Identification of BST2 as a conjunctival epithelial stem/progenitor cell marker.

The conjunctival epithelium consists of conjunctival epithelial cells and goblet cells derived from conjunctival epithelial stem/progenitor cells. However, the source of these cells is not well known because no specific markers for conjunctival epithelial stem/progenitor cells have been discovered. Therefore, to identify conjunctival epithelial stem/progenitor cell markers, we performed single-cell RNA sequencing of a conjunctival epithelial cell population derived from human-induced pluripotent stem cells (hiPSCs). The following conjunctival epithelial markers were identified: BST2, SLC2A3, AGR2, TMEM54, OLR1, and TRIM29. Notably, BST2 was strongly positive in the basal conjunctival epithelium, which is thought to be rich in stem/progenitor cells. Moreover, BST2 was able to sort conjunctival epithelial stem/progenitor cells from hiPSC-derived ocular surface epithelial cell populations. BST2-positive cells were highly proliferative and capable of successfully generating conjunctival epithelial sheets containing goblet cells. In conclusion, BST2 has been identified as a specific marker of conjunctival epithelial stem/progenitor cells.

Clinical Findings of Specular Microscopy Images in Cytomegalovirus Corneal Endotheliitis.

PURPOSE: The aim of this study is to evaluate the usefulness of specular microscopy as an alternative diagnostic tool for cytomegalovirus (CMV) corneal endotheliitis. DESIGN: A retrospective study. METHODS: One hundred and four patients with clinical manifestations of infectious corneal endotheliitis, iridocyclitis, and retinitis were included in this study. The presence of CMV deoxyribonucleic acid (DNA) was confirmed by multiplex polymerase chain reaction (PCR). Viral load was measured using real-time PCR. Corneal endothelium was observed by specular microscopy. The medical records and clinical manifestations of the patients were retrospectively reviewed and linked with the PCR results. RESULTS: Seventeen of 104 cases were CMV endotheliitis and/or iridocyclitis and had no history of intraocular surgery or corneal transplantation. There was a negative correlation between viral load and corneal endothelial cell counts. In 14 of 17 cases, owl's eye cells were observed by specular microscopy. The corneal endothelial cell counts were significantly reduced in the cases in which owl's eye cells were observed. CONCLUSIONS: In CMV endotheliitis, owl's eye cells were observed by specular microscopy with high probability (82%). Corneal endothelial cells significantly decreased when owl's eye cells were observed by specular microscopy. Specular microscopy represents a useful noninvasive auxiliary tool for diagnosing and monitoring CMV corneal endotheliitis.

Colonic Endoscopic Submucosal Dissection for a Granular Cell Tumor with Insufficient Endoscopic Manipulation in the Hepatic Flexure.

This report describes a granular cell tumor (GCT) with insufficient endoscopic manipulation in the hepatic flexure (HF) of the colon, which was treated by endoscopic submucosal dissection (ESD) using a splinting tube and the spring S-O clip traction method. A 44-year-old man presented with a 10 mm subepithelial tumor in the HF near the ascending colon on colonoscopy. The lesion had a smooth surface without erosion. The histology of biopsied specimen from the lesion was suspected as a GCT. Most GCTs are considered low-grade malignant, but ESD was chosen to treat the lesion due to the patient's insistence on endoscopic treatment. Because the lesion was located in the HF, it was assumed that the scope manipulation during ESD would be difficult. During ESD, a splinting tube was utilized to stabilize endoscopic manipulation and the spring S-O clip traction method to keep clear visualization of the submucosa, and the procedure was completed without adverse events. An 8 × 7 mm lesion with negative margins was removed by ESD. Hematoxylin and eosin staining showed atypical cells with round-to-oval nuclei and acidophilic vesicles, and immunohistochemical staining for S-100 protein was strongly positive with a Ki-67 labeling index of 5%. The lesion was pathologically confirmed as a GCT. This case showed the usefulness and safety of ESD for GCT with insufficient endoscopic manipulation in the HF.

Effects of cataract surgery on symptoms and findings of dry eye in subjects with and without preexisting dry eye.

PURPOSE: To compare dry eye symptoms and findings in post cataract surgery eyes' with and without preexisting dry eye. STUDY DESIGN: Prospective, observational case-control study. METHODS: Sixty-seven eyes that had undergone cataract surgery were included; 48 were classified into group D (preexisting dry eye) and 19 into group N (no preexisting dry eye). No subjects received perioperative treatment for dry eye. We evaluated between-group differences in symptom scores, corrected distance visual acuity (CDVA), tear film breakup time (BUT), tear film breakup pattern (BUP), and ocular surface fluorescein staining scores, at 1 week, 1 month, and 3 months postoperatively. RESULTS: Symptoms were unchanged in group N, but improved in group D (P < .001) postoperatively. CDVA was improved after surgery in both groups (P < .001). BUT was shorter preoperatively in group D than in group N although this difference was absent 1 month postoperatively. Fluorescein staining scores significantly increased at 1 month postoperatively in group N (P = .01), but did not change in group D. During the perioperative period, the predominant BUP was the random break pattern in both groups (≥ 85%). From 1 week to 3 months, dimple break patterns decreased in group D (P = .007), whereas spot break patterns increased (P = .01). CONCLUSIONS: Cataract surgery has an influence on tear film stability and the ocular surface. There was either a transient improvement or worsening of ocular surface wettability in some patients without preexisting dry eye.

Inflammatory marker and aryl hydrocarbon receptor-dependent responses in human macrophages exposed to emissions from biodiesel fuels.

Biodiesel or renewable diesel fuels are alternative fuels produced from vegetable oil and animal tallow that are being considered to help reduce the use of petroleum-based fuels and emissions of air pollutants including greenhouse gases. Here, we analyzed the gene expression of inflammatory marker responses and the cytochrome P450 1A1 (CYP1A1) enzyme after exposure to diesel and biodiesel emission samples generated from an in-use heavy-duty diesel vehicle. Particulate emission samples from petroleum-based California Air Resource Board (CARB)-certified ultralow sulfur diesel (CARB ULSD), biodiesel, and renewable hydro-treated diesel all induced inflammatory markers such as cyclooxygenase-2 (COX)-2 and interleukin (IL)-8 in human U937-derived macrophages and the expression of the xenobiotic metabolizing enzyme CYP1A1. Furthermore, the results indicate that the particle emissions from CARB ULSD and the alternative diesel fuel blends activate the aryl hydrocarbon receptor (AhR) and induce CYP1A1 in a dose- and AhR-dependent manner which was supported by the AhR luciferase reporter assay and gel shift analysis. Based on a per mile emissions with the model year 2000 heavy duty vehicle tested, the effects of the alternative diesel fuel blends emissions on the expression on inflammatory markers like IL-8 and COX-2 tend to be lower than emission samples derived from CARB ULSD fuel. The results will help to assess the potential benefits and toxicity from biofuel use as alternative fuels in modern technology diesel engines.

Clinical Findings of Anterior Segment Spectral Domain Optical Coherence Tomography Images in Cytomegalovirus Corneal Endotheliitis.

PURPOSE: To evaluate the morphological characteristics of posterior corneal regions including keratic precipitates in eyes with cytomegalovirus (CMV) corneal endotheliitis using anterior segment spectral domain optical coherence tomography (SD-OCT). METHODS: Thirteen eyes of 13 patients with polymerase chain reaction-proven CMV corneal endotheliitis were included in this study. Slit-lamp images and anterior segment SD-OCT images of the posterior cornea were obtained to analyze the clinical characteristics of corneal structures and keratic precipitates. Morphological changes in the posterior cornea throughout the course of an antiviral treatment were also investigated. RESULTS: Anterior SD-OCT images showed protruding structures at the posterior cornea. These protruding structures exhibited dendritic, dome-shaped, quadrangular, or saw-tooth appearance, and reflectivity of these structures was high. Reflectivity of posterior corneal images including the endothelium and deep stromal corneal regions were also high (76.9%). Because corneal inflammation and corneal edema improved, the protruding structures and high-intensity regions of posterior corneal images were resolved after a course of antiviral treatment. CONCLUSIONS: The anterior segment SD-OCT examination represents a useful noninvasive alternative to diagnose and monitor CMV corneal endotheliitis.

Increased expression of long non-coding RNA XIST predicts favorable prognosis of cervical squamous cell carcinoma subsequent to definitive chemoradiation therapy.

The present retrospective study aimed to examine the association between the expression of long non-protein-coding RNAs (lncRNAs) and clinical prognosis in the pretreatment formalin-fixed, paraffin-embedded (FFPE) tissue samples of cervical squamous cell carcinoma patients that underwent platinum-based chemoradiation therapy. Between 2001 and 2013, 49 consecutive patients with squamous cell cervical carcinoma were selected for the present study (median follow-up period, 44.1 months). The patients possessed an International Federation of Gynecology and Obstetrics stage of IB1/IIA1 (with pelvic lymph node metastasis), IB2 or IIA2-IVA, and had been treated with definitive chemoradiation therapy. The pretreatment FFPE tumor biopsies of the patients obtained diagnosis were used for analysis. Total RNAs were extracted from the FFPE tumor tissues and reverse transcription-quantitative polymerase chain reaction was performed to examine the expression level of lncRNAs. The expression level of X inactive-specific transcript (XIST) demonstrated a significant association with the overall survival rate (P=0.014). The 4-year overall survival rates were 87.1 and 54.4% in the high and low XIST expression groups, respectively. Since the expression of XIST is associated with the overall survival rate, this lncRNA has the potential to become a predictor for the prognosis of cervical squamous cell carcinoma patients that are treated with chemoradiation therapy. Additional studies are required to investigate the underlying mechanisms of XIST that are associated with prognosis.

Radiation-induced low-grade fibromyxoid sarcoma of the chest wall nine years subsequent to radiotherapy for breast carcinoma: A case report.

The present study reports a case of low-grade fibromyxoid sarcoma that occurred in a 62-year-old woman 9 years subsequent to whole breast irradiation for a carcinoma of the left breast, and 18 years following chemotherapy and radiotherapy (RT) for non-Hodgkin's lymphoma (NHL; diagnosed at the age of 43). The patient was 53 years of age when a cT2N0M0 stage IIA breast tumor was identified and excised. A 2.5 cm diameter nodule with dimpling in the upper-outer region of the left breast was detected. Pathological examination revealed that the tumor was an invasive ductal carcinoma, of a solid tubular type. The patient was treated with post-surgical whole breast RT. The left breast received 46 Gy in 23 fractions (2 Gy per fraction) for 4 weeks and 3 days, followed by a cone down boost of 14 Gy in 7 fractions (2 Gy per fraction); therefore a total dose of 60 Gy in 30 fractions was administered. In total, 9 years subsequent to RT, the patient observed a small lump in the left chest wall. The patient underwent excision of the tumor and pectoralis major fascia. Microscopically, the tumor consisted of atypical spindle cells with myxoid stroma. Pathologists concluded that the tumor was a low-grade fibromyxoid sarcoma. Since the tumor developed from tissue in a previously irradiated region, it was considered to be RT-induced, and was classified using the radiation-induced sarcoma (RIS) criteria as dictated by Cahan. Although the majority of RIS cases are angiosarcomas, a rare, low-grade fibromyxoid sarcoma was observed in the present study. The present study hypothesizes that there may have been an overlap region between the RT for supraclavicular nodes of NHL and the whole breast RT for primary breast cancer, due to the results of a retrospective dose reconstruction undertaken by the present study. The patient remained clinically stable for 4 years thereafter, until 2008 when the patient experienced a local relapse and underwent surgery. On 19 October 2011, the patient succumbed to RIS. The current study suggests that the RT history of a patient requires consideration due to the possible development of RIS, including the development of a low-grade fibromyxoid sarcoma, which may lead to a poor prognosis.

Details of recurrence sites after definitive radiation therapy for cervical cancer.

OBJECTIVE: This is a retrospective study aimed at clarifying the details of recurrence patterns and sites in patients with cervical cancer treated with definitive radiation therapy (RT). METHODS: Data were analyzed from consecutive patients, admitted to the University of Tokyo Hospital (Tokyo, Japan) between 2001 and 2013, who had received definitive RT, with or without chemotherapy, for International Federation of Gynecology and Obstetrics stages IB-IVA cervical cancer. RESULTS: One hundred and thirty-seven patients formed the patient cohort. The median follow-up period for surviving patients was 57.0 months. A complete response was achieved in 121 patients (88%). Of these, 36 (30%) developed a cancer recurrence during follow-up. The first sites of recurrence were located in intra-RT fields in nine, outside RT fields in 20, and both in seven patients. In the intra-RT field group, all patients showed a local recurrence, while no one experienced an isolated pelvic lymph node (PLN) recurrence. In the outside RT field group, the most frequent site of recurrence was lung (60%), and three-quarters of patients were free from intra-RT field recurrence until the last follow-up. Of the entire cohort, including 48 PLN-positive patients, only seven patients (5.1%) developed PLN persistence or recurrence, all in the common iliac, internal iliac, and/or obturator nodes, and all with another synchronous relapse. CONCLUSION: Local disease was a major type of intra-RT field recurrence, while PLN control was favorable even in initially PLN-positive patients. The predominance of outside RT field recurrence alone highlights issues concerning distant control, including the intensity enhancement of systematic therapy.

A study of high-dose-rate intracavitary brachytherapy boost for curative treatment of uterine cervical cancer.

PURPOSE: This study assessed the trial of boost intracavitary brachytherapy (ICBT) for the patients of uterine cervical cancer with residual malignant cells detected at the final stage of ICBT. MATERIAL AND METHODS: This is a retrospective analysis of 75 patients with cervical cancer treated radically with external beam radiotherapy and high-dose-rate intracavitary brachytherapy between August 2004 and December 2008. Eighteen patients (24%) out of 75 received one additional ICBT and five patients (7%) had two additional ICBT. All 75 patients were retrospectively analyzed. The median follow-up time was 30 months. The median age was 59 years (range 28-85 years). There were 12 patients (16%) in stage IB, 27 (36%) stage II, 22 (29%) stage III, and 14 (19%) stage IV. RESULTS: The 5-year overall survival rate was 65%. Non-hematological toxicities greater than grade 2 occurred in 12 patients (16%). Of these, only two patients received on additional ICBT. No significant difference was found in grade 3 toxicity between patients who did and did not receive additional ICBT (p = 0.8). CONCLUSIONS: The method to perform dose escalation should be examined depending on the treatment response.

Omission of axillary lymph node dissection for clinically node negative early-stage breast cancer patients.

BACKGROUND: For clinically node negative (N0) breast cancer patients, sentinel node (SN) biopsy (SNB) is a standard technique and complete axillary lymph node dissection (ALND) remains the standard treatment when the SN is positive. However, the American College of Surgeons Oncology Group Z0011 trial and the International Breast Cancer Study Group 23-01 trial showed that SNB without ALND can offer excellent regional control and equal survival compared with ALND for limited macrometastatic and micrometastatic SN involvement, respectively. We retrospectively evaluated axillary control rates in clinically N0 patients who had no axillary surgical treatment. METHODS: Data on 158 patients who underwent breast-conserving therapy without any axillary surgical procedure between 1994 and 2010 were extracted. The last follow-up was on May 2013, and the overall median follow-up period was 119.0 months. RESULTS: Of all 158 patients, 10 (6.3 %) and 3 (1.9 %) developed locoregional and axillary recurrences, respectively. The 10-year locoregional and axillary recurrence rates were 5.8 and 2.1 %, respectively. The 5- and 10-year overall survival rates were 94.0 and 84.8 %, respectively. Cases with axillary recurrence tended to have common risk factors for recurrence. CONCLUSION: Even if SNB and ALND were omitted, local and regional recurrence rates were very low among clinically N0 patients and were at the same levels shown in recent trials. This suggests that at least ALND might be safely avoided in clinically N0 patients without any obvious risk factors regardless of axillary nodal status after SNB.

Involved-field irradiation concurrently combined with nedaplatin/5-fluorouracil for inoperable esophageal cancer on basis of (18)FDG-PET scans: a phase II study.

PURPOSE: A prospective study was performed on chemoradiotherapy (CRT) for esophageal cancer using involved-field radiation therapy (IFRT) based on 18-fluorodeoxyglucose positron-emission tomography. The goal of this phase II study was to evaluate the efficacy of the IFRT procedure in newly diagnosed esophageal cancer. PATIENTS AND METHODS: Eligible patients were adults with newly diagnosed untreated, inoperable esophageal cancer in stages I-IV with lymph node metastases. Patients received nedaplatin 80mg/m(2) per day on day 1, 5-fluorouracil 800mg/m(2) on days 1-4 intravenously repeated every 28days for 2-4 cycles, and combined IFRT. Elective nodal irradiation was not performed. Irradiation was applied only to the primary tumor and positive lymph nodes. RESULTS: From September 2009 to July 2012, of the 63 patients enrolled, 58 were evaluable for response. The primary end point of isolated out-of-field loco-regional nodal recurrence was seen in only two patients. The expectant rate was assumed to be less than 5%. The threshold value was set as 10% to calculate the number of registrations. Progression-free and overall survival rates at 36months were 47.7% and 51.1%, respectively. The median progression-free survival was 34.6months, and overall survival was 38.4months. Salvage surgery was tried for 11 patients (17.5%) due to residual or recurrent disease. CONCLUSION: The primary end point of the trial was demonstrated, indicating the efficacy of IFRT in the treatment of inoperable esophageal cancer mostly of squamous cell carcinoma.

s-Afadin binds more preferentially to the cell adhesion molecules nectins than l-afadin.

l-Afadin was originally purified from rat brain as an actin filament (F-actin)-binding protein that was homologous to the AF-6 gene product. Concomitantly, s-afadin that did not show an F-actin-binding capability was copurified with l-afadin. Structurally, s-afadin lacks the C-terminal F-actin-binding domain but has two short sequences that were not present in l-afadin. The properties and roles of l-afadin have intensively been investigated, but those of s-afadin have poorly been understood. We show here an additional difference in their biochemical properties other than binding to F-actin between l-afadin and s-afadin. Both l-afadin and s-afadin bound to nectins, immunoglobulin-like cell adhesion molecules, whereas s-afadin more preferentially bound to nectins than l-afadin. The PDZ domain of l-afadin and s-afadin was essential for their binding to nectin-3. The dilute domain of l-afadin negatively regulated its binding to nectin-3, but the deletion of the C-terminal F-actin-binding domain of l-afadin did not increase the binding of l-afadin to nectin-3. These results indicate that the s-afadin-specific C-terminal inserts may be involved in its preference of binding to nectin-3 and raise the possibility that there are proteins other than nectins that more preferentially bind s-afadin than l-afadin.

Longitudinal assessments of quality of life and late toxicities before and after definitive chemoradiation for esophageal cancer.

OBJECTIVE: Definitive chemoradiotherapy is often considered for locally advanced esophageal cancer. We studied the effect of chemoradiotherapy treatment on patients' quality of life and late toxicities. METHODS: Patients undergoing definitive 5-fluorouracil and cis-diammine-glycolatoplatinum (nedaplatin) therapy concurrent with radiotherapy for esophageal cancer without operation adaptation completed standardized quality-of-life questionnaires before and after chemoradiotherapy and at regular times up to ∼5 years. We analyzed differences in a generic quality-of-life score questionnaire (Functional Assessment of Cancer Therapy-Esophageal scoring) over time by using a linear mixed-effects model. RESULTS: Longitudinal changes before the start of treatment were able to be evaluated in a total of 80 patients. The quality-of-life score before treatment was worse in patients with advanced stages than those with early stages. The quality-of-life score deteriorated once at the time of 2 or 3 months after starting chemoradiotherapy compared with pre-chemoradiotherapy and recovered and rose higher at 4 or 5 months than before starting chemoradiotherapy. After that, the recovery of quality of life was maintained up to the observation end. The score of physical functioning such as fatigue, nausea/vomiting, pain and dyspnea deteriorated at the time of 2 or 3 months after starting chemoradiotherapy compared with before chemoradiotherapy (80, 86, 94 and 89%). CONCLUSIONS: The quality-of-life score deteriorates once from before treatment due to acute complications by chemoradiotherapy, but recovers at 4 or 5 months and becomes better than before treatment.

Emissions of polycyclic aromatic hydrocarbons (PAHs) and nitro-PAHs from heavy-duty diesel vehicles with DPF and SCR.

UNLABELLED: In total, 24 polycyclic aromatic hydrocarbons (PAHs) in both gas and particle phases and 35 nitro-PAHs in particle phase were analyzed in the exhaust from heavy-duty diesel vehicles equipped with after-treatment for particulate matter (PM) and NO(x) control. The test vehicles were carried out using a chassis dynamometer under highway cruise, transient Urban Dynamometer Driving Schedule (UDDS), and idle operation. The after-treatment efficiently abated more than 90% of the total PAHs. Indeed, the particle-bound PAHs were reduced by > 99%, and the gaseous PAHs were removed at various extents depending on the type of after-treatment and the test cycles. The PAHs in gas phase dominated the total PAH (gas + particle phases) emissions for all the test vehicles and for all cycles; that is, 99% of the two-ring and 98% of the three-ring and 97% of the four-ring and 95% of the carcinogenic PAHs were in the gas-phase after a diesel particle filter (DPF) and not bound to the very small amount of particulate matter left after a DPF. Consequently, an evaluation of the toxicity of DPF exhaust must include this volatile fraction and cannot be based on the particle fraction only. The selective catalytic reduction (SCR) did not appear to promote nitration of the PAHs in general, although there might be some selective nitration of phenanthrene. Importantly the after-treatment reduced the equivalent B[a]P (B[a]Peq) emissions by > 95%, suggesting a substantial health benefit. IMPLICATIONS: This study demonstrated that after-treatments, including diesel particulate filters (DPF), diesel oxidation catalysts (DOC), and selective catalytic reduction (SCR), significantly reduce the emissions of PAHs from heavy-duty diesel engines. The gas-phase PAHs dominate the total PAH (gas + particle phases) emissions from heavy-duty diesel vehicles retrofitted with various DPFs and not bound to the very small amount of particulate matter left after a DPF. Consequently, an evaluation of the toxicity of DPF exhaust must also include this volatile fraction and cannot be based on the particle fraction only.

Osteoblast-like differentiation of cultured human coronary artery smooth muscle cells by bone morphogenetic protein endothelial cell precursor-derived regulator (BMPER).

Differentiation of vascular smooth muscle cells (SMCs) into osteoblast-like cells is considered to be a mechanism of vascular calcification. However, regulators of osteoblast-like differentiation of vascular SMCs are not fully elucidated. Here, we investigated the expression of bone morphogenetic protein (BMP)-binding endothelial cell precursor-derived regulator (BMPER), a vertebrate homologue of Drosophila crossveinless-2, in vascular SMCs and the role and mode of action of BMPER in osteoblast-like differentiation of human coronary artery SMCs (HCASMCs). BMPER was expressed in cultured human vascular SMCs, including HCASMCs. Silencing of endogenous BMPER expression by an RNA interference technique inhibited osteoblast-like differentiation of HCASMCs, as evaluated by up-regulation of osteoblast markers such as alkaline phosphatase (ALP) and runt-related transcription factor 2 (Runx2), by down-regulation of a SMC marker α-smooth muscle actin (αSMA), and by mineralization. Treatment with recombinant BMPER enhanced, whereas BMP-2 reduced osteoblast-like differentiation. BMPER antagonized BMP-2-induced phosphorylation of Smad 1/5/8, suggesting that the effect of BMPER was mediated by antagonizing the action of BMP. BMPER increased IκBα phosphorylation and NF-κB activity and specific NF-κB decoy oligonucleotides deteriorated osteoblast-like differentiation of HCASMCs by BMPER. In human coronary artery with atherosclerotic plaque containing calcification, the BMPER-positive signals were observed in the neointimal and medial SMCs in the vicinity of the plaque. These findings indicate that BMPER is a novel regulator of the osteoblast-like differentiation of HCASMCs.

Correlation between bladder volume and irradiated dose of small bowel in CT-based planning of intracavitary brachytherapy for cervical cancer.

OBJECTIVE: To quantify the effect of bladder volume on the dose distribution of intracavitary brachytherapy in computed tomography-based treatment planning for cervical cancer. METHODS: Ten patients with cervical cancer were treated with high-dose rate radiation brachytherapy. For the three-dimensional analysis, pelvic computed tomographic scans were obtained from patients with indwelling catheters in place and from patients who received 50, 100, 150 and 200 cc injections of sterile water into their bladders ('200 cc' was defined as a full bladder). Additionally, scans were made in the prone position with the full bladder. RESULTS: Bladder fullness significantly affected the dose to the small bowel and bladder. The median of maximal doses to the small bowel was significantly greater with an empty bladder in all factors of hot spot (480 vs. 256 cGy on D-2cc). Although dosimetry revealed lower doses for larger volumes of bladder (D-50 and V-25%), the median maximal dose to the bladder was significantly greater with a full bladder (420 vs. 775 cGy on D-2cc). The rectosigmoid doses were not affected by bladder distension (476 vs. 467 cGy on D-2cc). After changing to the prone position, the hot spot dose of small bowel did not change but that of the bladder significantly decreased, although this procedure was very difficult. CONCLUSIONS: An increase in bladder volume resulted in a significant reduction in the hot spot dose of the small bowel at the expense of an increase in that of the bladder without changing the dose distribution of the rectosigmoid.

Involvement of the interaction of afadin with ZO-1 in the formation of tight junctions in Madin-Darby canine kidney cells.

Tight junctions (TJs) and adherens junctions (AJs) are major junctional apparatuses in epithelial cells. Claudins and junctional adhesion molecules (JAMs) are major cell adhesion molecules (CAMs) at TJs, whereas cadherins and nectins are major CAMs at AJs. Claudins and JAMs are associated with ZO proteins, whereas cadherins are associated with beta- and alpha-catenins, and nectins are associated with afadin. We previously showed that nectins first form cell-cell adhesions where the cadherin-catenin complex is recruited to form AJs, followed by the recruitment of the JAM-ZO and claudin-ZO complexes to the apical side of AJs to form TJs. It is not fully understood how TJ components are recruited to the apical side of AJs. We studied the roles of afadin and ZO-1 in the formation of TJs in Madin-Darby canine kidney (MDCK) cells. Before the formation of TJs, ZO-1 interacted with afadin through the two proline-rich regions of afadin and the SH3 domain of ZO-1. During and after the formation of TJs, ZO-1 dissociated from afadin and associated with JAM-A. Knockdown of afadin impaired the formation of both AJs and TJs in MDCK cells, whereas knockdown of ZO-1 impaired the formation of TJs, but not AJs. Re-expression of full-length afadin restored the formation of both AJs and TJs in afadin-knockdown MDCK cells, whereas re-expression of afadin-DeltaPR1-2, which is incapable of binding to ZO-1, restored the formation of AJs, but not TJs. These results indicate that the transient interaction of afadin with ZO-1 is necessary for the formation of TJs in MDCK cells.

[Pulmonary edema due to laryngospasm immediately after extubation].

We describe a case of negative pressure pulmonary edema (NPPE) followed by laryngospasm occurred immediately after extubation. A 56-year-old man with a tumor at the site of ureteroneocystostomy underwent left ureterectomy and partial resection of the neobladder under general anesthesia. The tracheal intubation was difficult with glade 3 of Cormack classification. Anesthesia was maintained with sevoflurane, nitrous oxide, and oxygen. After fully awake extubation, the upper airway obstruction due to laryngospasm was observed. A nasal airway was inserted, but face mask ventilation was impossible. Ventilation became possible with SpO2 of around 40%, and spontaneous respiration appeared. The patient was nasally intubated with a fiberoptic bronchoscope. Furosemide was administered in ICU and mechanical ventilation with 5cmH2O PEEP was started. Seventeen hours later, the pulmonary edema disappeared and he was successfully extubated without any complications. It was warned that laryngospasm would occur even after the full emergence, leading to NPPE.