Optogenetic activation of axon guidance receptors controls direction of neurite outgrowth.

Growth cones of extending axons navigate to correct targets by sensing a guidance cue gradient via membrane protein receptors. Although most signaling mechanisms have been clarified using an in vitro approach, it is still difficult to investigate the growth cone behavior in complicated extracellular environment of living animals due to the lack of tools. We develop a system for the light-dependent activation of a guidance receptor, Deleted in Colorectal Cancer (DCC), using Arabidopsis thaliana Cryptochrome 2, which oligomerizes upon blue-light absorption. Blue-light illumination transiently activates DCC via its oligomerization, which initiates downstream signaling in the illuminated subcellular region. The extending axons are attracted by illumination in cultured chick dorsal root ganglion neurons. Moreover, light-mediated navigation of the growth cones is achieved in living Caenorhabditis elegans. The photo-manipulation system is applicable to investigate the relationship between the growth cone behavior and its surrounding environment in living tissue.

Novel scoring system and algorithm for classifying chronic rhinosinusitis: the JESREC Study.

BACKGROUND: Chronic rhinosinusitis (CRS) can be classified into CRS with nasal polyps (CRSwNP) and CRS without nasal polyps (CRSsNP). CRSwNP displays more intense eosinophilic infiltration and the presence of Th2 cytokines. Mucosal eosinophilia is associated with more severe symptoms and often requires multiple surgeries because of recurrence; however, even in eosinophilic CRS (ECRS), clinical course is variable. In this study, we wanted to set objective clinical criteria for the diagnosis of refractory CRS. METHODS: This was a retrospective study conducted by 15 institutions participating in the Japanese Epidemiological Survey of Refractory Eosinophilic Chronic Rhinosinusitis (JESREC). We evaluated patients with CRS treated with endoscopic sinus surgery (ESS), and risk of recurrence was estimated using Cox proportional hazard models. Multiple logistic regression models and receiver operating characteristics curves were constructed to create the diagnostic criterion for ECRS. RESULTS: We analyzed 1716 patients treated with ESS. To diagnose ECRS, the JESREC scoring system assessed unilateral or bilateral disease, the presence of nasal polyps, blood eosinophilia, and dominant shadow of ethmoid sinuses in computed tomography (CT) scans. The cutoff value of the score was 11 points (sensitivity: 83%, specificity: 66%). Blood eosinophilia (>5%), ethmoid sinus disease detected by CT scan, bronchial asthma, aspirin, and nonsteroidal anti-inflammatory drugs intolerance were associated significantly with recurrence. CONCLUSION: We subdivided CRSwNP in non-ECRS, mild, moderate, and severe ECRS according to our algorithm. This classification was significantly correlated with prognosis. It is notable that this algorithm may give useful information to clinicians in the refractoriness of CRS before ESS or biopsy.

Carotid intima-media thickness and cerebral white matter lesions are more advanced in acute ischemic stroke patients with renal dysfunction.

BACKGROUND: It has been shown that chronic kidney disease (CKD) is a risk factor for stroke, but there have been few studies on the relationship between CKD and stroke. The objective of this study was to investigate the relationship between renal dysfunction and cerebral white matter lesions or carotid plaque in patients with acute ischemic stroke. METHODS: Subjects were 202 consecutive patients with ischemic stroke who were admitted to the Stroke Center of Nippon Medical School Hospital from January 2007 to July 2008. The estimated glomerular filtration (eGFR) was calculated and the relationship of renal dysfunction to the subtype of ischemic stroke, cardiovascular risk factors, cerebral white matter lesions on brain magnetic resonance imaging (MRI), and maximum intima-media thickness (IMT) of the carotid artery was analyzed statistically. RESULTS: Among the 202 patients with ischemic stroke, 27.9% had an eGFR < 60 ml/min/1.73 m2 (eGFR < 60 ml group). Age was significantly higher and a history of hypertension, diabetes, and ischemic heart disease was significantly more frequent in this group than in the group with eGFR ≥ 60 ml/min/1.73 m2 (eGFR ≥ 60 ml group). Among the subtypes of ischemic stroke, atherothrombotic cerebral infarction was predominant and accounted for 41.1%, followed by cardiogenic cerebral infarction at 31.1%, lacunar infarction at 18.8%, and unclassified infarction at 8.9%. There was no significant difference in the distribution of ischemic stroke subtype between both groups. Deep and subcortical white matter hypertensity (DSWMH) and periventricular hyperintensity (PVH) were detected by brain MRI in 91.5% of the eGFR < 60 ml group. In the eGFR < 60 ml group, PVH was significantly more frequent than in the eGFR ≥ 60 ml group (p = 0.032) and DSWMH was also more frequent (p = 0.0519). The maximum IMT measured by carotid ultrasound was significantly larger in the eGFR < 60 ml group. CONCLUSION: In patients with acute ischemic stroke, the incidence of renal dysfunction was high like that of heart disease. In the eGFR < 60 ml group, carotid IMT was larger and the incidence of PVH was higher, so these patients presumably had more advanced atherosclerotic changes of the cerebral vessels.

KIT and platelet-derived growth factor receptor α gene expression in laryngeal small cell carcinoma.

OBJECTIVE: Small cell carcinoma has the worst prognosis of all laryngeal neoplasms. In order to further characterise this tumour, with a view to development of new therapeutic approaches, we report the results of KIT gene and platelet-derived growth factor receptor α gene expression analysis, for two extremely rare cases of primary small cell carcinoma of the larynx. METHOD: Case reports, including immunohistochemical study, and review of the literature. RESULTS: We present two patients with laryngeal small cell carcinoma, who died from tumour metastasis to the lungs and brain despite aggressive treatment. Immunohistochemical studies revealed positive reactions for KIT gene expression and platelet-derived growth factor α gene expression in patient one, and for KIT gene expression in patient two. Molecular genetic analysis, using polymerase chain reaction direct sequencing, identified no mutations of the KIT or platelet-derived growth factor receptor α genes. CONCLUSION: Although further investigation is necessary regarding KIT gene expression and platelet-derived growth factor receptor α gene expression in laryngeal small cell carcinoma, the reported results suggest that these genes may be significant in the development of molecular targeted therapy.

The effect of angiotensin-converting enzyme inhibitors of left atrial pressure in dogs with mitral valve regurgitation.

BACKGROUND: Despite many epidemiological reports concerning the efficacy of angiotensin-converting enzyme (ACE) inhibitors in dogs with mitral regurgitation (MR), the hemodynamic effects of ACE inhibitor administration have not been fully evaluated. OBJECTIVES: To document left atrial pressure (LAP) in dogs with MR administered ACE inhibitors, in order to obtain interesting information about daily LAP changes with administration of ACE inhibitors. ANIMALS: Five healthy Beagle dogs weighing 9.8 to 14.2 kg (2 males and 3 females; aged 2 years). METHODS: Experimental, crossover, and interventional study. Chordae tendineae rupture was induced, and a radiotelemetry transmitter catheter was inserted into the left atrium. LAP was recorded for 72 consecutive hours during which each of 3 ACE inhibitors--nalapril (0.5 mg/kg/d), temocapril (0.1 mg/kg/d), and alacepril (3.0 mg/kg/d)--were administered in a crossover study. RESULTS: Averaged diurnal LAP was significantly, but slightly reduced by alacepril (P = .03, 19.03 +/- 3.01-18.24 +/- 3.07 mmHg). The nightly drops in LAP caused by alacepril and enalapril were significantly higher than the daily drops (P = .03, -0.98 +/- 0.19 to -0.07 +/- 0.25 mmHg, and P = .03, -0.54 +/- 0.21-0.02 +/- 0.17 mmHg, respectively), despite the fact that the oral administrations were given in the morning. Systolic blood pressure (122.7 +/- 14.4-117.4 +/- 13.1 mmHg, P = .04) and systemic vascular resistance (5800 +/- 2685-5144 +/- 2077 dyne x s/cm5, P = .03) were decreased by ACE inhibitors. CONCLUSIONS AND CLINICAL IMPORTANCE: ACE inhibitors decrease LAP minimally, despite reductions in left ventricular afterload. ACE inhibitors should not be used to decrease LAP.

Maxillary haemangioma successfully resected by endoscopic approach.

OBJECTIVE: We report an extremely rare case of maxillary haemangioma. METHOD: Case report and review of the literature concerning haemangioma arising from the nasal cavity and paranasal sinuses. RESULTS: Maxillary haemangioma is rare and sometimes requires wider resection than nasal haemangioma if a large tumour is found. We present a case of maxillary haemangioma in a 37-year-old Japanese woman, which was completely resected by pre-operative embolisation and endoscopic sinus surgery. CONCLUSION: Our findings suggest that if a large maxillary haemangioma is diagnosed pre-operatively, the treatment of choice is pre-operative embolisation followed by endoscopic sinus surgery, in order to avoid the surgical complications associated with wide resection.

Expression of ecalectin, a novel eosinophil chemoattractant, in nasal polyps.

CONCLUSION: Ecalectin, which is produced in the mucosa of nasal polyps, seems to play an important role in the accumulation and activation of eosinophils in nasal polyps, regardless of the presence or absence of atopic predisposition. OBJECTIVE: Ecalectin is a recently discovered eosinophil chemoattractant which elongs to the galectin family. We investigated the expression of ecalectin in nasal polyp tissues associated with various nasal and paranasal diseases in order to clarify the pathogenesis of eosinophilia in nasal polyposis. MATERIAL AND METHODS: Nasal polyps were taken from 56 patients diagnosed as having chronic sinusitis with nasal polyposis. The surgically resected polyps and nasal turbinates were immunohistochemically stained using antibodies against EG2, human mast cell tryptase, CD3 and ecalectin. RESULTS: The number of EG2- and ecalectin-positive cells was significantly higher in nasal polyps than control turbinates. Ecalectin-positive cells were observed in the subepithelial layer, where many EG2-positive cells were present. The number of ecalectin-positive cells correlated significantly with the number of EG2-positive cells in nasal polyps. Many ecalectin mRNA-positive cells were also observed in nasal polyps with an accumulation of EG2-positive cells.

Eosinophil chemoattractants in the middle ear of patients with eosinophilic otitis media.

BACKGROUND: Patients with intractable otitis media associated with bronchial asthma have an extensive accumulation of eosinophils in the effusion and mucosa of the middle ear; this condition is called eosinophilic otitis media (EOM). It remained to be determined how eosinophils accumulate in the middle ear. OBJECTIVES: To clarify the pathogenesis of middle ear diseases, we measured the concentration of eosinophil chemoattractants in middle ear effusion (MEE), and carried out immunohistochemical studies of middle ear mucosa specimens to demonstrate the expression of eosinophil chemoattractants. METHODS: Middle ear effusion samples were obtained from 15 EOM patients with bronchial asthma and from six controls for the measurement of eosinophil cationic protein (ECP), IL-5, eotaxin and regulated on activation, normal T expressed and secreted concentrations. Middle ear mucosa samples were also taken from 14 EOM patients and 16 controls for immunohistochemical study. In 10 EOM patients, the numbers of immunoreactive cells as well as apoptotic cells were determined before and after the topical application of triamcinolone acetonide into the middle ear. RESULTS: In EOM, significantly higher ECP and IL-5 concentrations were detected in MEE than in serum, and ECP, IL-5 and eotaxin concentrations in MEE were higher in the EOM patients than in the controls. ECP concentration positively correlated with that of IL-5. Immunohistochemically, the numbers of cells positive for EG2 and ecalectin were significantly higher in the EOM patients than in the controls. After the topical application of triamcinolone acetonide, the numbers of infiltrating cells and immunoreactive cells distinctly decreased, whereas the number of apoptotic cells significantly increased. CONCLUSION: In EOM, locally produced IL-5 may play a crucial role in the accumulation of eosinophils in the middle ear. Chemokines such as ecalectin and eotaxin are also produced in the middle ear, and help activate and enhance the survival of eosinophils to induce the intractable condition in the middle ear. The topical application of triamcinolone acetonide induces the apoptosis of not only eosinophils but also eosinophil chemoattractant-producing cells, thereby improving the middle ear condition.

Mutations in the ST7/RAY1/HELG locus rarely occur in primary colorectal, gastric, and hepatocellular carcinomas.

Human cancers frequently show a loss of heterozygosity on chromosome 7q31, which indicates the existence of broad-range tumour-suppressor gene(s) at this locus. Truncating mutations in the ST7 gene at this locus are seen frequently in primary colon cancer and breast cancer cell lines. Therefore, the ST7 gene represents a novel candidate gene for the tumour suppressor at this locus. However, more recent studies have reported that ST7 mutations are infrequent or absent in primary cancer and cell lines. To ascertain the frequency of mutations of the ST7 gene in cancer cells, we examined mutations in the ST7 coding sequence in 48 colorectal, 48 gastric, and 48 hepatocellular carcinomas using polymerase chain reaction-single-strand conformational polymorphism and direct sequencing. We detected somatic mutations, which were located near the exon-intron junction in intron 8, in only three out of 144 cases. We conclude that mutations in the ST7 gene are rare in primary colorectal, gastric, and hepatocellular carcinomas.

[The role of staged Fontan operation on early and long-term outcome].

Of 185 patients with single ventricle physiology, 59 patients who underwent Fontan type operations between April 1970 and May 2002 served as subjects. Subjects displayed a median age of 5.2 years and a median body weight of 11.4 kg. In the first 22 years (group 1), 34 patients underwent concomitant right atrium-pulmonary artery (RA-PA) anastomosis, Björk procedure and total cavopulmonary connection (TCPC), while in the last 8 years (group 2), 25 patients underwent staged TCPC, where bidirectional cavopulmonary shunt (BCPS) and obliteration of additional pulmonary blood flow was performed previously. Four cases of early death (group 1:4 patients, group 2:0 patient) and 12 cases of late death (group 1:11 patients, group 2:1 patient) were encountered. Early mortality was 6.8% (group 1:12%, group 2:0%, p = 0.10) and late mortality was 21%. The 5-year survival rate was 85.2 +/- 4.9% (group 1:80.0 +/- 6.8%, group 2:93.8 +/- 6.1%, p = NS), and the 10-year survival rate was 79.6 +/- 6.0%. Staged TCPC, precedent BCPS with obliteration of additional pulmonary blood flow, seems to be beneficial for accurate patient selection for Fontan candidate.

Western blotting analysis of the beta-hexosaminidase alpha- and beta-subunits in cultured fibroblasts from cases of various forms of GM2 gangliosidosis.

OBJECTIVES: The GM2 gangliosidoses are a group of genetic disorders caused by the accumulation of ganglioside GM2 in neuronal cells. We examined the alpha- and beta-subunits of beta-hexosaminidases by a non-radioisotopes detecting system to evaluate whether it was a useful method for understanding of the pathophysiologies of GM2 gangliosidoses. MATERIALS AND METHODS: We investigated the alpha- and beta-subunits of beta-hexosaminidases in cultured fibroblasts from cases of various forms of GM2 gangliosidosis by means of Western blotting and a chemiluminescence detection system. RESULTS: In a patient with infantile Tay-Sachs disease [HEXA genotype, Int5-SA(g-1-->t)/Int5-SA(g-1-->t)], the mature alpha-subunit was undetectable. In a patient with infantile Sandhoff disease (HEXB genotype, C534Y/C534Y), the mature beta-subunit was deficient. However, a small amount of the mature beta-subunit was detected in a patient with adult Sandhoff disease (HEXB genotype, R505Q(+I207V)/R505Q(+I207V)), which may have resulted in the residual enzyme activity and mild clinical course. Normal amounts of alpha- and beta-subunits were detected in a patient with GM2 activator deficiency. CONCLUSION: This method is easy and sensitive for detecting target proteins, and is useful for clarification of the pathophysiologies of GM2 gangliosidoses.

Prognostic significance of dihydropyrimidine dehydrogenase expression in breast cancer.

We have investigated dihydropyrimidine dehydrogenase expression as a prognostic marker in breast cancer. A total of 119 women with breast cancer undergoing surgery between 1985 and 1996 were included in this study. Eighty-seven patients were treated with postoperative chemotherapy including 5-fluorouracil or 5-fluorouracil derivatives. Fifty-nine (50%) of 119 patients were determined to be immunostaining-positive for dihydropyrimidine dehydrogenase. There was no significant difference between dihydropyrimidine dehydrogenase staining and tumour size, lymph node status, clinical stage, oestrogen receptor status, histologic grade, or 5-fluorouracil administration. When evaluated in patients treated with 5-fluorouracil or 5-fluorouracil derivatives, patients with dihydropyrimidine dehydrogenase-positive tumours had a significantly (P<0.05) poorer disease-free survival compared to those with dihydropyrimidine dehydrogenase-negative tumour. No conclusion can be drawn about the prognostic impact of dihydropyrimidine dehydrogenase status in patients who were not treated with 5-fluorouracil regimes due to the small number of such cases in this series. Lymph node and dihydropyrimidine dehydrogenase status were independent prognostic factors for disease-free survival, and lymph node status for overall survival using multivariate analysis. In conclusion, dihydropyrimidine dehydrogenase is a possible prognostic factor in patients with breast cancer treated with 5-fluorouracil or 5-fluorouracil derivatives.

Predictive value of estrogen receptor status as assessed by ligand-binding assay in patients with early-stage breast cancer treated with breast conserving surgery and radiation therapy.

It is important to determine which factors are predictive for the prognosis of patients treated with breast conserving surgery (BCS) and radiation therapy (RT) in order to make a decision as to the adjuvant treatment. Although estrogen receptor (ER) is known to be a predictive marker for antiestrogens in breast cancer, the prognostic effect of hormone receptors has not been fully analyzed in Japanese breast cancer patients treated with BCS and RT. A total of 153 breast cancer patients having up to three positive nodes in the axilla as identified histologically and treated with both BCS and RT with or without systemic therapy were enrolled in this study. All tumors were measured for ER and progesterone receptor (PR) using ligand-binding assay (LBA). ER was inversely related to patients' age, however, PR was not related to any clinical features. When ER was classified into negative, weakly positive and strongly positive categories, with cut-off levels of zero and 50 fmol/mg protein, the relapse-free survival (RFS) was significantly better in patients with tumors having strongly positive ER than in patients with tumors having negative ER. Multivariate analysis revealed that ER as well as nodal status, was an independent predictive factor for RFS, however, PR was not. As a result, we believe that ER measured by LBA is valuable for predicting prognosis of early-stage breast cancer patients treated with BCS and RT.

Efficiency of an air filter at the drainage site in a closed circuit with a centrifugal blood pump: an in vitro study.

In a closed circuit with a centrifugal blood pump, one of the serious obstacles to clinical application is sucking of air bubbles into the drainage circuit. The goal of this study was to investigate the efficiency of an air filter at the drainage site. We used whole bovine blood and the experimental circuit consisted of a drainage circuit, two air filters, a centrifugal blood pump, a membrane oxygenator, a return circuit, and a reservoir. Air was injected into the drainage circuit with a roller pump, and the number and size of air bubbles were measured. The air filter at the drainage site could remove the air bubbles (>40 microm) by itself, but adding a vacuum removed more bubbles (>40 microm) than without vacuum. Our results suggest that an air filter at the drainage site could effectively remove air bubbles, and that adding the filter in a closed circuit with a centrifugal blood pump would be safer.

Intraoperative assessment of coronary artery bypass graft: transit-time flowmetry versus angiography.

BACKGROUND: Transit-time flowmetry has been used to assess graft status intraoperatively. This study examines the validity of this method by comparing its results with the findings of simultaneously performed graft angiography. METHODS: The left internal thoracic artery (LITA) anastomosed to the left anterior descending artery (LAD) was assessed intraoperatively with both transit-time flowmetry and graft angiography in 30 patients. The patients were stratified into two groups based on intraoperative angiographic findings. In 18 patients (group A), the LITA and the LAD were well filled with contrast medium and the anastomosis was widely patent. In the other 12 patients (group B), spastic LITA or LAD was observed. Postoperative angiography was also performed before discharge from the hospital. RESULTS: The mean graft flow was 44.0 +/- 25.4 mL/min in group A and 23.4 +/- 10.0 mL/min in group B (p = 0.0129). Diastolic-dominant flow pattern was observed in both groups, and the ratio of peak diastolic flow to peak systolic flow and the percent diastolic time-flow integral were not statistically different between the groups. The pulsatility index was almost the same between the two groups and was acceptable in both. Postoperative angiography revealed that all grafts were patent without spasm or anastomotic stenosis. CONCLUSIONS: LITA graft status is satisfactory when high graft flow with diastolic dominance is obtained. When there is vasospasm but no anastomotic problems, decreased graft flow with an acceptable pulsatility index and diastolic augmentation is observed.

[Surgical results of canal wall reconstruction tympanoplasty in middle ear cholesteatoma].

Materials were 236 ears of 213 patients with middle ear cholesteatoma undergoing canal wall reconstruction during 1993-1998. Subjects were followed up for at least 1 year after final operation. Of 236 ears, 147 (62%) underwent 1-stage operation and 89 ears (38%) required 2-stage operation. Hearing results were successful in 157 ears (67%) based on criteria proposed by the Otological Society of Japan. The success in ears undergoing 1-stage operation was 74% and 54% in ears undergoing 2-stage operation. Postoperative hearing and air-bone gap in the 1-stage group were significantly better than in the 2-stage group. For tympanoplasty, success was 97% in type I, 64% in type III, and 53% in type IV. The likelihood of undergoing 2-stage operation increased with the type of tympanoplasty, from type I to IV. Postoperative hearing was significantly worse in older age groups. Of the 89 ears, 13 (15%) had recurrent cholesteatoma and 29 (33%) had residual cholesteatoma at 2-stage operation. In the 135 in the 1-stage group, recurrent cholesteatoma was observed at follow-up in 13 ears (9.6%). When we analyzed clinical risk factors for both recurrent and residual cholesteatoma in age, gender, otorrhea, types of cholesteatoma, and types of tympanoplasty, no significant factors were seen for recurrent or residual cholesteatoma. These results indicate that canal wall reconstruction tympanoplasty for middle ear cholesteatoma yields relatively good hearing results. However, more effort is needed to reduce the incidence of recurrent and residual cholesteatoma.