performed linkage disequilibrium (LD) mapping of the locus using the HapMap database and conducted an association study of the locus with a total of 15 tag SNPs in two independent populations. We further evaluated correlations of genotypes with changes in expression of genes at the region in lymphoblastoid cell lines in the Japanese population and assessed the expression levels of the genes in nasal epithelium and various human tissues. Results We found a significant association between a total of five polymorphisms in the 17q21 asthma susceptibility locus, rs9303277, rs7216389, rs7224129, rs3744246, and rs4794820, and AR (minimum Pcombined = 0.00074, rs4794820). The expression level of the ORMDL3 transcript was significantly correlated with the genotype of rs12150079, rs7216389, rs3744246, and rs4794820 with P

Myeloid‐derived suppressor cells (MDSC) represent a heterogeneous population and have the potential to suppress immune responses via diverse mechanisms. In recent studies, a new subset of MDSC was identified by the markers CD14(+) and HLA‐DR (−) in the peripheral blood from cancer patients. In this study, we investigated the proportions and characteristics of CD14(+) HLA‐DR (−) cells in patients with squamous cell carcinoma of the head and neck (SCCHN). As expected, the percentage of CD14(+) HLA‐DR (−) cells was significantly elevated in patients relative to healthy donors and the sorted CD14(+) HLA‐DR (−) cells were able to suppress effectively both the proliferation and IFN‐γ production of anti‐CD3/anti‐CD28 stimulated T cells, suggesting that CD14(+) HLA‐DR (−) cells in patients with SCCHN contribute to the immune suppressive status. Furthermore, CD14(+) HLA‐DR (−) cells revealed a higher level of CD86 and PD‐L1 expression and transforming growth factor (TGF)‐β production than CD14(+) HLA‐DR (+) cells. Addition of anti‐CD86 mAb, anti‐PD‐L1 mAb and anti‐TGF‐β mAb partially restored T‐cell proliferation and IFN‐γ production, respectively, indicating that the suppressive effects of CD14(+) HLA‐DR (−) cells appear to be mediated by various molecules, including coinhibitory molecules and cytokines. Our data suggest that CD14(+) HLA‐DR (−) cells act as potent immunosuppressive cells and particularly contribute to tumor escape from the host immune system in patients with SCCHN. (Cancer Sci 2012; 103: 976–983)

Objective In practical guidelines for management of allergic rhinitis in Japan, pranlukast is a leukotriene receptor antagonist recommended for the treatment of pollinosis. However, the effect of pranlukast on nasal symptoms for cedar pollinosis has not been thoroughly investigated. The aim of this study is to examine this effect in a double-blind controlled crossover study using a pollen challenge chamber (the OHIO Chamber) developed in Japan. Research design and methods A total of 39 patients with cedar pollinosis were targeted. The subjects were exposed to a specific amount of cedar pollen (8000/m3) in the OHIO Chamber during the non-cedar pollen season. Efficacy of pranlukast for the treatment of artificially induced nasal symptoms was compared with that of a placebo using the crossover method. Pranlukast was administered orally for 3 days, after dinner on the day before cedar pollen exposure, after breakfast and after dinner on the day of cedar pollen exposure, and after breakfast on the following day. Pollen testing was carried out twice, with a 1-week wash-out interval. Clinical trial registration The University Hospital Medical Information Network in Japan (UMIN), number UMIN000001282. Main outcome measures The effect of pranlukast was evaluated using self-rating of nasal symptoms by the subjects. Results All 39 subjects demonstrated a positive skin reaction to cedar pollen by a positive CAP-RAST score (class 2 or higher) within the last 3 years, and experienced aggravated congestion during the cedar pollen season for more than 2 years. Nasal congestion was inhibited significantly in the pranlukast group compared to the placebo group during cedar pollen exposure. Furthermore, pranlukast significantly inhibited nasal congestion compared to the placebo on the day after exposure and on the following day. Conclusions The effect of pranlukast on cedar pollinosis indicates immediate action, and such an effect could take place continuously after cedar pollen exposure. These results demonstrate that pranlukast is effective for the relief of congestion due to pollinosis.

Background: In Japan, oral antihistamines are frequently used as the initial treatment for seasonal allergic rhinitis (SAR), and intranasal steroids are added when nasal symptoms worsen. This study aimed to evaluate whether starting treatment with fluticasone propionate nasal spray (FP) from the beginning of pollinosis symptoms and adding fexofenadine hydrochloride tablet (FEX) when SAR is aggravated could achieve improved amelioration of nasal symptoms throughout the pollen season in comparison with a treatment that involves starting with FEX and later adding FP. Methods: In this pragmatic, randomized, open-label, parallel-group trial, 51 Japanese cedar pollinosis patients (age, 16–85 years) were randomly divided and administered FP 100 mcg twice daily as an initial drug with FEX 60mg twice daily as an additional drug and the same treatment in the reverse order. Nasal symptoms were evaluated in a daily dairy using a 4-point scale. The primary outcome was area under curve of the line representing the daily total nasal symptom score in the pollen season on a graph. Results: Initial treatment with FP was significantly (P=0.0015) more effective than initial treatment with FEX in improving the primary outcome. The average daily total nasal symptom score in the initial treatment with FP group was better than that in the initial treatment with FEX group throughout the pollen season. Conclusions: Initiating treatment with FP and adding FEX might lead to improved outcomes for nasal symptoms in comparison with the same drugs administered in the reverse order.

Cancer immunotherapy is designed to activate and up-regulate the host immune responses against tumor cells and is currently in use or under investigation for cancer treatments. Although tumor cells can use a variety of mechanisms to create an immunosuppressive environment, many agents and strategies to reduce or eliminate tumor-induced immune suppression are either being developed or already in clinical trials. This review deals with recent advances in cancer immunotherapy, focusing on new strategies to enhance antitumor immunity. Numerous approaches have been tested for the development of therapeutic cancer vaccines. Recently, several promising reports suggest that clinical benefit has been shown in randomized phase III trials for vaccines against prostate cancer, follicular lymphoma, and melanoma. Moreover, in 2010, the first patient-specific immunotherapy (sipuleucel-T) was approved by the FDA for the treatment of prostate cancer. On the other hand, some chemotherapeutic agents such as cyclophosphamide and docetaxel, have direct effects on the immune system that is likely to contribute to an improved antitumor immunity. Therefore, cancer immunotherapy combined with chemotherapy may represent one approach for breaking tumor-induced immune suppression. To date, a number of monoclonal antibodies are in use or under evaluation. Among these antibodies, blocking negative immunoregulatory receptors, CTLA-4 and PD-1, also represent a new strategy to induce antitumor immunity by manipulation of the immune system. In clinical trials, these antibodies could increase anti-tumor T-cell immunity and result in objective tumor regression in some patients. Thus, accumulating data from recent cancer vaccine trials, the manipulation of immune suppression using chemotherapeutic agents, and the use of immune-stimulatory antibodies raise the hope that immunotherapy could become the fourth modality of cancer treatment. Based on these new findings, in our hospital, a clinical trial using dendritic cells combined with low dose cyclophosphamide and docetaxel in patients with relapsed and refractory head and neck squamous cell carcinoma is under way (UMIN000003725, ClinicalTrials.gov identifier, NCT01149902). In the near future, various immunotherapeutic modalities will most probably be applied as novel approaches in the treatment of head and neck cancer.

Objectives: The current study aimed to characterize comparatively the binding of imidafenacin to muscarinic receptors in the human bladder mucosa and detrusor muscle and parotid gland. Methods: The muscarinic receptor in homogenates of human tissues (bladder mucosa and detrusor muscle and parotid gland) was measured using a radioligand binding assay with [N-methyl-3H]scopolamine methyl chloride ([3H]NMS). Results: Imidafenacin competed with [3H]NMS for binding sites in the bladder mucosa and detrusor muscle and parotid gland, and its affinity was significantly (2.6–8.7 times) higher than that of oxybutynin. Also, the affinity of imidafenacin for muscarinic receptors was approximately two-fold higher in the parotid gland than bladder tissue. The affinity of imidafenacin in the mucosa was similar to that in the detrusor muscle, suggesting that this agent exhibits therapeutic effects by blocking muscarinic receptors in the mucosa as well as detrusor muscle. Scatchard analysis revealed that imidafenacin increased significantly (approximately four-fold) Kd values for [3H]NMS binding in the human detrusor muscle and parotid gland, with little effect on Bmax values. This observation indicates that imidafenacin binds to the muscarinic receptors in human tissues in a competitive and reversible manner. Conclusion: Imidafenacin binds to muscarinic receptors in the human bladder mucosa and detrusor muscle and parotid gland with high affinity. This agent was considered to exhibit therapeutic effects on the lower urinary tract symptoms due to an overactive bladder by blocking muscarinic receptors in the urothelium as well as detrusor muscle.

Background CD44 was identified previously as a surface marker in cancer stem cells (CSCs) of head and neck squamous cell carcinoma (HNSCC). Most cancer treatments have been linked to the activation of the apoptosis-signaling pathway; however, the resistance mechanisms to apoptosis in CSCs have not yet been fully elucidated. Methods The sensitivity of CD44+ cells to diverse apoptosis-inducing stimuli was compared with that of CD44- cells. Furthermore, cell cycle changes and the expression of anti-apoptosis-related genes were examined using flow cytometry and real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results CD44+ cells were resistant to various apoptosis-inducing stimuli. Moreover, CD44+ cells showed a higher proportion of cells in G2/M phase of the cell cycle and upregulation of Bcl-2 and inhibitor of apoptosis (IAP) family genes compared with CD44- cells. Conclusion Treatment resistance in CSCs seems to be regulated by various mechanisms, and, therefore, additional treatment strategies to target CSCs are required in patients with HNSCC. © 2011 Wiley Periodicals, Inc. Head Neck, 2012

It is well recognized that solid tumors are heterogeneous, composed of cells with different phenotypic characteristics, proliferative, differentiated, and/or with malignant potential. Recently, there has been overwhelming evidence that only a minority of cancer cells with stem cell properties, cancer stem cells (CSCs), are responsible for the maintenance and growth of the tumor. These CSC subpopulations show a capacity for highly tumorigenicity, self-renewal, and differentiation. To date, human CSCs have been identified and purified in a variety of malignancies, including those of the brain, breast, prostate, colon, pancreas, and head and neck.1–6 Moreover, several studies have demonstrated that drug or radiation treatment of tumor cells can enrich and maintain the CSC subpopulation in vitro and in vivo,7–10 suggesting that CSCs are responsible for tumor regeneration after conventional cancer treatments. Thus, CSCs have high malignant potential; however, the immunologic properties of CSCs remain unclear. In general, it has long been thought that antitumor immunity plays an important role in the protection against the development of malignancy. In earlier stages of cellular transformation, immunosurveillance can detect and eliminate tumor cells; however, with a developing tumor, tumor variants with reduced immunogenicity, and/or acquired various mechanisms to corrupt the host antitumor response and escape from the host immune system arise, survive and grow in the host.11,12 As a result, interactions between tumor cells and host immune cells in the tumor microenvironment create an immunosuppressive network in which CSCs might actively participate in inducing immunosuppressive ability and promoting evasion from immunosurveillance due to their malignant potential. Indeed, Levina et al13 have reported that lung CSCs selected by treatment with chemotherapeutic drugs produce higher levels of angiogenic and growth factors, such as vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, and granulocyte colony-stimulating factor (G-CSF), than a parental cell line. Moreover, CSCs expressed higher levels of cancer-associated antigens, including embryonic cancer antigens, which are often detected in malignantly transformed cells.13 On the other hand, Kawasaki et al14 found that CSCs derived from a prostate cancer cell line showed an increased expression of the glycoprotein CD200, which is involved in immunosuppression and immune tolerance. More recently, we have demonstrated the identification, expansion, and characterization of CD44+ cancer stem-like cells in a squamous cell carcinoma of the head and neck (SCCHN) cell line.15 Under serum-free medium culture conditions, we enriched a subpopulation of CD44+ cells that possess marked capacity for forming tumor spheres, proliferation, migration, and invasion in vitro. Furthermore, the CD44+ cell population that had been purified using immunomagnetic beads was significantly more resistant to various chemotherapeutic agents than the CD44− cell population. In the present study, we compared the immunologic properties of CD44+ cancer stem-like cells with CD44− cells. Our findings provide new insights into our understanding of immunosuppressive mechanisms in the tumor microenvironment and novel therapeutic approaches against CSCs in SCCHN.

Background: Evidence indicating that CD4+CD25+ regulatory T (Treg) cells play a crucial role in the maintenance of peripheral T cell tolerance to allergens has been accumulated. To explore the functional role of Treg cells in patients with Japanese cedar pollinosis, we performed an in vitro investigation of the regulation of immune responses to allergens by Treg cells. Methods: CD4+ and CD4+CD25– T cells obtained from 12 patients with Japanese cedar pollinosis were stimulated with Cry j 1 protein and Cry j 1-derived peptide. On day 6, T cells were tested for allergen-specific reactivity using a CFSE-based proliferation assay and cytokine ELISA assays. The frequency of Cry j 1-specific interleukin (IL)-10-producing Treg cells was assessed by ELISPOT assays. Results: The proportion of proliferated cells induced by allergen stimulation was similar in both CD4+ and CD4+CD25– cell cultures. The production of interferon (IFN)-γ, but not that of IL-5 was significantly enhanced in CD4+CD25– cell cultures compared to that in CD4+ cell cultures. Interestingly, the production of IL-10 was decreased in CD4+CD25– cell cultures. Moreover, Cry j 1-specific IL-10-producing Treg cells were detected in pollen-allergic patients. Conclusion: Our findings suggest that in pollen-allergic patients, Treg cells predominantly suppresses Th1 responses rather than Th2 responses, where allergen-specific IL-10-producing Treg cells may also be responsible for the downregulation of allergen-specific immune responses.

Background: The relationship between chronic rhinosinusitis (CRS) and asthma has been known for a long time. However, no large studies on the relationship between CRS and lower airway diseases have been reported to date in Japan. Additionally, eosinophilic chronic rhinosinusitis (ECRS) in Japan is considered to be a subgroup of CRS with nasal polyps (CRSwNP) characterized by eosinophil-dominant inflammation. However, the diagnostic criteria of ECRS have not been established. Methods: To investigate clinical and epidemiological features of patients with CRS from the aspect of their associations with lower airway diseases, 553 patients with CRS who visited one of six local university hospitals were examined and interviewed. Local eosinophilic infiltration was evaluated pathologically by exmining NPs. Results: The prevalences of olfactory dysfunction (OD) in the patients with nasal polyps (NPs) and those without NPs were 57.0% and 13.7%, respectively (p < 0.0001). The prevalence of asthma in all patients was 23.1%. Furthermore, the prevalences of NPs and OD in the patients with asthma and those without asthma were 81.0% and 50.1% (p < 0.0001) and 64.2% and 35.7% (p < 0.0001), respectively. 97.4% of the patients with asthma had >15% mucosal eosinophils, and 87.9% of the patients without asthma had

Background: Transforming growth factor-p (TGF-β) levels are elevated in the nasal mucosa in allergic rhinitis. However, because TGF-β is secreted extracellulary in latent complexes, it remains unclear whether the local TGF-β expression actually drives active signaling and affects the pathophysiology of allergic rhinitis. The objective of this study is to investigate whether TGF-β signaling is activated in allergic rhinitis and plays a role in the pathophysiology of allergic rhinitis. Methods: An ovabumin (OVA)-sensitized and -nasally challenged mouse model of allergic rhinitis was established and phosphorylation of Smad2 in the nasal mucosa was examined by immunohistochemistry. In addition, the effects of the pharmacological inhibition of endogenous TGF-β signaling on the allergic rhinitis model were histologically examined. Furthermore, phosphorylation of Smad2 in the nasal mucosa samples obtained from patients with allergic rhinitis was also evaluated. Results: In the mouse model of allergic rhinitis, OVA challenge induced phosphorylation of Smad2 predominantly in epithelial cells in the nasal mucosa. In addition, the administration of an inhibitor of TGF-β type I receptor kinase activity during OVA challenge suppressed goblet cell hyperplasia in the nasal mucosa. Furthermore, phosphorylated Smad2 expression increased in nasal epithelial cells in patients with allergic rhinitis. Conclusions: These results suggest that TGF-β signaling is activated in epithelial cells in the nasal mucosa in allergic rhinitis and may contribute to the development of goblet cell hyperplasia. KEY WORDS: allergic rhinitis, epithelial cells, Smad, TGF-p

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.

Background Allergen specific immunotherapy is highly effective, but adverse events may occur during treatment. Peptide-based immunotherapy has been proposed as one of new strategies for reduction of allergic adverse reactions. We examined the possibility of candidate peptides for the development of peptide-based immunotherapy for Japanese cedar pollinosis. Methods Twelve Cry j 1-specific T-cell lines were established from peripheral blood mononuclear cells (PBMC) of 12 patients with Japanese cedar pollinosis. Using these T-cell lines, 37 Cry j 1-derived overlapping peptides were assessed for their proliferative responses and cytokine production. Results Four peptides corresponding to the Cry j 1 sequence were able to induce proliferative responses to more than one T-cell line: p61-80 (3/12; 25.0%); p115–132 (2/12; 16.6%); p206–225 (4/12; 33.3%); and p337–353 (5/12; 41.7%). Furthermore, T-cell lines generated from 11 of 12 donors (91.7%) responded to at least one of these four peptides. On the other hand, the pattern of cytokine production from Cry j 1-specific T-cell lines varied. Moreover, cytokine production patterns by stimulation with Cry j 1 peptide did not reflect those by stimulation with Cry j 1 protein. Conclusions Our results suggest four Cry j 1-derived peptides (p61–80, p115–132, p206–225 and p337–353) may be considered to be the immunodominant T-cell epitopes of the Cry j 1 molecule, and can be useful for the design of peptide-based immunotherapy for the management of Japanese cedar pollinosis.

Anti-tumor immunity plays an important role in the development of and protection from malignancy. However, there is a lack of information regarding induction of CD4+ T helper responses in patients with squamous cell carcinoma (SCCHN). To explore anti-tumor immune responses against SCCHN, a permanent cell line, Gun-1 was established from a squamous cell carcinoma of the hypopharynx. In addition to its characterization, we performed mixed lymphocyte-tumor cell cultures (MLTC) using peripheral blood lymphocytes and autologous tumor cells. Furthermore, T cell responses to wild type (wt) p53-derived peptides were assessed. Gun-1 cells overexpressed p53 and were negative for HLA-A2 expression. No tumor-specific or wt p53-specific CD8+ CTL lines could be established from peripheral blood mononuclear cells (PBMCs) of this patient. Autologous tumor-specific HLA-DR-restricted CD4+ T helper clone was obtained by limiting dilutions using bulk populations from MLTC. This clone produced IFN-gamma but not IL-5 in response to autologous tumor cells. In addition, CD4+ T cells were generated from the patient's PBMCs which responded to two HLA-DP5-restricted wt p53-derived peptides. Our results suggest that the immune cells specific for autologous tumor as well as wt p53-derived epitopes are present in the peripheral circulation of this cancer patient. However, helper-type CD4+ T lymphocytes represent the predominant anti-tumor response.

Epithelial cell-derived thymic stromal lymphopoietin (TSLP) is a master switch for asthma or atopic dermatitis by inducing a dendritic cell-mediated Th2-type allergic inflammation. Allergic rhinitis is also pathologically characterized by Th2-type allergic inflammation. This study demonstrates that mast cells regulate the epithelial TSLP expression in allergic rhinitis. TSLP expression was found to be up-regulated predominantly in the nasal epithelium in the ovalbumin (OVA)-sensitized and -nasally challenged mouse model of allergic rhinitis, which was abolished in mast cell-deficient WBB6F1-W/W(v) in comparison with control WBB6F1-+/+ mice. Similarly, the epithelial TSLP expression was reduced in Fc receptor gamma chain (FcgammaR)-deficient mice, where the high-affinity IgE receptor (FcepsilonRI) is not expressed on mast cells, in comparison with control C57BL/6 mice. Furthermore, the administration of neutralizing TSLP antibody during the challenge phase of OVA inhibited the development of allergic rhinitis. These results suggest that the direct stimulation of epithelial cells by antigens alone may not be sufficient to induce TSLP expression in the nasal epithelium, and that mast cell regulation of epithelial TSLP expression, possibly via FcepsilonRI, plays an important role in the development of allergic rhinitis.