Microdosimetric Modeling of Biological Effectiveness for Boron Neutron Capture Therapy Considering Intra- and Intercellular Heterogeneity in 10B Distribution

We here propose a new model for estimating the biological effectiveness for boron neutron capture therapy (BNCT) considering intra- and intercellular heterogeneity in 10B distribution. The new model was developed from our previously established stochastic microdosimetric kinetic model that determines the surviving fraction of cells irradiated with any radiations. In the model, the probability density of the absorbed doses in microscopic scales is the fundamental physical index for characterizing the radiation fields. A new computational method was established to determine the probability density for application to BNCT using the Particle and Heavy Ion Transport code System PHITS. The parameters used in the model were determined from the measured surviving fraction of tumor cells administrated with two kinds of 10B compounds. The model quantitatively highlighted the indispensable need to consider the synergetic effect and the dose dependence of the biological effectiveness in the estimate of the therapeutic effect of BNCT. The model can predict the biological effectiveness of newly developed 10B compounds based on their intra- and intercellular distributions, and thus, it can play important roles not only in treatment planning but also in drug discovery research for future BNCT

Rab7-Mediated Endocytosis Establishes Patterning of Wnt Activity through Inactivation of Dkk Antagonism

Endocytosis has been proposed to modulate cell signaling activities. However, the role of endocytosis in embryogenesis, which requires coordination of multiple signaling inputs, has remained less understood. We previously showed that mouse embryos lacking a small guanosine triphosphate (GTP)-binding protein Rab7 implicated in endocytic flow are defective in gastrulation. Here, we investigate how subcellular defects associated with Rab7 deficiency are related to the observed developmental defects. Rab7-deficient embryos fail to organize mesodermal tissues due to defects in Wnt-β-catenin signaling. Visceral endoderm (VE)-specific ablation of Rab7 results in patterning defects similar to systemic Rab7 deletion. Rab7 mutants accumulate the Wnt antagonist Dkk1 in the extracellular space and in intracellular compartments throughout the VE epithelium. These data indicate that Rab7-dependent endocytosis regulates the concentration and availability of extracellular Dkk1, thereby relieving the epiblast of antagonism. This intercellular mechanism therefore organizes distinct spatiotemporal patterns of canonical Wnt activity during the peri-gastrulation stages of embryonic development

Subthreshold Photocoagulation Using Endpoint Management in the PASCAL® System for Diffuse Diabetic Macular Edema

We evaluated subthreshold photocoagulation using endpoint management (EPM) for the treatment of diabetic macular edema (DME). The study enrolled 10 eyes from 10 patients (6 men and 4 women) with DME. The entry criteria included central macular thickness (CMT) ≥ 300 μm and decimal visual acuity (VA) ≤ 0.5. The primary endpoints were VA (logMAR) and CMT at 6 months follow-up. Secondary endpoints included fundus autofluorescence, macular volume (MV), and macular sensitivity (MS). We used the PASCAL Streamline Yellow® (wavelength, 577 nm) system to perform grid pattern laser photocoagulation at 50% of the threshold (size, 100 μm; duration, 0.015 s; spacing, 0.5; and energy, 4.5–7.8 mJ). At 6 months posttreatment, CMT was significantly decreased, while there were no significant changes in macular sensitivity, mean BCVA (logMAR), or macular volume. Autofluorescence imaging revealed no changes after treatment in 6 of 10 eyes. No eyes exhibited subjective symptoms of scotoma after photocoagulation. Optical coherence tomography showed the complete resolution of macular edema in 4 eyes (40%) after a single treatment; MS was increased in all 4 of these eyes at 6 months posttreatment. In conclusion, subthreshold photocoagulation using EPM is safe and effective for DME treatment and preserves MS. This trial is registered with UMIN000012401

Self Running Droplet: Emergence of Regular Motion from Nonequilibrium Noise

Spontaneous motion of an oil droplet driven by chemical nonequilibricity is reported. It is shown that the droplet undergoes regular rhythmic motion under appropriately designed boundary conditions, whereas it exhibits random motion in an isotropic environment. This study is a novel manifestation on the direct energy transformation of chemical energy into regular spatial-motion under isothermal conditions. A simple mathematical equation including noise reproduces the essential feature of the transition from irregularity into periodic regular motion. Our results will inspire the theoretical study on the mechanism of molecular motors in living matter, working under significant influence of thermal fluctuation.Comment: 4 pages, 4 figure

Seleção de linhagens fúngicas isoladas de larvas de Simuliidae (Insecta: Diptera) para obtenção de enzimas de interesse industrial na Amazônia: Screening of fungal strains isolated from Simuliidae (Insecta: Diptera) larvae to obtain enzymes of industrial interest in the Amazon

As enzimas estão entre os principais alvos de pesquisas biotecnológicas pelo seu importante papel nos mecanismos celulares e aplicações em processos industriais. O objetivo do estudo foi selecionar linhagens de fungos, isoladas de larvas de Simuliidae, produtoras das enzimas amilase e celulase em meio de cultura sólido. Foram utilizadas 37 linhagens fúngicas cultivadas e armazenadas no INPA. Para o crescimento das diferentes linhagens fúngicas e indução das enzimas foi utilizada solução de Manachini, suplementada de substrato indutor. O cultivo das linhagens foi realizado com inóculo direto de 5x106 esporos/mL, mantidos sob agitação a 140 rpm em temperatura ambiente durante 72 horas. Ao término do período de incubação, as culturas foram filtradas e, em seguida foram adicionados 50µL do filtrado em meio de cultura sólido pela técnica de “Cup plate”. As placas foram envolvidas em papel alumínio e incubadas a 37º C durante 72 horas. Foram utilizadas soluções reveladoras de atividade enzimática, iodo 0,1 Mol/L e solução de vermelho congo a 0,1%), para amilases e celulases, respectivamente. A atividade enzimática foi detectada pelo diâmetro do halo. Do total de 37 linhagens analisadas, 60% apresentaram degradação de substrato tanto para amilase como para celulase e 40% não apresentaram atividade para estas enzimas. As espécies com maior nível de degradação do substrato foram Aspergillus japonicus Saito (halo= 7mm, celulase) e Trichoderma harzianum Rifai (halo= 5mm, amilase). Constata-se assim o potencial desses microrganismos para aplicação industrial, criando novas oportunidades para o desenvolvimento do mercado de enzimas na Amazônia

インシリコ及びインビトロスクリーニングによるA型インフルエンザウイルスの増殖阻害活性をもつ低分子化合物の同定

Influenza viruses have acquired resistance to approved neuraminidase-targeting drugs, increasing the need for new drug targets for the development of novel anti-influenza drugs. Nucleoprotein (NP) is an attractive target since it has an indispensable role in virus replication and its amino acid sequence is well conserved. In this study, we aimed to identify new inhibitors of the NP using a structure-based drug discovery algorithm, named Nagasaki University Docking Engine (NUDE), which has been established especially for the Destination for GPU Intensive Machine (DEGIMA) supercomputer. The hit compounds that showed high binding scores during in silico screening were subsequently evaluated for anti-influenza virus effects using a cell-based assay. A 4-hydroxyquinolinone compound, designated as NUD-1, was found to inhibit the replication of influenza virus in cultured cells. Analysis of binding between NUD-1 and NP using surface plasmon resonance assay and fragment molecular orbital calculations confirmed that NUD-1 binds to NP and could interfere with NP-NP interactions essential for virus replication. Time-of-addition experiments showed that the compound inhibited the mid-stage of infection, corresponding to assembly of the NP and other viral proteins. Moreover, NUD-1 was also effective against various types of influenza A viruses including a clinical isolate of A(H1N1)pdm09 influenza with a 50% inhibitory concentration range of 1.8?2.1 μM. Our data demonstrate that the combined use of NUDE system followed by the cell-based assay is useful to obtain lead compounds for the development of novel anti-influenza drugs.長崎大学学位論文 学位記番号:博(医歯薬)甲第997号 学位授与年月日:平成29年9月20日Author: Juliann Nzembi Makau, Ken Watanabe, Takeshi Ishikawa, Satoshi Mizuta, Tsuyoshi Hamada, Nobuyuki Kobayashi, Noriyuki NishidaCitation: PLOS ONE, 12(3), e0173582; 2017Nagasaki University (長崎大学)課程博

Induction of Non-Targeted Stress Responses in Mammary Tissues by Heavy Ions

Purpose Side effects related to radiation exposures are based primarily on the assumption that the detrimental effects of radiation occur in directly irradiated cells. However, several studies have reported over the years of radiation-induced non-targeted/ abscopal effects in vivo that challenge this paradigm. There is evidence that Cyclooxygenase-2 (COX2) plays an important role in modulating non-targeted effects, including DNA damages in vitro and mutagenesis in vivo. While most reports on radiation-induced non-targeted response utilize x-rays, there is little information available for heavy ions. Methods and Materials Adult female transgenic gpt delta mice were exposed to an equitoxic dose of either carbon or argon particles using the Heavy Ion Medical Accelerator in Chiba (HIMAC) at the National Institute of Radiological Sciences (NIRS) in Japan. The mice were stratified into 4 groups of 5 animals each: Control; animals irradiated under full shielding (Sham-irradiated); animals receiving whole body irradiation (WBIR); and animals receiving partial body irradiation (PBIR) to the lower abdomen with a 1 x 1 cm2 field. The doses used in the carbon ion group (4.5 Gy) and in argon particle group (1.5 Gy) have a relative biological effectiveness equivalent to a 5 Gy dose of x-rays. 24 hours after irradiation, breast tissues in and out of the irradiated field were harvested for analysis. Induction of COX2, 8-hydroxydeoxyguanosine (8-OHdG), phosphorylated histone H2AX (γ-H2AX), and apoptosis-related cysteine protease-3 (Caspase-3) antibodies were examined in the four categories of breast tissues using immunohistochemical techniques. Analysis was performed by measuring the intensity of more than 20 individual microscopic fields and comparing the relative fold difference. Results In the carbon ion group, the relative fold increase in COX2 expression was 1.01 in sham-irradiated group (p > 0.05), 3.07 in PBIR (p 0.05), 11.31 in PBIR (p 0.05), 8.41 in PBIR (p < 0.05) and 10.59 in WBIR (p < 0.05). Results for the argon particle therapy group showed a similar magnitude of changes in the various biological endpoints examined. There was no statistical significance observed in Caspase-3 expression among the 4 groups. Conclusions Our data show that both carbon and argon ions induced non-targeted, out of field induction of COX2 and DNA damages in breast tissues. These effects may pose new challenges to evaluate the risks associated with radiation exposure and understanding radiation-induced side effects

A novel in vitro survival assay of small intestinal stem cells after exposure to ionizing radiation

The microcolony assay developed by Withers and Elkind has been a gold standard to assess the surviving fraction of small intestinal stem cells after exposure to high (?8 Gy) doses of ionizing radiation (IR), but is not applicable in cases of exposure to lower doses. Here, we developed a novel in vitro assay that enables assessment of the surviving fraction of small intestinal stem cells after exposure to lower IR doses. The assay includes in vitro culture of small intestinal stem cells, which allows the stem cells to develop into epithelial organoids containing all four differentiated cell types of the small intestine. We used Lgr5-EGFP-IRES-CreERT2/ROSA26-tdTomato mice to identify Lgr5+ stem cells and their progeny. Enzymatically dissociated single crypt cells from the duodenum and jejunum of mice were irradiated with 7.25, 29, 101, 304, 1000, 2000 and 4000 mGy of X-rays immediately after plating, and the number of organoids was counted on Day 12. Organoid-forming efficiency of irradiated cells relative to that of unirradiated controls was defined as the surviving fraction of stem cells. We observed a significant decrease in the surviving fraction of stem cells at ?1000 mGy. Moreover, fluorescence-activated cell sorting analyses and passage of the organoids revealed that proliferation of stem cells surviving IR is significantly potentiated. Together, the present study demonstrates that the in vitro assay is useful for quantitatively assessing the surviving fraction of small intestinal stem cells after exposure to lower doses of IR as compared with previous examinations using the microcolony assay