Strategies for Preclinical Studies Evaluating the Biological Effects of an Accelerator-based BNCT System

This review discusses the strategies of preclinical studies intended for accelerator-based (AB)-boron neutron capture therapy (BNCT) clinical trials, which were presented at the National Cancer Institute (NCI) Workshop on Neutron Capture Therapy held from April 20 to 22, 2022. Clinical studies of BNCT have been conducted worldwide using reactor neutron sources, with most targeting malignant brain tumors, melanoma, or head and neck cancer. Recently, small accelerator-based neutron sources that can be installed in hospitals have been developed. AB-BNCT clinical trials for recurrent malignant glioma, head and neck cancers, high-grade meningioma, melanoma, and angiosarcoma have all been conducted in Japan. The necessary methods, equipment, and facilities for preclinical studies to evaluate the biological effects of AB-BNCT systems in terms of safety and efficacy are described, with reference to two examples from Japan. The first is the National Cancer Center, which is equipped with a vertical downward neutron beam, and the other is the University of Tsukuba, which has a horizontal neutron beam. The preclinical studies discussed include cell-based assays to evaluate cytotoxicity and genotoxicity, in vivo cytotoxicity and efficacy of BNCT, and radioactivation measurements

On the Food Habits of the Dragonets, Genus Callionymus : I. Food Habits of Callionymus punctatus LANGSDORFF

ネズミゴチの食性を明らかにするために,瀬戸内海中部の小型底曳網漁獲物の中から採集したネズミゴチ46個体の消化管内容物を調査し,さらに瀬戸内海中部の仙酔島の砂場および藻場のベントスとそこで採集した23個体のネズミゴチの消化管内容物をそれぞれ調査して次のような結果を得た。 1) ネズミゴチの餌生物は甲殻類のCumacea, Ostracoda, カニ類, Amphipoda, Copepoda,エビ類, Anomura, オキアミ類, Branchiopoda, Mycidaceaと巻貝,二枚貝および多毛類であるが,調査したネズミゴチの半数以上の個体に捕食されていたCumacea, Ostracodaおよび巻貝が重要な餌生物である。 2) ネズミゴチの成魚の餌生物組成は雌雄の間で差異がなく,また体長110～129mmと130～164mmとの間についても差異がない。 3) 仙酔島の砂場のベントスの組成と藻場のベントスの組成とは著しく相違しているのに対し,砂場と藻場の両者において,それぞれのベントスとそこで採集したネズミゴチの消化管内容物の組成は極めてよく類似していた。すなわち個体数において,砂場ではウミホタルが卓越してベントスの53%を占め,ネズミゴチの消化管内容物の中では79%を占めた。次いで砂場では貝類が多数出現して,それはベントスの28%を占めたのに対し,消化管内容物では8%を占めた。藻場ではホトトギスガイが卓越してベントスの87%を占め,消化管内容物の中では59%を占めた。 4) 藻場に生息していたネズミゴチの消化管内容物の59%を占め,ネズミゴチの最も重要な餌生物の一つであるホトトギスガイは仙酔島の藻場に比較的多く出現したアサヒアナハゼ,クジメおよびアイナメには殆んど捕食されていなかった。しかし藻場のネズミゴチに殆んど捕食されていないGammarideaはこれら3種の魚類によってしばしば捕食されており,それらの最も重要な餌生物であることがわかった。したがってネズミゴチは藻場ではアサヒアナハゼ,クジメおよびアイナメと食物について競合関係がない。The dragonets, genus Callionymus, is very common and found frequently with other demersal fish in the catch of a small trawl in the Seto Inland Sea. In this study, the food habits of "Nezumigochi", Callionymus punctatus LANGSDORFF, have been investigated through analysis of their gut contents and benthos of their habitat, Sensui-shima in the central region of the Seto Inland Sea (Fig. 1). The material of 46 individuals were caught by a small trawl net. They ranged from 110 mm to 163 mm in standard length (Fig. 2). To compair the diet of this fish with the benthos of their habitat, 14 individuals in the sand area and 9 individuals in the Zostra zone were sampled by spear, the benthos of the area were also sampled by quardrat method (Fig. 4 ). The general conclusions reached in this study are as follows: 1) The food organisms occurred in the guts were crustacea, bivalvia, gastropod a, and poly-chaeta. The crustacea contained the following organisms: branchiopoda, ostracoda, copepoda, mysidacea, cumacea, amphipoda, euphausiacea, macurura, anomura, brachyuran and stomatopoda. The main food organisms were cumacea for 72 % of the 46 stomachs, gastropoda for 65 %and ostracoda for 61 %. On the contrary, euphausiacea, macurura and anomura seemed to be preied on only occasionally, and branchiopoda, mysidacea and stomatopoda still more rarely (Table 1). 2) The difference between the diet of the male and female adult could not be statistically determined (Fig. 3). The diets of the specimens of 110-129mm and those of 130-164 mm were very similar. 3) The benthos of the sand area and that of the Zostera zone were clearly different in composition. Namely, in the benthos of the sand area Cypridina hilgendorfii (ostracoda) occupied 53%, but in the benthos of the Zostera zone Musculus (Musculista) senhousia (bivalvia) 87%. The composition of the food organisms and benthos sampled in each habitat were very similar to each other (Table 3). In the sand area, Cypridina hilgendorfii occupied 53% of the benthos but 79% in the food organisms. In the Zostera zone, Musculus (Musculista) senhousia occupied 87% of the benthos, but 59% in the food organisms (Table 2). 4) Musculus (Musculista) senhousia, the main food organisms of C. punctatus in the Zostera zone, were rarely taken in by the common inhabitant of the Zostera zone around Sensui-shima: Pseudoblennius cottoides, Agrammus agrammus and Hexagrammos otakii. But gammaridae that occupied only 6% in the food organisms of C. punctatus in the Zostera zone were often eaten by these 3 species mentioned above (Fig. 5). Accordingly, it is considered that C. punctatus is not in competition with these 3 species for prey

Isolation of a Drosophila gene coding for a protein containing a novel phosphatidylserine-binding motif

金沢大学医薬保健研究域薬学系To elucidate the molecular basis of the binding of proteins to the membrane phospholipid phosphatidylserine (PS), we characterized PS-binding peptides isolated from a phage display library. Amino acid sequences deduced from the nucleotide sequences of over 60 phage clones isolated revealed that there was no common primary structure among these peptides, but all peptides were rich in basic amino acid residues. In particular, 15 clones encoded peptides that contained contiguous arginine residues. Characterization of two such peptides in more detail showed that they bound to PS, and to a much lower extent to other phospholipids, including phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine. Unlike other Ca2+-dependent PS-binding proteins, these peptides did not require Ca2+ for binding to PS, and the addition of Ca2+ did not alter the phospholipid specificity. Substitution of one of the two RR sequences in one peptide by alanine had no effect, but that of both sequences completely abolished the activity. Furthermore, we identified a Drosophila gene coding for a presumed nuclear protein that shares an amino acid sequence, including a RR residue, with one of the two PS-binding peptides. This protein bound to PS partly depending on the presence of the RR residue. These results allowed us to conclude that an amino acid sequence including contiguous arginine residues is a novel motif that defines Ca2+-independent PS-binding activity. © 2005 The Japanese Biochemical Society

Risk factors associated with late aneurysmal sac expansion after endovascular abdominal aortic aneurysm repair

PURPOSEWe aimed to identify the risk factors associated with late aneurysmal sac expansion after endovascular abdominal aortic aneurysm repair (EVAR).METHODSWe retrospectively reviewed contrast-enhanced computed tomography (CT) images of 143 patients who were followed for ≥6 months after EVAR. Sac expansion was defined as an increase in sac diameter of 5 mm relative to the preoperative diameter. Univariate and multivariate analyses were performed to identify associated risk factors for late sac expansion after EVAR from the following variables: age, gender, device, endoleak, antiplatelet therapy, internal iliac artery embolization, and preprocedural variables (aneurysm diameter, proximal neck diameter, proximal neck length, suprarenal neck angulation, and infrarenal neck angulation).RESULTSUnivariate analysis revealed female gender, endoleak, aneurysm diameter ≥60 mm, suprarenal neck angulation >45°, and infrarenal neck angulation >60° as factors associated with sac expansion. Multivariate analysis revealed endoleak, aneurysm diameter ≥60 mm, and infrarenal neck angulation >60° as independent predictors of sac expansion (P < 0.05, for all).CONCLUSIONOur results suggest that patients with small abdominal aortic aneurysms (<60 mm) and infrarenal neck angulation ≤60° are more favorable candidates for EVAR. Intraprocedural treatments, such as prophylactic embolization of aortic branches or intrasac embolization, may reduce the risk of sac expansion in patients with larger abdominal aortic aneurysms or greater infrarenal neck angulation

限局性腎細胞癌における腫瘍浸潤性リンパ球と関連サイトカインの予後因子としての意義

Renal cell carcinoma (RCC) is an immunogenic tumor and pathological specimen generally contain large quantities of tumor-infiltrating lymphocytes (TILs). Numerous cell types and cytokines could affect the immune escape mechanism of tumor cells. The aim of the present study was to investigate the prognostic impact of TILs and the associated circulating cytokines on localized clear cell RCC following radical nephrectomy. A total of 87 patients who had undergone radical nephrectomy and were pathologically diagnosed with localized clear cell RCC were included. The present study evaluated the profile of TILs with immunohistochemical analysis of tumor specimens using a panel of antibodies [cluster of differentiation (CD)-4, CD8, CD80, CD86, CD276, and Forkhead box p3 (Foxp3)]. Counts of each TIL were compared with clinicopathological variables. Based on the results of immunohistochemical analyses, putative cytokines, including interleukin (IL)-6, IL-10, IL-17, interferon-γ, tumor necrosis factor (TNF)-α, and transforming growth factor (TGF)-β, were selected, and their levels in preoperative serum were measured by ELISA. The levels were compared with TIL counts in tumor specimens. High counts of the CD276+ and Foxp3+ TILs were identified as independent factors for poor prognosis for metastasis and local recurrence following radical nephrectomy (P=0.033 and 0.006, respectively). A high CD276+ TIL count was associated with preoperative serum levels of TNF-α and IFN-γ (P=0.027 and P=0.035, respectively), whereas a high count of Foxp3+ TILs was associated with preoperative serum levels of TGF-β (P=0.021). High levels of TNF-α and TGF-β were associated with recurrence-free survival (P=0.035 and P=0.031, respectively). Topical intra-tumoral immunoreaction and systemic immune status may be associated with patients with localized RCC. The topical induction of the CD276+ and Foxp3+ TILs was suggested to be associated with high levels of serum TNF-α and IFN-γ. Preoperative serum levels of TNF-α and TGF-β could be simple and non-invasive biomarkers for risk stratification before radical surgery.博士（医学）・甲第741号・令和2年3月16日Copyright © Spandidos Publications 2019. All rights reserved.Articles from Oncology Letters are provided here courtesy of Spandidos Publications

非小細胞肺がんと非小細胞肺がんから発生した転移性脳腫瘍における光線力学的測定によりPEPT1の発現は5-ALAから代謝されるプロトポルフィリンⅨの蓄積において正の相関をする

BACKGROUND: Recently, 5-aminolevulinic acid (5-ALA)-induced protoporphyrin IX fluorescence was reported to be a useful tool during total surgical resection of high-grade gliomas. However, the labeling efficacy of protoporphyrin IX fluorescence is lower in metastatic brain tumors compared to that in high-grade gliomas, and the mechanism underlying protoporphyrin IX fluorescence in metastatic brain tumors remains unclear. Lung cancer, particularly non-small cell lung cancer (NSCLC), is the most common origin for metastatic brain tumor. Therefore, we investigated the mechanism of protoporphyrin IX fluorescence in NSCLC and associated metastatic brain tumors. METHODS: Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) was employed to evaluate the protein and mRNA levels of five transporters and enzymes involved in the porphyrin biosynthesis pathway: peptide transporter 1 (PEPT1), hydroxymethylbilane synthase (HMBS), ferrochelatase (FECH), ATP-binding cassette 2 (ABCG2), and heme oxygenase 1 (HO-1). The correlation between protein, mRNA, and protoporphyrin IX levels in NSCLC cells were evaluated in vitro. Immunohistochemistry was used to determine proteins that played a key role in intraoperative protoporphyrin IX fluorescence in clinical samples from patients with NSCLC and pathologically confirmed metastatic brain tumors. RESULTS: A significant correlation between PEPT1 expression and protoporphyrin IX accumulation in vitro was identified by western blotting (P = 0.003) and qRT-PCR (P = 0.04). Immunohistochemistry results indicated that there was a significant difference in PEPT1 between the intraoperative protoporphyrin IX fluorescence-positive and protoporphyrin IX fluorescence-negative groups (P = 0.009). CONCLUSION: Expression of PEPT1 was found to be positively correlated with 5-ALA-induced protoporphyrin IX accumulation detected by photodynamic reaction in metastatic brain tumors originating from NSCLC.博士（医学）・甲第714号・令和元年年6月26日Copyright © 2019 Elsevier B.V. All rights reserved

尿路上皮癌微小環境内におけるDisabled Homolog 2 (DAB2) は腫瘍細胞上皮間葉転換を介して遊走能・浸潤能を高める

Disabled homolog-2 (DAB2) has been reported to be a tumor suppressor gene. However, a number of contrary studies suggested that DAB2 promotes tumor invasion in urothelial carcinoma of the bladder (UCB). Here, we investigated the clinical role and biological function of DAB2 in human UCB. Immunohistochemical staining analysis for DAB2 was carried out on UCB tissue specimens. DAB2 expression levels were compared with clinicopathological factors. DAB2 was knocked-down by small interfering RNA (siRNA) transfection, and then its effects on cell proliferation, invasion, and migration, and changes to epithelial-mesenchymal transition (EMT)-related proteins were evaluated. In our in vivo assays, tumor-bearing athymic nude mice subcutaneously inoculated with human UCB cells (MGH-U-3 or UM-UC-3) were treated by DAB2-targeting siRNA. Higher expression of DAB2 was associated with higher clinical T category, high tumor grade, and poor oncological outcome. The knock-down of DAB2 decreased both invasion and migration ability and expression of EMT-related proteins. Significant inhibitory effects on tumor growth and invasion were observed in xenograft tumors of UM-UC-3 treated by DAB2-targeting siRNA. Our findings suggested that DAB2 expression was associated with poor prognosis through increased oncogenic properties including tumor proliferation, migration, invasion, and enhancement of EMT in human UCB.博士（医学）・甲第768号・令和3年3月15日© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/)

Forward photon energy spectrum at LHC 7 TeV p-p collisions measured by LHCf

Abstract The LHCf experiment is one of the LHC forward experiments. The aim is to measure the energy and the transverse momentum spectra of photons, neutrons and π 0 's at the very forward region (the pseudo-rapidity range of η > 8.4 ), which should be critical data to calibrate hadron interaction models used in the air shower simulations. LHCf successfully operated at s = 900 GeV and s = 7 TeV proton–proton collisions in 2009 and 2010. We present the first physics result, single photon energy spectra at s = 7 TeV proton–proton collisions and the pseudo-rapidity ranges of η > 10.94 and 8.81 η 8.9 . The obtained spectra were compared with the predictions by several hadron interaction models and the models do not reproduce the experimental results perfectly