Nagasaki University

This study aimed to compare the dosimetric parameters of the simultaneous integrated boost (SIB) technique using volumetric modulated arc therapy (VMAT) guided by 99mTc-labeled diethylenetriamine pentaacetate-galactosyl human serum albumin (99mTc-GSA) single-photon emission computed tomography (SPECT) image with those of conventional three-dimensional conformal radiotherapy (3D-CRT). Five patients with diffuse liver metastases underwent 99mTc-GSA SPECT and computed tomography (CT) imaging before RT planning. A SPECT threshold was used for determining the functional liver structure (FLS). The 3D-CRT plan included anterior-posterior opposed beams. The prescribed dose was 8 Gy in one fraction, with a goal of 95% of the planning target volume (PTV), D95, receiving at least 95% of the prescribed dose. The absorbed dose received by 2 cm3 of organ at risk (OAR) volume (D2cc) was below 10 Gy. The VMAT plan comprised two or three arcs. The goals for the PTV and OARs matched those of 3D-CRT. The PTV dose in SIB (PTVboost) defined according to FLS was optimized at an isodose line [~80% of the maximum dose (Dmax)] received at D95 of PTVboost. Two plans were compared using the dose-volume histogram parameters of PTV, PTVboost, FLS, and OARs. Dosimetric parameters including Dmean, Dmax, V110%, and V120% for PTV were significantly higher in the VMAT plan than in the 3D-CRT plan. Likewise, for PTVboost, the VMAT plan had significantly higher values of dosimetric parameters (D95, Dmean, Dmax, V110%, and V120%) than the 3D-CRT plan. The FLS and OAR parameters did not significantly differ between the two plans, except for the spinal cord. 99mTc-GSA SPECT image-guided VMAT planning using SIB allows selective target dose increase in palliative radiotherapy for diffuse liver metastases.

Current diagnostics for tuberculosis (TB) rely on sputum or site-specific specimen collection, posing critical limitations in certain populations. Urine has emerged as an alternative specimen, as demonstrated by urine lipoarabinomannan (LAM) testing. Beyond LAM, no urine-based tests are established for clinical use, and evidence for non-LAM urine antigen tests and urine-based nucleic acid amplification tests (NAATs) remains inconsistently synthesized. This scoping review aimed to comprehensively map current evidence and identify gaps in urine-based diagnostic tests for TB. We conducted a scoping review across three databases (PubMed, Scopus, and Embase) of studies evaluating urine-based diagnostic tests for TB and reporting diagnostic performance metrics, extracting data on urine test type, country, TB disease type, study population, and HIV status. Findings were summarized descriptively and visualized to illustrate trends. We also extracted technical approaches for urine-based NAATs and urine concentration methods. In total, 208 publications reporting 274 urine-based tests published between 1995 and 2024 were included. Most studies were conducted in high-burden settings and focused on pulmonary TB (44.7%), adult populations (62.5%), and people living with HIV (31.3%). LAM-based tests dominated the literature, accounting for 49.3% (n = 135) of all tests, primarily in lateral flow formats. Non-LAM urine antigen tests were evaluated less frequently (n = 26), typically in small cohorts and laboratory-based assays. Urine-based NAATs showed wide variability in technical approaches. Only a few studies (n = 10) evaluated urine concentration methods. Evidence for urine-based diagnostic tests for TB remains limited for extrapulmonary TB, children, and non-HIV individuals, despite substantial unmet need. Although urine LAM is the most established assay, evaluations of non-LAM urine antigen tests and urine-based NAATs remain exploratory, with few studies, small cohorts, and predominantly laboratory-based assays. Further research is needed to identify and validate reliable, broadly applicable urine-based diagnostic tests to address these gaps.

Tunnel linings are critical structural components of underground infrastructure, and their seismic performance plays a decisive role in maintaining the serviceability and safety of tunnels. Under dynamic loading, excessive deformation and damage of the lining may reduce the effective cross-sectional capacity and threaten the minimum safety clearance required for tunnel operation. Therefore, it is essential to investigate the deformation behavior and failure mechanisms of tunnel linings subjected to seismic excitation and to evaluate the effectiveness of reinforcement measures. In this study, a coupled numerical framework combining the finite difference method (FLAC3D) and the discrete element method (PFC3D) is developed to analyze the dynamic response of tunnel lining systems. The surrounding rock mass is modeled in FLAC3D to simulate stress wave propagation and global deformation, while the tunnel lining is represented in PFC3D using bonded particles to capture crack initiation, propagation, and post-peak failure behavior. The proposed FLAC3D–PFC3D coupled approach provides an effective tool for evaluating the seismic performance of reinforced tunnel linings and offers a practical basis for the design and assessment of seismic strengthening measures in underground engineering.

This study aimed to determine the effects of hydroxychloroquine [HCQ]-induced autophagy suppression prior to irradiation on radiosensitivity of the thyroid gland and compared it with that observed in the liver. Briefly, 6-week-old Wistar rats were divided into non-treated (saline) and treated groups (HCQ 200 mg/kg administered thrice orally). The treated group was further divided into irradiated (4 Gy of whole-body X-ray) and non-irradiated (NR) groups. The liver and thyroid tissues were resected from the NR and irradiated (at 3, 6 and 24 h) group rats. Changes in apoptosis (number of TUNEL-positive cells), proliferation (number or rate of Ki-67-positive cells), DNA damage response (number of 53BP1 nuclear foci and phospho-p53Ser15 expression), and autophagy-related molecules (LC3 and p62 expression and autophagy-related gene expression) in the liver and thyroid were examined. More TUNEL-positive cells were present in the liver of HCQ-treated group than in that of non-treated group at each time point, but no changes in thyroid were observed between groups at any time point. Fewer Ki-67-positive cells were observed in the liver and thyroid of HCQ-treated group than in non-treated group after irradiation. The number of 53BP1 nuclear foci in the thyroid increased slightly in the HCQ-treated group at 3 h compared with that in non-treated and NR group. Increased p62 expression was observed at 3 and 6 h in the liver, but not in the thyroid, in HCQ-treated group. The punctate staining for LC3 and p62 in the thyroid cytoplasm of HCQ-treated rats was observed in NR and irradiation groups. A lower expression of five autophagy machinery component genes and 12 autophagy regulatory genes was observed in the HCQ-treated irradiated rats than in non-treated irradiated rats at 24 h. HCQ administration before irradiation reduced cell proliferation in the thyroid gland with a slight increase in DNA damage response and decrease in autophagy-related gene expression. However, no change was observed in apoptosis, unlike in the liver. In the liver, radiation exposure after HCQ administration induced apoptosis, eliminating the damaged cells. In contrast, in the thyroid, damaged cells might continue to survive as they are not eliminated by apoptosis.

Mechanical compression of the periodontal ligament (PDL) is a key trigger of orthodontically induced root resorption (OIRR). Although PDL compression during the early phase of orthodontic force application correlates strongly with OIRR severity, associated cellular events remain insufficiently defined. This study aimed at elucidating mechanisms underlying the initiation of OIRR, focusing on PDL compression and associated cellular responses, and identifying cellular targets through which lithium exerts inhibitory effects. Thirty-seven 10-week-old male Wistar rats were assigned to lithium-treated and saline groups. Daily intraperitoneal injections were administered, and a 25 cN mesial orthodontic force was applied to the maxillary first molar for 14 days. PDL thickness was evaluated using microcomputed tomography. Histological analyses quantified the OIRR area, apoptotic cell count, and odontoclast number. In both groups, early PDL compression ratios showed strong positive correlations with OIRR area and early apoptotic cell counts. Lithium significantly reduced OIRR but did not alter PDL compression compared with saline. At comparable PDL compression ratios, the lithium group exhibited consistently smaller OIRR areas and fewer early apoptotic cells. The early odontoclast numbers remained low and showed no intergroup differences. These findings suggest that the protective effect of lithium was primarily mediated by the suppression of early apoptosis, cellular response to PDL compression. Early PDL compression and apoptosis play pivotal roles in OIRR initiation and are potential predictive markers. Lithium attenuated OIRR by suppressing early apoptosis without altering PDL compression, highlighting early apoptosis as a promising therapeutic target for OIRR prevention.

The GNAS locus produces multiple transcripts, including the maternally derived, GNAS-H, and GNAS-NESP55 and the paternally derived GNAS-XL, GNAS-A/B, and GNAS-AS, all of which are expressed in specific tissues. GNAS-Gsα is biparentally expressed in most tissues and imprinted in several tissues, specifically, the proximal tubules, thyroid, gonads, and pituitary. These imprinted transcripts are regulated by five differentially methylated regions (GNAS-DMRs), and hypomethylation in the GNAS-A/B:TSS-DMR causes resistance to hormones, including parathyroid hormone, leading to pseudohypoparathyroidism type 1B (PHP1B). Sporadic PHP1B shows broad methylation defects in the GNAS-DMRs, whereas most familial PHP1B cases show localized methylation defects at the GNAS-A/B:TSS-DMR and GNAS-AS2:TSS-DMR. We identified an inherited maternal exon H variant that causes a GNAS methylation pattern typically observed in sporadic PHP1B. To determine the underlying genetic cause in this family, we conducted long-read sequencing (LRS). LRS revealed a maternal exon H variant together with methylation defects of CpGs in the GNAS-DMRs. To evaluate the association of the variant with this familial PHP1B, we established patient-derived induced pluripotent stem cells (iPSCs). Reverse-transcription PCR (RT-PCR) and quantitative RT-PCR in patient-derived iPSCs showed no expression of GNAS-NESP55 or GNAS-H and increased expression of GNAS-AS, together with hypermethylation of the GNAS-NESP:TSS-DMR and hypomethylation of the GNAS-AS1:TSS-DMR and GNAS-XL:Ex1-DMR. RNA sequencing revealed no abnormal GNAS transcripts in iPSCs established from patients. To determine whether loss of GNAS-H expression itself causes PHP1B, or whether methylation defects of the GNAS-DMRs, followed by loss of GNAS-H expression, result in PHP1B in this family, we conducted RT-PCR after demethylation treatment in patient-derived iPSCs. We detected biallelic expressed GNAS-NESP55 transcript and only paternally expressed GNAS-H transcript. These findings indicated impaired maternal GNAS-H transcription in the patients, regardless of the methylation levels in the GNAS-DMRs. Furthermore, amplicon LRS spanning the region from exon H to GNAS-AS exon 1 in 40 sporadic PHP1B patients showed that genomic variants in this region are infrequent. Loss of maternal GNAS-H transcript precedes the onset of abnormal methylation imprinting at the GNAS-DMRs and contributes to its establishment. We also revealed that LRS is useful for diagnosing and researching imprinting disorders.