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Potassium octatitanate (K 2 O·8TiO 2 , POT) fibers are used as an alternative to asbestos. Their shape and biopersistence suggest that they are possibly carcinogenic. However, inhalation studies have shown that respired POT fibers have little carcinogenic potential. We conducted a short-term study in which we administered POT fibers, and anatase and rutile titanium dioxide nanoparticles (a-nTiO 2 , r-nTiO 2 ) to rats using intra-tracheal intra-pulmonary spraying (TIPS). We found that similarly to other materials, POT fibers were more toxic than non-fibrous nanoparticles of the same chemical composition, indicating that the titanium dioxide composition of POT fibers does not appear to account for their lack of carcinogenicity. The present report describes the results of the 3-week and 52-week interim killing of our current 2-year study of POT fibers, with MWCNT-7 as a positive control and r-nTiO2 as a non-fibrous titanium dioxide control. Male F344 rats were administered 0.5 ml vehicle, 62.5 µg/ml and 125 µg/ml r-nTiO 2 and POT fibers, and 125 µg/ml MWCNT-7 by TIPS every other day for 2 weeks (eight doses: total doses of 0.25 and 0.50 mg/rat). At 1 year, POT and MWCNT-7 fibers induced significant increases in alveolar macrophage number, granulation tissue in the lung, bronchiolo-alveolar cell hyperplasia and thickening of the alveolar wall, and pulmonary 8-OHdG levels. The 0.5 mg POT- and the MWCNT-7-treated groups also had increased visceral and parietal pleura thickness, increased mesothelial cell PCNA labeling indices, and a few areas of visceral mesothelial cell hyperplasia. In contrast, in the r-nTiO2-treated groups, none of the measured parameters were different from the controls.

We developed a Magnetic Resonance elastography (MRE) technique using a conventional magnetic resonance imaging (MRI), which allows a simultaneous elastography of the supraspinatus and trapezius muscles, by designing a new wave transducer (vibration pad) and optimizing the mechanical vibration frequency. Five healthy volunteers underwent an MRE. In order to transmit the mechanical vibration (pneumatic vibration) to the supraspinatus and trapezius muscles, a new vibration pad was designed using a three-dimensional (3D) printer. The vibration pad was placed on the skin 2 cm medial and 2 cm cephalad the deltoid tubercle. MRE acquisition was performed with a multi-slice gradient-echo type multi-echo MR sequence, which allows MREs even in a conventional MRI; two oblique axial images of the supraspinatus and trapezius muscles were obtained simultaneously. Vibration frequencies were set at 50-150 Hz, with a 25 Hz step. Wave image quality in each frequency was analyzed using a phase-to-noise ratio (PNR) and clarity of propagating wave that was assessed by two readers qualitatively. In the supraspinatus muscle, the wave images were of good quality especially at frequencies >75 Hz. In the trapezius muscle, the wave images were of better quality at low frequencies (50 and 75 Hz) compared with high frequencies (100-150 Hz). The PNR of both muscles were higher at low frequencies. The mean stiffness in the trapezius muscle (7.26 ± 2.13 kPa at 75 Hz) was larger than those in the supraspinatus muscle (4.16 ± 0.50 kPa at 75 Hz). The results demonstrated that our MRE technique allows simultaneous assessment of the stiffness in the supraspinatus and trapezius muscles using a conventional MRI, and that optimal vibration frequency for simultaneous MRE of these muscles is 75 Hz. This technique provides a new means for early detection of abnormality in the shoulder., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Very little is known about the in vivo toxicity of inhaled double-walled carbon nanotubes (DWCNTs). In the present study, we compared the pulmonary toxicity of DWCNT to MWCNT-7, a well-known multi-walled carbon nanotube. Rats were divided into six groups: untreated, vehicle, low-dose DWCNT, high-dose DWCNT, low-dose MWCNT-7, and high-dose MWCNT-7. The test materials were administered by intra-tracheal intra-pulmonary spraying (TIPS) every other day for 15 days: the low-dose and high-dose groups were administered final total doses of 0.25 and 0.50 mg/rat of the test material. The animals were sacrificed 1 and 6 weeks after the final TIPS administration. Six weeks after the final TIPS administration, rats administered MWCNT-7 had high levels of macrophage infiltration into the lung with dense alveolar wall fibrous thickening throughout the lung; significant elevation of lactate dehydrogenase activity, alkaline phosphatase activity, and total protein concentration in the bronchioalveolar lavage fluid; an increase in the pulmonary cell PCNA index; slightly elevated levels of 8-OHdG DNA adducts in lung tissue DNA; a small but significant increase in protein concentration in the pleural cavity lavage fluid and an increase in the visceral mesothelial cell PCNA index. None of these parameters was increased in rats administered DWCNT. The primary lesion in rats administered DWCNT was scattered formation of granulation tissue containing internalized DWCNT fibers. Our data indicate that DWCNT has lower pulmonary and pleural toxicity than MWCNT-7.

The purpose of this study was to investigate an influence of vibration waveform on magnetic resonance elastography (MRE). MRE is an innovative imaging technique for the non-invasive quantification of the elasticity of soft tissues through the direct visualization of propagating shear waves in vivo using a special phase-contrast magnetic resonance imaging sequence. Since the elasticity of soft tissue calculates from the wavelength of propagating shear waves, it is necessary to propagate sine-wave-shape shear wave at the target soft tissue. However, due to the various factors; i.e. overload of vibration generator, poor contact between imaging object and vibration pad, etc.; it may be difficult to generate a simple sine wave. This work was focused on change vibration waveforms; i.e. square wave, triangle wave, saw-tooth wave; which is induced by the various factors. Phantom experimental results demonstrated that when square and saw-tooth waveforms of 25 Hz vibration frequency, into the phantom, the waveform of propagating wave was not similar to sine waveform. It may influence on the MRE that in case of the waveforms has low frequency and square or saw-tooth like waveforms.

Diffusion-magnetic resonance elastography (dMRE) is an emerging practical technique that can acquire diffusion magnetic resonance imaging and MRE simultaneously. However, a signal loss attributable to intravoxel phase dispersion (IVPD) interferes with the calculation of the apparent diffusion coefficient (ADC). This study presents an approach to dMRE that reduces the influence of IVPD by introducing a new pulse sequence. The existing and proposed techniques were performed using a phantom comprising five rods with different elasticities at 60 Hz vibration to investigate the accuracy of previous and proposed dMRE techniques. The measures of ADC and stiffness, obtained by using both dMRE techniques, were compared with conventional spin-echo (SE) diffusion and SE-MRE. Then, we evaluated those differences by using the mean of absolute differences (MAD) in each rod within the phantom. The results of the MAD of the stiffness from both dMRE techniques showed almost no difference. In contrast, the value of the ADC MAD (MAD ≒ 0.16 × 10 -3  mm 2 /s), obtained in the soft region within the phantom with the previous dMRE technique, was large. This value was about 2.7 times that of the value produced by the proposed dMRE technique. This difference must reflect the degree of influence of IVPD in both techniques. These results demonstrate that our dMRE technique is a robust method for addressing the signal loss attributable to IVPD., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Potassium octatitanate fibers (K 2 O·8TiO 2 , POT fibers) are widely used as an alternative to asbestos. We investigated the pulmonary and pleural toxicity of POT fibers with reference to 2 non-fibrous titanium dioxide nanoparticles (nTiO 2 ), photoreactive anatase (a-nTiO 2 ) and inert rutile (r-nTiO 2 ). Ten-week-old male F344 rats were given 0.5 mL of 250 μg/mL suspensions of POT fibers, a-nTiO 2 , or r-nTiO 2 , 8 times (1 mg/rat) over a 15-day period by trans-tracheal intrapulmonary spraying (TIPS). Rats were killed at 6 hours and at 4 weeks after the last TIPS dose. Alveolar macrophages were significantly increased in all treatment groups at 6 hours and at 4 weeks. At week 4, a-nTiO 2 and r-nTiO 2 were largely cleared from the lung whereas a major fraction of POT fibers were not cleared. In the bronchoalveolar lavage, alkaline phosphatase activity was elevated in all treatment groups, and lactate dehydrogenase (LDH) activity was elevated in the a-nTiO 2 and POT groups. In lung tissue, oxidative stress index and proliferating cell nuclear antigen (PCNA) index were elevated in the a-nTiO 2 and POT groups, and there was a significant elevation in C-C motif chemokine ligand 2 (CCL2) mRNA and protein in the POT group. In pleural cavity lavage, total protein was elevated in all 3 treatment groups, and LDH activity was elevated in the a-nTiO 2 and POT groups. Importantly, the PCNA index of the visceral mesothelium was increased in the POT group. Overall, POT fibers had greater biopersistence, induced higher expression of CCL2, and provoked a stronger tissue response than a-nTiO 2 or r-nTiO 2 ., (© 2018 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

We developed a facile one-pot fabrication process for magnetic iron oxide-calcium phosphate (IO-CaP) composite nanoparticles via coprecipitation in labile supersaturated CaP solutions containing IO nanocrystals. All the source solutions used were clinically approved for injection, including water and magnetic IO nanocrystals (ferucarbotran, used as a negative magnetic resonance imaging (MRI) contrast agent). This ensured that the resulting nanoparticles were pathogen- and endotoxin-free. The dispersants used were clinically approved heparin sodium (heparin) or adenosine triphosphate disodium hydrate (ATP), which were added to the IO-containing labile supersaturated CaP solutions. Both heparin and ATP coprecipitated with CaP and ferucarbotran to form heparin- and ATP-modified IO-CaP nanoparticles, respectively, with a hydrodynamic diameter of a few hundred nanometers. Both the resulting nanoparticles exhibited relatively large negative zeta potentials, caused by the negatively charged functional groups in heparin and ATP, which improved the particle dispersibility when compared to non-modified IO-CaP nanoparticles. The heparin-modified IO-CaP nanoparticles were effectively ingested by murine macrophages (RAW264.7) without showing significant cytotoxicity but barely ingested by non-phagocytotic human umbilical vein endothelial cells, indicating the potential of these nanoparticles for targeted delivery to macrophages. The heparin-modified IO-CaP nanoparticles exhibited a negative contrast enhancing ability for MRI. Our results show that IO-CaP nanoparticles have potential as delivery and MRI contrast agents for macrophages., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Short-term alternatives to traditional 2-year carcinogenic studies in rodents are being actively pursued. Recently, a 26-week short-term carcinogenicity study using CB6F1-Tg rasH2@Jcl (rasH2) mice has become a worldwide standard for the evaluation of chemical carcinogenesis. However, an acceptable short-term carcinogenic study model for dermally applied products is still lacking. To investigate the suitability of using the rasH2 mouse to test carcinogenic potential, 1,2-dichloroethane (1,2-DCE) was dermally applied to rasH2 mice: 1,2-DCE is a known carcinogen that causes lung bronchiolo-alveolar adenomas and adenocarcinomas when administered topically, orally, or by inhalation exposure; 1,2-DCE at a dose level of 126 mg/mouse in 200 μl acetone or acetone alone (vehicle control) was applied to the dorsal skin of 10 mice of each sex 3 times a week for 26 weeks. As a positive control, 10 mice of each sex received a single intraperitoneal injection of 75 mg/kg of N-methyl- N-nitrosourea. Bronchiolo-alveolar adenomas and adenocarcinomas were significantly increased in 1,2-DCE-treated rasH2 mice of both sexes, and bronchiolo-alveolar hyperplasias were significantly increased in female mice. Overall, almost all mice of each sex developed adenomas and/or adenocarcinomas with 100% of female rasH2 mice developing bronchiolo-alveolar adenocarcinomas.

Magnetic resonance elastography (MRE) can measure tissue stiffness quantitatively and noninvasively. Supraspinatus muscle injury is a significant problem among throwing athletes. The purpose of this study was to develop an MRE technique for application to the supraspinatus muscle by using a conventional magnetic resonance imaging (MRI). MRE acquisitions were performed with a gradient-echo type multi-echo MR sequence at 100Hz pneumatic vibration. A custom-designed vibration pad was used as a pneumatic transducer in order to adapt to individual shoulder shapes. In a gradient-echo type multi-echo MR sequence, without motion encoding gradient (MEG) that synchronizes with vibrations, bipolar readout gradient lobes achieved a similar function to MEG (MEG-like effect). In other words, a dedicated MRE sequence (built-in MEG) is not always necessary for MRE. In this study, 7 healthy volunteers underwent MRE. We investigated the effects of direction of the MEG-like effect and selected imaging planes on the patterns of wave propagation (wave image). The results indicated that wave images showed clear wave propagation on a condition that the direction of the MEG-like effect was nearly perpendicular to the long axis of the supraspinatus muscle, and that the imaging plane was superior to the proximal supraspinatus muscle. This limited condition might be ascribed to specific features of fibers in the supraspinatus muscle and wave reflection from the boundaries of the supraspinous fossa. The mean stiffness of the supraspinatus muscle was 10.6±3.17kPa. Our results demonstrated that using MRE, our method can be applied to the supraspinatus muscle by using conventional MRI., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Multiwalled carbon nanotubes (MWCNT) have a fibrous structure and physical properties similar to asbestos and have been shown to induce malignant mesothelioma of the peritoneum after injection into the scrotum or peritoneal cavity in rats and mice. For human cancer risk assessment, however, data after administration of MWCNT via the airway, the exposure route that is most relevant to humans, is required. The present study was undertaken to investigate the carcinogenicity of MWCNT-N (NIKKISO) after administration to the rat lung. MWCNT-N was fractionated by passing it through a sieve with a pore size of 25 μm. The average lengths of the MWCNT were 4.2 μm before filtration and 2.6 μm in the flow-through fraction; the length of the retained MWCNT could not be determined. For the present study, 10-week-old F344/Crj male rats were divided into five groups: no treatment, vehicle control, MWCNT-N before filtration, MWCNT-N flow-through and MWCNT-N retained groups. Administration was by the trans-tracheal intrapulmonary spraying (TIPS) method. Rats were administered a total of 1 mg/rat during the initial 2 weeks of the experiment and then observed up to 109 weeks. The incidences of malignant mesothelioma and lung tumors (bronchiolo-alveolar adenomas and carcinomas) were 6/38 and 14/38, respectively, in the three groups administered MWCNT and 0/28 and 0/28, respectively, in the control groups. All malignant mesotheliomas were localized in the pericardial pleural cavity. The sieve fractions did not have a significant effect on tumor incidence. In conclusion, administration of MWCNT to the lung in the rat induces malignant mesothelioma and lung tumors., (© 2016 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

In cartilage regenerative medicine, autologous chondrocyte implantation (ACI) has been applied clinically for partial defects of joint cartilage or nasal augmentation. To make treatment with ACI more effective and prevalent, modalities to evaluate the quality of transplanted constructs noninvasively are necessary. In this study, we compared the efficacy of several noninvasive modalities for evaluating the maturation of tissue-engineered auricular cartilage containing a biodegradable polymer scaffold. We first transplanted tissue-engineered cartilage consisting of human auricular chondrocytes, atelocollagen gel, and a poly-l-lactic acid (PLLA) porous scaffold subcutaneously into the back of athymic nude rats. Eight weeks after transplantation, the rats were examined by magnetic resonance imaging (MRI), X-ray, and ultrasound as noninvasive modalities. Then, the excised constructs were examined by histological and biochemical analysis including toluidine blue (TB) staining, glycosaminoglycans content, and enzyme-linked immunosorbent assay of type II collagen. Among the modalities examined, transverse relaxation time (T2) and apparent diffusion coefficient of MRI showed quite a high correlation with histological and biochemical results, suggesting that these can effectively detect the maturation of tissue-engineered auricular cartilage. Since these noninvasive modalities would realize time-course analysis of the maturation of tissue-engineered auricular cartilage, this study provides a substantial insight for improving the quality of tissue-engineered cartilage, leading to improvement of the quality and technique in cartilage regenerative medicine.

Palpation is a standard clinical tool to diagnose abnormal stiffness changes in soft tissues. However, it is difficult to palpate the supraspinatus muscle because it locates under the trapezius muscle. The magnetic resonance elastography (MRE) uses harmonic mechanical excitation to quantitatively measure the stiffness (shear modulus) of both the superficial and deep tissues. The purpose of this study was to build a vibration system for applying the MRE to the supraspinatus muscle. In this study, a power amplifier and a pneumatic pressure generator were used to supply vibrations to a vibration pad. Six healthy volunteers underwent MRE. We investigated the effects of position (the head of the humerus and the trapezius muscle) of the vibration pad on the patterns of wave propagation (wave image). When the vibration pad was placed in the trapezius muscle, the wave images represented clear wave propagation. On the other hand, when the vibration pad was placed in the head of the humerus, the wave images represented unclear wave propagation. This result might be caused by wave interferences resulting from the vibrations from bones and an intramuscular tendon of the supraspinatus muscle. The mean shear modulus also was 8.12 ± 1.83 (mean ± SD) kPa, when the vibration pad was placed in the trapezius muscle. Our results demonstrated that the vibration pad should be placed in the trapezius muscle in the MRE of the supraspinatus muscle.

To demonstrate the feasibility of a novel MR elastography (MRE) technique based on a conventional gradient-echo type multi-echo MR sequence which does not need additional bipolar magnetic field gradients (motion encoding gradient: MEG), yet is sensitive to vibration. In a gradient-echo type multi-echo MR sequence, several images are produced from each echo of the train with different echo times (TEs). If these echoes are synchronized with the vibration, each readout's gradient lobes achieve a MEG-like effect, and the later generated echo causes a greater MEG-like effect. The sequence was tested for the tissue-mimicking agarose gel phantoms and the psoas major muscles of healthy volunteers. It was confirmed that the readout gradient lobes caused an MEG-like effect and the later TE images had higher sensitivity to vibrations. The magnitude image of later generated echo suffered the T2 decay and the susceptibility artifacts, but the wave image and elastogram of later generated echo were unaffected by these effects. In in vivo experiments, this method was able to measure the mean shear modulus of the psoas major muscle. From the results of phantom experiments and volunteer studies, it was shown that this method has clinical application potential., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Background: Transmission and tomographic X-ray measurements are useful in assessing bone structures, but only a few studies have examined cartilage growth because of the poor contrast in conventional X-ray imaging., Objective: In this study, we attempted to use the linear attenuation coefficient (LAC) as a metric of tissue-engineered cartilage development, which would be useful in high-throughput screening of cartilage products., Methods: Assuming that the LAC is related to the amount of extracellular matrix (ECM) in terms of the density and its atomic components, we measured X-ray absorption through tissue-engineered cartilage constructs. Characteristic X-ray beams from a molybdenum microfocus X-ray tube were employed to avoid beam hardening. The correlation of the LAC with mechanical properties was analyzed for verification., Results: The LAC was higher for chondrocyte constructs and lower for fibroblast-dominant constructs and was consistent with the quantification of toluidine blue staining, which is a proof of ECM production. The LAC was positively correlated with the bending modulus but negatively correlated with the dynamic elastic modulus and stiffness, possibly because of the remaining scaffold., Conclusions: The LAC has the potential to be used as a metric of development of tissue-engineered cartilage. However, the calcified regions should be excluded from analysis to avoid decreasing the correlation between the LAC and the amount of ECM.

Surfactant proteins (SPs), originally known as human lung surfactants, are essential to respiratory structure and function. There are 4 subtypes, SP-A, SP-B, SP-C and SP-D, with SP-A and SP-D having immunological functions, and SP-B and SP-C having physicochemical properties that reduce the surface tension at biological interfaces. In this experiment, the expressions of SP-A, SP-B, SP-C and SP-D in lung neoplastic lesions induced by N-bis (2-hydroxypropyl) nitrosamine (DHPN) and inflammatory lesions due to quartz instillation were examined and compared immunohistochemically. Formalin fixed paraffin embedded (FFPE) lung samples featuring inflammation were obtained with a rat quartz instillation model, and neoplastic lesions, hyperplasias and adenomas, were obtained with the rat DHPN-induced lung carcinogenesis model. In the rat quartz instillation model, male 10-week old F344 rats were exposed by intratracheal instillation (IT) to quartz at a dose of 2 mg/rat suspended in saline (0.2 ml) on day 0, and sacrificed on day 28. Lung tumorigenesis in F344 male rats was initiated by DHPN in drinking water for 2 weeks, and the animals were then sacrificed in week 30. Lung proliferative lesions, hyperplasias and adenomas, were observed with DHPN, and inflammation was observed with quartz. The expressions of SP-A, SP-B, SP-C and SP-D were examined immunohistochemically. SP-B and SP-C showed strong expression in lung hyperplasias and adenomas, while SP-A and SP-D were observed in mucus or exudates in inflammatory alveoli. These results suggest the possibility that SP-B and SP-C are related to lung tumorigenesis.

Multiwalled carbon nanotubes (MWCNT) have a fibrous structure similar to asbestos, raising concern that MWCNT exposure may lead to asbestos-like diseases. Previously we showed that MWCNT translocated from the lung alveoli into the pleural cavity and caused mesothelial proliferation and fibrosis in the visceral pleura. Multiwalled carbon nanotubes were not found in the parietal pleura, the initial site of development of asbestos-caused pleural diseases in humans, probably due to the short exposure period of the study. In the present study, we extended the exposure period to 24 weeks to determine whether the size and shape of MWCNT impact on deposition and lesion development in the pleura and lung. Two different MWCNTs were chosen for this study: a larger sized needle-like MWCNT (MWCNT-L; l = 8 μm, d = 150 nm), and a smaller sized MWCNT (MWCNT-S; l = 3 μm, d = 15 nm), which forms cotton candy-like aggregates. Both MWCNT-L and MWCNT-S suspensions were administered to the rat lung once every 2 weeks for 24 weeks by transtracheal intrapulmonary spraying. It was found that MWCNT-L, but not MWCNT-S, translocated into the pleural cavity, deposited in the parietal pleura, and induced fibrosis and patchy parietal mesothelial proliferation lesions. In addition, MWCNT-L induced stronger inflammatory reactions including increased inflammatory cell number and cytokine/chemokine levels in the pleural cavity lavage than MWCNT-S. In contrast, MWCNT-S induced stronger inflammation and higher 8-hydroxydeoxyguanosine level in the lung tissue than MWCNT-L. These results suggest that MWCNT-L has higher risk of causing asbestos-like pleural lesions relevant to mesothelioma development., (© 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.)

Two types of nanosized titanium dioxide, anatase (anTiO2) and rutile (rnTiO2), are widely used in industry, commercial products and biosystems. TiO2 has been evaluated as a Group 2B carcinogen. Previous reports indicated that anTiO2 is less toxic than rnTiO2, however, under ultraviolet irradiation anTiO2 is more toxic than rnTiO2 in vitro because of differences in their crystal structures. In the present study, we compared the in vivo and in vitro toxic effects induced by anTiO2 and rnTiO2. Female SD rats were treated with 500 ?g/ml of anTiO2 or rnTiO2 suspensions by intra-pulmonary spraying 8 times over a two week period. In the lung, treatment with anTiO2 or rnTiO2 increased alveolar macrophage numbers and levels of 8-hydroxydeoxyguanosine (8-OHdG); these increases tended to be lower in the anTiO2 treated group compared to the rnTiO2 treated group. Expression of MIP1??mRNA and protein in lung tissues treated with anTiO2 and rnTiO2 was also significantly up-regulated, with MIP1??mRNA and protein expression significantly lower in the anTiO2 group than in the rnTiO2 group. In cell culture of primary alveolar macrophages (PAM) treated with anTiO2 and rnTiO2, expression of MIP1??mRNA in the PAM and protein in the culture media was significantly higher than in control cultures. Similarly to the in vivo results, MIP1??mRNA and protein expression was significantly lower in the anTiO2 treated cultures compared to the rnTiO2 treated cultures. Furthermore, conditioned cell culture media from PAM cultures treated with anTiO2 had less effect on A549 cell proliferation compared to conditioned media from cultures treated with rnTiO2. However, no significant difference was found in the toxicological effects on cell viability of ultra violet irradiated anTiO2 and rnTiO2. In conclusion, our results indicate that anTiO2 is less potent in induction of alveolar macrophage infiltration, 8-OHdG and MIP1??expression in the lung, and growth stimulation of A549 cells in vitro than rnTiO2.

Zinc oxide (ZnO) is known to induce lung toxicity, including terminal bronchiolar epithelial hyperplasia, which gives rise to concerns that nanosized ZnO (nZnO) might lead to lung carcinogenesis. We studied the tumor promoting activity of nZnO by an initiation-promotion protocol using human c-Ha-ras proto-oncogene transgenic rats (Hras128 rats). The rats were given 0.2 % N-nitrosobis(2-hydroxypropyl)amine (DHPN) in the drinking water for 2 weeks and then treated with 0.5 ml of 250 or 500 μg/ml nZnO suspension by intra-pulmonary spraying once every 2 weeks for a total of 7 times. Treatment with nZnO particles did not promote DHPN-induced lung carcinogenesis. However, nZnO dose-dependently caused epithelial hyperplasia of terminal bronchioles (EHTB) and fibrosis-associated interstitial pneumonitis (FAIP) that were independent of DHPN treatment. Tracing the fate of EHTB lesions in wild-type rats indicated that the hyperplastic lesions almost completely disappeared within 12 weeks after the last nZnO treatment. Since nZnO particles were not found in the lung and ZnCl2 solution induced similar lung lesions and gene expression profiles, the observed lesions were most likely caused by dissolved Zn(2+). In summary, nZnO did not promote carcinogenesis in the lung and induced EHTB and FAIP lesions that regressed rapidly, probably due to clearance of surplus Zn(2+) from the lung.

The purpose of this study was to develop a new technique for a powerful compact MR elastography (MRE) actuator based on a pneumatic ball-vibrator. This is a compact actuator that generates powerful centrifugal force vibrations via high speed revolutions of an internal ball using compressed air. This equipment is easy to handle due to its simple principles and structure. Vibration frequency and centrifugal force are freely adjustable via air pressure changes (air flow volume), and replacement of the internal ball. In order to achieve MRI compatibility, all parts were constructed from non-ferromagnetic materials. Vibration amplitudes (displacements) were measured optically by a laser displacement sensor. From a bench test of displacement, even though the vibration frequency increased, the amount of displacement did not decrease. An essential step in MRE is the generation of mechanical waves within tissue via an actuator, and MRE sequences are synchronized to several phase offsets of vibration. In this system, the phase offset was detected by a four-channel optical-fiber sensor, and it was used as an MRI trigger signal. In an agarose gel phantom experiment, this actuator was used to make an MR elastogram. This study shows that the use of a ball actuator for MRE is feasible., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The aim of our study was to simulate an effective diffusion-weighted imaging (DWI) pulse sequence and to evaluate the best b value in skeletal muscle. The evaluated pulse sequences were spin echo (SE), stimulated echo (STE), and gradient-recalled echo (GRE). The signal intensity changed in some DWI pulse sequences when the b value was changed. Moreover, the static magnetic field of 1.5 T was compared with that of 3.0 T. Next, diffusion tensor imaging of the human skeletal muscle was measured. The experimental subjects were healthy male volunteers (n = 7; age 21.8 ± 1.1 years). We changed the b value in steps of 100 s/mm(2) from 0 to 2000 s/mm(2), and the diffusion values [fractional anisotropy (FA), λ(1), λ(2), and λ(3)] were calculated based on the data. The STE method could maintain signals up to b values of 1300 s/mm(2), but the SE and GRE methods suffered from high noise. In the human study, the mean FA (±SD) was 0.41 ± 0.02 in the tibialis anterior muscle (TA) and 0.31 ± 0.02 in the soleus muscle (SOL) at a b value of 1000 s/mm(2). The correlation of the FA with the b value coefficient and the P value by Student's t test were r = 0.981 and P < 0.001 for the TA and r = 0.982, P < 0.001 for the SOL. However, the FA became high with a low b value. In conclusion, STE was very useful for DWI in a short T (2)-value tissue. Moreover, a minimum b value of 800 s/mm(2) was necessary for evaluation of human skeletal muscle.

The modifying potential on tumor development of arachidonate-enriched triglyceride oil (ARA-oil) containing approximately 40% arachidonic acid was investigated in a medium-term multi-organ carcinogenesis bioassay using male and female F344 rats. The animals were sequentially given five carcinogens with different target sites in the first 4 weeks, and then administered ARA-oil for 24 weeks at dietary levels of 0% (control), 1.25%, 2.5% or 5.0%. No statistically significant differences in incidences and multiplicities of hyperplastic and neoplastic lesions were showed in the large intestine in either sex. In the liver, kidney, and lung in both sexes, and the mammary gland and uterus in females, tumor promoting potential was not evident with ARA-oil treatment. ARA-oil did not affect the quantitative data for glutathione S-transferase placental form positive foci of the liver. Increased induction of hyperplastic or neoplastic lesions in the urinary bladder and thyroid in ARA-oil-treated groups was without dose dependence. In addition, a second experiment with ARA-oil only administration for 8-week revealed no effects on cellular proliferation in the urinary bladder or thyroid in either sex. These results indicate that ARA-oil has no tumor promoting potential in any organs or tissues initiated with the five carcinogens applied in the present study., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objective: Using an anthropomorphic phantom, we have investigated the feasibility of digital tomosynthesis (DT) of flat-panel detector (FPD) radiography to reduce radiation dose for sinonasal examination compared to multi-detector computed tomography (MDCT)., Materials and Methods: A female Rando phantom was scanned covering frontal to maxillary sinus using the clinically routine protocol by both 64-detector CT (120 kV, 200 mAs, and 1.375-pitch) and DT radiography (80 kV, 1.0 mAs per projection, 60 projections, 40° sweep, and posterior-anterior projections). Glass dosimeters were used to measure the radiation dose to internal organs including the thyroid gland, brain, submandibular gland, and the surface dose at various sites including the eyes during those scans. We compared the radiation dose to those anatomies between both modalities., Results: In DT radiography, the doses of the thyroid gland, brain, submandibular gland, skin, and eyes were 230 ± 90 μGy, 1770 ± 560 μGy, 1400 ± 80 μGy, 1160 ± 2100 μGy, and 112 ± 6 μGy, respectively. These doses were reduced to approximately 1/5, 1/8, 1/12, 1/17, and 1/290 of the respective MDCT dose., Conclusion: For sinonasal examinations, DT radiography enables dramatic reduction in radiation exposure and dose to the head and neck region, particularly to the lens of the eye., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Magnetic resonance elastography (MRE) is designed for imaging the mechanical properties of soft tissues. However, the interpretation of shear modulus distribution is often confusing and cumbersome. For reliable evaluation, a common practice is to specify the regions of interest and consider regional elasticity. Such an experience-dependent protocol is susceptible to intrapersonal and interpersonal variability. In this study we propose to remodel shear modulus distribution with piecewise constant level sets by referring to the corresponding magnitude image. Optimal segmentation and registration are achieved by a new hybrid level set model comprised of alternating global and local region competitions. Experimental results on the simulated MRE data sets show that the mean error of elasticity reconstruction is 11.33% for local frequency estimation and 18.87% for algebraic inversion of differential equation. Piecewise constant level set modeling is effective to improve the quality of shear modulus distribution, and facilitates MRE analysis and interpretation., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: 18q-Syndrome is a chromosomal disorder exhibiting various symptoms arising from the central nervous system. Brain magnetic resonance imaging (MRI) of patients with this syndrome usually demonstrates abnormal white matter intensities. This is widely believed to be due to impaired myelin formation because this syndrome involves the deletion of the myelin basic protein (MBP) gene in 18q23. However, this hypothesis has not been confirmed by actual pathology because early death is unusual and autopsy rarely performed., Patient: A 6-year-old boy with ring chromosome 18 syndrome was examined by genetic analysis for the MBP gene, brain MRI, and autopsy., Results: Haploinsufficiency of the MBP gene was confirmed. T(2)-weighted MRI revealed diffuse high intensities throughout the cerebral white matter. Pathological examination showed the cerebral white matter to be uniformly stained by Klüver-Barrera and MBP immunohistochemical staining. Oligodendrocytes were immunoreactive for proteolipid protein and ferritin but not MBP. Electron microscopy revealed clusters of axons wrapped in compact myelin sheaths with distinct major dense lines. Holzer and immunohistochemical staining for glial fibrillary acidic protein showed extensive staining of the white matter and an increased number of glial filaments., Conclusions: This pathological study demonstrated that in this disorder, the brain was well myelinated, contrary to established hypotheses about this disorder. The MRI signal abnormalities in 18q-syndrome could be attributed to gliosis and not to dysmyelination., (Copyright © 2011 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Applying tissue-engineered cartilage in a clinical setting requires noninvasive evaluation to detect the maturity of the cartilage. Magnetic resonance imaging (MRI) of articular cartilage has been widely accepted and applied clinically in recent years. In this study, we evaluated the negative fixed-charge density (nFCD) of tissue-engineered cartilage using gadolinium-enhanced MRI and determined the relationship between nFCD and biomechanical properties. To reconstruct cartilage tissue, articular chondrocytes from bovine humeral heads were embedded in agarose gel and cultured in vitro for up to 4 weeks. The nFCD of the cartilage was determined using the MRI gadolinium exclusion method. The equilibrium modulus was determined using a compressive stress relaxation test, and the dynamic modulus was determined by a dynamic compression test. The equilibrium compressive modulus and dynamic modulus of the tissue-engineered cartilage increased with an increase in culture time. The nFCD value--as determined with the [Gd-DTPA(2-)] measurement using the MRI technique--increased with culture time. In the regression analysis, nFCD showed significant correlations with equilibrium compressive modulus and dynamic modulus. From these results, gadolinium-enhanced MRI measurements can serve as a useful predictor of the biomechanical properties of tissue-engineered cartilage.

This study was designed to evaluate any adverse effect of a hot water extract of black soybeans (Glycine max (L.) Merr.), when administered to both sexes of Crj:CD(SD)IGS rats at dietary levels of 0 (control), 0.5, 1.5 and 5.0% (6 rats/sex/group). During the study, the treatment had no adverse effects on clinical signs, survival, body weights, and food and water consumption, or on findings of ophthalmology, urinalysis, hematology, or blood biochemistry. Organ weights, gross pathology and histopathology exhibited no differences of toxicological significance between control and treated rats. Thus, the no-observed-adverse-effect level (NOAEL) of black soybean extract was concluded to be 5.0% (3,618 mg/kg body weight/day for males and 4,066 mg/kg body weight/day for females) from the present study.

High b-value diffusion magnetic resonance imaging (MRI) enables us to detect far smaller architectures, by using q-space analysis, than the resolution in conventional MRI. Average displacement, one of the q-space parameters, quantitatively reflects architecture size and is very useful in observing small changes in microstructures in vivo (e.g., neurodegeneration, tumor heterogeneity, and others). Diffusion-weighted imaging (DWI) is performed by a two-dimensional (2D) multislice method; however, due to finite slice thickness and slice gap, there is a partial-volume effect that makes it difficult to detect the net q-space signal. On the other hand, three-dimensional (3D) MRI, having the advantages of very thin slice thickness and no slice gap (contiguous slices), allows volumetric evaluation acquired in a small isotropic voxel, as compared to 2D multislice imaging. Little is known about the isotropic high-resolution 3D DWI application to q-space analysis. In this study, we have developed and implemented a high b-value 3D DWI sequence, applied q-space analysis to study the reliability of high b-value 3D DWI and obtained a microscopic analytical map with isotropic high resolution and less contamination.

For security, boards containing lead are commonly used to partition off rooms adjacent to radiation facilities. From the viewpoints of environmental safety and of material recycling during the process of remodeling or repair, however, the trend is toward avoiding the use of lead-containing radiation-protection board. Consequently, we developed a lead-free board(W 910xH 1820xD 12.5 mm)composed of a mixed material incorporating calcium sulfate and barium sulfate to be used as a partition at a diagnostic X-ray institution. The mass attenuation coefficients of the lead-free board were lower by about 12% and 60% for energy ranges of 25 approximately 37 keV and 38 approximately 41 keV, respectively, as compared with those of lead. Lead equivalents for the newly developed board(12.5 mm thickness)were 0.87+/-0.01 mmPb at 100 kV tube voltage, 0.60+/-0.01 at 125 kV, and 0.47+/-0.01 at 150 kV. A double-layer lead-free board proved to have enough shielding ability even at usual photographic frequencies.

The application of tissue-engineered cartilage in a clinical setting requires a noninvasive method to assess the biophysical and biochemical properties of the engineered cartilage. Since articular cartilage is composed of 70-80% water and has dense extracellular matrixes (ECM), it is considered that the condition of the water molecules in the tissue is correlated with its biomechanical property. Therefore, magnetic resonance imaging (MRI) represents a potential approach to assess the biophysical property of the engineered cartilage. In this study, we test the hypothesis that quantitative MRI can be used as a noninvasive assessment method to assess the biophysical property of the engineered cartilage. To reconstruct a model of cartilaginous tissue, chondrocytes harvested from the humeral head of calves were embedded in an agarose gel and cultured in vitro up to 4 weeks. Equilibrium Young's moduli were determined from the stress relaxation tests. After mechanical testing, MRI-derived parameters (longitudinal relaxation time T1, transverse relaxation time T2, and water self-diffusion coefficient D) were measured. The equilibrium Young's modulus of the engineered cartilage showed a tendency to increase with an increase in the culture time, whereas T1 and D decreased. Based on a regression analysis, T1 and D showed a strong correlation with the equilibrium Young's modulus. The results showed that T1 and D values derived from the MRI measurements could be used to noninvasively monitor the biophysical properties of the engineered cartilage.

In this work we report on the development of a novel technique for fat-saturated three-dimensional (3D) diffusion-weighted (DW) MRI sequence based upon 3D magnetization-prepared rapid gradient-echo (3D-MP-RAGE). In order to saturate fat, two kinds of procedures were competed CHESS-DW-3D-MP-RAGE sequence (CHESS-3D-DWI) and DW-3D-MP-WE-RAGE sequence (WE-3D-DWI) "chemical shift selective: CHESS method vs. water-excitation: WE method". The CHESS-3D-DWI sequence and WE-3D-DWI sequence were compared in terms of their degree of fat suppression. In CHESS-3D-DWI sequence a preparation phase with a "CHESS-90 degrees RF-motion probing gradient: MPG-180 degrees RF-MPG-90 degrees RF" pulse-train was used to sensitize the magnetization to fat-saturated diffusion. In contrast, WE-3D-DWI sequence a RAGE-excitation pulse with a "binominal-pulse 1-1 or 1-2-1" was selected to water-excited (fat-saturated) diffusion imaging. These imaging were done during in vivo studies using an animal experiment. From experimental results obtained with a phantom, the effect of diffusion weighting and the effect of fat-saturation were confirmed. Fat-saturation was much better in the WE-3D-DWI sequence than CHESS-3D-DWI sequence. From rat experimental results, fat-saturated diffusion-weighted image data were obtained. This sequence was useful for in vivo imaging.

Purpose: Applying regenerated cartilage in a clinical setting requires noninvasive evaluation to detect the maturity of cartilage tissue. Magnetic resonance (MR) imaging of articular cartilage is well accepted and has been applied clinically in recent years. We attempt to establish a noninvasive method to evaluate the maturity of regenerated cartilage tissue using gadolinium-enhanced MR imaging., Methods: To reconstruct cartilaginous tissue, we embedded articular chondrocytes harvested from bovine humeral head in agarose gel and cultured the cells in vitro up to 4 weeks. The fixed charge density (FCD) of the cartilage was determined using MRI gadolinium exclusion method. The sulfated glycosaminoglycan (sGAG) content was determined by dimethylmethylene blue dye-binding assay., Results: The sGAG content and FCD of the regenerated cartilage increased with duration of culture. In the T1Gd maps, the [Gd-DTPA(2-)] in the specimen decreased, and the boundary between the sample disk and the bath solution of phosphate buffered saline (PBS) became clearer as time in culture increased. In the linear regression analysis, FCD and sGAG content correlated significantly., Conclusion: Gadolinium-enhanced MR imaging measurements can be useful predictors of the degree of cartilaginous tissue formation.

The purpose of this study was to examine the potential of diffusion-weighted (DW) three-dimensional (3D) MP-RAGE MRI for diffusion-tensor mapping of the rat brain in vivo. A DW-3D-MP-RAGE (3D-DWI) sequence was implemented at 2.0 T using six gradient orientations and a b value of 1000 s/mm2. In this sequence, the preparation sequence with a "90 degrees RF-motion proving gradient (MPG): MPG-180 degrees RF-MPG-90 degrees RF" pulse train (DW driven equilibrium Fourier transform) was used to sensitize the magnetization to diffusion. A centric k-space acquisition order was necessary to minimize saturation effects (T1 contamination) from tissues with short relaxation time. The image matrix was 128x128x128 (interpolated from 64x64x64 acquisitions), which resulted in small isotropic DW image data (voxel size: 0.273x0.273x0.273 mm3). Moreover, 3D-DWI-derived maps of the fractional anisotropy (FA), relative anisotropy (RA) and main-diffusion direction were completely free of susceptibility-induced signal losses and geometric distortions. Two well-known commissural fibers, the corpus callosum and anterior commissure, were indicated and shown to be in agreement with the locations of these known stereotaxic atlases. The experiment took 45 min, and shorter times should be possible in clinical application. The 3D-DWI sequence allows for in vivo 3D diffusion-tensor mapping of the rat brain without motion artifacts and susceptibility to distortion.

The advantages of three-dimensional (3D) acquisition are that you obtain thinner and more slices with better profiles, and better signal-to-noise ratio for an equivalent slice thickness. Three-dimensional acquisition is preferable for obtaining contiguous thin-slice MR images. However, the acquisition time extends compared with the two-dimensional acquisition because the second phase-encode axis is applied by the 3D acquisition. Therefore, 3D acquisition should be a high-speed imaging method. In this paper, a new diffusion-sensitive 3D magnetization-prepared rapid gradient-echo (3D MP-RAGE) sequence was studied. In this sequence, a preparation phase with a 90 degrees RF-motion proving gradient (MPG): MPG-180 degrees RF-MPG-90 degrees RF pulse train (diffusion-weighted driven-equilibrium Fourier transform) was used to sensitize the magnetization to diffusion. Centric k-space acquisition order is necessary to minimize saturation effects from tissues with short relaxation times. From phantom experimental results, the effect of the diffusion weighting was changed by the centric vs. sequential k-space acquisition order. The effect of centric k-space acquisition order was larger than the effect of sequential k-space acquisition order. The contrast of centric k-space acquisition order became equal to the contrast of conventional diffusion-weighted spin echo. From rat experimental results, small isotropic diffusion-weighted image data (voxel size: 0.625 x 0.625 x 0.625 mm3) were obtained. This sequence was useful in vivo.

Kanagawa Occupational Health Promotion Center conducted a survey on how the MSDS is utilized at workplaces with more than 50 employees handling chemical substances, and what measures are taken to help employees to thoroughly understand information in the Material Safety Data Sheet (MSDS). Questionnaires were sent out to 265 enterprises in Kanagawa prefecture, putting questions to industrial physicians and industrial hygiene supervisors. The objective of the survey was to find out how MSDS is adopted in the system to manage occupational health, what improvements the survey respondents want in MSDS and what expectations the respondents have of our center. 193 enterprises (72.8%) returned answers to the questionnaire. The major findings are as follows. (1) In many companies, information on hazardous/toxic materials is "controlled by a division using such materials", and roughly half of the companies have compiled a common list shared throughout the company. (2) For the most part suppliers submit to the MSDS. Larger companies have a higher rate of posting up or filing the MSDS at their workplaces. Only 25.8% of the companies "rewrite the MSDS so that workers can understand it." (3) Companies that carry out a hazard/toxicity assessment before introducing a new chemical substance account for 72.1%, which is higher than we expected. It indicates that even though the companies don't manage the MSDS adequately, they are highly concerned about hazard control of chemical substances. (4) The rate of answering that "the current MSDS is not easy to understand" is higher among large-sized enterprises and lower among enterprises with fewer than 300 employees. (5) Asked what improvement needs to be made on the MSDS, the industrial physicians and industrial hygiene supervisors gave same answers such as "Workers find the terminology difficult to understand." and "Levels of toxicity can't be clearly identified." (6) The respondents expect our center to provide information for the MSDS. In conclusion, it is considered that in order to prepare understandable MSDSs to workers in enterprises, the role of our center to provide information and education on the MSDS to respondents was important.

Data from the large-scale biological monitoring program in Japan were assembled and analyzed and the following results were obtained. All workers handling lead and eight kinds of major organic solvents received physical examinations and biological monitoring at the same time. Therefore, the number of workers handling industrial chemicals and that received physical examinations and the number of workers been examined by biological monitorings were similar to each other. The total number of cases examined from 1989 to 1994 was about 661,000 for lead in the blood and about 4,173,000 for the urinary metabolites of eight organic solvents. The results were classified into three categories and category 3 consists of workers having exposure concentrations above the 1988-1989 biological exposure indices of the ACGIH with the exception of lead concentration in the blood where the limit in Japan was set at 40 micrograms/100 ml. The percentage of exposed workers in category 3 was 1.4% for blood lead and 0.2-2.4% for the urinary metabolites of the eight organic solvents. The percentage of exposed workers in category 3 for blood lead, delta-aminolevulinic acid, urinary mandelic acid, N-methylformamide and 2,5-hexanedione in the urine has decreased with time. In ambient monitoring, the percentage of workplaces in classification 3 for lead and styrene also has decreased with time.