Utility of T2 mapping and dGEMRIC for evaluation of cartilage repair after allograft chondrocyte implantation in a rabbit model.

OBJECTIVE: To investigate the effectiveness of quantitative Magnetic resonance imaging (MRI) for evaluating the quality of cartilage repair over time following allograft chondrocyte implantation using a three-dimensional scaffold for osteochondral lesions. DESIGN: Thirty knees from 15 rabbits were analyzed. An osteochondral defect (diameter, 4 mm; depth, 1 mm) was created on the patellar groove of the femur in both legs. The defects were filled with a chondrocyte-seeded scaffold in the right knee and an empty scaffold in the left knee. Five rabbits each were euthanized at 4, 8, and 12 weeks and their knees were examined via macroscopic inspection, histological and biochemical analysis, and quantitative MRI (T2 mapping and dGEMRIC) to assess the state of tissue repair following allograft chondrocyte implantation with a three-dimensional scaffold for osteochondral lesions. RESULTS: Comparatively good regenerative cartilage was observed both macroscopically and histologically. In both chondrocyte-seeded and control knees, the T2 values of repair tissues were highest at 4 weeks and showed a tendency to decrease with time. ΔR1 values of dGEMRIC also tended to decrease with time in both groups, and the mean ΔR1 was significantly lower in the CS-scaffold group than in the control group at all time points. ΔR1 = 1/r (R1post - R1pre), where r is the relaxivity of Gd-DTPA(2-), R1 = 1/T1 (longitudinal relaxation time). CONCLUSION: T2 mapping and dGEMRIC were both effective for evaluating tissue repair after allograft chondrocyte implantation. ΔR1 values of dGEMRIC represented good correlation with histologically and biochemically even at early stages after the implantation.

Enhancement of denitrification in a down-flow hanging sponge reactor by effluent recirculation.

A down-flow hanging sponge reactor, constructed by connecting three identical units in series, was applied to the treatment of artificial wastewater containing phenol and ammonia under high salinity conditions (10.9 g-Cl(-)/L). The theoretical hydraulic retention time (HRT) of each unit was 4 h (total HRT = 12 h). To enhance denitrification by effluent recirculation, the effluent recirculation ratio was increased in increments ranging from 0.0 to 2.0. The concentration of total ammonia nitrogen (TAN), NO2-N, and NO3-N in the final effluent as a proportion of the TAN in the influent was determined to calculate the unrecovered, or denitrification, proportion. The denitrification proportion of the reactor was equivalent to 19.1 ± 14.1% with no effluent recirculation; however, this was increased to 58.6 ± 6.2% when the effluent recirculation ratio was increased to 1.5. Further increasing the effluent recirculation ratio to 2.0 resulted in a decrease in the denitrification proportion to 50.9 ± 9.3%. Activity assays of nitrification and denitrification, as well as 16S rRNA gene sequence analysis, revealed that denitrification occurred primarily in the upper sections of the reactor, while nitrification increased in the lower sections of the reactor. Gene sequence analysis revealed that denitrification by Azoarcus-like species using phenol as an electron donor was dominant.

Radiation and cancer risk in atomic-bomb survivors.

With the aim of accurately assessing the effects of radiation exposure in the Japanese atomic-bomb survivors, the Radiation Effects Research Foundation has, over several decades, conducted studies of the Life Span Study (LSS) cohort, comprising 93 000 atomic-bomb survivors and 27 000 controls. Solid cancer: the recent report on solid cancer incidence found that at age 70 years following exposure at age 30 years, solid cancer rates increase by about 35%  Gy(-1) for men and 58% Gy(-1) for women. Age-at-exposure is an important risk modifier. In the case of lung cancer, cigarette smoking has been found to be an important risk modifier. Radiation has similar effects on first-primary and second-primary cancer risks. Finally, radiation-associated increases in cancer rates appear to persist throughout life. Leukaemia: the recent report on leukaemia mortality suggests that radiation effects on leukaemia mortality persisted for more than 50 years. Moreover, significant dose-response for myelodysplastic syndrome was observed in Nagasaki LSS members even 40-60 years after radiation exposure. Future perspective: given the continuing solid cancer increase in the survivor population, the LSS will likely continue to provide important new information on radiation exposure and solid cancer risks for another 15-20 years, especially for those exposed at a young age.

[Lung adenocarcinoma with mixed subtypes which had been followed up for 5-years].

A 63-year-old female was admitted to our hospital for investigation of serum elevation of carcinoembryonic antigen (CEA). She underwent high anterior resection for a rectal cancer 5-years ago. Chest computed tomography (CT) obtained 5-years ago showed a nodule in the right S10, measuring 1.3 x 0.8 cm in size. The nodule was assessed as benign. Chest CT on admission showed the enlarged nodule with a pleural indentation, measuring 2.2 x 1.6 cm in size. Definitive diagnosis could not be established. Since it was difficult to exclude the possibility of malignancy, video-assisted partial resection was performed. Histological examination of the nodule revealed primary adenocarcinoma in frozen sections. Lobectomy with lymph node dissection was performed. The ultimate diagnosis was adenocarcinoma with mixed subtypes. The tumor was classified as stage IA with T1bN0M0. We reported this case because it was a rare slow-growing adenocarcinoma that had a 5-years clinical history before operation.

[Detection of the point of communication by pneumoperitoneum at the surgery for pleuroperitoneal communication].

A 53-year-old male was admitted to our hospital because of dyspnea. Chest radiograph showed a massive right-sided hydrothorax. He was suffering from chronic renal failure and had undergone continuous ambulatory peritoneal dialysis (CAPD) for 8 months. The diagnosis of pleuroperitoneal communication (PPC) was made using injection of indigocarmine into the peritoneal cavity with subsequent pleural detection by thoracocentesis. Injection of contrast media into the peritoneal cavity showed a dome shaped radio-opaque shadow which is located on the diaphragmatic dome followed by the movement of contrast media into the thoracic cavity. Video-assisted thoracic surgery (VATS) was performed under general anesthesia. To identify the point of communication, the method of detecting air leakage was employed. A bleb like lesion on which the hole existed was observed at the center of the diaphragm, and air leakage was identified by filling the thoracic cavity with saline. The pressure in the peritoneal cavity was maintained at 10 mmHg by continuous CO2 inflation. Direct closure was performed to repair the PPC, which succesfully stopped the air leakage. CAPD could be restarted immediately after surgery. No recurrence of hydrothorax has been detected for more than 14 months after surgery.

Theanine, gamma-glutamylethylamide, a unique amino acid in tea leaves, modulates neurotransmitter concentrations in the brain striatum interstitium in conscious rats.

Theanine (gamma-glutamylethylamide) is one of the major amino acid components in green tea and can pass through the blood-brain barrier. Recent studies suggest that theanine affects the mammalian central nervous system; however, the detailed mechanism remains unclear. In this study, we demonstrated the effect of theanine on neurotransmission in the brain striatum by in vivo brain microdialysis. Theanine injection into the rat brain striatum did not increase the concentration of excitatory neurotransmitters in the perfusate. On the other hand, theanine injection increased the concentration of glycine in the perfusate. Because it has been reported that theanine promotes dopamine release in the rat striatum, we investigated the glycine and dopamine concentrations in the perfusate. Co-injection of glycine receptor antagonist, strychnine, reduced theanine-induced changes in dopamine. Moreover, AMPA receptor antagonist, which regulates glycine and GABA release from glia cells, inhibited these effects of theanine and this result was in agreement with the known inhibitory effect of theanine at AMPA receptors.

A series of magnon crystals appearing under ultrahigh magnetic fields in a kagomé antiferromagnet.

Geometrical frustration and a high magnetic field are two key factors for realizing unconventional quantum states in magnetic materials. Specifically, conventional magnetic order can potentially be destroyed by competing interactions and may be replaced by an exotic state that is characterized in terms of quasiparticles called magnons, the density and chemical potential of which are controlled by the magnetic field. Here we show that a synthetic copper mineral, Cd-kapellasite, which comprises a kagomé lattice consisting of corner-sharing triangles of spin-1/2 Cu2+ ions, exhibits an unprecedented series of fractional magnetization plateaus in ultrahigh magnetic fields of up to 160 T. We propose that these quantum states can be interpreted as crystallizations of emergent magnons localized on the hexagon of the kagomé lattice.

Bacillus cereus nosocomial infection from reused towels in Japan.

It was noticed that there was an increase in Bacillus cereus nosocomial infections in the summer from 2000 to 2005. In 2005, five bloodstream infections occurred in five patients related to catheter use. The causative strains were distinct from each other and belonged to novel multilocus sequence types (ST): ST365, ST366, ST367 and ST368. Two ST365 strains from two patients were further distinguished by pulsed-field gel electrophoresis. B. cereus contamination was observed with reused (dried and steamed) towels (>10(6)cfu/towel) and washing machines in hospital linen rooms. B. cereus strains from towels belonged to ST167, ST365, ST380 and ST382, and a proportion of these were the same, or similar, to strains from patients. All the hospital strains of B. cereus were distinct from those from food-poisoning strains (ST26, ST142, ST381). Ciprofloxacin resistance was observed only in hospital strains. Neither emetic toxin nor cytotoxin K gene, usually present in food poisoning strains, were found in the hospital strains, except for one patient isolate. The data suggest that specific B. cereus strains are circulating within a hospital, with genotypes, antibiotic susceptibilities and virulence gene patterns generally distinct from those of food poisoning, and that in Japan, towels are an important source of contamination, especially in summer.

Randomized controlled trial for an effect of green tea-extract powder supplementation on glucose abnormalities.

OBJECTIVE: We examined whether green tea-extract powder supplementation improves glucose abnormality. METHODS: The study was conducted for volunteers who resided in eastern communities of Shizuoka Prefecture and who had fasting blood glucose levels of >or=6.1 mmol/l or nonfasting blood glucose levels of >or=7.8 mmol/l in a recent health check-up. Sixty subjects aged 32-73 years (49 males and 11 females) participated in the trial. The Early intervention group consumed a packet of green tea-extract powder containing 544 mg polyphenols (456 mg catechins) daily for the first 2 months and then entered the 2-month nonintervention period. The Later intervention group was observed for the first 2 months and then consumed green tea-extract powder as described above for the subsequent 2 months. Using the two-period crossover design, we analyzed the changes in fasting hemoglobin A1c level and other biomarkers in blood samples collected at baseline, 2 months and 4 months. RESULTS: A significant reduction in hemoglobin A1c level and a borderline significant reduction in diastolic blood pressure were associated with the intervention. The intervention caused no significant changes in weight, body mass index, body fat, systolic blood pressure, fasting serum glucose level, homeostasis model assessment index, serum lipid level or hypersensitive C-reactive protein. CONCLUSION: Daily supplementary intake of green tea-extract powder lowered the hemoglobin A1c level in individuals with borderline diabetes.

Ferroelectric and magnetic properties of multiferroic BiFeO(3)-based composite films.

BiFeO(3)-based composite films were fabricated onto the Pt/Ti/SiO(2)/Si(100) substrates by a chemical solution deposition (CSD) method using the precursor solutions with various excess iron composition followed by annealing at 923 K for 30 minutes under oxygen gas flow. Coexistence of spontaneous magnetization and remanent polarization could be obtained in the BiFeO(3)-based composite films with high excess iron composition. The remanent magnetization of almost 20 emu/cm(3) and the magnetic coercive field of 1.5 kOe were obtained at the iron composition ratio of Fe/Bi = 1.25. In this specimen, the remanent polarization at 90 K was approximately 10 microC/cm(2) at the electric field of 1500 kV/cm. Structural analysis suggested that the remanent polarization has a possibility to increase by suppressing the formation of the secondary phases of Bi(2)Fe(4)O(9) and alpha-Fe(2)O(3), these are the nonferroelectric material as well as antiferromagnetic phase.

[Non-chondromatous hamartoma].

A 69-year-old female visited our hospital because of dyspnea. Chest X-ray showed an abnormal shadow in the right lower lung field. Chest computed tomography (CT) showed a round, well-circumscribed, homogenous subpleural nodle of 8 mm in diameter in the right lower lobe, which had no calcification and no pleural indentation. Bronchofiber scope, abdominal CT, brain magnetic resonance imaging (MRI), bone scintigraphy could not establish definitive diagnosis. Scince the possibility of malignancy could not be excluded throughout, video-assisted thoracoscopic surgery was performed to obtain confirmed diagnosis. Pathological examination revealed non-chondromatous hamartoma of the lung. Non-chondromatous hamartoma should be considered in the differential diagnosis of pulmonary nodles. We report a rare case of non-chondromatous hamartoma.

Imaging Differences between Neuromyelitis Optica Spectrum Disorders and Multiple Sclerosis: A Multi-Institutional Study in Japan.

BACKGROUND AND PURPOSE: Both clinical and imaging criteria must be met to diagnose neuromyelitis optica spectrum disorders and multiple sclerosis. However, neuromyelitis optica spectrum disorders are often misdiagnosed as MS because of an overlap in MR imaging features. The purpose of this study was to confirm imaging differences between neuromyelitis optica spectrum disorders and MS with visually detailed quantitative analyses of large-sample data. MATERIALS AND METHODS: We retrospectively examined 89 consecutive patients with neuromyelitis optica spectrum disorders (median age, 51 years; range, 16-85 years; females, 77; aquaporin 4 immunoglobulin G-positive, 93%) and 89 with MS (median age, 36 years; range, 18-67 years; females, 68; relapsing-remitting MS, 89%; primary-progressive MS, 7%; secondary-progressive MS, 2%) from 9 institutions across Japan (April 2008 to December 2012). Two neuroradiologists visually evaluated the number, location, and size of all lesions using the Mann-Whitney U test or the Fisher exact test. RESULTS: We enrolled 79 patients with neuromyelitis optica spectrum disorders and 87 with MS for brain analysis, 57 with neuromyelitis optica spectrum disorders and 55 with MS for spinal cord analysis, and 42 with neuromyelitis optica spectrum disorders and 14 with MS for optic nerve analysis. We identified 911 brain lesions in neuromyelitis optica spectrum disorders, 1659 brain lesions in MS, 86 spinal cord lesions in neuromyelitis optica spectrum disorders, and 102 spinal cord lesions in MS. The frequencies of periventricular white matter and deep white matter lesions were 17% and 68% in neuromyelitis optica spectrum disorders versus 41% and 42% in MS, respectively (location of brain lesions, P < .001). We found a significant difference in the distribution of spinal cord lesions between these 2 diseases (P = .024): More thoracic lesions than cervical lesions were present in neuromyelitis optica spectrum disorders (cervical versus thoracic, 29% versus 71%), whereas they were equally distributed in MS (46% versus 54%). Furthermore, thoracic lesions were significantly longer than cervical lesions in neuromyelitis optica spectrum disorders (P = .001), but not in MS (P = .80). CONCLUSIONS: Visually detailed quantitative analyses confirmed imaging differences, especially in brain and spinal cord lesions, between neuromyelitis optica spectrum disorders and MS. These observations may have clinical implications.

Exploratory Analysis of Associations Between Postmarketing Safety Events and Approved Doses of New Drugs in Japan.

While efficient and less onerous for the industry, the globalization of clinical drug development may lead to limited efforts to optimize drugs for regional conditions. We examined the association between clinical development pathways, approved doses, and postmarketing safety risks in Japan for 135 new molecular entities approved between 2004 and 2011. The risk of drug-related deaths seemed higher when pharmaceutical companies chose exactly the same dose as in the United States, even after conducting Japanese dose-ranging studies. We also found a positive association with drug-related deaths when the review process was expedited and when Japanese dose-ranging studies were not conducted for nonexpedited drugs. Our findings suggest that the decisions on regional dose settings and the choice of global clinical development pathways are associated in ways that may influence the postmarketing outcomes in the target populations.

Cardioprotective effect of edaravone against ischaemia-reperfusion injury in the rabbit heart before, during and after reperfusion treatment.

The free radical scavenger edaravone is able to stimulate prostacyclin release and inhibit the lipoxygenase pathway in the arachidonic acid cascade. The effect of edaravone administration on myocardial damage in rabbit hearts subjected to ischaemia-reperfusion was examined at different times relative to reperfusion. All rabbits underwent sustained coronary artery occlusion for 30 min followed by 3 h of reperfusion. Rabbits were divided into the following groups: control; early (3 mg/kg edaravone IV 10 min before reperfusion); immediate (3 mg/kg edaravone IV immediately after the start of reperfusion); and late (3, 6 or 10 mg/kg edaravone IV 5 min after the start of reperfusion). Single bolus administration of edaravone 10 min before reperfusion or immediately upon initiation of reperfusion appears to be associated with reductions in infarction size and the percentage of apoptotic cells, but treatment with edaravone 5 min after initiation of reperfusion does not appear to have this protective effect.

Decrease in herbicide concentrations and affected factors in lagoons located around Lake Biwa.

The contamination levels and changes in the concentrations in four lagoons around Lake Biwa of paddy-use herbicide were studied. Four lagoons, Sone-numa (52 days of HRT (hydraulic residence time) estimated from the lagoon volume and the average discharge at the outlet, 21 ha area), Yanagihira-ko (40 days, 5.0 ha), Noda-numa (11 days, 6.0 ha), and Iba-naiko (2 days, 55.5 ha), were selected as monitoring sites. Intensive water sampling was carried out once a week from May to June at the outlet of each lagoon. Although twelve of the monitored herbicides were detected, the maximum concentrations did not exceed the guidelines for water-supply law in Japan. The relation between half-lives in herbicide concentrations and characteristics of a lagoon such as HRT and chlorophyll-a concentrations were examined. The shorter half-lives of herbicide concentrations in lagoons with shorter HRT means that replacement by influent water effectively decreased the pesticide concentrations. Shorter half-lives in lagoons with high chlorophyll-a concentrations between the lagoons with similar HRT suggest that biological degradation during the residence time worked more efficiently in the lagoon with high chlorophyll-a concentrations.

Discordant gene and surface expression of the T-cell receptor/CD3 complex in adult T-cell leukemia cells.

Cell surface expression of the T-cell receptor (TCR)/CD3 complex on the cells from 11 acute type adult T-cell leukemia (ATL) and 4 lymphoma type ATL patients was examined by flow cytometry. Cells from 10 of 11 acute ATL patients were TCR alpha beta+ and CD3+, and their mean fluorescence intensities were low (TCR alpha beta, 25.3-84.6; CD3, 22.8-87.8). Cells from two of four lymphoma type ATL did not express this complex, and the other two were CD3+, TCR alpha beta-. In contrast, the mean fluorescence intensity of the TCR/CD3 complex in cells from a patient with T4 chronic lymphocytic leukemia was not low (TCR alpha beta, 129.9; CD3, 117.1). mRNA expressions of the TCR alpha, beta, and CD3 gamma, delta, epsilon, and zeta chains were examined by Northern blots. ATL cells from two acute and two lymphoma types expressed amounts of this complex equal to or greater than those expressed by T4 chronic lymphocytic leukemia. CD3 delta and TCR beta mRNA in ATL and T4 chronic lymphocytic leukemia cells were equally stable to actinomycin D treatment. The synthesis of CD3 zeta protein by ATL cells was detected by Western blotting assay. On the basis of these findings, we discuss the possible involvement of the TCR/CD3 complex in activation of ATL cells.

Down-regulation of promoter 1.3 activity of the human aromatase gene in breast tissue by zinc-finger protein, snail (SnaH).

Aromatase (estrogen synthetase) is expressed in breast cancer tissue, and in situ expression of the enzyme stimulates breast cancer growth. Promoter I.3 is one of the major promoters that control the expression of aromatase in breast cancer tissue. Using the yeast one-hybrid approach to screen a human breast tissue hybrid cDNA expression library, we found that the zinc-finger transcriptional factor Snail (SnaH) interacted with a regulatory region near promoter I.3 of the human aromatase gene. DNA mobility shift assays and mutation analyses using recombinant SnaH protein expressed in Escherichia coli have revealed that this protein interacts with a segment, 5'-CTGATGAAGT-3', which is between 66 and 76 bp upstream from the transcriptional start site of promoter I.3. Using mammalian cell transfection experiments, SnaH was found to act as a repressor of promoter I.3 activity. Site-directed mutagenesis experiments have revealed that the NH2-terminal SNAG domain is important for the repressor activity of SnaH. To demonstrate the inhibitory activity against aromatase expression, a stable SnaH-expressing MDA-MB-231 breast cancer cell line was generated, and the aromatase RNA messages in the SnaH-transfected cell line were found to be 30% of those in the vector-transfected cell line. Reverse transcription-PCR analysis on RNAs isolated from 12 cell lines has confirmed that SnaH is expressed at a higher level in normal breast epithelial cell and stromal fibroblast cell lines than in breast cancer cell lines. In addition, SnaH mRNA was detected in only 16 of 55 breast cancer specimens. On the other hand, aromatase mRNA was detected in 54 of the 55 specimens. Our results indicate that SnaH acts as a repressor that down-regulates the expression of aromatase in normal breast tissue by suppressing the function of promoter I.3. A reduction of the expression of SnaH in breast cancer tissue further suggests a cancer-protective role for this protein in normal breast tissue.

Pressure dependence of pyrene excimer fluorescence in human erythrocyte membranes.

The intensity of pyrene excimer fluorescence in human erythrocyte membranes and in sonicated dispersions of the membrane lipid (liposomes) was examined as a function of pressure (1-2080 bar) and temperature (5-40 degrees C). Higher pressure or lower temperature decreased the excimer/monomer intensity ratios. A thermotropic transition was detected in both membranes and liposomes by plots of the logarithm of the excimer/monomer intensity ratio versus 1/K. The transition temperature of the membranes was 19-21 degrees C at 1 bar and 28-31 degrees C at 450 bar, a shift with pressure of approx. 20-22 K per kbar. Corresponding transition temperatures of the liposomes were 21 degrees C at 1 bar and 33 degrees C at 450 bar, a shift of approx. 27 K per kbar. The observed pressure dependence of the thermotropic transition temperature is similar to that reported for phospholipid bilayers and greatly exceeds that of protein conformation changes. In concert with the liposome studies the results provide direct evidence for a lipid transition in the erythrocyte membrane.

Epigallocatechin gallate and gallocatechin gallate in green tea catechins inhibit extracellular release of Vero toxin from enterohemorrhagic Escherichia coli O157:H7.

We studied the effects of six catechin derivatives (catechin, epigallocatechin, epicatechin, epicatechin gallate, epigallocatechin gallate (EGCg) and gallocatechin gallate (GCg)) in green tea on the production and extracellular release of Vero toxins (VTs) from enterohemorrhagic Escherichia coli (EHEC) cultured at 37 degrees C for 24 h. EGCg and GCg in the culture medium markedly inhibited extracellular VTs release from EHEC cells into the culture supernatant fluid at concentrations of 0.05 mg/ml or higher, as estimated by both the reversed passive latex agglutination assay and cytotoxic assay using Vero cells. Production and extracellular release of maltose binding protein, a periplasmic protein, into the culture supernatant were also inhibited by EGCg and GCg, indicating that their inhibitory effect on release from periplasm into the outer milieu is not specific to VTs, but general to the proteins accumulated in EHEC periplasm.

p53 gene mutation and loss of heterozygosity are associated with increased risk of disease progression in adult T cell leukemia.

Although the prototype of adult T cell leukemia (ATL) is an aggressive T cell neoplasm, ATL manifests four major clinical subtypes, acute, lymphoma, chronic, and smoldering. We studied the relationship between p53 gene alteration and clinical features in 34 patients with ATL, 14 acute type, 15 chronic type, and five crisis type transformed from chronic type. Using a polymerase chain reaction/single strand conformation polymorphism (PCR/SSCP) assay, followed by nucleotide sequencing, we detected mutations of the p53 gene in six of the 14 acute type patients, two of the five crisis type, and one of the 15 chronic type patients. Gene dosage studies, using PCR amplification and Southern blotting, showed loss of heterozygosity (LOH) of the p53 gene in four of the 14 acute type patients, two of the five crisis type, and one of 14 chronic type patients examined. These observations indicated that the frequency of p53 gene alterations in the acute and crisis types of ATL was markedly higher than that in chronic type, suggesting that p53 gene alteration plays a role in the disease progression of ATL.