Large round cell granulomas induced by murine sarcoma virus (Moloney) in mouse brains

The effect of murine sarcoma virus of Moloney strain on central nervous system was examined morphologically in Swiss mice of different age. A single intracranial inoculation of cell-free virus solution resulted in the induction of characteristic intracerebral granulomas in 82.8% of the newborn to 5 day-old group, in 71.4% of the 6 to 10 day-old group, and in 68.0% of the 11 to 20 day-old group. The mean latency periods to tumor recognition were 16.5, 21.1, and 33.5 days, respectively. The granuloma consisted of inflammatory cell infilrations, reactive gliosis, and richly developed blood vessels. The lesions consistently contained numerous characteristic large round cells. In cases of long-survival, the findings included reparative changes, such as extensive gliosis, withdrawal of inflammation, and a decrease in the numbers of large round cells and blood vessels. These lesions were tentatively designated as &#34;large round cell granuloma.&#34; The early foci of the granoloma were composed of proliferating glial cells and large round cells at the subependymal regions. Electron microscopically these large round cells had abundant intracytoplasmic fibrils quite similar to gliofibrils. Numerous C-type virus particles were present in the intercellular nad perivascular spaces, and occasionally budded from cell membranes of the large round cells and vascular endothelia. The large round cells were considered to be reactive astrocytes activated by biral infection. It was conclided that MSV-M was not a sarcomogenic but a granulomogenic virus in mice. Control animals showed no pathological changes.</p

Development of Numerical Removal Model of Arsenic from Groundwater

Abstract There is a tremendous demand for developing efficient methods for arsenic removal from drinking waters due to its severe toxicity effect on living beings. In order to predict an appropriate technique for arsenic removal from groundwater a numerical model that is able to describe the removal of arsenic and other background species from the groundwater considering chemical and biochemical processes has been developed in the present paper. Based on the adsorption/co-precipitation of arsenic with iron(III) hydroxides, the behavior of arsenic and iron under oxidation dominant environment including advection, dispersion, molecular diffusion and the microbially mediated chemical processes are also presented. The arsenic transport equation and the balance equations are solved numerically using the finite difference method and method of characteristics (MOC), subject to prescribed initial and boundary conditions are also presented. The kinetic sub model describes the heterotrophic metabolisms of several groups of microorganisms. Microbial growth is assumed to follow Monod type kinetics. The results of the removal simulation model demonstrated that availability of substrate (e.g. organic carbon), maximum growth rate, yield coefficient of bacteria, the precipitation rate of iron-hydroxide and the adsorption/precipitation rate of arsenic with iron hydroxide were important parameters which affected the removal of arsenic from groundwater. Among all the parameters organic carbon concentration proved as the most important

Bio-geochemical simulation for solute transport in Piyungan landfill, Yogyakarta Special Province, Indonesia

Piyungan Landfill is the largest in Bantul Regency. According to water quality sampling taken from a leachate pond, there are significant contaminant issues resulting from landfill leachate. The objectives of this research were achieved by applying a two-dimensional bacteria mediated reduction numerical model was applied. Method of characteristic was applied to solve the advection part of the solute transport equation. Three bacteria (X1, X2, and X3) groups were defined in the redox model. In the conceptual model, bacterial X1 utilizes oxygen under aerobic conditions and nitrate, NO-3 under aerobic conditions as electron acceptors. Consequently, under aerobic conditions bacteria X2, and X3 utilize MnO2, and Fe(OH)3 respectively as electron acceptors. In the redox model organic carbon which was defined as CH2O was considered as the electron donor for all bacteria mediated reduction reactions. The results of research are to improve the understanding of biogeochemical processes in aquifer

Magnetic characterization change by solvents of magnetic nanoparticles in liquid-phase magnetic immunoassay

Liquid-phase magnetic immunoassay (MIA) using magnetic nano-particles (MNPs) has been studied as a more rapid method compared to optical methods for inspecting proteins and viruses. MIA can estimate the number of conjugated antibodies without being washed differently from conventional optical immunoassay. However, in the case of the liquid phase, it is considered that the magnetic properties of MNPs are affected by physical properties such as viscosity and impurity substances such as biological substances contained in the blood. In this study, the effect of sodium chloride (NaCl) in buffer and serum solution was evaluated to reveal the effect of serum because the sodium (Na+) and chloride (Cl-) ions in the serum dominate ion balance of blood. The measurement results of AC magnetic susceptibility and a dynamic light scattering (DLS) showed that the aggregation of MNPs was largely affected by the concentration of NaCl. This effect of the NaCl could be explained by shielding of the surface charge of MNPs by ions in the solution. Although the concentrations of NaCl in the buffer and serum solution were almost same, we found that MNPs were aggregated more in their size for those in the serum solution because of other impurities, such as proteins. These results suggest evaluation of effects of the contaminants in serum and optimization of polymer coatings of MNPs could be important factors to realize measurements of magnetic immunoassay with high accuracy. (C) 2019 Author(s)

SIMULATION OF BIO-GEOCHEMICAL PROCESSES IN A COASTAL AQUIFER

Numerical models which discuss about the bio-geochemical processes which take place in coastal aquifers are still limited. Bacteria mediated reduction is one of the main causes which directly contributes to the groundwater quality of coastal aquifers. In this study a two-dimensional numerical model was developed to simulate the bacteria mediated reduction processes which are occurring in the coastal aquifers. The model was applied to a cross section of Kujyukurihama coast in Chiba prefecture, Japan, where a reduced environment has been formed. The objective of this paper is not to provide an orthodox numerical simulation practice, but to convince the numerical modellers; the possibility of the simulation of bacterial reduction processes in coastal subsurface environments. This paper provides a preliminary numerical study of the reduction of MnO 2 and Fe(OH) 3 under anaerobic bacteria mediation. Further researches are needed to improve the numerical simulations on the reduction environments in coastal aquifers

Melanin pigments in the melanocytic nevus regress spontaneously after inactivation by high hydrostatic pressure

We report a novel treatment for giant congenital melanocytic nevi (GCMN) that involves the reuse of resected nevus tissue after high hydrostatic pressurization (HHP). However, the remaining melanin pigments in the inactivated nevus tissue pose a problem; therefore, we performed a long-term observation of the color change of inactivated nevus tissue after HHP. Pressurized nevus specimens (200 MPa group, n = 9) and non-pressurized nevus tissues (control group, n = 9) were subcutaneously implanted into nude mice (BALB/c-nu) and then harvested 3, 6, and 12 months later. Color changes of the nevus specimens were evaluated. In the 200 MPa group, the specimen color gradually regressed and turned white, and brightness values were significantly higher in the 200 MPa group than in the control group after 6 months. This indicated that melanin pigments in the pressurized nevus tissue had spontaneously degraded and regressed. Therefore, it is not necessary to remove melanin pigments in HHP-treated nevus tissue

Laser monitoring of dynamic behavior of magnetic nanoparticles in magnetic field gradient

Manipulation of magnetic nanoparticles (MNP) by an external magnetic field has been widely studied in the fields of biotechnology and medicine for collecting and/or reacting biomaterials in the solutions. Here, dynamic behaviors of MNP in solution under changing gradient magnetic field were investigated using our newly developed laser transmission system (LTS) with a variable magnetic field manipulator. The manipulator consists of a moving permanent magnet placed beside the optical cell filled with MNP solution. A laser beam was focused on the cell and the transmitted laser beam was detected by a silicon photodiode, so that the localized concentration of the MNP at the focused area could be evaluated by the intensity of transmitted laser beam. In this study, the LTS was applied to evaluate dynamic behaviors of MNP in serum solution. Dispersion and aggregation of MNP in the solution were evaluated. While time evolution of dispersion depends on the serum concentration, the behavior during aggregation by the magnetic field was independent of the serum concentration. A series of measurements for zeta-potentials, distributions of particle size, and magnetization distributions was carried out to understand this difference in the behavior. The results indicated that a Brownian motion was main force to distribute the MNP in the solution; on the other hand, the magnetic force to the MNP mainly affected the behavior during aggregation of the MNP in the solution

コロナ禍における2020年度前期開講科目「ジョギング・ウォーキング」の実践

本報告は、長崎国際大学にて開講された2020（令和2）年度前期科目『ジョギング・ウォーキング』の実践記録である。本報告の目的は新型コロナウイルス感染症の発症が日常的になることが予測される中で、これからの実技系科目のより良い実践方法の創出に寄与することである。本報告では、授業方法の詳細と受講生の感想を分析した結果を掲載する。分析方法は、KH Coder によるテキストマイニングを用いている。受講生の取り組み意欲を促進させたと考えられるのが、スマートフォンアプリでの運動実践記録、記録を写真として編集し SNS に投稿できる機能の活用であろう。ほとんどの学生がスマートフォンを所有し、SNS もまたほとんどの学生が利用している中で、身近なものを教材化して利用したことが、今回の遠隔授業を展開する上での鍵であった。新型コロナウイルス感染症の流行によりオンライン授業となった『ジョギング・ウォーキング』の実践を通して学習効果を高める3つの特性が浮かび上がった。すなわち、即時的なフィードバック、相互的なやり取り、そして共有することであった。それらを踏まえることが、オンライン授業における学生の学習意欲の促進や満足度の向上に繋がると考えられる

The Alteration of the Epidermal Basement Membrane Complex of Human Nevus Tissue and Keratinocyte Attachment after High Hydrostatic Pressurization

We previously reported that human nevus tissue was inactivated after high hydrostatic pressure (HHP) higher than 200 MPa and that human cultured epidermis (hCE) engrafted on the pressurized nevus at 200 MPa but not at 1000 MPa. In this study, we explore the changes to the epidermal basement membrane in detail and elucidate the cause of the difference in hCE engraftment. Nevus specimens of 8 mm in diameter were divided into five groups (control and 100, 200, 500, and 1000 MPa). Immediately after HHP, immunohistochemical staining was performed to detect the presence of laminin-332 and type VII collagen, and the specimens were observed by transmission electron microscopy (TEM). hCE was placed on the pressurized nevus specimens in the 200, 500, and 1000 MPa groups and implanted into the subcutis of nude mice; the specimens were harvested at 14 days after implantation. Then, human keratinocytes were seeded on the pressurized nevus and the attachment was evaluated. The immunohistochemical staining results revealed that the control and 100 MPa, 200 MPa, and 500 MPa groups were positive for type VII collagen and laminin-332 immediately after HHP. TEM showed that, in all of the groups, the lamina densa existed; however, anchoring fibrils were not clearly observed in the 500 or 1000 MPa groups. Although the hCE took in the 200 and 500 MPa groups, keratinocyte attachment was only confirmed in the 200 MPa group. This result indicates that HHP at 200 MPa is preferable for inactivating nevus tissue to allow its reuse for skin reconstruction in the clinical setting

Haplotypes and a Novel Defective Allele of CES2 Found in a Japanese Population

ABSTRACT: Human carboxylesterase 2 (hCE-2) is a member of the serine esterase superfamily and is responsible for hydrolysis of a wide variety of xenobiotic and endogenous esters. hCE-2 also activates an anticancer drug, irinotecan (7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin, CPT-11), into its active metabolite, 7-ethyl-10-hydroxycamptothecin (SN-38). In this study, a comprehensive haplotype analysis of the CES2 gene, which encodes hCE-2, in a Japanese population was conducted. Human carboxylesterases are members of the serine esterase superfamily and are responsible for hydrolysis of a wide variety of xenobiotic and endogenous esters. They metabolize esters, thioesters, carbamates, and amides to yield soluble acids and alcohols or amines Although both hCE-1 and hCE-2 show broad substrate specificities, hCE-2 is relatively specific for heroin, cocaine (benzoyl ester), 6-acetylmorphine, procaine, and oxybutynin 1865 camptothecin (SN-38), a topoisomerase inhibitor, by carboxylesterases Previously, 12 exons and their flanking regions of CES2 were sequenced from 153 Japanese subjects, who received irinotecan or steroidal drugs, and 12 novel SNPs, including the nonsynonymous SNP, 100CϾT (Arg 34 Trp), and the SNP at the splice acceptor site of intron 8 (IVS8-2AϾG) were found Materials and Methods Chemicals. Irinotecan, SN-38, and SN-38G were kindly supplied by Yakult Honsha Co. Ltd. (Tokyo, Japan). Patients. A total of 262 Japanese subjects analyzed in this study consisted of 85 patients with allergies who received steroidal drugs and 177 patients with cancer who received irinotecan. The ethical review boards of the National Cancer Center, National Center for Child Health and Development, and National Institute of Health Sciences approved this study. Written informed consent was obtained from all participants. DNA Sequencing. Total genomic DNA was extracted from blood leukocytes or Epstein-Barr virus-transformed lymphocytes and used as a template in the polymerase chain reaction (PCR). Sequence data of the CES2 gene from 72 patients and 81 cancer patients were described previously Linkage Disequilibrium and Haplotype Analyses. LD analysis was performed by the SNPAlyze software (version 5.1; Dynacom Co., Yokohama, Japan), and a pairwise two-dimensional map between SNPs was obtained for the DЈ and rho square (r 2 ) values. All allele frequencies were in HardyWeinberg equilibrium. Some haplotypes were unambiguously assigned in the subjects with homozygous variations at all sites or a heterozygous variation at only one site. Separately, the diplotype configurations (combinations of haplotypes) were inferred by LDSUPPORT software, which determines the posterior probability distribution of the diplotype configuration for each subject on the basis of estimated haplotype frequencies Administration of Irinotecan and Pharmacokinetic Analysis. The demographic data and eligibility criteria for 177 cancer patients who received irinotecan in the National Cancer Center Hospitals (Tokyo and Chiba, Japan) were described elsewhere Each patient received a 90-min i.v. infusion at doses of 60 to 150 mg/m 2 , which varied depending on regimens/coadministered drugs: i.e., irinotecan dosages were 100 or 150 mg/m 2 for monotherapy and combination with 5-FU, 150 mg/m 2 for combination with mitomycin C (MMC), and 60 (or 70) mg/m 2 for combination with platinum anticancer drugs. Heparinized blood was collected before administration of irinotecan and at 0 min (end of infusion), 20 min, 1 h, 2 h, 4 h, 8 h, and 24 h after infusion. Plasma concentrations of irinotecan, SN-38, and SN-38G were determined as described previously Expression of Wild-Type and Variant CES2 Proteins in COS-1 Cells. Expression of wild-type and variant CES2 proteins in COS-1 cells was examined as described previously and ZERO-Dscan software (Raytest, Straubenhardt, Germany). The relative expression levels are shown as the means Ϯ S.D. of three separate transfection experiments. Determination of CES2 mRNA by Real-Time RT-PCR. Total RNA was isolated from transfected COS-1 cells using the RNeasy Mini Kit (QIAGEN, Tokyo, Japan). After RNase-free DNase treatment of samples to minimize plasmid DNA contamination, first-strand cDNA was prepared from 1 g of total RNA using the High-Capacity cDNA Archive Kit (Applied Biosystems, Foster City, CA) with random primers. Real-time PCR assays were performed with the ABI7500 Real Time PCR System (Applied Biosystems) using the TaqMan Gene Expression Assay for CES2 (Hs01077945_m1; Applied Biosystems) according to the manufacturer&apos;s instructions. The relative mRNA levels were determined using calibration curves obtained from serial dilutions of the pooled wild-type CES2 cDNA. Samples without reverse transcriptase were routinely included in the RT-PCR reactions to measure possible contributions of contaminating DNA, which was usually less than 1% of the mRNA-derived amplification. Transcripts of ␤-actin were quantified as internal controls using TaqMan ␤-Actin Control Reagent (Applied Biosystems), and normalization of CES2 mRNA levels were based on ␤-actin concentrations. Enzyme Assay. CPT-11 hydrolyzing activity of the postmitochondrial supernatants (microsomal fraction plus cytosol) was assayed over the substrate concentration range of 0.25 to 50 M as described previously Statistical Analysis. Statistical analysis of the differences in the AUC ratios among CES2 diplotypes, coadministered drugs. or irinotecan dosages was performed using the Kruskal-Wallis test, Mann-Whitney test, or Spearman rank correlation test (Prism 4.0, GraphPad Software, Inc., San Diego, CA). The t test (Prism 4.0) was applied to the comparison of the average values of protein expression and mRNA levels between wild-type and variant CES2. Results CES2 Variations Detected in a Japanese Population. Previously, the promoter region, all 12 exons, and their flanking introns of the CES2 gene were sequenced from 72 allergic patients and 81 cancer patients and resulted in the identification of 12 novel SNPs The nonsynonymous SNP 424GϾA (V142M) reported by our group LD and Haplotype Analysis. Using the detected SNPs, LD analysis was performed, and the pairwise values of r 2 and DЈ were obtained. A perfect linkage (r 2 ϭ 1.00) was observed between SNPs Ϫ363CϾG and IVS10-87GϾA. A close association (r 2 ϭ 0.85) was found between SNPs IVS10-108GϾA and 1749AϾG. Other associations were much lower (r 2 Ͻ 0.1). Therefore, the entire CES2 gene was analyzed as one LD block. The determined/inferred haplotypes are summarized i