Identification of heparin-binding proteins expressed on Trichosporon asahii cell surface.

Trichosporon asahii is a pathogenic yeast that cause trichosporonosis. T. asahii exhibits several colony morphologies, such as white (W)- or off-white (O)-type, which may affect virulence. In this study, we compared the expression pattern of heparin-binding proteins in various colony morphologies and identified heparin-binding protein in T. asahii. Surface plasmon resonance analysis revealed that cell surface molecules attached more strongly to heparin in W- than O-type cells. We purified and identified a heparin-binding protein strongly expressed in W-type cells using heparin-Sepharose beads, named it heparin-binding protein 1 (HepBP1), and expressed Flag-tagged HepBP1 in mammalian cells. The heparin-binding ability of Flag-tagged HepBP1 was confirmed by pulldown assay using heparin-Sepharose beads. Thus, HepBP1 is a heparin-binding protein on T. asahii cell surface. These results suggest that several T. asahii cell surface proteins interact with glycosaminoglycans; therefore, they could contribute to infection.

Interaction of Host Proteins with Cell Surface Molecules of the Pathogenic Yeast Trichosporon asahii.

Trichosporon asahii is an invasive pathogenic yeast that infects immunocompromised hosts. Several virulence factors contribute to the fungal infection; however, the factors that contribute to the occurrence of T. asahii infections remain unclear. Since adhesins are typical virulence factors reported for pathogenic fungi, we looked for host proteins that interact with the T. asahii cell surface. T. asahii and Candida albicans were used for screening using a pull-down assay with fetal bovine serum. Serum albumin and elongation factor 2 were identified as the yeast-binding serum proteins. Additionally, we investigated the interactions of the cell surface-associated molecules (CSM) of T. asahii with vitronectin (VTN), fibronectin, fetuin-A, and alpha-1antitrypsin (AAT). The surface plasmon resonance (SPR) method was used to examine the interaction between CSM and human proteins. On the other hand, the pull-down assay was used to examine the interaction between human proteins and the T. asahii cell surface. Serum albumin, AAT, and VTN were found to interact with T. asahii in both SPR and pull-down assays. This study identified several proteins that interact with T. asahii, suggesting that these proteins play a role in infection mechanisms.

A disposable, ultra-fine endoscope for non-invasive, close examination of the intraluminal surface of the peritoneal dialysis catheter and peritoneal cavity.

The ability to visualize intraluminal surface of peritoneal dialysis (PD) catheter and peritoneal cavity could allow elucidation of the cases of outflow problems, and provide information on changes to the peritoneal membrane leading to encapsulating peritoneal sclerosis. A non-invasive examination that allows those monitoring in need is desirable. We have developed a disposable ultra-fine endoscope that can be inserted into the lumen of the existing PD catheter, allowing observation of the luminal side of the catheter and peritoneal cavity from the tip of the PD catheter, with minimum invasion in practice. In a pre-clinical study in pigs and a clinical study in 10 PD patients, the device provided detailed images, enabling safe, easy observation of the intraluminal side of the entire catheter, and of the morphology and status of the peritoneal surface in the abdominal cavity under dwelling PD solution. Since this device can be used repeatedly during PD therapy, clinical application of this device could contribute to improved management of clinical issues in current PD therapy, positioning PD as a safer, more reliable treatment modality for end-stage renal disease.

Efficacy of Percutaneous Endoscopic Gastrostomy for Patients With Esophageal Cancer During Preoperative Therapy.

BACKGROUND/AIM: This study aimed to clarify the benefits and disadvantages of percutaneous endoscopic gastrostomy (PEG) for patients with esophageal cancer (EC) during preoperative therapy. MATERIALS AND METHODS: We retrospectively reviewed 92 patients who underwent esophagectomy for EC after preoperative therapy. Patients were divided into the PEG group (n=14) and the control group (n=78) and compared regarding patient characteristics, nutritional status, operative variables, and postoperative complications. RESULTS: In the PEG group first nutritional status and tumor stage were significantly worse, but changes of nutritional status from first visit to operation were significantly better. According to the intraoperative thermal imaging, there was no patient with blood flow disturbance in the gastric conduit due to PEG. Short-term surgical outcomes did not significantly differ. CONCLUSION: PEG has less adverse effects on gastric tube production in esophagectomy and may be considered in highly selective patients during preoperative therapy.

Sentinel Node Navigation Surgery for Early Gastric Cancer: Analysis of Factors Which Affect Direction of Lymphatic Drainage.

BACKGROUND AND PURPOSE: We started performing sentinel node navigation surgery (SNNS) for patients with early gastric cancer (EGC) using infrared ray electronic endoscopy (IREE) with indocyanine green injection from year 2000. The EGCs usually have complex lymphatic drainage, unidirectional or multidirectional lymphatic flow. In this study, we investigated and clarified factors that affect the direction of gastric lymphatic drainage. PATIENTS AND METHOD: Consecutive 60 patients with EGC who underwent SNNS by IREE from year 2006 to 2014 were enrolled to this study. Patients' age, gender, location of tumors, operative method, previous treatment by endoscopic submucosal dissection (ESD), presence of pathological ulcerative scar and maximum tumor diameter were enrolled as parameters which may affect direction of lymphatic drainage and analyzed. RESULT: Bivariate analysis demonstrated that the presence of pathological ulcerative scar (P = 0.01), tumor location (g.c vs. a.w vs. p.w vs. l.c, P = 0.01), and maxim tumor diameter (P = 0.0003) were relevant to direction of gastric lymphatic drainage. Multivariate analysis showed that tumor location (g.c/a.w/p.w vs. l.c, odds ratio 8.227, P = 0.011) and the maximum tumor diameter (odds ratio 1.057, P = 0.037) are independent factors that affect direction of gastric lymphatic flow. Of tumors, 78% located at lesser curvature had unidirectional lymphatic drainage, and 93% of tumors whose diameter was 40 mm and more had multidirectional lymphatic drainage. CONCLUSION: Our investigation revealed that the tumor location and tumor diameter were the key factors which affect the direction of lymphatic drainage, which is useful fact to understand the complexity of gastric lymphatic drainage.

Comparison of Short-term and Long-term Clinical Outcomes Between Laparoscopic and Open Total Gastrectomy for Patients With Gastric Cancer.

BACKGROUND AND PURPOSE: Validation of laparoscopic total gastrectomy (LTG) for patients with gastric cancer has not been fully investigated. In particular, the technique for esophagojejunostomy remains controversial. We performed 103 cases of LTG for patients with gastric cancer between 2007 and 2013, in which all esophagojejunostomy reconstruction was performed with intracorporeal circular stapling esophagojejunostomy using the OrVil system except for the first 3 cases. The purpose of this study is to retrospectively analyze the clinical usefulness of LTG with intracorporeal circular stapling esophagojejunostomy using the OrVil system and oncological feasibility of LTG as compared with open total gastrectomy (OTG). PATIENTS AND METHOD: We retrospectively analyzed clinical course of consecutive 100 operations with LTG in comparison with consecutive 53 operations with OTG for patients with gastric cancer. As an estimation of short-term outcome, operative time, blood loss, postoperative hospital days and postoperative data of blood and drain examination were included. Moreover, relapse-free survival time and overall survival time stratified by each stage were calculated by log-rank test as an estimation of prognostic relevance. RESULTS: Blood loss and postoperative hospital stay of LTG were significantly less than that of OTG. Postoperative complications were equivalent between the 2 groups and no patient died within 1 month post-LTG. Only 1 patient had recurrence and died for carcinomatosa peritonitis 50 months after LTG (median follow-up period: 44 mo). CONCLUSIONS: Our experience revealed that LTG with intracorporeal circular stapling esophagojejunostomy using the OrVil system could be performed safely and with acceptable oncological outcome for patients with gastric cancer.

Early Response of Esophageal Cancer to Neoadjuvant Chemotherapy with Docetaxel-Cisplatin-5-Fluorouracil Represents Sensitivity: A Phase II Study.

AIM: The aim of this study was to assess the response rate and find improvements. PATIENTS AND METHODS: Fifty-five patients with esophageal cancer were enrolled. Neoadjuvant chemotherapy (one or two courses) consisted of 60 mg/m(2)docetaxel on day 1, 70 mg/m(2)cisplatin on day 1 and continuous infusion of 5-fluorouracil at 600 mg/m(2)/day on day 1-5. All patients were examined for clinical response by computed tomography and endoscopy at each course. RESULTS: Grade 3/4 hematological toxicity was observed in 63.6% and grade 3/4 non-hematological toxicity in 41.8% of patients. The clinical response rate was 71% and histological complete response rate was 7.8%. We divided patients into three groups by clinical response to the first course of chemotherapy: partial response (PR), incomplete response (IR) and stable disease (SD). The final clinical response rate in those with SD to the first course was significantly lower (vs. those with PR p<0.001, vs. IR p<0.001). CONCLUSION: A high response rate was obtained and tolerability was good. Moreover, the presence of sensitivity to therapy was reflected in the initial clinical response.

Comparative Analysis of Extracellular Polymeric Substances from Cryptococcus gattii VGIIa Strain Isolated for the First Time in Japan.

Cryptococcus gattii and C. neoformans are pathogenic yeasts that cause meningoencephalitis. C. gattii has four molecular types: VGI, VGII, VGIII, and VGIV. Furthermore, three genotypes have been reported for VGII, and a high pathogenicity of the VGIIa genotype has been proposed. The VGIIa strain has been isolated from a patient in Japan, but little is known about the characteristics of the polysaccharides in this strain. In this study we examined the induction of interleukin-8（IL-8）transcriptional activation and compared the nuclear magnetic resonance（NMR）spectra of extracellular polymeric substances（EPSs）, mainly polysaccharides, from the VGIIa, VGIIb, and VGIIc genotypes. The induction of IL-8 by C. gattii EPSs was weaker than that by C. neoformans EPSs. The anomeric proton signals in the NMR spectra of EPSs obtained from VGII isolates were similar, and the polysaccharides were mainly mannose, xylose, galactose, and glucuronic acid. These results suggest that the extracellular polysaccharides from the VGIIa strain isolated in Japan are almost the same as those from other VGII strains.

Short-term surgical outcomes of reduced port surgery for esophageal achalasia.

PURPOSE: To clarify the feasibility and utility of reduced port surgery (RPS) for achalasia. METHODS: Between September 2005 and June 2013, 359 patients with esophageal achalasia, excluding cases of reoperation, underwent laparoscopic Heller myotomy and Dor fundoplication (LHD) according to our clinical pathway. Three-hundred and twenty-seven patients underwent LHD with five incisions (conventional approach), while the other 32 patients underwent RPS, including eight via SILS. The clinical data were collected in a prospective fashion and retrospectively reviewed. We selected 24 patients matched for gender, age and morphologic type with patients in the RPS group from among the 327 patients (C group). The surgical outcomes were compared between the C and RPS groups. RESULTS: There were no significant differences between the two groups in the duration of symptoms, dysphagia score, chest pain score, shape of the distal esophagus and esophageal clearance. The operative time was significantly longer in the RPS group than in the C group (p < 0.001). There were no significant differences between the two groups in the length of postoperative hospital stay or rates of bleeding, mucosal injury of the esophagus and/or stomach and postoperative complications. The symptom scores significantly improved after surgery in both groups (p < 0.001). Furthermore, there were no significant differences between the C group and RPS group in terms of the postoperative symptom scores or satisfaction scores after surgery. CONCLUSIONS: The surgical outcomes of RPS for achalasia are comparable to those obtained with the conventional method.

Prognostic significance of ZNF217 expression in gastric carcinoma.

BACKGROUND: The zinc finger protein ZNF217 is a candidate oncogene in breast cancer and ovarian clear cell cancer. The purpose of the present study was to clarify the significance of this protein's expression in gastric carcinoma and to evaluate the outcome of these patients. MATERIALS AND METHODS: Using paraffin-embedded specimens from 84 patients with gastric cancer, ZNF217 protein was detected using an anti-ZNF217 goat polyclonal antibody. We evaluated the ZNF217 protein expression in relation to patient outcome and clinicopathological parameters. RESULTS: The ZNF217 protein was expressed in 34 (40.5%) tumor sections. Patients with ZNF217-negative tumors had better relapse-free survival (RFS) and overall survival (OS) than those with ZNF217-positive tumors by the log-rank test. Notably, multivariate analysis indicated that ZNF217 was an independent prognostic factor for RFS. CONCLUSION: ZNF217 expression seems to be a novel prognostic biomarker in gastric cancer.

Prediction of lymph node metastasis in patients with submucosa-invading early gastric cancer.

BACKGROUND AND AIM: Early gastric cancer (EGC), with wall invasion limited to the submucosa, has approximately 15 to 20% chance of lymph node metastasis. The purpose of this study is to clarify the parameters which affect lymph node metastasis and survey whether lymph node metastasis can be predicted preoperatively. PATIENTS AND METHODS: We retrospectively analyzed 145 consecutive patients with EGC using multivariate analysis and developed a formula which predicts lymph node metastasis by linear discriminant analysis. In addition, we prospectively validated this formula in another subset of 106 consecutive patients with EGC and compared the predicted with the actual pathological lymph node metastasis. RESULTS: Multivariate analyses revealed that independent factors, which affect lymph node metastasis for EGC, were lymphatic system invasion (p=0.00002, odds ratio 3.11) and venous system invasion (p=0.039, odds ratio 2.44). In addition, we developed the lymph node metastasis-predicting formula using these two factors by linear discriminant analysis. The formula is as follows: Y=0.12 × (venous system invasion: 0, 1, 2 or 3) + 0.19 × (lymphatic system invasion: 0, 1, 2, or 3) - 0.14. If Y>0, we judge that a patient with gastric cancer is susceptible lymph node metastasis. The result of this prospective study showed that the sensitivity and specificity rates were 70% and 61.6%, respectively. CONCLUSION: We developed a formula which can predict lymph node metastasis using linear discriminant analysis. This formula seems useful in predicting for lymph node metastasis in patients with EGC.

High UBCH10 protein expression as a marker of poor prognosis in esophageal squamous cell carcinoma.

BACKGROUND/AIM: Ubiquitin-conjugating enzyme H10 (UBCH10) is required in the cell-cycle transition from metaphase to anaphase. Therefore, we investigated whether its expression level in cancerous esophageal lesions affected prognosis of patients with esophageal squamous-cell carcinoma. MATERIALS AND METHODS: Paraffin-embedded tissue samples from 121 patients with esophageal squamous cell carcinoma were stained with antibody to UBCH10 for immunohistochemical analysis. RESULTS: UBCH10 was expressed in cancerous and dysplastic lesions, but not in normal tissue. Patients were grouped according to expression: High (N=33) or low (N=88), depending on the staining pattern. There were significant differences between the groups in terms of invasion into lymphatic vessels, number of metastatic lymph nodes, TNM classification, and stages, as well as in survival: the 50% survival rate in the high expression group was 2.3 years, whereas it was 9.9 years for the low-expression group (p<0.0001). Even with multivariate adjusting for stage 0 to stage IV using the Cox proportional hazard model, patients belonging to the high-expression group had a poor prognosis (Hazard ratio=2.5; 95% Confidence Interval=1.3-4.5; p=0.004). CONCLUSION: High protein expression of UBCH10 is a marker of poor prognosis in esophageal squamous cell carcinoma.

Effect of laparoscopic esophagomyotomy on chest pain associated with achalasia and prediction of therapeutic outcomes.

BACKGROUND: The effect of myotomy for achalasia on chest pain has not been clarified. The current study aimed to investigate the therapeutic effect of laparoscopic myotomy on chest pain associated with achalasia and to identify prognostic factors for outcomes. METHOD: Between March 2005 and September 2008, 95 patients were available for detailed interviews and for assessment of clearance by timed barium esophagogram (TBE) before and after surgery. Of the 95 patients, 47 (24 men; mean age, 42.9 ± 13.5 years) who experienced chest pain before surgery were studied. The subjects were asked in detail about dysphagia and chest pain before surgery and 6 months after surgery. The frequency and severity of the symptoms were graded on a scale of 0 to 4. In addition, the values obtained by multiplying the grade for frequency by the grades for severity of the two symptoms were defined as the dysphagia score and the chest pain score, respectively. The patients with chest pain scores of 0 after surgery were defined as group A and those with scores smaller than their preoperative scores as group B. The remaining patients with other scores were defined as group C. The background factors and clinical conditions of the three groups were compared. RESULTS: The mean chest pain score decreased from 5.0 ± 3.2 to 1.0 ± 1.6 (p < 0.001). The score after surgery was 0 for 27 patients and showed a decrease for 15 patients. Although the three groups did not differ in their characteristics, differences were noted in postoperative TBE factors (i.e., groups A and B had significantly shorter barium columns than group C at 1 and 5 min after surgery (p = 0.001). CONCLUSION: Laparoscopic myotomy had a therapeutic effect on chest pain associated with achalasia, and improvement in postoperative esophageal clearance may influence the therapeutic effect.

Misinsertion of an esophageal bougie between the mucosa and muscular layer of the lower esophagus during operation in a case of achalasia.

Laparoscopic fundoplication is the current standard operation for gastroesophageal reflux disease and achalasia. An esophageal bougie is often placed for constructing a wrap or performing myotomy easily during the operation. The surgical complications reportedly include hemorrhage, esophageal perforation, gastric perforation, and vagus nerve damage. However, to the best of our knowledge, there is no report of any case of esophageal bougie misinsertion into the lower esophageal mucosa. The patient was a woman in her 50s who underwent laparoscopic Heller-Dor operation for achalasia in April 2007. During myotomy, we encountered difficulty in inserting a 56Fr-bougie and eventually realized that we had misinserted the Endolumina through the pyriform sinus into the esophageal mucosa. As the pyriform sinus injury was limited to the mucosa and there was no evidence of mediastinal perforation as assessed by intraoperative endoscopic examination, we continued the laparoscopic operation. On postoperative day 1, the patient complained of mild chest pain and pharyngeal pain, but there were only few signs of inflammation. Chest computed tomography revealed findings suggestive of a hematoma extending from the neck to the lower chest, most prominently in the right esophageal wall, however, the patient was followed up conservatively as there was no clear indication of perforation. Subsequently, both the chest pain and pharyngeal pain subsided. On postoperative day 5, the patient was allowed oral water intake, as a gastrografin swallow study did not indicate any problems. On postoperative day 6, the patient was allowed semiliquid foods orally. She was discharged from the hospital on day 11, without any particular problems. Currently, 10 months postsurgery, she is doing well, without any recurrence of symptoms such as dysphagia.

[Role of Malassezia colonization in cutaneous immune response].

Malassezia yeasts are part of the cutaneous microflora and are also associated with a number of skin diseases such as pityriasis versicolor, seborrheic dermatitis, and atopic dermatitis (AD). Among organisms of the Malassezia species, M. globosa and M. restricta are highly associated with AD. However, their precise role in AD has remained uncertain. We first attempted to identify major allergens from M. globosa using a proteomics analysis. Immunoblotting showed that IgE-reactive components with molecular masses of 40-45 kDa proteins were detected by 100% (28 of 28) of sera from AD patients. The IgE-reactive allergens corresponding to the 42 kDa protein (MGp42) were identified by two-dimensional immunoblotting, and partially sequenced by MALDI-TOF MS with post source decay (PSD) of the peptide digest. Comparison of sequences with known protein sequences revealed that MGp42 showed similarity to the heat shock protein (hsp) family. Our studies have also demonstrated that human keratinocytes responded to the two Malassezia species with different Th2-type cytokine profiles, i.e. M. globosa induced IL-5, IL-10, and IL-13 secretion from the keratinocytes, whereas M. restricta induced IL-4 secretion. These findings suggest that M. globosa and M. restricta play a synergistic role in triggering or exacerbating AD by stimulating the Th2 immune response.

Identification of the major allergen of Malassezia globosa relevant for atopic dermatitis.

BACKGROUND: Malasseziaglobosa constitutes a part of the normal flora of human skin, but may induce IgE production in atopic dermatitis (AD). However, information on M.globosa allergens is scant. OBJECTIVE: To identify the major M. globosa allergens by using proteomic analysis. METHODS: Immunoglobulin E (IgE) immunoblotting and cross-inhibition tests for M. globosa allergens were performed using sera from AD patients and control subjects. These allergens were identified and characterized using the proteomics approach involving a combination of two-dimensional (2D) electrophoresis, mass spectrometry, and bioinformatics tools. We cloned the cDNA of this allergen using sequences obtained by 5'- and 3'-rapid amplification of cDNA ends polymerase chain reaction. RESULTS: The sera of the AD patients had IgE-reactive 40-45-kDa protein components. By 2D immunoblotting, we detected a 42-kDa protein spot with an isoelectric point (pI) of 4.8; the protein was highly reactive to IgE and was designated MGp42. Full-length MGp42 cDNA contained a 1908-bp open reading frame encoding 635 amino acid residues (calculated molecular mass, 69.7kDa; pI, 6.02). The N-terminal MGp42 sequence started from the 250th residue (Asp-250) of the deduced amino acid sequence and consisted of 386 amino acid residues; these results are consistent with those of 2D immunoblotting. MGp42 showed sequence similarity to members of the heat shock protein 70 (hsp70) family. Immunoblot inhibition tests revealed no IgE cross-reactivity between MGp42 and human HSP70. CONCLUSIONS: MGp42 may be a cleavage product of intact HSP70. This novel M. globosa allergen could be useful for the diagnosis of AD.

Pre-pyloric site gastric cancer after pylorus-preserving gastrectomy (PPG): a case report.

Pylorus-preserving gastrectomy (PPG) is a function-preserving surgical procedure which is now applied to treat early gastric cancer in the mid-portion of the stomach. We report a patient who developed a pre-pyloric site gastric cancer after PPG. To our knowledge, this is the first report on the development of pre-pyloric site gastric cancer after PPG in the English literature using PubMed.