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The influence of chromophore solubility enhancements on photorefractive grating rise time and device lifetime is investigated. Three azo chromophores differing primarily in compatibility with a polyvinylcarbazole host polymer were synthesized. Aromatic substitutions to the chromophore increased the device lifetime from several days to years although electric-field-induced poling experiments indicated that chromophore orientational mobility is severely hindered, resulting in photorefractive grating rise times approaching several hours. The incorporation of a flexible butyl chain to the aromatic substituted chromophores significantly enhanced the orientational mobility. These chromophores could be loaded as high as 60 wt % with no degradation in transparency for one year following fabrication. [ABSTRACT FROM AUTHOR]

BACKGROUND: Erdheim-Chester disease is a rare histiocytic disease entity related to juvenile xanthogranuloma. It is a systemic condition, usually occurs in adult, characterized by infiltration of foamy histiocytes within the bone and soft tissues. METHODS AND RESULTS: We report a case of 13-year-old female patient who first presented with multiple osteolytic lesions of the jaws followed by bilateral symmetrical bone lesions affecting the lower extremities, as well as brain and abdominal involvement. Histological findings of the jaw lesions showed lipid-storing CD68 (+), CD1a (-) histiocytes with Touton type giant cells. CONCLUSION: To the best of our knowledge, this is the first case of Erdheim-Chester disease with jaw bone lesions occurring as initial presenting symptom. [ABSTRACT FROM AUTHOR]

In this study, we aimed to conduct a survey on the current clinical practice of, staffing for, commissioning of, and staff training for online adaptive radiotherapy (oART) in the institutions that installed commercial oART systems in Japan, and to share the information with institutions that will implement oART systems in future. A web-based questionnaire, containing 107 questions, was distributed to nine institutions in Japan. Data were collected from November to December 2023. Three institutions each with the MRIdian (ViewRay, Oakwood Village, OH, USA), Unity (Elekta AB, Stockholm, Sweden), and Ethos (Varian Medical Systems, Palo Alto, CA, USA) systems completed the questionnaire. One institution (MRIdian) had not performed oART by the response deadline. Each institution had installed only one oART system. Hypofractionation, and moderate hypofractionation or conventional fractionation were employed in the MRIdian/Unity and Ethos systems, respectively. The elapsed time for the oART process was faster with the Ethos than with the other systems. All institutions added additional staff for oART. Commissioning periods differed among the oART systems owing to provision of beam data from the vendors. Chambers used during commissioning measurements differed among the institutions. Institutional training was provided by all nine institutions. To the best of our knowledge, this was the first survey about oART performed using commercial systems in Japan. We believe that this study will provide useful information to institutions that installed, are installing, or are planning to install oART systems., (© 2024. The Author(s), under exclusive licence to Japanese Society of Radiological Technology and Japan Society of Medical Physics.)

Purpose: Measurement of beam alignment, including an isocenter, is a fundamental part of commissioning and quality assurance for a medical linear accelerator (linac). An alignment shift is caused by the focal spot displacement from the beam axis. In this study, we present a procedure for the analysis of the focal spot positions in the radiation alignment using a newly developed quality control tool (QCT) device and analysis software., Methods: The device has the function to perform the light/radiation field coincidence test, and the isocenter test at the same time. First, the light/radiation field was evaluated by using cross-slits on the surface of the QCT device. Second, the focal spot positions of beam alignment were measured using QCT, and the acquired images were collectively analyzed using the developed analysis software. Finally, measurements of focal spot positions with different energies were made using the accelerators at the 2 facilities., Results: The focal spot position was changed with a gantry angle. The focal spot displacement was 0.15-1.37 mm. With the validation of 2 accelerators at 2 facilities, the displacement of the focal spot position could be measured using the QCT phantom., Conclusion: The focal spot position measurement using the QCT test captured the change in the beam alignment between the beam axes without being affected by the jaw collimator. This study suggested that focal spot position measurement using a QCT device is useful for evaluating beam alignment at linac.

Background/aim: The aim of this study was to evaluate the mechanical performance and the effect on dose distribution and deliverability of volumetric modulated arc therapy (VMAT) plans for prostate cancer created with the commercial knowledge-based planning (KBP) system (RapidPlan™)., Materials and Methods: Three institutions, A, B, and C were enrolled in this study. Each institution established and trained a KBP model with their own cases. CT data and structures for 45 patients at institution B were utilized to validate the dose-volume parameters (D 2(%) , D 95(%) , and D 98(%) for target, and V 50(%) , V 75(%) , and V 90(%) for rectum and bladder), and the following mechanical performance parameters and gamma passing rates of each KBP model: leaf sequence variability (LSV), aperture area variability (AAV), total monitor unit (MU), modulation complexity score for VMAT (MCSv), MU/control point (CP), aperture area (AA)/CP, and MU×AA/CP., Results: Significant differences (p<0.01) in dosimetric parameters such as D 2 and D 98 for target and V 50 , V 75 , and V 90 for bladder were observed among the three institutions. The means and standard deviations of MCSv were 0.31±0.03, 0.29±0.02, and 0.32±0.03, and the angles of maximum and minimum MU×AA/CP were 269° and 13°, 269° and 13°, and 273° and 153° at institutions A, B, and C, respectively. The mean gamma passing rate (1%/1 mm.) was >95% for all cases in each institution. Dose distribution and mechanical performance significantly differed between the three models., Conclusion: Each KBP model had different dose distributions and mechanical performance but could create an acceptable plan for deliverability regardless of mechanical performance., (Copyright© 2022, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

In the present study, by using double self-activated CsI detectors, the development of a neutron dosemeter system whose response indicates better agreement with the International Commission on Radiological Protection-74 rem-response was carried out to simply evaluate the neutron dose with high accuracy. The present double neutron dosemeter system, using a slow-neutron dosemeter (thermal to 10 keV) and a fast-neutron dosemeter (above 10 keV), consists of CsI scintillators wrapped with two types of neutron energy filtering materials: polyethylene and B4C silicon rubber. After optimization of each filter thickness, to confirm the validity of our method, the neutron ambient dose equivalents under several operating conditions of medical linear accelerators (Linacs) were evaluated using a Monte Carlo simulation and an experiment with the present dosemeter. From these results, the present dosimetry system has enabled a more accurate neutron dose evaluation than our conventional dosemeter, and the present dosemeter was suitable for the neutron dosimetry for 10 MV Linac environments., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

MTG8 (RUNX1T1) is a fusion partner of AML1 (RUNX1) in the leukemic chromosome translocation t(8;21). The AML1-MTG8 fusion gene encodes a chimeric transcription factor. One of the highly conserved domains of MTG8 is TAFH which possesses homology with human TAF4 [TATA-box binding protein-associated factor]. To obtain specific inhibitors of the AML1-MTG8 fusion protein, we isolated RNA aptamers against the MTG8 TAFH domain using systematic evolution of ligands by exponential enrichment. All TAF aptamers contained guanine-rich sequences. Analyses of a TAF aptamer by NMR, CD, and mutagenesis revealed that it forms a parallel G-quadruplex structure in the presence of K + . Furthermore, the aptamer could bind to the AML1-MTG8 fusion protein and dissociate the AML1-MTG8/DNA complex, suggesting that it can inhibit the dominant negative effects of AML1-MTG8 against normal AML1 function and serve as a potential therapeutic agent for leukemia., (© 2020 Federation of European Biochemical Societies.)

This study aimed to evaluate the performance of a single-energy metal artifact reduction (SEMAR) algorithm for radiation therapy treatment using phantom cases with metal inserts, assess improvements in computed tomography (CT) number accuracy, and investigate its effects on treatment planning dosimetry. A standard electron density phantom was scanned with and without metal inserts. The numbers of tissue-equivalent materials on both uncorrected and SEMAR-corrected CT images were compared. Treatment planning accuracy was evaluated by comparing dose distributions computed using true density images (without metal inserts), uncorrected images (with metal inserts), and SEMAR-corrected images (with metal inserts) using three-dimensional gamma analysis. The numbers of the true density and uncorrected and SEMAR-corrected CT images in a muscle plug with unilateral inserts were 25.9 HU, - 281.8 HU, and 26.1 HU, respectively. A similar tendency was obtained for other tissue-equivalent materials, and the numbers on CT images were improved with the SEMAR algorithm. In cases involving 1 portal irradiation, 10-MV X-ray, and the Acuros XB algorithm, the pass ratio between the true density and uncorrected images was 89.89%, while that between the true density and SEMAR-corrected images was 95.03%. Improvements in dose distribution were evident using the SEMAR algorithm. Similar trends were found for different irradiation methods and dose calculation algorithms. The SEMAR algorithm can significantly reduce metal artifacts on CT images used for radiation treatment planning. This aspect influenced dosimetry in the region of the artifact and dose distribution was significantly improved with use of the SEMAR-corrected images.

Background: Insulin-derived amyloidosis is a skin-related complication of insulin therapy that interferes with insulin therapy. Although toxicities of in vitro-formed insulin amyloid fibrils have been well studied, the toxicity of insulin-derived amyloidosis remains to be clarified., Case Presentation: A 58-year-old man with type 2 diabetes mellitus underwent a lower limb amputation due to diabetic gangrene. Several antibiotics including minocycline were administered for infection and sepsis. A hard mass at the insulin injection sites in the lower abdomen was discovered by chance four months later. Although no abnormal findings in the surface skin of the mass were observed, necrotic tissue was seen around the mass when a biopsy was performed. Histological and toxicity studies were performed for this patient and four other patients with abdominal masses at insulin injection sites. Histological and immunohistochemical studies showed that the masses had typical characteristics of amyloid deposits in all cases, whereas necrotic findings were seen adjacent to the amyloid deposit only in the case presented. Toxicity studies indicated that the amyloid tissue from the present case had significant cell toxicity compared to the control skin tissue or the amyloid tissues from the other four cases., Conclusions: This report showed that toxic insulin-derived amyloidosis can occur. In addition, this report suggested that toxic insulin-derived amyloidosis may cause necrosis in the surrounding tissue. Although the toxic amyloid deposit of insulin-derived amyloidosis was found in only one patient, no structural differences between toxic and non-toxic deposits were seen on histological and immunohistochemical studies.

The present study compares dosimetric parameters between volumetric-modulated arc therapy (VMAT) and 3D conformal radiation therapy (3D-CRT) in lung tumors adjacent to the chest wall treated with stereotactic body radiation therapy (SBRT). The study focused on the radiation dose to the chest wall of 16 patients who had developed radiation-induced rib fractures (RIRF) after SBRT using 3D-CRT. The targets in all patients were partially overlapping with the fractured ribs, and the median overlapping rib-PTV distance was 0.4 cm. Stereotactic body radiation therapy was re-planned for all patients. The prescribed dose was 48 Gy in four fractions to cover at least 95% of the planning target volume (PTV). Evaluated dosimetric factors included D98% and the conformation number (CN) of the PTV, the D2cm3, V40 and V30 of the fractured ribs, the V30 of the chest wall, and the Dmean, V20 and V5 of the lung. A comparison of 3D-CRT with the VMAT plan for PTV revealed that CN was significantly improved in the VMAT plan, whereas D98% did not significantly differ between the two plans. Regarding organs at risk (OARs), the D2cm3, V40 and V30 of fractured ribs, the V30 of the chest wall, and the Dmean, V20 and V5 of the lung, were significantly decreased in the VMAT plan. We concluded that the dose to OARs such as ribs and chest wall could be reduced with improved target conformity using VMAT instead of 3D-CRT for SBRT to treat peripheral lung tumors.

Purpose: To analyze the uncertainties of the rectum due to anisotropic shape variations by using a statistical point distribution model (PDM)., Materials and Methods: The PDM was applied to the rectum contours that were delineated on planning computed tomography (CT) and cone-beam CT (CBCT) at 80 fractions of 11 patients. The standard deviations (SDs) of systematic and random errors of the shape variations of the whole rectum and the region in which the rectum overlapped with the PTV (ROP regions) were derived from the PDMs at all fractions of each patient. The systematic error was derived by using the PDMs of planning and average rectum surface determined from rectum surfaces at all fractions, while the random error was derived by using a PDM-based covariance matrix at all fractions of each patient., Results: Regarding whole rectum, the population SDs were larger than 1.0 mm along all directions for random error, and along the anterior, superior, and inferior directions for systematic error. The deviation is largest along the superior and inferior directions for systematic and random errors, respectively. For ROP regions, the population SDs of systematic error were larger than 1.0 mm along the superior and inferior directions. The population SDs of random error for the ROP regions were larger than 1.0 mm except along the right and posterior directions., Conclusions: The anisotropic shape variations of the rectum, especially in the ROP regions, should be considered when determining a planning risk volume (PRV) margins for the rectum associated with the acute toxicities., (Copyright © 2018 Associazione Italiana di Fisica Medica. Published by Elsevier Ltd. All rights reserved.)

To develop a high-affinity aptamer against AML1 Runt domain, two aptamers were conjugated based on their structural information. The newly designed aptamer Apt14 was generated by the conjugation of two RNA aptamers (Apt1 and Apt4) obtained by SELEX against AML1 Runt domain, resulting in improvement in its binding performance. The residues of AML1 Runt domain in contact with Apt14 were predicted in silico and confirmed by mutation and NMR analyses. It was suggested that the conjugated internal loop renders additional contacts and is responsible for the enhancement in the binding affinity. Conjugation of two aptamers that bind to different sites of the target protein is a facile and robust strategy to develop an aptamer with higher performance., (© The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.)

Purpose: The purpose of this study was to evaluate the effect of prostate matching on dose distribution using kilovolt cone beam computed tomography (kV-CBCT) with image guided radiation therapy for prostate cancer., Materials and Method: Sixteen prostate cancer patients were treated with intensity modulated radiation therapy to 76 Gy at 2 Gy per fraction in 38 fractions. Daily target localization was performed using "bone matching" and "prostate matching" based on planning CT and kV-CBCT. Prostate dose coverage was assessed by the proportion of the CTV fully encompassed by 95%, 98% isodose lines, and mean dose lines. As for rectal and bladder, dose coverage was assessed by volumes which received 40 Gy, 60 Gy, 70 Gy, 75 Gy and mean dose at treatment. And we calculated the tumor control probability (TCP) and normal tissue complication probability (NTCP), accordingly. They were compared to the bone and prostate matching image., Result: Our study found an improvement in dose usage in CTV and bladder which enabled us to compare the bone matching image and the prostate matching image. However, it did not improve dose usage in the rectal. Then we chose patients who were a large shift from bone matching image to prostate matching image. As a result, rectal dose and NTCP were reduced., Discussion: Prostate matching is useful and safe when compared to bone matching because of improving CTV dose usage and reducing dose rectal and bladder.

The applicability of the activation of an NaI scintillator for neutron monitoring at a clinical linac was investigated experimentally. Thermal neutron fluence rates are derived by measurement of the I-128 activity generated in an NaI scintillator irradiated by neutrons; β-rays from I-128 are detected efficiently by the NaI scintillator. In order to verify the validity of this method for neutron measurement, we irradiated an NaI scintillator at a research reactor, and the neutron fluence rate was estimated. The method was then applied to neutron measurement at a 10-MV linac (Varian Clinac 21EX), and the neutron fluence rate was estimated at the isocenter and at 30 cm from the isocenter. When the scintillator was irradiated directly by high-energy X-rays, the production of I-126 was observed due to photo-nuclear reactions, in addition to the generation of I-128 and Na-24. From the results obtained by these measurements, it was found that the neutron measurement by activation of an NaI scintillator has a great advantage in estimates of a low neutron fluence rate by use of a quick measurement following a short-time irradiation. Also, the future application of this method to quasi real-time monitoring of neutrons during patient treatments at a radiotherapy facility is discussed, as well as the method of evaluation of the neutron dose.

The aim of this study was to evaluate the influence of metal markers on dose distributions and dose evaluation indices in intensity modulated radiation therapy (IMRT) plans for prostate cancer. The dose distribution calculation in the prostate IMRT was performed in a virtual phantom with and without insertion of the metal markers. The deviations of Dmax, Dmin, homogeneity index (HI), Dmean, D2, D98, and D95 of clinical target volume (CTV) and planning target volume (PTV) were obtained for estimation of the influence on the dose evaluation indices. Analytical anisotropic algorithm (AAA) and Acuros external beam (AXB) algorithms were employed for calculating the dose distributions. There were no deviations in any dose evaluation indices in dose distributions calculated by using AAA, whereas the maximum deviations for CTV and PTV by using AXB were +7.93% and +6.43% for Dmax, -16.61% and -1.77% for Dmin, +29.46% and +8.34% for HI, +0.15% and +0.02% for Dmean, +1.50% and +0.24% for D2, respectively. Additional data were -0.20% in D98 (CTV) and -0.27% in D95 (PTV). This study suggests that local dose changes, which were produced around metal markers, affected dose distributions and the dose evaluation indices.

In postoperative radiotherapy for seminoma, control of the testicular absorbed dose is important, since exposure of the testis can lead to temporary or permanent infertility. In this case, instead of using a dog-leg-shaped field, treatment using a field focused near the aorta was provided in several disease stages of seminoma. However, the precise need for testicular shielding during treatment and dose of testis exposure was not clear. We examined these questions by measuring the testicular absorbed dose with and without a testicular shield using two clinical treatment plans and a phantom. The distance from the testis phantom and the lower end of the irradiation field was varied. Where the total dose for the tumor was 20 Gy, the testicular absorbed dose was below 0.1 Gy, the threshold dose for temporary infertility. At this dosage, the distance between the testis phantom and the edge of the irradiation field was 14.6 cm without the shield and 9.99 cm with the shield. Using a testes shield, it was thus possible to reduce the dose by 58.5%.

CyberKnife(®) provides continuous guidance through radiography, allowing instantaneous X-ray images to be obtained; it is also equipped with 6D adjustment for patient setup. Its disadvantage is that registration is carried out just before irradiation, making it impossible to perform stereo-radiography during irradiation. In addition, patient movement cannot be detected during irradiation. In this study, we describe a new registration system that we term "Machine Vision," which subjects the patient to no additional radiation exposure for registration purposes, can be set up promptly, and allows real-time registration during irradiation. Our technique offers distinct advantages over CyberKnife by enabling a safer and more precise mode of treatment. "Machine Vision," which we have designed and fabricated, is an automatic registration system that employs three charge coupled device cameras oriented in different directions that allow us to obtain a characteristic depiction of the shape of both sides of the fetal fissure and external ears in a human head phantom. We examined the degree of precision of this registration system and concluded it to be suitable as an alternative method of registration without radiation exposure when displacement is less than 1.0 mm in radiotherapy. It has potential for application to CyberKnife in clinical treatment.

The purpose of this study was to verify the dose absorbed by an implantable cardioverter defibrillator (ICD) from an (192)Ir sealed source during uterine intracavitary brachytherapy, and to confirm its immunity to radiation effects. First, prior to treatment, the doses around the ICD position of an anthromorphic phantom were evaluated. Next, we also measured the dose at the ICD position using a fluorescent glass dosimeter and silicon diode dosimeter during the treatment of intracavitary brachytherapy of a patient implanted with an ICD. The results of the phantom study showed the dose percentage at the ICD location, 2 cm deep, to be 0.074% of the prescribed dose. The results of a treatment study similarly showed the dose, measured using a fluorescent glass dosimeter in the ICD position, to be 0.071% of the prescribed dose. During the application of the total prescribed dose, 30 Gy/5 fraction, the dose at the surface of the ICD position was estimated to be 21.2 mGy, well below the 1 Gy maximum recommended in the JASTRO guidelines. We regard dose verification and monitoring during treatment to be both necessary and useful in the treatment of individual cases.

AML1/RUNX1 is an essential transcription factor involved in the differentiation of hematopoietic cells. AML1 binds to the Runt-binding double-stranded DNA element (RDE) of target genes through its N-terminal Runt domain. In a previous study, we obtained RNA aptamers against the AML1 Runt domain by systematic evolution of ligands by exponential enrichment and revealed that RNA aptamers exhibit higher affinity for the Runt domain than that for RDE and possess the 5'-GCGMGNN-3' and 5'-N'N'CCAC-3' conserved motif (M: A or C; N and N' form Watson-Crick base pairs) that is important for Runt domain binding. In this study, to understand the structural basis of recognition of the Runt domain by the aptamer motif, the solution structure of a 22-mer RNA was determined using nuclear magnetic resonance. The motif contains the AH(+)-C mismatch and base triple and adopts an unusual backbone structure. Structural analysis of the aptamer motif indicated that the aptamer binds to the Runt domain by mimicking the RDE sequence and structure. Our data should enhance the understanding of the structural basis of DNA mimicry by RNA molecules.

AML1 (RUNX1) is a key transcription factor for hematopoiesis that binds to the Runt-binding double-stranded DNA element (RDE) of target genes through its N-terminal Runt domain. Aberrations in the AML1 gene are frequently found in human leukemia. To better understand AML1 and its potential utility for diagnosis and therapy, we obtained RNA aptamers that bind specifically to the AML1 Runt domain. Enzymatic probing and NMR analyses revealed that Apt1-S, which is a truncated variant of one of the aptamers, has a CACG tetraloop and two stem regions separated by an internal loop. All the isolated aptamers were found to contain the conserved sequence motif 5'-NNCCAC-3' and 5'-GCGMGN'N'-3' (M:A or C; N and N' form Watson-Crick base pairs). The motif contains one AC mismatch and one base bulged out. Mutational analysis of Apt1-S showed that three guanines of the motif are important for Runt binding as are the three guanines of RDE, which are directly recognized by three arginine residues of the Runt domain. Mutational analyses of the Runt domain revealed that the amino acid residues used for Apt1-S binding were similar to those used for RDE binding. Furthermore, the aptamer competed with RDE for binding to the Runt domain in vitro. These results demonstrated that the Runt domain of the AML1 protein binds to the motif of the aptamer that mimics DNA. Our findings should provide new insights into RNA function and utility in both basic and applied sciences.

During upper endoscopic screening, a 71-year-old asymptomatic woman was found to have a small, yellowish, superficial elevated lesion in the upper third of her stomach, without any signs of atrophic mucosa. The patient underwent endoscopic follow-up once a year for approximately five years; however, changes in the tumor were barely detectable. Endoscopic mucosal resection was performed, and a histological examination confirmed the diagnosis of gastric adenocarcinoma with chief cell differentiation (GA-CCD). GA-CCD is rare; therefore, its clinicopathological features remain unknown. This case suggests that only barely detectable endoscopic changes may be observed in GA-CCD during long-term follow-up.

The monitor unit (MU) was calculated by pencil beam convolution (inhomogeneity correction algorithm: batho power law) [PBC (BPL)] which is the dose calculation algorithm based on measurement in the past in the stereotactic lung irradiation study. The recalculation was done by analytical anisotropic algorithm (AAA), which is the dose calculation algorithm based on theory data. The MU calculated by PBC (BPL) and AAA was compared for each field. In the result of the comparison of 1031 fields in 136 cases, the MU calculated by PBC (BPL) was about 2% smaller than that calculated by AAA. This depends on whether one does the calculation concerning the extension of the second electrons. In particular, the difference in the MU is influenced by the X-ray energy. With the same X-ray energy, when the irradiation field size is small, the lung pass length is long, the lung pass length percentage is large, and the CT value of the lung is low, and the difference of MU is increased.

We previously studied dose distributions of stereotactic radiotherapy (SRT) for lung cancer. Our aim is to compare in combination pencil beam convolution with the inhomogeneity correction algorithm of Batho power low [PBC (BPL)] to the anisotropic analytical algorithm (AAA) by using the dose evaluation indexes. There were significant differences in D95, PTV mean dose, homogeneity index, and conformity index, V10, and V5. The dose distributions inside the PTV calculated by PBC (BPL) were more uniform than those of AAA. There were no significant differences in V20 and mean dose of total lung. There was no large difference for the whole lung. However, the surrounding high-dose region of PTV became smaller in AAA. The difference in dose evaluation indexes extended between PBC (BPL) and AAA that as many as low CT value of lung. When the dose calculation algorithm is changed, it is necessary to consider difference dose distributions compared with those of established practice.

Bone morphogenetic protein (BMP)-2 plays an important role in bone growth and regeneration; however, BMP-2 is easily lost by diffusion through body fluid and has some inhibitory pathways. To address this problem, we previously immobilized recombinant human BMP-2 (rhBMP-2) on succinylated type I atelocollagen. Here, we examined the effect of immobilized rhBMP-2 in vitro and vivo. In ST2, MC3T3-E1, and C2C12 cells, alkaline phosphatase activity, which is a marker of osteoblast differentiation, was enhanced more by immobilized than nonimmobilized rhBMP-2. In addition, the phosphorylation of receptor-activated Smads, part of the signaling pathway activated by BMP-2, was prolonged by immobilized rhBMP-2 in these cells. Furthermore, implantation of immobilized rhBMP-2 into the backs of rats promoted the formation of mature bone-like structure. These results demonstrate that immobilized rhBMP-2 has higher bioactivity than nonimmobilized rhBMP-2, and, therefore, immobilization of rhBMP-2 can prolong BMP signaling., ((c) 2008 Wiley Periodicals, Inc.)

The mechanism underlying airway hyperresponsiveness (AHR), a characteristic feature of asthma, remains obscure. We attempted to elucidate the mechanism responsible for the different degrees of AHR in two mouse strains, BALB/c and C57BL/6, following exposure to an anaphylactic trigger. When ovalbumin (OVA)-sensitized mice were challenged daily with OVA for up to three consecutive days, the BALB/c mice showed a higher degree of airway responsiveness to methacholine than did C57BL/6. Following the OVA challenge, eosinophils and macrophages in bronchoalveolar lavage fluid (BALF) from BALB/c increased significantly in number compared to those from C57BL/6. BALB/c mice also exhibited a higher serum IgE level than that of C57BL/6 after OVA challenge. The enhanced AHR and eosinophilic infiltration in BALF were significantly reduced by pretreatment with a selective cysteinyl-leukotriene type 1 receptor (cysLT(1)R) antagonist, montelukast. In the in vitro study, cysLT production was significantly lower in the dissected lung tissue from BALB/c than in tissue from C57BL/6 when both groups were stimulated with saline. The lungs from BALB/c generated significantly larger amounts of cysLTs on incubation with OVA rather than with saline, while the lungs from C57BL/6 did not show any significant increase in cysLTs with antigen stimulation. Significant upregulation of cysLT(1)R and cysLT(2)R mRNA expression was induced by OVA challenge in the lungs of BALB/c, but not in those of C57BL/6. It is suggested that, after an anaphylactic reaction, the degree of AHR is dependent on the genetic background and that cysLTs play an important role in the mechanism involved.

Although the yeast amber suppressor tRNA(Tyr) is a good candidate for a carrier of unnatural amino acids into proteins, slight misacylation with lysine was found to occur in an Escherichia coli protein synthesis system. Although it was possible to restrain the mislysylation by genetically engineering the anticodon stem region of the amber suppressor tRNA(Tyr), the mutant tRNA showing the lowest acceptance of lysine was found to accept a trace level of glutamine instead. Moreover, the glutamine-acceptance of various tRNA(Tyr) transcripts substituted at the anticodon stem region varied in reverse proportion to the lysine-acceptance, similar to a 'seesaw'. The introduction of a C31-G39 base pair at the site was most effective for decreasing the lysine-acceptance and increasing the glutamine-acceptance. When the same substitution was introduced into E.coli tRNA(Lys) transcripts, the lysine-accepting activity was decreased by 100-fold and faint acceptance of glutamine was observed. These results may support the idea that there are some structural element(s) in the anticodon stem of tRNA, which are not shared by aminoacyl-tRNA synthetases that have similar recognition sites in the anticodon, such as E.coli lysyl- and glutaminyl-tRNA synthetases.

Through an exhaustive search for Escherichia coli aminoacyl-tRNA synthetase(s) responsible for the misacylation of yeast suppressor tRNA(Tyr), E. coli lysyl-tRNA synthetase was found to have a weak activity to aminoacylate yeast amber suppressor tRNA(Tyr) (CUA) with L-lysine. Since our protein-synthesizing system for site-specific incorporation of unnatural amino acids into proteins is based on the use of yeast suppressor tRNA(Tyr)/tyrosyl-tRNA synthetase (TyrRS) pair as the "carrier" of unusual amino acid in E. coli translation system, this misacylation must be repressed as low as possible. We have succeeded in effectively repressing the misacylation by changing several nucleotides in this tRNA by genetic engineering. This "optimized" tRNA together with our mutant TyrRS should serve as an efficient and faithful tool for site-specific incorporation of unnatural amino acids into proteins in a protein-synthesizing system in vitro or in vivo.

Because T7 RNA polymerase has a strong preference for particular sequences to initiate transcription, some RNAs having pyrimidine-rich sequences at their 5'-end (yeast tRNA(Tyr), for example) are hardly transcribed by this enzyme. To circumvent this inconvenience, we have developed an efficient method for in vitro preparation of such tRNAs. The RNA of interest is first transcribed as a precursor form that has purine-rich extra sequences at its 5'-end, then processed with RNase P to generate the objective tRNAs. By using this protocol, we were able to prepare easily and efficiently yeast tRNA(Tyr) transcript and its mutants harboring base substitutions within the anticodon loop and/or acceptor stem regions. Aminoacylation analyses of these tRNA transcripts with yeast tyrosyl-tRNA synthetase revealed that the replacement of G34 by C34 (mutation to amber suppressor) severely impaired the aminoacylation, whereas the replacement of the U4:G69 wobble base-pair in the acceptor stem region by C4:G69 normal Watson-Crick type base-pair improved it.

Repair of bone defects remains a major concern in plastic and maxillofacial surgery. Based on modern concepts of tissue engineering, periosteum has gained attention as a suitable osteogenic material. We tested the hypothesis that surgically released and immediately repositioned periosteum would exhibit high osteogenic capacity upon grafting in a rat calvarial defect. Seven days after periosteum was released from the tibia and immediately repositioned, the "primed periosteum graft" (PPG; n = 15) was placed into a critical-sized defect of rat calvaria and the process of bone formation was evaluated histologically, immunohistologically, and radiographically at 7, 14, and 21 days after grafting. Findings were compared with a nonprimed periosteal graft (NPG; n = 15). Endochondral ossification was observed in both the PPG and NPG. The PPG showed higher expression of proliferative cell nuclear antigen, bone morphogenetic protein, and vascular endothelial growth factor than the NPG. Three-dimensional radiographic examination revealed significantly increased bone formation in the PPG than in the NPG (P < 0.01). These findings suggested that surgical stimulation of the periosteum enhanced the osteogenic potential of periosteal cells. This method may be suitable for the clinical repair of bone defects.

Vessel invasion is an important step in cartilage replacement that leads to bone formation, and vascular endothelial growth factor (VEGF) has been implicated as a key player in this process. Although grafted periosteum undergoes endochondral ossification, little is known about the role of VEGF in this process. In the current study the authors investigated by immunohistochemical, histochemical, and ultrastructural techniques the localization of VEGF during bone formation in periosteal grafts. At day 14 after grafting the tibias of Japanese white rabbits, periosteal cells in the grafted tissue had differentiated into chondrocytes to form cartilage. Some chondrocytes were immunopositive for VEGF expression, and subsequent vessel invasion occurred predominantly in these VEGF-positive areas. At day 45, the cartilage invaded by blood vessels had been replaced by newly formed bone. These findings suggest that VEGF is associated with the process of blood vessel invasion into cartilage before bone replacement in endochondral ossification from grafted periosteum.

While several reports have documented immunosuppressant-induced osteoporosis, the exact mechanism of the pathological change of the joint remains to be clarified. In the present study, we have demonstrated the pathological change of the articular cartilage in the mandibular head of five Sprague-Dawley rats administered with the immunosuppressant FK 506 for 28 days. Three-dimensional micro-computed tomography of the mandibular heads in treated rats showed a significant decrease in trabecular bone volume compared to control rats. Histological observation revealed atrophic change of the articular cartilage. Immunohistological observation using anti-proliferative cell nuclear antibody (PCNA), type I, II, and type X collagen antibodies showed significantly decreased proliferation and differentiation of chondrocytes in the articular cartilage compared with the control group (p<0.05). Tartrate-resistant acid phosphatase (TRAP) staining revealed no significant difference in the numbers of osteoclasts at the chondro-osseous junction. Thus, FK 506 administration inhibited chondrogenic cell proliferation and differentiation and might cause osteoporotic change of subcartilage trabecular bone that subsequently forms in the mandibular head.

After organ transplantation, severe osteoporosis is occasionally seen, and the use of immunosuppressants is thought to be one of the causes of such osteoporosis. In the present study, we investigated the effects of FK506 monotherapy on bones and determined the mechanism of onset of osteoporosis, both by assessing chronological changes in bone metabolism and by identifying factors that facilitate bone resorption. In 8-week-old male Sprague-Dawley rats, FK506 (1 mg/kg) was injected intraperitoneally every day for 5 weeks (FK506-treated group), and for comparison, physiological saline was administered in the same manner in a control group of rats. Serum and urine samples were collected at weeks 0, 1, 3, and 5 of administration. The femur and tibia were collected within 24 h of the final administration. When compared to the control group, findings on three-dimensional micro-computed tomography of the femur for the FK506-treated group showed a significant decrease in trabecular bone volume. The level of serum osteocalcin in the FK506-treated group at week 1 of administration was significantly higher than the control. Throughout the administration period, the sum of urinary pyridinoline (PYD) and deoxypyridinoline (Dpd) was significantly higher in the FK506-treated group. Of the various bone resorption factors tested, the level of serum parathyroid hormone (PTH) in the FK506-treated group was significantly higher than the control at week 3 of administration. The results of the present study confirmed that FK506 monotherapy in rats induced high-turnover osteoporosis. Soon after the start of FK506 administration, bone formation and resorption were elevated, and PTH appeared to have been involved in the maintenance of the elevated bone resorption., (Copyright 2004 Springer-Verlag)

Purpose: To clarify the involvement of bone morphogenetic proteins (BMPs) in the proliferation and differentiation of osteo/chondrogenic cells during the process of bone formation from grafted periosteum., Material and Methods: Tibial periosteum of young Japanese white rabbits was grafted into suprahyoid muscles and removed after 7, 9, 14 or 21 days. BMP-2, -4, proliferative cell nucleus antigen (PCNA) immunoreaction and Alcian blue staining in grafted periosteum was then sought microscopically., Results: PCNA positive cells in the grafted periosteum expressed BMP-2 at 7 days. These cells differentiated into chondroblasts that expressed BMP-2 and Alcian blue at 9 days. After 14 days, cartilage formation was seen, and BMP-2 and -4 expressions were observed in mature and hypertrophic chondrocytes. Endochondral ossification was observed at 21 days and osteoblasts showed both BMP-2 and -4 expression., Conclusion: Both BMP-2 and -4 appear to play regulatory roles in the process of endochondral ossification from grafted periosteum, due to their involvement in the proliferation and differentiation into chondrogenic and osteogenic cells.

Introduction: The pathogenetic features of rheumatoid arthritis of the temporomandibular joint (TMJ) are not well defined. In this paper the histological features of TMJs affected by rheumatoid arthritis, and the detection of secondary amyloidosis and macrophage populations in the TMJs of two patients with progressive rheumatoid arthritis are described., Methods: In two patients (64-year-old man and 61-year-old woman) with rheumatoid arthritis total TMJ replacement were performed. The surgical specimens were studied histologically., Results: It was found that the articular cartilage had been completely replaced by proliferating fibrous tissue. Congo red staining and polarizing microscopy revealed amyloid deposition in the connective tissue of the joint space. Immunohistochemical staining showed CD 68 positive macrophages around the amyloid deposition in the proliferating soft tissue., Conclusion: TMJ involvement in rheumatoid arthritis followed the same destructive pathway as in other joints. Amyloid deposition and macrophage populations were detected in two TMJs affected by rheumatoid arthritis.

Grafted periosteum has a rich potential to induce heterotopic bone formation. In the current study the authors investigate whether autogenous periosteal grafts can regenerate the mandibular head in a rabbit model. They removed the mandibular head of Japanese white rabbits and grafted tibial periosteum to the cut surface of the mandible. Grafted periosteum was observed histologically and radiographically at day 7, 14, 21, and 45 after surgery. At day 7 after grafting, grafted tissue showed remarkable cell proliferation. By 14 days these cells had differentiated into chondrocytes to form cartilage, and endochondral ossification took place after 21 days. At 45 days after surgery, soft X-ray findings showed a newly formed mandibular head, which was similar histologically to that of a normal mandibular head. The cut mandible without periosteal graft showed no regeneration. These findings indicate that grafted periosteum can regenerate the mandibular head without special procedures such as bone fixation in a rabbit model, and suggest that this technique may be useful clinically.

Cisplatin (CDDP) is a useful drug for the treatment of malignant solid tumors of the head and neck. Because CDDP includes the heavy metal platinum as a component, it is thought metallothionein (MT) may be involved in CDDP-resistance. However, functional differences between the four MT isoforms (MT-I, II, III and IV) remain unclear. The aim of this study was to investigate the relationship between MT isoform expression and CDDP-resistance. Two human tongue squamous cell carcinoma cell lines not exposed to anticancer chemotherapy were studied. The cell lines were subjected to reverse transcriptase-polymerase chain reaction (RT-PCR) analysis before and after CDDP-treatment. Both cell lines expressed MT-I/II and MT-IV isoforms but not the MT-III isoform. Following CDDP treatment, MT-I/II mRNA levels were induced only in the CDDP-resistant cell line. Our results showed that expression of the MT I/II isoform was induced by CDDP treatment, and may play an important role in CDDP-resistance in squamous cell carcinoma of the human tongue.

To determine whether grafted young periosteum can induce new bone formation in elderly patients, this preliminary study evaluated cell proliferation and differentiation in xenogeneic periosteal grafts in old rats radiographically, histologically, and immunohistochemically. Periosteum harvested from the tibia of young Japanese white rabbits were grafted into old Sprague-Dawley rats with or without administration of 1.0 mg per kilogram per day immunosuppressant FK506. Autogenous old periosteal tissue grafts were also evaluated as a control. Grafted tissue was extirpated after 7, 14, 21, and 45 days. In the xenogeneic group, proliferative cell nuclear antigen-positive cells were observed 7 days after surgery, which differentiated into chondroblasts with bone morphogenetic protein-2 expression and finally formed cartilage by 14 days. Endochondral ossification was observed at 21 days, and bone replacement was completed by 45 days. No osteogenic cell activity was observed in the two other groups. Xenogeneic young periosteum thus maintained its osteogenic/chondrogenic potentiality in older rats.