Usefulness of immunocytochemistry using a Breast Marker antibody cocktail targeting P63/cytokeratin7/18/cytokeratin5/14 for fine needle aspiration of the breast: a retrospective cohort study of 139 cases.

OBJECTIVE: The Breast Marker Cocktail from Biocare Medical comprises five antibodies recognising p63, and cytokeratins (CKs) 7, 18, 5 and 14. Immunohistochemistry using this cocktail is useful for diagnosing proliferative intraductal breast lesions. However, cytology using the cocktail has not been reported. METHODS: We report 139 cases of mammary samples collected by fine needle aspiration (FNA) for which histological diagnoses were available. After cell transfer, immunocytochemistry was performed using the cocktail, and clusters of cells were classified. A cluster with no or limited CK5/14 expression (<20% of cells) was classified as a monotonous cluster. One with more than 20% of cells showing CK5/14 expression was defined as a mosaic cluster. When at least one p63-positive cell was present, we defined it as a cluster with p63. We also evaluated background p63-positive myoepithelial cell densities. RESULTS: The diagnostic sensitivity and specificity for carcinomas were 97.8% (89/91) and 91.7% (11/12), respectively, using the criterion of two or more monotonous clusters lacking p63. Two false-negative cases were triple-negative cancers; one false-positive was an apocrine papilloma. The numbers of monotonous clusters with p63 differed significantly between benign lesions, ductal carcinoma in situ (DCIS)/lobular carcinoma in situ (LCIS) and invasive carcinomas (P < 0.001). The background myoepithelial cell density was significantly higher in fibroepithelial tumours than in other lesions (P < 0.001). CONCLUSIONS: Immunocytochemistry using this antibody cocktail showed good sensitivity and specificity for diagnosing breast cancers. Thus, this method is useful for mammary cytology using FNA.

Occlusal trauma accelerates attachment loss at the onset of experimental periodontitis in rats.

BACKGROUND AND OBJECTIVE: Occlusal trauma is an important factor that influences the progression of periodontitis, but it is unclear whether occlusal trauma influences periodontal destruction at the onset of periodontitis. We established an experimental periodontitis model with both site-specific loss of attachment and alveolar bone resorption. The purpose of the present study was to investigate the effects of occlusal trauma on periodontal destruction, particularly loss of attachment, at the onset of experimental periodontitis. MATERIAL AND METHODS: Sixty rats were used in the present study. Forty-eight rats immunized with lipopolysaccharide (LPS) intraperitoneally were divided into four groups. In the trauma (T) group, occlusal trauma was induced by placing an excessively high metal wire in the occlusal surface of the mandibular right first molar. In the inflammation (I) group, periodontal inflammation was induced by topical application of LPS into the palatal gingival sulcus of maxillary right first molars. In the trauma + inflammation (T+I) group, both trauma and periodontal inflammation were simultaneously induced. The PBS group was administered phosphate-buffered saline only. Another 12 nonimmunized rats (the n-(T+I) group) were treated as described for the T+I group. All rats were killed after 5 or 10 d, and their maxillary first molars with surrounding tissues were observed histopathologically. Loss of attachment and osteoclasts on the alveolar bone crest were investigated histopathologically. To detect immune complexes, immunohistological staining for C1qB was performed. Collagen fibers were also observed using the picrosirius red-polarization method. RESULTS: There were significant increases in loss of attachment and in the number of osteoclasts in the T+I group compared with the other groups. Moreover, widespread distribution of immune complexes was observed in the T + I group, and collagen fibers oriented from the root surface to the alveolar bone crest had partially disappeared in the T, T+I and n-(T+I) groups. CONCLUSION: When inflammation was combined with occlusal trauma, immune complexes were confirmed in more expanding areas than in the area of the I group without occlusal trauma, and loss of attachment at the onset of experimental periodontitis was increased. Damage of collagen fibers by occlusal trauma may elevate the permeability of the antigen through the tissue and result in expansion of the area of immune-complex formation and accelerating inflammatory reaction. The periodontal tissue destruction was thus greater in the T+I group than in the I group.

Correlation of circulating dehydroepiandrosterone with activated protein C generation and carotid intima-media thickness in male patients with type 2 diabetes.

AIMS: Dehydroepiandrosterone exerts a protective effect against cardiovascular diseases. However, the relationship of dehydroepiandrosterone with the anticoagulant factor activated protein C, generated by the thrombin-thrombomodulin complex on vascular endothelial cells, remains unknown. This study aimed at studying the relationship between dehydroepiandrosterone and activated protein C generation in patients with Type 2 diabetes. METHODS: Sixty-two male patients with Type 2 diabetes were enrolled in this study. Data obtained from 40 healthy male subjects were used as controls. The plasma levels of dehydroepiandrosterone, the activated protein C-protein C inhibitor complex, high-sensitivity C-reactive protein and monocyte chemoattractant protein-1 were measured by enzyme immunoassays. Carotid intima-media thickness was measured by ultrasonography. RESULTS: The plasma levels of dehydroepiandrosterone (5.15 ± 2.81 vs. 3.76 ± 2.16 ng/ml; P < 0.005) and the activated protein C-protein C inhibitor complex (1.90 ± 1.07 vs. 1.02 ± 0.51 ng/ml; P < 0.001) were significantly lower in patients with diabetes than in normal subjects. Univariate analysis showed a significant correlation of the plasma level of dehydroepiandrosterone with that of the activated protein C-protein C inhibitor complex (r = 0.48, P < 0.001), high-sensitivity C-reactive protein (r = -0.30, P < 0.05) and with the mean intima-media thickness (r = -0.28, P < 0.05) in patients with diabetes. Stepwise multiple regression analysis showed that the plasma level of dehydroepiandrosterone is significantly correlated with the plasma levels of the activated protein C-protein C inhibitor complex (F = 18.06) and high-sensitivity C-reactive protein (F = 4.94). There was no correlation between the plasma levels of dehydroepiandrosterone and monocyte chemoattractant protein-1. CONCLUSIONS: These results suggest that lower circulating levels of dehydroepiandrosterone are associated with decreased activated protein C generation and higher intima-media thickness in patients with Type 2 diabetes.

Development of a field test for evaluating aerobic fitness.

The purpose of this study was to investigate the reproducibility and utility of a standardised and externally paced field test (15-m Incremental Shuttle Walk and Run Test [15 mISWRT]) to assess aerobic fitness in middle-aged adults. 14 middle-aged participants performed the 15-m ISWRT 3 times within one week (Test 1, Test 2, Test 3). Reproducibility of the 15-m ISWRT was tested by comparing 15-m ISWRT performance (distance completed), HRmax, and VO 2max for each test. The utility of the 15-m ISWRT for evaluating VO 2max over a wide range in middle-aged adults was tested by comparing the range of VO 2max obtained from the portable expired gas analyzer with the VO 2max reference values and ranges for health promotion published by Japan's Ministry of Health, Labour and Welfare. A multiple comparison of distance completed in the 15-m ISWRT Test 1, Test 2, and Test 3 found no significant difference between Test 2 and Test 3. The ICC was 0.99 for Test 2 vs. Test 3. VO 2max measured from the 15-m ISWRT in Test 3 had a minimum value of 22.8 ml/kg/min and a maximum value of 38.7 ml/kg/min. In conclusion, the 15-m ISWRT is reliable and useful for evaluating VO 2max in middle-aged adults.

Numerical and Experimental Study to Fabricate the New Type Compact NMR Device Using Stacked HTS Bulks.

In fabrication of a compact NMR (Nuclear Magnetic Resonance) magnet which consists of a stacked HTS (High Temperature Superconducting) bulk annuli, generally there are three key issues: spatial homogeneity, temporal stability, strength of trapped magnetic fields. This paper presents a study on the effects of axial gap length between stacked HTS bulks on the three key issues of a bulk HTS magnet for compact HTS NMR applications. The HTS bulk magnet of which the ID and OD are 20 and 60 mm respectively has a 50 and 80 mm heights depending on the axial gap lengths between HTS bulks. The gap length between each HTS bulk varied from 0 mm to 10 mm and were used as parameters to optimize, analytically as well as experimentally, the overall field homogeneity of the HTS bulk magnet. The optimized axial gap length was obtained by analytical results, and the better magnetic field homogeneity and temporal stability of trapped magnetic field were achieved by lower magnetization field. The improved spatial homogeneity and strength of generated magnetic field by a new compact NMR magnet will be presented.

Diversity and characterization of oxytetracycline-resistant bacteria associated with non-native species, white-leg shrimp (Litopenaeus vannamei), and native species, black tiger shrimp (Penaeus monodon), intensively cultured in Thailand.

AIMS: This study aimed at surveying prevalence of oxytetracycline (OTC)-resistant bacteria in the white-leg shrimp Litopenaeus vannamei, and the black tiger shrimp Penaeus monodon, intensively cultured in Thailand. We investigated the phylogenetic diversity of the bacterial isolates, as well as the minimum inhibitory concentration (MIC) of OTC, the occurrence of major OTC-resistant genes and multiple-antibiotic resistance in the isolates. METHODS AND RESULTS: Shrimps were collected from culture ponds, and the homogenates of whole bodies were plated on tryptic soy agar supplemented with or without OTC. Percentages of OTC-resistant bacteria were 0·3-52·1% in white-leg samples and 0·008-22·3% in black tiger samples. Analyses of 16S rDNA sequences indicated that most OTC-resistant isolates were closely related to Aeromonas spp. and Lactococcus garvieae. MICs of OTC were 4-128 µg ml(-1) in the OTC-resistant aeromonads and 128-256 µg ml(-1) in OTC-resistant L. garvieae. OTC resistance was found to be conferred by the genes tet(A), tet(C), tet(D), tet(E), tet(M) and tet(S), detected either singly or in pairs. No resistance to ceftazidime, imipenem or chloramphenicol was observed in any isolate. CONCLUSIONS: Both species of shrimp are associated with OTC-resistant bacteria, occasionally at high densities exceeding 10(6) cfu g(-1). The associated bacteria, predominantly Lactococcus and Aeromonas genera, are potential pathogens and are reservoirs of a variety of OTC-resistant genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Cultured shrimps can be vehicle to carry OTC-resistant bacteria to domestic and foreign consumers via the food chain. Very low populations of OTC-resistant bacteria observed in the several ponds suggest that levels of the resistant bacteria are artificially high and should be reduced in farmed shrimps.

Magnetospheric vortex formation: self-organized confinement of charged particles.

A magnetospheric configuration gives rise to various peculiar plasma phenomena that pose conundrums to astrophysical studies; at the same time, innovative technologies may draw on the rich physics of magnetospheric plasmas. We have created a "laboratory magnetosphere" with a levitating superconducting ring magnet. Here we show that charged particles (electrons) self-organize a stable vortex, in which particles diffuse inward to steepen the density gradient. The rotating electron cloud is sustained for more than 300 s. Because of its simple geometry and self-organization, this system will have wide applications in confining single- and multispecies charged particles.

Protective role of thrombin activatable fibrinolysis inhibitor in obstructive nephropathy-associated tubulointerstitial fibrosis.

BACKGROUND: Thrombin-activatable fibrinolysis inhibitor (TAFI) has been reported to affect wound healing and fibrotic processes, but its role in renal tubulointerstitial fibrosis remains unknown. OBJECTIVE: To study its potential role, we compared TAFI-deficient and wild-type mice for the degree of renal fibrosis caused by unilateral ureteral obstruction (UUO). METHODS: The grade of tubulointerstitial fibrosis, the activity of plasmin, MMP-2 and MMP-9 were evaluated on days 4 and 9 after UUO. RESULTS: The renal content of hydroxyproline and the activity of plasmin, MMP-2 and MMP-9 were significantly increased in kidneys with UUO from TAFI-deficient mice compared with those from wild-type mice. These differences disappeared when animals with UUO from both groups were treated with the plasmin inhibitor tranexamic acid. The renal concentrations of fibrogenic cytokines were also significantly elevated in kidneys with UUO from TAFI-deficient mice compared with those from wild-type mice. CONCLUSION: The results of this study suggest that increased renal activity of plasmin in TAFI-deficient mice causes increased renal interstitial fibrosis in obstructive nephropathy.

Detection of Kobe-type Babesia microti associated with Japanese human babesiosis in field rodents in central Taiwan and southeastern mainland China.

Field rodent surveys for Babesia infection were performed from 2002 to 2005 in the vicinities of human babesiosis occurrences in Taiwan and mainland China. Babesia microti was identified by microscopical examination and/or PCR in 1 Rattus coxinga and 1 Crocidura horsfieldii in central Taiwan and in 13 Niviventer confucianus and 1 Apodemus agrarius in Zhejiang and Fujian Provinces of southeastern China. Of 15 B. microti samples detected by PCR, all except 1 were shown to be the Kobe-type, the aetiological small subunit rRNA gene-type of the first Japanese patient; the exception was also a Kobe-related type. The Kobe-type had been found in rodents only in a few places including the human infection occurrence place in Japan. The internal transcribed spacer 1 to 2 sequences of the Taiwanese and Chinese Kobe-types were very similar to each other but considerably different (approx. 94% pairwise identities) from that of the Japanese Kobe-type. A Taiwanese Kobe-type strain was serologically differentiated from the Kobe strain originating from the Japanese first patient. The distribution of the Kobe-type in the vicinities of human babesiosis occurrences in Taiwan and China as well as in Japan is suggestive of involvement of the Kobe-type in Asian human babesiosis.

The enhancement effect of three sugar alcohols on the fungicidal effect of benzethonium chloride toward Candida albicans.

OBJECTIVE: The purpose of this study was to evaluate if the sugar alcohols erythritol, xylitol and sorbitol enhance the fungicidal effect of benzethonium chloride (BTC) toward in vitro candidal biofilms. METHODS: An in vitroCandida albicans biofilm was formed on a plastic coverslip coated with type I collagen. The enhancement of the fungicidal effect was evaluated using microbial assay after treating the biofilm with the test solutions (sugar alcohols: erythritol, xylitol, and sorbitol, each containing BTC solution). RESULTS: No fungicidal effect was observed with sugar alcohols without BTC. The fungicidal effect of erythritol-containing BTC increased with the erythritol concentration. Further, the level of enhancement of erythritol was the highest in three sugar alcohols. CONCLUSION: Sugar alcohols, especially erythritol, enhanced the fungicidal effect of BTC toward in vitro candidal biofilms.

New superconducting electron cyclotron resonance ion source for RIKEN RI beam factory project.

For RIKEN radio isotope beam project, we started to construct the new superconducting electron cyclotron resonance ion source (SC-ECRIS), which has an operational frequency of 28 GHz, in 2007. The main feature of this ion source is that we can produce large size of resonance zone with six sets of solenoid coils. Before starting, we intensively studied the effect of several key parameters of ECRIS (magnetic field configuration, microwave power density, negatively biased disk) on the plasma. In this article, we describe the effect of key parameters on the plasma and detailed structure of the new SC-ECRIS.

Production of U beam from RIKEN 18 GHz electron cyclotron resonance ion source.

For the RIKEN radio isotope factory (RIBF) project, we produced the multicharged uranium beam with two methods. To produce lower charge state U ion beams (14+-20+) we used the UF(6) gas as an ionized gas. The typical beam intensity of U(14+-20+) was 2-1 particle microA at the extraction voltage of 14 kV. To produce higher charge state U ion beam (U(35+)), we chose the sputtering method. The beam intensity was 70 particle nA at the extraction voltage of 5.4 kV. Using this method, we successfully produced multicharged U beam continuously for one month without break for RIBF commissioning.

Arsenic speciation for the phytoremediation by the Chinese brake fern, Pteris vittata.

Arsenic (As) speciation for the phytoremediation by the Chinese brake fern was studied. In particular, the mechanism of how plants induce compounds containing thiol (SH) and proteins by As exposure in terms of the relationship between As and phosphate uptaken into plant cells was examined. Pteris vittata callus could efficiently reduce As(V) to As(III) by the rapid introduction of reductase and synthesize thiols leading to phytochelatins production. Furthermore, Pteris vittata could control phosphate concentration in the cells corresponding to the concentration of arsenite and arsenate. To our best knowledge, this is the first report to show the mechanisms of such high As tolerance of Pteris vittata using their callus in terms of in vitro approach for the analysis of As speciation and metabolism route.

Microwave oscillator using piezoelectric thin-film resonator aiming for ultraminiaturization of atomic clock.

We developed a microwave oscillator and a micro electromechanical systems-based rubidium cell for the miniaturization of atomic clocks. A thin-film bulk acoustic resonator (FBAR) having a resonant frequency of the fundamental mode in the 3.5 GHz band was employed instead of a crystal resonator. It delivers a clock transition frequency of Rb atoms of 3.417 GHz without the need for a complicated frequency multiplication using a phase-locked loop. This topology considerably reduces the system scale and power consumption. For downsizing the atomic clock system toward the chip level as well as mass production, a microfabricated gas cell containing Rb and N2 gases was also developed. These microcomponents were incorporated into an atomic clock test bench, resulting in a clock operation with a short-term frequency instability of 2.1 × 10-11 at 1 s. To the best of our knowledge, this is the first report of a coherent population trapping clock operation using an FBAR-based microwave oscillator as well as a microfabricated gas cell.

Coexpression of glucose transporters and glucokinase in Xenopus oocytes indicates that both glucose transport and phosphorylation determine glucose utilization.

A Xenopus oocyte expression system was used to examine how glucose transporters (GLUT 2 and GLUT 3) and glucokinase (GK) activity affect glucose utilization. Uninjected oocytes and low rates of both glucose transport and phosphorylation; expression of GLUT 2 or GLUT 3 increased glucose phosphorylation approximately 20-fold by a low Km, endogenous hexokinase at glucose concentrations < or = 1 mM, but not at higher glucose concentrations. Coexpression of functional GK isoforms with GLUT 2 or 3 increased glucose utilization approximately an additional two- to threefold primarily at the physiologic glucose concentrations of 5-20 mM. The Km for glucose of both the hepatic and beta cell isoforms of GK, determined in situ, was approximately 5-10 mM when coexpressed with either GLUT 2 or GLUT 3. The increase in glucose utilization by coexpression of GLUT 3 and GK was dependent upon glucose phosphorylation since two missense GK mutations linked with maturity-onset diabetes, 182: Val-->Met and 228:Thr-->Met, did not increase glucose utilization despite accumulation of both a similar amount of immunoreactive GK protein and glucose inside the cell. Coexpression of a mutant GK and a normal GK isoform did not interfere with the function of the normal GK enzyme. Since the coexpression of GK and a glucose transporter in oocytes resembles conditions in the hepatocyte and pancreatic beta cell, these results indicate that increases in glucose utilization at glucose concentrations > 1 mM depend upon both a functional glucose transporter and GK.

Risk evaluation for staphylococcal food poisoning in processed milk produced with skim milk powder.

The growth of S. aureus and the production of staphylococcal enterotoxin A (SEA) in skim milk concentrates stored at inappropriate temperatures in a recovery milk tank (tank for excess concentrated skim milk) used in the manufacture of skimmed milk powder were investigated. Also, it was estimated if a possible outbreak of food poisoning would occur if the contaminated skimmed milk powder was used in the manufacture of processed milk. Skim milk concentrates with milk solid content of 15, 25, and 35% were inoculated with S. aureus at 1-2 log CFU/ml and incubated at 15, 25, or 35 degrees C for 0 to 24 h with or without shaking. Bacterial growth and the level of SEA production were measured. At 35 degrees C with shaking, there was a significant difference (p<0.05) in one way layout analysis of variance, and it was demonstrated that the growth of S. aureus and SEA production could be milk solid content-dependent. Shaking accelerated the growth of S. aureus and SEA production at 35 degrees C. Generally, skim milk powder is produced by mixing a set percentage of skim milk concentrates (recovery milk) from the recovery milk tank into raw milk. If recovery milk contaminated with S. aureus at levels of 1-2 log CFU/ml is kept at 15 to 35 degrees C due to a power failure, it was estimated that processed milk consumption of 670-1200 ml, 420-1500 ml and 18-83 ml would trigger the onset of food poisoning symptoms when skim milk concentrates (recovery milk) are stored at 25 degrees C for 24 h, 35 degrees C for 10 h, and 35 degrees C for 24 h, respectively, during the production of the skim milk powder. Based on these consumption levels, it was concluded that, if recovery milk cannot be refrigerated and is stored at room temperature (25 to 35 degrees C), it must be used within 8 h and preferably within 6 h.

Electro-optic probe measurements of electric fields in plasmas.

The direct measurements of high-frequency electric fields in a plasma bring about significant advances in the physics and engineering of various waves. We have developed an electro-optic sensor system based on the Pockels effect. Since the signal is transmitted through an optical fiber, the system has high tolerance for electromagnetic noises. To demonstrate its applicability to plasma experiments, we report the first result of measurement of the ion-cyclotron wave excited in the RT-1 magnetosphere device. This study compares the results of experimental field measurements with simulation results of electric fields in plasmas.

Risk factor analysis for ascending aorta and aortic arch repair using selective cerebral perfusion with open technique: role of open-stent graft placement.

AIM: The present study was designed to identify risk factors that may induce adverse outcome defined as permanent neurological dysfunction and mortality after aortic arch surgery using selective cerebral perfusion by logistic regression analysis and to reveal the role of open stent-graft placement. METHODS: One hundred and nineteen consecutive patients underwent ascending aorta and/or aortic arch operation with open technique between 1995 and 2005 were examined. Ascending aorta and/or hemiarch was replaced in 28 patients, total arch in 75 patients, and proximal or distal aortic arch replacement in 16 patients. Open stent-graft placement was used in 25 patients. RESULTS: The in-hospital mortality rate was 9.2%. Permanent neurological dysfunction occurred in 10 patients (8.4%). Thoracotomy (P=0.0331) and cardiopulmonary bypass time (P=0.0238) were significant risk factors for permanent neurological dysfunction. Preoperative shock (P=0.0266) was significant independent risk factor for mortality. Emergent operation (P=0.0454), thoracotomy (P=0.0232), and cardiopulmonary bypass time (P=0.0379) were significant independent risk factors for adverse outcome. The duration of selective cerebral perfusion was not associated with adverse outcome. Open stent-graft placement has no need of thoracotomy for aneurysm extending descending thoracic aorta and time variables concerning the operation were significantly shorter in the patients with open stent-graft placement than in patients with standard operation for total arch replacement. RESULTS: Thoracotomy was significant risk factor for adverse outcome after aortic arch repair using selective cerebral perfusion. Total arch replacement with open stent-graft placement can avoid the need of thoracotomy and reduce time variables concerning the operation to improve the surgical

Point mutation of ornithine decarboxylase gene in human hepatocellular carcinoma.

Ornithine decarboxylase (ODC) plays an important role in cell growth, and its activity is regulated by many mechanisms. The biochemical characteristics of ODC in malignant cells differ from those of ODC in normal cells. To determine whether novel changes occur in ODC in neoplastic tissue, we compared the nucleotide sequence of ODC cDNA obtained from human hepatoma tissue as determined by reverse transcriptase-PCR with that of ODC cDNA obtained from nontumorous tissue in the same patients. There were three point mutations accompanied by replacements of amino acids in hepatoma tissue with other amino acids or a stop codon. In one poorly differentiated hepatoma, codon 415, CAA was converted to TAA, resulting in replacement of Gln-415 by a stop codon. The mutated ODC protein produced by translation in a reticulocyte-lysate protein synthesizing system was truncated and stabilized in an ATP antizyme-dependent degradation system. These findings suggest that formation of a truncated and stabilized ODC protein due to point mutation is one reason why ODC activity is high in human hepatoma tissue.