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Purpose: Stereotactic body radiotherapy (SBRT) boost is a promising treatment for cervical cancer patients who are ineligible for intracavitary brachytherapy (ICBT). The aim of this multicenter, single-arm, phase I/II study was to prospectively evaluate the efficacy and toxicity of SBRT boost., Materials and Methods: ICBT-ineligible patients with untreated cervical cancer were enrolled. Patients underwent whole-pelvic radiotherapy (45 Gy in 25 fractions) with SBRT boost to the primary lesion. In the phase I dose-escalation cohort (3 + 3 design), patients were treated with SBRT boost of 21 or 22.5 Gy in three fractions. Although dose-limiting toxicity was not confirmed, a dose of 21 Gy was selected for the phase II cohort because it was difficult to reproduce the pelvic organs position in two patients during the phase I trial. The primary endpoint was 2-year progression-free survival., Results: Twenty-one patients (phase I, n = 3; phase II, n = 18) were enrolled between April 2016 and October 2020; 17 (81%) had clinical stage III-IV (with para-aortic lymph node metastases) disease. The median (range) follow-up was 40 (10-84) months. The initial response was complete response in 20 patients and partial response in one patient. The 2-year locoregional control, progression-free survival, and overall survival rates were 84%, 67%, and 81%, respectively. Grade ≥ 3 toxicity was confirmed in one patient each in the acute (diarrhea) and late (urinary tract obstruction) phases., Conclusion: These findings suggested that a SBRT boost is more effective than the conventional EBRT boost and can be an important treatment option for ICBT-ineligible patients with cervical cancer., Study Registration: This study was registered at the University Hospital Medical Information Network Clinical Trials Registry (UMIN000036845)., (© 2024. The Author(s).)

Purpose: The optimal method for the second course of stereotactic body radiotherapy (SBRT) for spinal metastases remains poorly established. This single-center, single-arm, phase II trial was conducted to propose a safe and effective salvage spine SBRT., Methods: The patients initially treated with SBRT for spine-targeted protocol treatment, or for areas adjacent to the spine, were enrolled. The second SBRT dose was 30 Gy delivered in five fractions; the spinal cord dose constraint was 15.5 Gy at the maximum point dose. The brachial or lumbosacral plexuses were dose-constrained to <30 Gy if the boundary between the nerves and tumors was detected. The primary endpoint was dose-limiting toxicity (DLT) (grade ≥ 3 severe radiation-related toxicity) within a year after the second SBRT., Results: The second SBRT was administered to the same spinal level in 12 patients and to an adjacent spinal level in 8 patients. SBRT2 was performed for 14 painful lesions, 10 MESCC, and 6 oligometastases, with some lesions having multiple indications. The median interval between SBRT sessions was 21 months (range: 6-51 months). The median follow-up duration was 14 months. No radiation myelopathy or local failure was reported during the follow-up period. DLT was confirmed in two patients (10%) within a year, both of whom developed grade 3 lumbosacral plexopathy. These two patients received SBRT twice to the S1-2 and S1-5 vertebrae, respectively, and both experienced paralysis of the tibialis anterior muscle (L5 level). Grade 3 late adverse effects (including lumbosacral plexopathy and vertebral compression fracture) were observed in 25% of the patients throughout the entire follow-up period., Conclusions: The second spine SBRT achieved good local control without causing myelopathy. However, one-quarter of the patients experienced grade 3 late adverse effects, suggesting that the treatment protocol carries a risk of toxicity.

Topological gels possess structures that are cross-linked only via physical constraints; ideally, no attractive intermolecular interactions act between their components, which yields interesting physical properties. However, most reported previous topological gels were synthesized based on supramolecular interlocked structures such as polyrotaxane, for which attractive intermolecular interactions are essential. Here, we synthesize a water-soluble "molecular net" (MN) with a large molecular weight and three-dimensional network structure using poly(ethylene glycol). When a water-soluble monomer (N-isopropylacrylamide) is polymerized in the presence of the MNs, the extending polymer chains penetrates the MNs to form an ideal topological MN gel with no specific attractive interactions between its components. The MN gels show unique physical properties as well a significantly high degree of swelling and high extensibility due to slipping of the physical cross-linking. We postulate this method to yield a new paradigm in gel science with unprecedented physical properties., (© 2024 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Natural monoclinic zirconia (baddeleyite) was irradiated with 340 MeV Au ions, and the irradiation-induced nanostructures (i.e., ion tracks and nanohillocks) were observed using transmission electron microscopy. The diameter of the nanohillocks was approximately 10 nm, which was similar to the maximum molten region size calculated using the analytical thermal spike model. Ion tracks were imaged as strained regions that maintained their crystalline structure. The cross-sections of most of the ion tracks were imaged as rectangular contrasts as large as 10 nm. These results strongly indicated that the molten region was recrystallized anisotropically, reflecting the lattice structure. Furthermore, low-density track cores were formed in the center of the ion tracks. The formation of low-density track cores can be attributed to the ejection of molten matter toward the surface. A comparison of the ion tracks in the synthetic zirconia nanoparticles and those in larger natural zirconia samples showed that the interface between the strained track contrast and the matrix was less clear in the former than in the latter. These findings suggest that the recrystallization process was affected by the size of the irradiated samples., Competing Interests: The authors declare no conflict of interest.

Background/aim: The purpose of this study was to evaluate the impact of recent radiotherapy on mortality from heart disease in Asians or Pacific islanders with breast cancer using the Surveillance, Epidemiology, and End Results (SEER) database., Patients and Methods: Asians or Pacific islanders with stage 0 or I (AJCC 6th) breast cancer between 2000 and 2015 were analyzed. The impact of radiotherapy for mortality from heart disease after treatment was evaluated by comparing patients who received radiotherapy for left-sided breast cancer, patients who received radiotherapy for right-sided breast cancer and patients who did not receive radiotherapy., Results: In 25,684 Asians or Pacific islanders, the incidence of cardiac death was higher in patients who did not receive radiotherapy than in patients who received radiotherapy. Among patients who received external beam irradiation, the incidence of cardiac death was 2.00% for patients with left-sided breast cancer and 1.69% for patients with right-sided breast cancer, with no significant difference (chi-square test, p=0.427). In the period from 2000 to 2008, there was no significant difference between the cumulative heart-related death rates in patients who received radiotherapy and in patients who did not receive radiotherapy (Tarone-Ware test, p=0.406); however, in 2009-2015, the cumulative heart-related death rate in patients who did not receive radiotherapy was significantly higher than that in patients who received radiotherapy (log-rank test, p<0.001)., Conclusion: Heart-related death after treatment for breast cancer is relatively rare in Asians or Pacific islanders. Since at least 2000, the cardiac impact of postoperative radiotherapy has not been significant., (Copyright © 2023 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Most studies of vertebral compression fractures (VCF) caused by stereotactic body radiotherapy (SBRT) do not discuss the symptoms of this complication. In this paper, we aimed to determine the rate and prognostic factors of painful VCF caused by SBRT for spinal metastases. Spinal segments with VCF in patients treated with spine SBRT between 2013 and 2021 were retrospectively reviewed. The primary endpoint was the rate of painful VCF (grades 2-3). Patient demographic and clinical characteristics were evaluated as prognosticators. In total, 779 spinal segments in 391 patients were analyzed. The median follow-up after SBRT was 18 (range: 1-107) months. Sixty iatrogenic VCFs (7.7%) were identified. The rate of painful VCF was 2.4% (19/779). Eight (1.0%) VCFs required surgery for internal fixation or spinal canal decompression. The painful VCF rate was significantly higher in patients with no posterolateral tumor involvement than in those with bilateral or unilateral involvement (50% vs. 23%; p = 0.042); it was also higher in patients with spine without fixation than in those with fixation (44% vs. 0%; p < 0.001). Painful VCFs were confirmed in only 2.4% of all the irradiated spinal segments. The absence of posterolateral tumor involvement and no fixation was significantly associated with painful VCF.

A lateral overflow integration capacitor (LOFIC) complementary metal oxide semiconductor (CMOS) image sensor can realize high-dynamic-range (HDR) imaging with combination of a low-conversion-gain (LCG) signal for large maximum signal electrons and a high-conversion-gain (HCG) signal for electron-referred noise floor. However, LOFIC-CMOS image sensor requires a two-channel read-out chain for LCG and HCG signals whose polarities are inverted. In order to provide an area-efficient LOFIC-CMOS image sensor, a one-channel read-out chain that can process both HCG and LCG signals is presented in this paper. An up/down double-sampling circuit composed of an inverting amplifier for HCG signals and a non-inverting attenuator for LCG signals can reduce the area of the read-out chain by half compared to the conventional two-channel read-out chain. A test chip is fabricated in a 0.18 μm CMOS process with a metal-insulator-metal (MIM) capacitor, achieving a readout noise of 130 μVrms for the HCG signal and 1.19 V for the LCG input window. The performance is equivalent to 103 dB of the dynamic range with our previous LOFIC pixel in which HCG and LCG conversion gains are, respectively, 160 μV/e- and 10 μV/e-.

This prospective phase I trial aimed to determine the recommended dose of 3-day total marrow and lymphoid irradiation (TMLI) for a myeloablative conditioning regimen by increasing the dose per fraction. The primary end-point of this single-institution dose escalation study was the recommended TMLI dose based on the frequency of dose-limiting toxicity (DLT) ≤100 days posthematopoietic stem cell transplantation (HSCT); a 3 + 3 design was used to evaluate the safety of TMLI. Three dose levels of TMLI (14/16/18 Gy in six fractions over 3 days) were set. The treatment protocol began at 14 Gy. Dose-limiting toxicities were defined as grade 3 or 4 nonhematological toxicities. Nine patients, with a median age of 42 years (range, 35-48), eight with acute lymphoblastic leukemia and one with chronic myeloblastic leukemia, received TMLI followed by unrelated bone marrow transplant. The median follow-up period after HSCT was 575 days (range, 253-1037). Three patients were enrolled for each dose level. No patient showed DLT within 100 days of HSCT. The recommended dose of 3-day TMLI was 18 Gy in six fractions. All patients achieved neutrophil engraftment at a median of 19 days (range, 14-25). One-year overall and disease-free survival rates were 83.3% and 57.1%, respectively. Three patients experienced relapse, and no nonrelapse mortality was documented during the observation period. One patient died due to disease relapse 306 days post-HSCT. The recommended dose of 3-day TMLI was 18 Gy in six fractions. The efficacy evaluation of this regimen is currently being planned in a phase II study., (© 2022 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.)

Background and Objectives: Intensity-modulated radiation therapy (IMRT) helps achieve good radiation dose conformity and precise dose evaluation. We conducted a single-centre prospective study to assess the safety and feasibility of total body irradiation with IMRT (IMRT-TBI) using helical tomotherapy in allogeneic haematopoietic stem cell transplantation (allo-HSCT)., Patients and Methods: Thirty-nine adult patients with haematological malignancy (acute lymphoblastic leukaemia [ n  = 21], chronic myeloid leukaemia [ n  = 6], mixed phenotype acute leukaemia [ n  = 5], acute myeloid leukaemia [ n  = 4], and malignant lymphoma [ n  = 3]) who received 12 Gy IMRT-TBI were enrolled with a median follow-up of 934.5 (range, 617-1254) d. At the time of transplantation, 33 patients (85%) achieved complete remission. The conditioning regimen used IMRT-TBI (12 Gy in 6 fractions twice daily, for 3 d) and cyclophosphamide (60 mg/kg/d, for 2 d), seven patients were combined with cytarabine, and five with etoposide. We set dose constraints for the lungs, kidneys and lens as the organs at risk., Results: The mean doses for the lungs and kidneys were 7.50 and 9.11 Gy, respectively. The mean maximum dose for the lens (right/left) was 5.75/5.87 Gy. The 2-year overall survival (OS), disease-free survival (DFS), cumulative incidence of relapse (CIR) and non-relapse mortality (NRM) were 69, 64, 18 and 18%, respectively. Thirty-six patients developed early adverse events (AEs) (including four patients with Grade 3/4 toxicities), most of which were reversible oral mucositis and may partially have been related to IMRT-TBI. However, the incidence of toxicity was comparable to conventional TBI-based conditioning transplantation. None of the patients developed primary graft failure, or Grade III-IV acute graft-versus-host disease (GVHD). In late complications, chronic kidney disease was observed in six patients, a lower incidence compared to conventional TBI-based conditioning transplantation. No radiation pneumonitis or cataracts were observed in any of the patients., Conclusions: IMRT-TBI is safe and feasible for haematological malignancies with acceptable clinical outcomes.KEY MESSAGESIMRT-TBI-helical tomotherapy aids in accurate dose calculation and conformity.It could be used without any considerable increase in the rate of TBI-related AEs.Allo-HSCT with IMRT-TBI may be an alternative to conventional TBI for clinical use.

(1) Background: The superiority of stereotactic body radiotherapy (SBRT) over conventional external beam radiotherapy (cEBRT) in terms of pain palliation for bone metastases remains controversial. (2) Methods: This propensity score-matched study compared the overall pain response (OR) 3 months after radiotherapy among patients with painful (≥2 points on a 0-to-10 scale) non-spine bone metastases. Patients with lesions that were treated with SBRT or cEBRT and whose pain scores were evaluated 3 months after radiotherapy were included in this study. Pain response was evaluated according to the International Consensus Criteria. (3) Results: A total of 234 lesions (SBRT, n = 129; cEBRT, n = 105) were identified in our institutional database. To reduce the confounding effects, 162 patients were selected using a propensity score-matched analysis (n = 81 for each treatment). The OR rate at 3 months after SBRT was significantly higher than that after cEBRT (76.5% vs. 56.8%; p = 0.012). A noteworthy finding of our study is that the same trend was observed even after 6 months (75.9% vs. 50.0%; p = 0.011). The 1-year local failure rates after SBRT and cEBRT were 10.2% and 33.3% (p < 0.001), respectively. (4) Conclusions: Our findings suggest that SBRT is superior to cEBRT for pain palliation in patients with non-spine bone metastases., Competing Interests: Kei Ito was a member of the advisory board and has received honorariums from Varian Medical Systems K. K. The other authors declare no conflict of interest.

Objective: Stereotactic body radiotherapy is used to treat spinal metastases; however, 10% of patients experience local failure. We aimed to clarify the outcomes of the second course of stereotactic body radiotherapy for spinal metastases with a uniform fractionation schedule at our institution., Methods: Data of patients treated with a second salvage stereotactic body radiotherapy course at the same spinal level or adjacent level from July 2018 to December 2020 were retrospectively reviewed. The initial prescribed dose was 24 Gy in two fractions, and the second dose 30 or 35 Gy in five fractions. The spinal cord dose constraint at the second course was 15.5 Gy at the maximum point dose. The endpoints were local failure and adverse effects. Local failure was defined as tumor progression using imaging., Results: We assessed 19 lesions in 17 patients, with radioresistant lesions in 14 (74%) cases, the second stereotactic body radiotherapy to the same/adjacent spinal level in 13/6 cases, the median interval between stereotactic body radiotherapy of 23 (range, 6-52) months, and lesions compressing the cord in 5 (26%) cases. The median follow-up period was 19 months. The 12- and 18-month local failure rates were 0% and 8%, respectively. Radiation-induced myelopathy, radiculopathy and vertebral compression fractures were observed in 0 (0%), 4 (21%) and 2 (11%) lesions, respectively. Three patients with radiculopathy experienced almost complete upper or lower limb paralysis., Conclusions: The second course of salvage stereotactic body radiotherapy for spinal metastases achieved good local control with a reduced risk of myelopathy. However, a high occurrence rate of radiation-induced radiculopathy has been confirmed., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: This study investigated the outcomes and prognostic factors of fractionated stereotactic radiosurgery (FSRS) for treatment of brain metastases (BMs) ≥20 mm and determined whether FSRS could replace surgery, the primary treatment for large BMs., Methods: Patients with BMs ≥20 mm treated with FSRS were retrospectively examined. Patients who underwent FSRS postoperatively were excluded. Local failure, intracranial failure, and adverse events were evaluated., Results: Overall, 116 lesions in 105 patients were evaluated. The performance status was 0-1, 2-4, and unknown for 86, 28, and 2 patients, respectively. The median maximum tumor diameter was 25 mm, and the median prescribed dose was 35 Gy in 3 fractions. The median follow-up period after FSRS was 8 months. The 1-year local failure, intracranial failure, and overall survival rates were 12.5%, 56.6%, and 49.0%, respectively. A maximum dose of ≥135 Gy (biological equivalent dose [α/β = 10 Gy]) and good performance status were independent favorable prognostic factors for local control. After FSRS, 21 (20%) patients were treated with whole-brain radiotherapy because of multiple intracranial recurrences, and 4 (3.4%) patients underwent surgery because of local recurrence., Conclusions: FSRS for BMs ≥20 mm achieved good local control. Only 3.4% of patients required surgery after FSRS, suggesting that FSRS is a potential alternative to surgery. For FSRS, a higher maximum tumor dose was useful for local control., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Stereotactic body radiotherapy is a new treatment modality for long bone metastasis and has not been discussed in literature. We aimed to clarify stereotactic body radiotherapy outcomes for long bone metastases., Methods: Data of patients receiving stereotactic body radiotherapy for long bone metastases (July 2016-November 2020) were retrospectively reviewed. The prescribed dose was 30 or 35 Gy in five fractions. The endpoints were local failure and adverse effects. Local failure was defined as radiological tumor growth within the irradiation field. Adverse effects were evaluated according to the National Cancer Institute Common Terminology Criteria for Adverse Events, version 5., Results: Nineteen osseous lesions in 17 patients were assessed. The target lesions included 13 femoral, 4 humeral and 2 radial lesions. The median follow-up duration was 14 (range, 3-62) months. The 12- and 18-month local failure rates were 0 and 11%, respectively. Following 2 and 46 months of stereotactic body radiotherapy, two lesions (11%) resulted in painful femoral fractures (grade 3). Both patients underwent bipolar hip arthroplasty and could walk normally after surgery. In the late phase, one patient developed radiculopathy (almost complete paralysis of grasp) and another developed grade 2 limb edema. Other grade 2 or more severe acute and late toxicities were not observed during the follow-up period., Conclusions: Stereotactic body radiotherapy for long bone metastases achieved excellent local control and caused two femoral fractures. We argue that stereotactic body radiotherapy for curative intent should not be contraindicated in long bone oligometastasis because fractures do not directly contribute to life expectancy., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Airborne personal protective equipment is required for healthcare workers when performing aerosol-generating procedures on patients with infectious diseases. Chest compressions, one of the main components of cardiopulmonary resuscitation, require intense and dynamic movements of the upper body. We aimed to investigate the protective effect of tight-fitting powered air-purifying respirators (PAPRs) during chest compressions., Methods: This single-center simulation study was performed from February 2021 to March 2021. The simulated workplace protection factor (SWPF) is the concentration ratio of ambient particles and particles inside the PAPR mask; this value indicates the level of protection provided by a respirator when subjected to a simulated work environment. Participants performed continuous chest compressions three times for 2 min each time, with a 4-min break between each session. We measured the SWPF of the tight-fitting PAPR during chest compression in real-time mode. The primary outcome was the ratio of any failure of protection (SWPF <500) during the chest compression sessions., Results: Fifty-four participants completed the simulation. Overall, 78% (n = 42) of the participants failed (the measured SWPF value was less than 500) at least one of the three sessions of chest compressions. The median value and interquartile range of the SWPF was 4304 (685-16,191). There were no reports of slipping down of the respirator or mechanical failure during chest compressions., Conclusions: Although the median SWPF value was high during chest compressions, the tight-fitting PAPR did not provide adequate protection., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Purpose: The present multicenter, single-arm, phase 2 study aimed to prospectively evaluate the palliative efficacy of stereotactic body radiation therapy (SBRT) in patients with painful nonspine bone metastases., Methods and Materials: Patients with painful (≥2 points on a 0-to-10 scale) nonspine bone metastases from a solid tumor were enrolled. The prescribed dose was 35 Gy in 5 fractions. The primary endpoint was overall pain response rate (complete response [CR] and partial response [PR]) as measured per the International Consensus on Palliative Radiotherapy Endpoints guideline 6 months after SBRT., Results: Forty-one osseous lesions in 38 patients were registered between June 2018 and June 2019. All lesions satisfied the inclusion criteria, and the patients completed the protocol treatment. Patients most commonly had lung cancer (22%), followed by prostate cancer, uterus cancer, and renal cell carcinoma (15%, 15%, and 12%, respectively). Bone metastases were most commonly located in coxal bones (56%). The median duration of follow-up after registration was 8 months (range, 1-19 months). Among evaluable lesions at 3 and 6 months after SBRT, the 3- and 6-month pain response rates were 78% and 75%, respectively. The local control rate at 6 months was 92%. Seven patients (17%) experienced bone fracture after irradiation, and 3 patients (7%) experienced grade 2 limb edema. One patient had regrowth of coxal bone metastases, and the tumor penetrated the sigmoid colon. Soft tissue abscess around the tumor and osteonecrosis of coxal bones were confirmed. The patient died of infection 4 months after SBRT and was determined to be a possible treatment-related death., Conclusions: This prospective clinical trial showed that SBRT for nonspine bone metastases was effective in terms of pain palliation. These findings warrant a larger randomized controlled trial to compare SBRT with conventional radiation therapy. Additionally, operation history should be considered as a risk factor for edema., (Copyright © 2020 American Society for Radiation Oncology. Published by Elsevier Inc. All rights reserved.)

Objective: We aimed to clarify the outcomes of re-irradiation stereotactic body radiotherapy for spinal metastases with a uniform dose fractionation schedule at our institution., Methods: Data of patients treated with re-irradiation stereotactic body radiotherapy for spinal metastases (September 2013-March 2020) were retrospectively reviewed. The prescribed dose was 24 Gy in two fractions. The spinal cord dose constraint and dose for previously irradiated cases ≥50 Gy in 25 fractions were 12.2 Gy (maximum dose) and 11 Gy, respectively. The endpoints were pain control, local failure and adverse effects. Pain status was measured on a scale of 0-10 using the patients' self-reports and pain response was defined using international consensus criteria. Local failure was defined as tumor progression on imaging evaluations., Results: We assessed 133 lesions in 123 patients, where 70 (52.6%) had three or more spinal levels treated, 58 (43.6%) had previous irradiation doses of 40 Gy or more and 53 (39.8%) had the targets compressing the cord. The median follow-up was 12 months and the 3-, 6- and 12-month pain response rate was 75, 64 and 59%, respectively. The 1-year local failure rate was 25.8%. Previous irradiation dose was not correlated with local failure rate (P = 0.13). Radiation-induced myelopathy, radiculopathy and vertebral compression fractures were observed in 4 (3.0%), 2 (1.5%) and 17 (13.8%) lesions, respectively. A trend towards an association between any toxicity and previous irradiation dose was not observed., Conclusions: Re-irradiation spine stereotactic body radiotherapy achieved good local control and pain control, with reduced risk of radiation myelopathy., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Objective: To clarify the clinical outcomes of stereotactic body radiotherapy for colorectal cancer-derived bone metastases and identify factors predicting treatment failure., Methods: Patients treated with stereotactic body radiotherapy for bone metastases from colorectal cancer between September 2013 and June 2019 were retrospectively reviewed. The prescribed dose for spine and non-spine bone metastases was 24 Gy in two fractions and 35 Gy in five fractions, respectively. The end point was local failure, which was defined as tumour progression on imaging evaluations. In addition, various treatment- and tumour-specific factors were evaluated to determine predictors of local failure., Results: This study included 43 lesions in 38 patients, with solitary bone metastases in 18 lesions (42%), re-irradiation stereotactic body radiotherapy in 28 lesions (65%) and postoperative stereotactic body radiotherapy due to spinal cord compression in 10 lesions (23%). The median follow-up after stereotactic body radiotherapy was 12 (range, 2-60) months. The 1-year LF rate was 44%. In the univariate analysis, sacral metastases (P = 0.02) were found to be significantly correlated with LF, and multiple-course systemic therapy before stereotactic body radiotherapy (P= 0.06) and large target volume (P = 0.07) showed a trend towards an association with LF. However, these factors were not independent predictors of LF in the multivariate analysis., Conclusion: More than 40% of the lesions treated with stereotactic body radiotherapy for bone metastases from colorectal cancer showed LF within 1 year. No poor prognostic factors could be identified statistically. The poor outcomes in all groups indicate that the treatment intensity of the stereotactic body radiotherapy was insufficient to control colorectal cancer bone metastases., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

Total body irradiation using intensity-modulated radiation therapy total body irradiation (IMRT-TBI) by helical tomotherapy in allogeneic hematopoietic stem cell transplantation (allo-HSCT) allows for precise evaluation and adjustment of radiation dosage. We conducted a single-center pilot study to evaluate the safety of IMRT-TBI for allo-HSCT recipients. Patients with hematological malignancies in remission who were scheduled for allo-HSCT with TBI-based myeloablative conditioning were eligible. The primary endpoint was the incidence of adverse events (AEs). Secondary endpoints were engraftment rate, overall survival, relapse rate, non-relapse mortality, and the incidence of acute and chronic graft-versus-host disease (aGVHD and cGVHD, respectively). Between July 2018 and November 2018, ten patients were recruited with a median observation duration of 571 days after allo-HSCT (range, 496-614). D80% for planning target volume (PTV) in all patients was 12.01 Gy. Average D80% values for lungs, kidneys and lenses (right/left) were 7.50, 9.03 and 4.41/4.03 Gy, respectively. Any early AEs (within 100 days of allo-HSCT) were reported in all patients. Eight patients experienced oral mucositis and gastrointestinal symptoms. One patient experienced Bearman criteria grade 3 regimen-related toxicity (kidney and liver). All cases achieved neutrophil engraftment. There was no grade III-IV aGVHD or late AE. One patient died of sinusoidal obstruction syndrome 67 days after allo-HSCT. The remaining nine patients were alive and disease-free at final follow-up. Thus, IMRT-TBI was well tolerated in terms of early AEs in adult patients who underwent allo-HSCT; this warrants further study with longer observation times to monitor late AEs and efficacy., (© The Author(s) 2020. Published by Oxford University Press on behalf of The Japanese Radiation Research Society and Japanese Society for Radiation Oncology.)

Background: The role and impact of radiation therapy (RT) on the development of herpes zoster (HZ) has not been well studied. The objective of this study was to investigate the association between RT and HZ., Methods: A propensity score-matched, retrospective cohort study was conducted using institutional cancer registry data and medical records from 2011 to 2015. The risk of developing HZ in the RT and non-RT groups was compared using a Cox proportional hazards model. Associations also were explored between the RT field and the anatomic location of HZ in patients who developed HZ after RT. The expected number of HZ events within the radiation field was calculated according to the RT received by each patient; then, this number was compared with the observed number of in-field events., Results: Of 17,655 patients, propensity score matching yielded 4350 pairs; of these, 3891 pairs were eligible for comparison. The cumulative incidence of HZ in the RT group (vs the non-RT group) during the first 5 years after the index date was 2.1% (vs 0.7%) at 1 year, 3.0% (vs 1.0%) at 2 years, 3.4% (vs 1.3%) at 3 years, 4.1% vs 1.7% at 4 years, and 4.4% vs 1.8% at 5 years. The RT group showed a significantly higher risk of HZ than the non-RT group (hazard ratio, 2.59, 95% CI, 1.84-3.66). In the 120 patients who developed HZ after RT, HZ events were observed significantly more frequently within the RT field than expected (74 vs 43.8 events; P < .001)., Conclusions: Patients with cancer who received RT showed a significantly higher risk of HZ, which was commonly observed within the radiation field., (© 2020 American Cancer Society.)

Background: The clinical role of sputum Gram stain (SGS) in community-acquired pneumonia (CAP) diagnosis remains controversial. A 1996 meta-analysis of the diagnostic accuracy of SGS reported heterogeneous results. To update the available evidence, we performed a systematic review and a Bayesian standard and latent-class model meta-analysis., Methods: We searched Medline, Embase, and Cochrane Central by 23 August 2018 to identify studies reporting on the diagnostic accuracy, yield (percentage of patients with any pathogen[s] correctly identified by SGS), and clinical outcomes of SGS in adult patients with CAP. Two reviewers extracted the data. We quantitatively synthesized the diagnostic accuracy and yield, and descriptively analyzed other outcomes., Results: Twenty-four studies with 4533 patients were included. The methodological and reporting quality of the included studies was limited. When good-quality sputum specimens were selected, SGS had a summary sensitivity of 0.69 (95% credible interval [CrI], .56-.80) and specificity of 0.91 (CrI, .83-.96) for detecting Streptococcus pneumoniae, and a sensitivity of 0.76 (CrI, .60-.87) and specificity of 0.97 (CrI, .91-.99) for Haemophilus influenzae. Adjusted analyses accounting for imperfect reference standards provided higher-specificity estimates than the unadjusted analyses. Bacterial pathogens were identified in 73% (CrI, 26%-96%) of good-quality specimens, and 36% (CrI, 22%-53%) of all specimens regardless of quality. Evidence on other bacteria was sparse., Conclusions: SGS was highly specific to diagnose S. pneumoniae and H. influenzae infections in patients with CAP. With good-quality specimens, SGS can provide clinically actionable information for pathogen-directed antibiotic therapies., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Contact lens users very often become patients of allergic conjunctivitis, which is caused by protein and bacteria adsorption to the eye, because contact lenses easily adsorb proteins and bacteria. However, even if contact lens users develop eye diseases such as allergic conjunctivitis, most of them continue to use contact lenses to avoid interference to daily life or a decrease in their quality of life. If novel contact lenses able to prevent and additionally cure eye diseases can be manufactured, they could improve the quality of life of contact lens users worldwide. Thus, we aim to develop a novel material for contact lenses to prevent diseases by incorporating a zwitterionic polymer with the ability to suppress protein and bacteria adsorption. In addition, we also aim to effectively introduce and release a drug against allergic conjunctivitis from the contact lens material. Because the poorly water-soluble drug for allergic conjunctivitis (pranoprofen) forms a rigid crystal structure, we developed the novel "hot-melt press method" to construct a contact lens able to effectively release it. In the present study, polymer sheets containing carboxymethyl betaine (a kind of zwitterionic monomer), 2-hydroxyethyl methacrylate, and 1-vinyl-2-pyrrolidone were prepared using three different procedures. The sheets were hydrophilic and showed a strong resistance against protein and bacteria adsorption. The sheets prepared by the hot-melt press method were transparent and seemed to have potential as a material for contact lenses. In addition, the drug introduced into the sheets during preparation was observed to release at a practically appropriate dose. Therefore, it is expected that the sheets could possibly be used as a material for contact lenses which not only protect against the development of eye trouble due to protein and bacterial adsorption, but also heal allergic conjunctivitis., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Treatment of bone metastasis using stereotactic body radiotherapy (SBRT) is being widely used in clinical practice. The reported clinical advantages of SBRT include high pain and local control rates, high response rates against bone metastasis from radio-resistant tumors, and safe re-irradiations. Although most reports in the literature use local control as the primary treatment endpoint, this endpoint is not appropriate because local control does not relate directly to patient benefit. Herein, we proposed five pathophysiology-based patient groups, as well as appropriate endpoints for each group., (© 2019 Greater Poland Cancer Centre. Published by Elsevier B.V. All rights reserved.)

Objective: Some patients are ineligible for intracavitary brachytherapy (ICBT) for locally advanced cervical cancer. Stereotactic body radiotherapy (SBRT) could be a good treatment option for such patients. This phase I clinical trial aimed to determine the recommended SBRT boost dose for ICBT-ineligible cervical cancer patients., Methods: Patients with untreated uterine cervical cancer (clinical stages IB1-IIIB) who were ineligible for ICBT were enrolled. Radiotherapy consisted of whole-pelvis radiotherapy (45 Gy in 25 fractions) followed by SBRT. Three dose levels of SBRT (19.5/21/22.5 Gy in three fractions) were set; the treatment protocol began at 21 Gy (level 2). The 'rolling-six' design study was used to establish the recommended dose of SBRT. Each dose level covered three or six patients. The primary endpoint included dose-limiting toxicity (DLT), defined as the occurrence of grade 3 (or worse) non-hematologic adverse effects within 6 months after SBRT., Results: The median follow-up after registration was 17 (range, 8-32) months. Three patients were enrolled in study level 2 (SBRT of 21 Gy); none of the patients exhibited DLT within 6 months after treatment completion. In study level 3 (SBRT of 22.5 Gy), three patients did not exhibit DLT. Although all six patients achieved locoregional control during follow-up, one patient treated with level 2 SBRT experienced distant metastases 14 months after registration., Conclusions: The recommended dose of SBRT boost was 22.5 Gy in three fractions. We plan to conduct a phase II multi-center clinical trial using the methodology obtained from the current study., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objectives: The clinical role of sputum Gram stain for rapid etiologic pathogen diagnosis in patients with community-acquired pneumonia (CAP) remains an unresolved controversy. Variability in protocols and reporting of diagnostic performance in different studies has hampered assessments of clinical utility and interpretation. Since the last meta-analysis published in 1996, several reports and resources to accurately evaluate the diagnostic accuracy of sputum Gram stain have become available. Therefore, we will conduct a systematic review and meta-analysis of the clinical validity and utility of sputum Gram stain., Methods: We will search PubMed, Ovid MEDLINE, Embase, and The Cochrane Controlled Register of Trials (CENTRAL) databases from inception through July 30, 2018, with no language restriction and perform a full-text evaluation of potentially relevant articles. We will include prospective and retrospective studies that assess sputum Gram stain in adults (aged ≥18 years) with CAP. Two reviewers will independently extract data and rate each study's validity with standard quality assessment tools. We will subsequently perform standard and latent-class random-effects model meta-analyses to quantitatively synthesize the diagnostic accuracy and yield. Finally, we will assess the totality of evidence by the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach for diagnostic tests and strategies., Results: Results of the analysis will be submitted for publication in a peer-reviewed journal., Conclusions: This systematic review and meta-analysis will provide a 30-year synopsis of clinical evidence on sputum Gram stain in patients with CAP., Competing Interests: The authors report no conflicts of interest in this work.

Purpose: Stereotactic body radiotherapy (SBRT) is expected to achieve safe and effective re-irradiation for painful bone metastases. This study aimed to clarify the efficacy of re-irradiation using SBRT for painful bone metastases., Methods: Prospective database at our institution for the period between September 2013 and December 2017 were retrospectively reviewed for patients with: (1) painful bone metastases; (2) history of radiotherapy to the metastasis; and (3) SBRT performed as re-irradiation. Pain response, pain failure-free duration, analgesics medications, and adverse events were evaluated. Pain was evaluated using the Numerical Rating Pain Score, and pain response was evaluated based on International Consensus Pain Response Endpoints. Best response during follow-up was noted. Patients with complete or partial response were defined as showing pain response, and patients with pain progression were defined as showing pain failure. Adverse events were evaluated based on the RTOG/EORTC Late Radiation Morbidity Scoring Schema., Results: Sixty-six patients selected from our database showed: median age, 65 years (range, 33-82 years); ECOG performance status, 0-1/2/3/4, 51/10/3/2; lesion histopathology, rectal/lung/renal/thyroid/other cancer, 13/11/9/5/28; median previous irradiated dose, 30 Gy (range, 8-70.4 Gy); median interval from latest irradiation, 21 months (range, 4-192 months); prescribed dose for SBRT, 24 Gy in 2 fractions/30 Gy in 5 fractions/35 Gy in 5 fractions, 51/13/2. Median follow-up after SBRT was 10 months (range, 1-37 months). Fifty-seven patients achieved pain response (86%). The 1-year pain failure-free rate was 55%. Median pain failure-free duration was 13 months (range, 1-24 months). Grade 4 adverse events were observed in six patients (vertebral compression fracture, n = 5; radiation myelopathy, n = 1). No other toxicities of Grade 3 or greater were encountered., Conclusions: Re-irradiation SBRT has potential to achieve good response and long-term pain control for painful bone metastases. Prospective analysis is necessary to confirm the safety and efficacy of SBRT as re-irradiation.

Controlling the formation of Fe-N-C catalytic sites is crucial to activate the oxygen reduction reaction (ORR) for realization of non-precious electrocatalysts in proton exchange membrane fuel cells (PEMFCs). We present a quantitative study on the effect of a newly obtained thermal history on the formation of Fe-N-C catalytic sites. A short and repeated heating process is employed as the new thermal history, where short heating (1 min) followed by quenching is applied to a sample with arbitrary repetition. Through electrochemical quantitative analysis, it is found that the new process effectively increases the Fe-N-C mass-based site density (MSD) to almost twice that achieved using a conventional continuous heating process, while the turn-over frequency (TOF) is independent of the process. Elemental analysis shows that the new process effectively suppresses the thermal desorption of Fe and N atoms during the initial formation stage and consequently contributes to an increase in the Fe-N-C site density. The resultant catalytic activity (gravimetric kinetic current density (0.8 V vs. RHE)) is 1.8 times higher than that achieved with the continuous heating process. The results indicate that fine control of the thermal history can effectively increase the catalytic activity and provide guidelines for further activation of non-precious ORR electrocatalysts for PEMFCs., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

OBJECTIVE This study aimed to clarify the outcomes of postoperative re-irradiation using stereotactic body radiotherapy (SBRT) for metastatic epidural spinal cord compression (MESCC) in the authors' institution and to identify factors correlated with local control. METHODS Cases in which patients with previously irradiated MESCC underwent decompression surgery followed by spine SBRT as re-irradiation between April 2013 and May 2017 were retrospectively reviewed. The surgical procedures were mainly performed by the posterior approach and included decompression and fixation. The prescribed dose for spine SBRT was 24 Gy in 2 fractions. The primary outcome was local control, which was defined as elimination, shrinkage, or no change of the tumor on CT or MRI obtained approximately every 3 months after SBRT. In addition, various patient-, treatment-, and tumor-specific factors were evaluated to determine their predictive value for local control. RESULTS Twenty-eight cases were identified in the authors' institutional databases as meeting the inclusion criteria. The histology of the primary disease was thyroid cancer in 7 cases, lung cancer in 6, renal cancer in 3, colorectal cancer in 3, and other cancers in 9. The most common previous radiation dose was 30 Gy in 10 fractions (15 cases). The mean interval since the most recent irradiation was 16 months (range 5-132 months). The median duration of follow-up after SBRT was 13 months (range 4-38 months). The 1-year local control rate was 70%. In the analysis of factors related to local control, Bilsky grade, number of vertebral levels in the treatment target, the interval between the latest radiotherapy and SBRT, recursive partitioning analysis (RPA), the prognostic index for spinal metastases (PRISM), and the revised Tokuhashi score were not significantly correlated with local control. The favorable group classified by the Rades prognostic score achieved a significantly higher 1-year local control rate than the unfavorable group (1-year local control rate: 100% vs 33%; p < 0.01). Radiation-induced myelopathy and vertebral compression fracture were observed in 1 and 3 patients, respectively. No other grade 3 or greater toxicities were encountered. CONCLUSIONS The results indicate that spine SBRT as postoperative re-irradiation was effective, and it was especially useful for patients classified as having a good survival prognosis according to the Rades score.

Purpose: Although stereotactic body radiation therapy is one of the standard treatments for stage I nonsmall cell lung cancer, in the case of central tumors it carries the risk of severe adverse events for serial organs. Accelerated hypofractionated radiotherapy is considered a reasonable alternative to treat central tumors. We have been treating central tumors with accelerated hypofractionated radiotherapy using a 75 Gy/25 fr/5 weeks regimen, and we compared the results with those of stereotactic body radiation therapy using 48 Gy/4 fr/1 week., Methods: Patients with central tumors and/or unfit for 1-hour fixation were candidates for accelerated hypofractionated radiotherapy. Based on the proximity to the biologically effective dose at 10 Gy, above accelerated hypofractionated radiotherapy regimen was adopted., Results: From October 2003 to December 2010, 159 patients, who received either accelerated hypofractionated radiotherapy (103 cases) or stereotactic body radiation therapy (56 cases), were included in the analysis. In the accelerated hypofractionated radiotherapy group, 40 (39%) cases were central tumors, whereas all cases were peripheral tumors in the stereotactic body radiation therapy group. Overall 5-year local control and survival rates were 81.9% (95% confidence interval 73.6%-90.1%) and 46.5% (95% confidence interval 36.7%-56.2%), respectively for the accelerated hypofractionated radiotherapy group, and 75.4% (95% confidence interval 63.0%-87.8%) and 44.6% (95% confidence interval 31.6%-57.7%), respectively for the stereotactic body radiation therapy group (n.s.). Among central tumors, ultracentral tumors (21 cases) and the remaining central tumors (19 cases) were similar in both local control and survival. On multivariate analysis, hazard ratios for accelerated hypofractionated radiotherapy versus stereotactic body radiation therapy were <1 for both local control and survival. Pulmonary toxicity was similar in both groups. No serial organ toxicity was observed for central tumors., Conclusions: Accelerated hypofractionated radiotherapy with a 75 Gy/25 fr/5 weeks regimen is promising in that it can obtain similar local control and survival results to stereotactic body radiation therapy, and it can control both central and peripheral tumors without any serial organ toxicities. Based on these results, prospective multicenter trials are worth conducting, especially for ultracentral tumors.

Object: This study aimed to clarify the outcomes of stereotactic body radiotherapy for spinal metastases with a uniform dose fractionation schedule in our institution., Materials and Methods: Patients treated with spine stereotactic body radiotherapy were retrospectively reviewed. The prescribed dose was 24 Gy in 2 fractions. End points were local control, pain control, and adverse events. Local control was defined as elimination, shrinkage, or stable disease in the tumor on imaging evaluations. Pain status was measured on a scale of 0 to 10 by patients' self-reports, and pain response was defined as the time at which pain scale score decreased by 2 or more from the baseline score without increase in analgesics. In addition, various treatment- and tumor-specific factors were evaluated to determine predictive values for local and pain control., Results: This study included 134 lesions in 131 patients, with: lesion histopathology, lung/colorectal/thyroid/renal/breast/prostate/sarcoma/other cancer, 24/22/18/14/12/10/6/25; reirradiation stereotactic body radiotherapy, 82 (61.2%) cases; and postoperative stereotactic body radiotherapy for epidural spinal cord compression, 45 (33.6%) cases. Median follow-up after stereotactic body radiotherapy was 9 months. The 1-year local control rate was 72.3%. Seventy (79.5%) of the 88 cases with pain from spinal metastases achieved pain response. The 1-year pain progression-free rate was 61.7%. Regarding metastases from colorectal cancer, local and pain control rates at 1 year were significantly lower compared with other cancer types (local control rate, 34.1% vs 81.8%; P < .01; pain progression-free rate, 36.9% vs 69.9%; P = .02). On multivariate analysis, colorectal cancer metastases and radiation history were identified as independent predictors of lower local and pain control rates. Radiation-induced myelopathy, radiculopathy, and vertebral compression fractures were observed in 0, 2 (1.5%), and 16 (11.9%) cases, respectively., Conclusions: This study showed that spine stereotactic body radiotherapy achieved good local and pain control, with a clinically acceptable safety profile. However, stereotactic body radiotherapy may be less effective against spinal metastases from colorectal cancer.

Purpose: To analyze the detailed pattern of intraosseous failure after stereotactic body radiation therapy (SBRT) for coxal bone metastasis., Methods and Materials: Patients treated with SBRT to coxal bone metastasis were identified by retrospective chart review. The SBRT doses were 30 Gy or 35 Gy in 5 fractions. A margin of 5 to 10 mm was added to the gross tumor volume to create the clinical target volume. We evaluated the presence or absence of intraosseous recurrence using magnetic resonance imaging. Intraosseous recurrences were assessed as "in-field" or "marginal/out-of-field." In addition, we measured the distance between the center of the recurrent tumor and the nearest edge of the initial bone metastasis in cases of marginal/out-of-field recurrence., Results: Seventeen patients treated for 17 coxal bone metastases were included. Median age was 64 years (range, 48-79 years). Coxal lesions involved the ilium in 14 cases, pubis in 3, and ischium in 4 (3 lesions crossed over multiple regions). Patients most commonly had renal cell carcinoma (29.4%), followed by lung, hepatic cell, and colorectal cancers (23.5%, 11.8%, and 11.8%, respectively). Median follow-up after SBRT was 13 months (range, 2-44 months). Among all 17 cases, 7 cases developed 8 intraosseous recurrences, including in-field recurrence in 1 case and marginal/out-of-field recurrences in 7 cases. Median time to intraosseous recurrence was 10 months (range, 2-35 months). Among 7 cases with marginal/out-of-field recurrence, mean distance to the center of the recurrent tumor from the nearest edge of the initial bone metastasis was 34 mm (range, 15-55 mm)., Conclusions: Most recurrences were observed out-of-field in the same coxal bone. These results suggest that defining the optimal clinical target volume in SBRT for coxal bone metastasis to obtain sufficient local tumor control is difficult., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: Carcinogens are known to cause swelling of the mammalian cell nucleus. However, the mechanism of the swelling and its toxicological significance have not been fully elucidated. Since nuclear swelling (NS hereafter) has been frequently observed in chromosomal aberration (CA hereafter) tests (in vitro), the relationship between NS and CAs was investigated in this study., Results: In a short-term CA test using the fibroblast CHL cell line, the appearance of NS increased in a dose-dependent manner after exposure to six types of clastogens (mitomycin C, methyl methane sulfonate, 1-methyl-3-nitro-1-nitrosoguanidine, benzo[a]pyrene, cyclophosphamide monohydrate, and 9,10-dimethyl-2-benzanthracene), and a strong correlation was found between NS (%) and CAs (%) at each dosage. Therefore, we hypothesized that clastogens cause NS in cultured mammalian cells, since the mouse lymphoma L5178Y cell line is known to have a similar sensitivity to clastogens. Thus, we measured NS for 14 compounds (clastogens) that are known to induce structural CAs, 4 aneugens, and 12 non-mutagenes. Almost all clastogens caused NS of more than 5 %, which increased in a dose-dependent manner. Among the aneugens, colchicine, and diethylstilbestrol caused the same level of NS % as the clastogens, while carbendazim and trichlorfon caused a similar level of NS % as the clastogens only at higher levels of cytotoxicity. Almost all the non-mutagens caused less than 5 % NS., Conclusions: These results strongly suggest that NS is mainly caused by structural aberrations in the nucleus during interphase of the cell cycle.

Aim: To investigate the impact of telaprevir-based triple therapy on the serum alpha-fetoprotein (AFP) level of chronic hepatitis C patients., Methods: A total of 210 patients with chronic hepatitis C genotype 1 of high viral load (baseline serum hepatitis C virus RNA > 5.0 log10 IU/mL) were divided into two groups by type of treatment: triple therapy with telaprevir, pegylated-interferon-α (PEG-IFNα), and ribavirin (RBV) for 24 wk (n = 88), or dual therapy with PEG-IFNα and RBV for 48 wk (n = 122). The relationship between virological response and the change in the serum AFP level from baseline to 24 wk after the end of treatment was examined., Results: No significant difference in mean baseline AFP level was found between the triple and dual therapy groups (8.8 ng/mL vs 7.8 ng/mL). Triple therapy produced significant declines in the AFP level in sustained virological response (SVR) and non-SVR patients (7.8 ng/mL at baseline to 3.5 ng/mL at 24 wk after the end of treatment, P < 0.001 and 14.3 ng/mL to 9.5 ng/mL, P = 0.004, respectively). In contrast, dual therapy resulted in a significant decline in AFP level only in SVR patients (4.7 ng/mL to 2.8 ng/mL, P < 0.001), but not in non-SVR patients (10.2 ng/mL to 10.1 ng/mL). Among patients with a high-baseline AFP level (≥ 10 ng/mL), the decline in the AFP level was significantly higher in the triple therapy than in the dual therapy group (15.9 ng/mL vs 1.6 ng/mL, P = 0.037)., Conclusion: Regardless of virological response, telaprevir-based triple therapy reduced the serum AFP level.

In the methane production from waste activated sludge (WAS), complex bacterial interactions in WAS have been known as a major contribution to methane production. Therefore, the influence of bacterial community changes toward methane production from WAS was investigated by an application of antibiotics as a simple means for it. In this study, azithromycin (Azm) as an antibiotic was mainly used to observe the effect on microbial changes that influence methane production from WAS. The results showed that at the end of fermentation, Azm enhanced methane production about twofold compared to control. Azm fostered the growth of acid-producing bacterial communities, which synthesized more precursors for methane formation. DGGE result showed that the hydrolysis as well as acetogenesis stage was improved by the dominant of B1, B2 and B3 strains, which are Clostridium species. In the presence of Azm, the total population of archaeal group was increased, resulting in higher methane productivity achievement.

We developed a detection method that uses quantitative real-time PCR (qPCR) and the TaqMan system to easily and rapidly assess the population of aniline-degrading bacteria in activated sludge prior to conducting a biodegradability test on a chemical compound. A primer and probe set for qPCR was designed by a multiple alignment of conserved amino acid sequences encoding the large (α) subunit of aniline dioxygenase. PCR amplification tests showed that the designed primer and probe set targeted aniline-degrading strains such as Acidovorax sp., Gordonia sp., Rhodococcus sp., and Pseudomonas putida, thereby suggesting that the developed method can detect a wide variety of aniline-degrading bacteria. There was a strong correlation between the relative copy number of the α-aniline dioxygenase gene in activated sludge obtained with the developed qPCR method and the number of aniline-degrading bacteria measured by the Most Probable Number method, which is the conventional method, and a good correlation with the lag time of the BOD curve for aniline degradation produced by the biodegradability test in activated sludge samples collected from eight different wastewater treatment plants in Japan. The developed method will be valuable for the rapid and accurate evaluation of the activity of inocula prior to conducting a ready biodegradability test., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Synthesis-dependent strand-annealing (SDSA)-mediated homologous recombination replaces the sequence around a DNA double-strand break (DSB) with a copy of a homologous DNA template, while maintaining the original configuration of the flanking regions. In somatic cells at the 4n stage, Holliday-junction-mediated homologous recombination and nonhomologous end joining (NHEJ) cause crossovers (CO) between homologous chromosomes and deletions, respectively, resulting in loss of heterozygosity (LOH) upon cell division. However, the SDSA pathway prevents DSB-induced LOH. We developed a novel yeast DSB-repair assay with two discontinuous templates, set on different chromosomes, to determine the genetic requirements for somatic SDSA and precise end joining. At first we used our in vivo assay to verify that the Srs2 helicase promotes SDSA and prevents imprecise end joining. Genetic analyses indicated that a new DNA/RNA helicase gene, IRC20, is in the SDSA pathway involving SRS2. An irc20 knockout inhibited both SDSA and CO and suppressed the srs2 knockout-induced crossover enhancement, the mre11 knockout-induced inhibition of SDSA, CO, and NHEJ, and the mre11-induced hypersensitivities to DNA scissions. We propose that Irc20 and Mre11 functionally interact in the early steps of DSB repair and that Srs2 acts on the D-loops to lead to SDSA and to prevent crossoverv.

A novel protease secreted by Brevibacillus sp. KH3 isolated from excess sludge at 50 °C and used as a sludge-lysing strain was investigated in this study. Sludge reduction was minimized by protease inhibitors and a 40-kDa protease, which significantly contributed to this sludge-reducing activity, was purified as the target protein. The final purified protease demonstrated 92-fold higher specific activity than the initial crude extracts. The sludge-reducing efficiency deteriorated relative to decreased protease activity triggered by EDTA; thus, the purified protease was a causative agent in reducing excess sludge. The 40-kDa protease was a serine metalloprotease and showed the highest activity at 50 °C and pH 8.0, and the activity was enhanced in the presence of calcium ions, indicating that the purified protease contained calcium ion. Furthermore, this 40-kDa protease inhibited biofilm formation in excess sludge. These results imply that sludge reduction is because of reduction of biofilm formation in excess sludge., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Escherichia coli DNA-unwinding protein RecQ has roles in the regulation of general recombination and the processing of stalled replication forks. In this study, we found that knockout of the recQ gene in combination with xonA xseA recJ mutations, which inhibit methyl-directed mismatch repair (MMR), caused about 100-fold increase in sensitivity to a purine analog 2-aminopurine (2AP). Intriguingly, inactivation of a MMR initiator due to the either mutation mutS or uvrD completely suppressed the 2AP sensitivity caused by recQ xonA xseA recJ mutations, suggesting that RecQ helicase might act on the DNA structures that are generated by the processing of DNA by the MutSLH complex and UvrD helicase. Moreover, the recQ gene knockout in combination with xonA xseA recJ mutations enhanced 2AP-induced filament formation, and increased by twofold the rate of spontaneous forward mutations in the thyA locus but did not increase the rate of rifampicin-resistant mutations. We discuss about the possible interplay between E. coli RecQ helicase and mismatch recognition factors., (Copyright 2010. Published by Elsevier Inc.)

The development of a facile technology for utilizing effectively and/or reducing excess sludge is one of the urgent problems since a large quantity of sewage sludge is formed by activated sludge processes. Excess sludge containing 50mM sucrose was fermented at 50 degrees C using endogenous bacteria in excess sludge, resulting in a high lactic acid production (8.45 g/L) and in an increased sludge reduction (38.2%). Conversion rate to lactic acid was up to 106.0% by standard fermentation at 50 degrees C compared to 43.8% at 30 degrees C and this phenomenon that conversion rate was higher was observed only at 50 degrees C as the fermentation at less or more than 50 degrees C had lower conversion rate than that at 50 degrees C. Lactic acid bacteria increased at 50 degrees C during 1-d fermentation whereas the number of total viable bacteria only increased slightly, indicating that lactic acid bacteria in sludge at 50 degrees C were preferentially able to utilize the sucrose for producing lactic acid. Finally, pH-vibration fermentation at 50 degrees C enabled to completely consume residual sucrose in the normal fermentation, resulting in the maximum production of lactic acid. Lactate fermentation by a purely cultured lactic acid bacterium TS1 with autoclaved excess sludge containing 50mM sucrose had more than 100% of conversion rate to lactic acid, indicating that a part of sludge was converted into lactic acid during the fermentation. Our technique is useful as a facile engineering for reducing excess sludge concomitantly with producing lactic acid by lactate fermentation.

A strain of sludge-lysing bacteria was isolated from waste activated sludge (WAS) in this study. The result of 16S rRNA gene analysis demonstrated that it was a species of new genus Brevibacillus (named Brevibacillus sp. KH3). The strain could release the protease with molecule weight of about 40 kDa which could enhance the efficiency of sludge thermophilic aerobic digestion. During the sterilized sludge digestion experiment inoculated with Brevibacillus sp. KH3, the maximum protease activity was 0.41 U/ml at pH 8 and 50 degrees C, and maximum TSS removal ratio achieved 32.8% after 120 h digestion at pH 8 and 50 degrees C. In the case of un-sterilized sludge digestion inoculated with Brevibacillus sp. KH3, TSS removal ratio in inoculated-group was 54.8%, increasing at 11.86% compared with un-inoculation (46.2%). The result demonstrated that inoculation of Brevibacillus sp. KH3 could help to degrade the EPS and promote the collapse of cells and inhibit the growth of certain kinds of microorganisms. It indicated that Brevibacillus sp. KH3 strain had a high potential to enhance WAS-degradation efficiency in thermophilic aerobic digestion.

A water-soluble derivative of N-confused porphyrin (NCP: 5,10,15,20-tetraaryl-2-aza-21-carbaporphyrin) was synthesized by introducing glucamine groups at the para-position of meso-aryl groups. The tetraglucamine-appended NCP (TG-NCP) exists as monocation in aqueous solution containing 6 mM sodium dodecyl sulfate (SDS) but exists as freebase to form aggregates in pure water. These properties are distinct from those of corresponding regular porphyrin, which exists as freebase in the micellar solution and practically insoluble in water.

Anaerobic bacteria have been used to produce 2,4-dihydroxylamino-nitrotoluene (2,4DHANT), a reductive metabolite of 2,4,6-trinitrotoluene (TNT). Here, an aerobic TNT biodegrader Pseudomonas sp. strain TM15 produced 2,4DHANT as evidenced by the molecular ion with m/z of 199 identified from LC-TOFMS analyses. TNT biodegradation with a high cell concentration (10(9) cells/ml) led to a significant accumulation of 2,4DHANT in the culture medium, as well as hydroxylamino-dinitrotoluenes (HADNTs), although these products were not accumulated when a low cell concentration was used; also, the accumulation of diamino-nitrotoluene and of an unidentified metabolite were observed in the culture medium with the high cell concentration (10(10) cells/ml). 2,4DHANT overproduction was a function of the aeration speed since cultures with low aeration speeds (30 rpm) had a 19-fold higher DHANT productivity than those aerated with high speeds (180 rpm); this indicates that molecular oxygen was related to the formation of 2,4DHANT. The quantification of dissolved oxygen (DO) in the media demonstrated that the productivity of 2,4DHANT was increased at low DO values. Moreover, supplying oxygen to the culture media produced a remarkable decrease of 2,4DHANT accumulation; these results clearly indicate that high 2,4DHANT production was a consequence of the oxygen deficit in the culture medium. This finding is useful for understanding the TNT biodegradation (bioremediation technology) in an anoxic environment.

Two kinds of hydroxylamino-dinitrotoluenes, 2-hydroxylamino-4, 6-dinitrotoluene (2HADNT) and 4-hydroxylamino-2, 6-dinitrotoluene (4HADNT), are known to be major metabolites produced from 2,4,6-trinitrotoluene (TNT) by bacteria. The amounts of 2HADNT and 4HADNT in distilled water are found to spontaneously decrease with time, albeit abiotic conditions, and many white precipitates are educed in the processes of this phenomenon; however, how these compounds are converted is unclear. We evinced the mystery of this phenomenon by using thin-layer chromatography (TLC) and laser time-of-flight mass spectrometry (TOFMS). TLC analyses in the spontaneous conversion processes of 2HADNT, 4HADNT, and 2HADNT plus 4HADNT demonstrats that three novel spots emerge on the TLC plate, respectively. These products are individually extracted into acetonitrile by collecting each spot. The purity of these extracts, which have retention times of 14.0, 17.7, and 15.4 min, is approximately 98%, judging from the results of high-performance liquid chromatographic analyses. The spontaneous conversion products of 2HADNT, 4HADNT, and 2HADNT plus 4HADNT are identified as 4,4',6,6'-tetranitro-2,2'-azoxytoluene (2,2'AZT), 2,2',6,6'-tetranitro-4,4'-azoxytoluene (4,4'AZT), and 4,2',6, 6'-tetranitro-2,4'-azoxytoluene (2,4'AZT) by obtaining their mass spectra with laser TOFMS. It is confirmed that most of the spontaneous conversion products are 2,2'AZT, 4,4'AZT, or 2,4'AZT, judging from the results of mass balance in the spontaneous conversion processes of 2HADNT, 4HADNT, and 2HADNT plus 4HADNT.

Although many studies have reported that nitroaromatics such as 2,4,6-trinitrotoluene (TNT) have strong mutagenicity, the mechanism of mutagenic activity in these compounds has not yet been reported. We examined the mutagenicity versus reactivity and biodegradation by bacteria using TNT and its analogs (1,3,5-trinitrobenzene [TNB], 2,4,6-trinitroaniline [TNA], 2,4,6-trinitro-phenol [TNP], N,2,4,6-tetranitro-N-methyl-aniline [tetryl], 2,4-dinitrotoluene [24DNT], and 2,6-dinitrotoluene [26DNT]). Aromatic compounds harboring three nitro groups (except TNP) have high mutagenicity, judging from the results of the umu test using luminescent bacteria (Salmonella typhimurium strain TA1535/pTL210). Single-electron reduction potentials for these chemicals were -530, -555, -565, -575, -640, -674, and -764 mV for TNA, tetyl, TNT, TNB, 24DNT, 26DNT, and TNP, respectively, indicating that trinitro-aromatics (except TNP) were more reducible than other compounds. Pseudomonas sp. strain TM15, which was isolated from TNT-contaminated soils in the Yamada Green Zone, Kitakyushu City, Japan, could efficiently biotransform TNT, TNB, TNA, and tetryl; 24DNT, 26DNT, and TNP were less biodegradable. This strain converted all TNT analogs into reduction products; nitro groups were reduced to amino groups. We revealed that the mutagenicity of nitroaromatics correlate with reactivity and biodegradability. This finding may contribute to the elucidation of mutagenic expression of nitroaromatic compounds in organisms.

A rapid spectrophotometric determination for 2,4,6-trinitrotoluene (TNT) is significant because this method is suitable for simultaneous analyses of the numerous samples. We found one problem that TNT reduction products interfere with the TNT detection in this assay, and we overcame this problem by heating the samples at 95 degrees C, resulting in the production of compounds that did not interfere.

Two kinds of hydroxylamino-dinitrotoluenes (HADNTs), 2-hydroxylamino-4,6-dinitrotoluene (2HADNT) and 4-hydroxylamino-2,6-dinitrotoluene (4HADNT), are known to be major metabolites produced from 2,4,6-trinitrotoluene (TNT) by bacteria. These chemicals could not be identified as TNT metabolites produced by Pseudomonas sp. strain TM15 because the mass spectra of these chemicals could not be obtained by liquid chromatography-mass spectrometry (MS) or gas chromatography-MS, which are the classic methods for identifying the metabolites of xenobiotics. However, these problems are overcome by isolating 2HADNT and 4HADNT from TNT metabolites with one-step thin-layer chromatography using dichloromethane as the developing solvent, and individually extracting them into acetonitrile by collecting spots of 2HADNT and 4HADNT. The purity of each HADNT was approximately 98%, based on the results of high-performance liquid chromatographic analyses. 2HADNT and 4HADNT are identified by obtaining their mass spectra with laser time-of-flight MS. 2HADNT and 4HADNT dissolve in distilled water and are spontaneously broken down with time. Also, heat treatment (increasing temperatures) and dissolved oxygen accelerate the destruction of HADNTs. This technique may be applicable for the identification and exact quantitative analysis of unstable and fragile compounds such as HADNTs.

The product of the BLM gene, which is mutated in Bloom syndrome in humans, and the Saccharomyces cerevisiae protein Sgs1 are both homologous to the Escherichia coli DNA helicase RecQ, and have been shown to be involved in the regulation of homologous recombination. Mutations in these genes result in genome instability because they increase the incidence of deletions and translocations. We present evidence for a genetic interaction between SGS1 and YKU70, which encodes the S. cerevisiae homologue of the human DNA helicase Ku70. In a yku70 mutant background, sgs1 mutations increased sensitivity to DNA breakage induced either by treatment with camptothecin or by the expression of the restriction enzyme EcoRI. The yku70 mutation caused a fourfold increase in the rate of double-strand break (DSB)-induced target integration as that seen in the sgs1 mutant. The combination of yku70 and sgs1 mutations additively increased the rate of the targeted integration, and this effect was completely suppressed by deletion of RAD51. Interestingly, an extra copy of YKU70 partially suppressed the increase in targeted integration seen in the sgs1 single mutant. These results suggest that Yku70 modulates the repair of DSBs associated with homologous recombination in a different way from Sgs1, and that the inactivation of RecQ and Ku70 homologues may enhance the frequency of gene targeting in higher eukaryotes.