Waste Management in a Hot Laboratory of Japan Atomic Energy Agency – 3: Volume Reduction and Stabilization of Solid Waste

In the Japan Atomic Energy Agency, three types of experimental research, advanced reactor fuel reprocessing, radioactive waste disposal, and nuclear fuel cycle technology, have been carried out at the Chemical Processing Facility. The facility has generated high level radioactive liquid and solid wastes in hot cells. The high level radioactive solid waste is divided into three main categories, a flammable waste, a non-flammable waste, and a solid reagent waste. A plastic product is categorized into the flammable waste and molten with a heating mantle. The non-flammable waste is cut with a band saw machine for reducing the volume. Among the solid reagent waste, a used adsorbent after the experiments is heated, and an extractant is decomposed for its stabilization. All high level radioactive solid wastes in the hot cells are packed in a high level radioactive solid waste can. The high level radioactive solid waste can is transported to the 2nd High Active Solid Waste Storage in the Tokai Reprocessing Plant in the Japan Atomic Energy Agency.

Characterisation of Wind-Driven Ventilation in Complex Terrain Conditions

The physical effects of upstream flow obstructions such as vegetation on cross-ventilation phenomena of a building are important for issues such as indoor thermal comfort. Modelling such effects in Computational Fluid Dynamics simulations may also be challenging. The aim of this work is to establish the cross-ventilation jet behaviour in such complex terrain conditions as well as to provide guidelines on the implementation of CFD numerical simulations in order to model complex terrain features such as vegetation in an efficient manner. The methodology consists of onsite measurements on a test cell coupled with numerical simulations. It was found that the cross-ventilation flow is highly turbulent despite the very low velocities encountered internally within the test cells. While no direct measurement of the jet direction was made, the measurements indicate that flow tends to be reversed from the leeward to the windward side. Modelling such a phenomenon proves challenging and is strongly influenced by how vegetation is modelled. A solid vegetation tends to predict better the direction and magnitude of the flow than a porous vegetation approach. A simplified terrain model was also shown to provide good comparisons with observation. The findings have important implications on the study of cross-ventilation in complex terrain conditions since the flow direction does not remain trivial, as with the traditional isolated building case.

Immunolabeling of TGF-β during Muscle Regeneration

Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular and cellular mechanisms are still unclear. Cytokines are believed to play fundamental role in the different stages of muscle regeneration. They are secreted by many cell populations, but the predominant producers are macrophages and helper T cells. On the other hand, it has been shown that adipose tissue derived stromal/stem cell (ASC) injection could improve muscle regeneration. Stem cells probably induce the coordinated modulations of gene expression in different macrophage cells. Therefore, we investigated the patterns and timing of changes in gene expression of different cytokines occurring upon stem cells loading. Muscle regeneration was studied in an irradiated muscle of minipig animal model in presence or absence of ASC treatment (irradiated and treated with ASCs, IRR+ASC; irradiated not-treated with ASCs, IRR; and non-irradiated no-IRR). We characterized macrophage populations by immunolabeling in the different conditions. In our study, we found mostly M2 and a few M1 macrophages in the IRR+ASC samples. However, only few M2b macrophages were noticed in the IRR muscles. In addition, we found intensive fibrosis in the IRR samples. With in situ hybridization and immunolabeling, we analyzed the cytokine expression of the different macrophages and we showed that M2d macrophage are the most abundant in the IRR+ASC samples. By in situ hybridization, strong expression of the transforming growth factor β (TGF-β) was observed in the IRR+ASC but very week in the IRR samples. But when we analyzed TGF-β level with immunolabeling the expression was very different: many M2 macrophages showed week expression in IRR+ASC and few cells expressing stronger level in IRR muscles. Therefore, we investigated the MMP expressions in the different muscles. Our data showed that the M2 macrophages of the IRR+ASC muscle expressed MMP2 proteins. Our working hypothesis is that MMP2 expression of the M2 macrophages can decrease fibrosis in the IRR+ASC muscle by capturing TGF-β.

Parametric Analysis of Water Lily Shaped Split Ring Resonator Loaded Fractal Monopole Antenna for Multiband Applications

A coplanar waveguide (CPW) feed is presented, and comprising a split ring resonator (SRR) loaded fractal with water lily shape is used for multi band applications. The impedance matching of the antenna is determined by the number of Koch curve fractal unit cells. The antenna is designed on a FR4 substrate with a permittivity of εr = 4.4 and size of 14 x 16 x 1.6 mm3 to generate multi resonant mode at 3.8 GHz covering S band, 8.68 GHz at X band, 13.96 GHz at Ku band, and 19.74 GHz at K band with reflection coefficient better than -10 dB. Simulation results show that the antenna exhibits the desired voltage standing wave ratio (VSWR) level and radiation patterns across the wide frequency range. The fundamental parameters of the antenna such as return loss, VSWR, good radiation pattern with reasonable gain across the operating bands are obtained.

CRLH and SRR Based Microwave Filter Design Useful for Communication Applications

CRLH (composite right/left-handed) based and SRR (split-ring resonator) based filters have been designed at microwave frequency which can provide better performance compared to conventional edge-coupled band-pass filter designed around the same frequency, 2.45 GHz. Both CRLH and SRR are unit cells used in metamaterial design. The primary aim of designing filters with such structures is to realize size reduction and also to realize novel filter performance. The CRLH based filter has been designed in microstrip transmission line, while the SRR based filter is designed with SRR loading in waveguide. The CRLH based filter designed at 2.45 GHz provides an insertion loss of 1.6 dB with harmonic suppression up to 10 GHz with 67 % size reduction when compared with a conventional edge-coupled band-pass filter designed around the same frequency. One dimensional (1-D) SRR matrix loaded in a waveguide shows the possibility of realizing a stop-band with sharp skirts in the pass-band while a stop-band in the pass-band of normal rectangular waveguide with tailoring of the dimensions of SRR unit cells. Such filters are expected to be very useful for communication systems at microwave frequency.

Liquid Crystal Based Reconfigurable Reflectarray Antenna Design

This paper presents the design and analysis of Liquid Crystal (LC) based tunable reflectarray antenna with slot embedded patch element configurations within X-band frequency range. The slots are shown to modify the surface current distribution on the patch element of reflectarray which causes the resonant patch element to provide different resonant frequencies depending on the slot dimensions. The simulated results are supported and verified by waveguide scattering parameter measurements of different reflectarray unit cells. Different rectangular slots on patch element have been fabricated and a change in resonant frequency from 10.46GHz to 8.78GHz has been demonstrated as the width of the rectangular slot is varied from 0.2W to 0.6W. The rectangular slot in the center of the patch element has also been utilized for the frequency tunable reflectarray antenna design based on K-15 Nematic LC. For the active reflectarray antenna design, a frequency tunability of 1.2% from 10GHz to 9.88GHz has been demonstrated with a dynamic phase range of 103° provided by the measured scattering parameter results. Time consumed by liquid crystals for reconfiguration, which is one of the drawback of LC based design, has also been disused in this paper.

Utilization of Whey for the Production of β-Galactosidase Using Yeast and Fungal Culture

Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.

T Cell Immunity Profile in Pediatric Obesity and Asthma

The mechanisms underlying the association between obesity and asthma may be related to a decreased immunological tolerance induced by a defective function of regulatory T cells (Tregs). The aim of this study is to establish the potential link between these diseases and CD4+, CD25+ FoxP3+ Tregs as well as T helper cells (Ths) in children. This is a prospective case control study. Obese (n:40), asthmatic (n:40), asthmatic obese (n:40) and healthy children (n:40), who don't have any acute or chronic diseases, were included in this study. Obese children were evaluated according to WHO criteria. Asthmatic patients were chosen based on GINA criteria. Parents were asked to fill up the questionnaire. Informed consent forms were taken. Blood samples were marked with CD4+, CD25+ and FoxP3+ in order to determine Tregs and Ths by flow cytometric method. Statistical analyses were performed. p≤0.05 was chosen as meaningful threshold. Tregs exhibiting anti-inflammatory nature were significantly lower in obese (0,16%; p≤0,001), asthmatic (0,25%; p≤0,01) and asthmatic obese (0,29%; p≤0,05) groups than the control group (0,38%). Ths were counted higher in asthma group than the control (p≤0,01) and obese (p≤0,001) groups. T cell immunity plays important roles in obesity and asthma pathogeneses. Decreased numbers of Tregs found in obese, asthmatic and asthmatic obese children may help to elucidate some questions in pathophysiology of these diseases. For HOMA-IR levels, any significant difference was not noted between control and obese groups, but statistically higher values were found for obese asthmatics. The values obtained in all groups were found to be below the critical cut off points. This finding has made the statistically significant difference observed between Tregs of obese, asthmatic, obese asthmatic and control groups much more valuable. These findings will be useful in diagnosis and treatment of these disorders and future studies are needed. The production and propagation of Tregs may be promising in alternative asthma and obesity treatments.

In vitro Cytotoxic and Genotoxic Effects of Arsenic Trioxide on Human Keratinocytes

Although, arsenic trioxide has been the subject of toxicological research, in vitro cytotoxicity and genotoxicity studies using relevant cell models and uniform methodology are not well elucidated. Hence, the aim of the present study was to evaluate the cytotoxicity and genotoxicity induced by arsenic trioxide in human keratinocytes (HaCaT) using the MTT [3-(4, 5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide] and alkaline single cell gel electrophoresis (Comet) assays, respectively. Human keratinocytes were treated with different doses of arsenic trioxide for 4 h prior to cytogenetic assessment. Data obtained from the MTT assay indicated that arsenic trioxide significantly reduced the viability of HaCaT cells in a dose-dependent manner, showing an IC50 value of 34.18 ± 0.6 μM. Data generated from the comet assay also indicated a significant dose-dependent increase in DNA damage in HaCaT cells associated with arsenic trioxide exposure. We observed a significant increase in comet tail length and tail moment, showing an evidence of arsenic trioxide -induced genotoxic damage in HaCaT cells. This study confirms that the comet assay is a sensitive and effective method to detect DNA damage caused by arsenic.

Extractive Fermentation of Ethanol Using Vacuum Fractionation Technique

A vacuum fractionation technique was introduced to remove ethanol from fermentation broth. The effect of initial glucose and ethanol concentrations were investigated for specific productivity. The inhibitory ethanol concentration was observed at 100 g/L. In order to increase the fermentation performance, the ethanol product was removed as soon as it is produced. The broth was boiled at 35oC by reducing the pressure to 65 mBar. The ethanol/water vapor was fractionated for up to 90 wt% before leaving the column. Ethanol concentration in the broth was kept lower than 25 g/L, thus minimized the product inhibition effect to the yeast cells. For batch extractive fermentation, a high substrate utilization rate was obtained at 26.6 g/L.h and most of glucose was consumed within 21 h. For repeated-batch extractive fermentation, addition of glucose was carried out up to 9 times and ethanol was produced more than 8-fold higher than batch fermentation.

An Interference Reduction Strategy for TDD-OFDMA Cellular Systems

Downlink/Uplink (DL/UL) time slot allocation (TSA) in time division duplex (TDD) systems is generally uniform for all the cells. This TSA however is not efficient in case of different traffic asymmetry ratios in different cells. We first propose a new 3-coordinate architecture to identify cells in an orthogonal frequency division multiple access (OFDMA) system where each cell is divided into three sectors. Then, this coordinate system is used to derive a TSA for symmetric traffic. Mathematical analysis and simulations are used to show that the proposed TSA outperforms the traditional all uniform type of TSA in terms of total intercellular interference, even under uniform symmetrical traffic. Two adaptation strategies are further proposed to adjust the proposed TSA to asymmetrical traffic with different DL/UL traffic ratios in different cells. Further simulation results show that the adaptation strategies also yield higher signal-to-interference ratio (SIR).

Inhibition Effect of Brazilin to Human Bladder Cancer Cell Line T24

The inhibition effect of brazilin to human bladder tumor cell line T24 in vitro and in vivo was studied. The results of the in vitro experiments showed that brazilin has strong inhibition activity on the target cells. The inhibition ratio of 100 μg/mL brazilin and 100 μg/mL mitomycin to the target cells was 90.90 % and 63.24 % respectively, which showed that brazilin has higher inhibition activity than mitomycin under the same concentration. Brazilin could induce cell apoptosis in T24 cells. Significant antitumor activity of brazilin was also showed in the animals experiments. The life extention rate of 200 mg/mL, 300 mg/kg, and 400 mg/kg brazilin intraperitoneally injected into Balb/c-nu-nu nude mice that with human bladder cancer were 51.50 %, 56.90 %, and 58.42 %(P

Application of Whole Genome Amplification Technique for Genotype Analysis of Bovine Embryos

In recent years, there has been an increasing interest toward the use of bovine genotyped embryos for commercial embryo transfer programs. Biopsy of a few cells in morulla stage is essential for preimplantation genetic diagnosis (PGD). Low amount of DNA have limited performing the several molecular analyses within PGD analyses. Whole genome amplification (WGA) promises to eliminate this problem. We evaluated the possibility and performance of an improved primer extension preamplification (I-PEP) method with a range of starting bovine genomic DNA from 1-8 cells into the WGA reaction. We optimized a short and simple I-PEP (ssI-PEP) procedure (~3h). This optimized WGA method was assessed by 6 loci specific polymerase chain reactions (PCRs), included restriction fragments length polymorphism (RFLP). Optimized WGA procedure possesses enough sensitivity for molecular genetic analyses through the few input cells. This is a new era for generating characterized bovine embryos in preimplantation stage.

Rice cDNA Encoding PROLM is Capable of Rescuing Salt Sensitive Yeast Phenotypes G19 and Axt3K from Salt Stress

Rice seed expression (cDNA) library in the Lambda Zap 11® phage constructed from the developing grain 10-20 days after flowering was transformed into yeast for functional complementation assays in three salt sensitive yeast mutants S. cerevisiae strain CY162, G19 and Axt3K. Transformed cells of G19 and Axt3K with pYES vector with cDNA inserts showed enhance tolerance than those with empty pYes vector. Sequencing of the cDNA inserts revealed that they encode for the putative proteins with the sequence homologous to rice putative protein PROLM24 (Os06g31070), a prolamin precursor. Expression of this cDNA did not affect yeast growth in absence of salt. Axt3k and G19 strains expressing the PROLM24 were able to grow upto 400 mM and 600 mM of NaCl respectively. Similarly, Axt3k mutant with PROLM24 expression showed comparatively higher growth rate in the medium with excess LiCl (50 mM). The observation that expression of PROLM24 rescued the salt sensitive phenotypes of G19 and Axt3k indicates the existence of a regulatory system that ameliorates the effect of salt stress in the transformed yeast mutants. However, the exact function of the cDNA sequence, which shows partial sequence homology to yeast UTR1 is not clear. Although UTR1 involved in ferrous uptake and iron homeostasis in yeast cells, there is no evidence to prove its role in Na+ homeostasis in yeast cells. Absence of transmembrane regions in Os06g31070 protein indicates that salt tolerance is achieved not through the direct functional complementation of the mutant genes but through an alternative mechanism.

Characterization and Evaluation of the Activity of Dipeptidyl Peptidase IV from the Black-Bellied Hornet Vespa basalis

Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).

Oleate Induces Apoptosis in 3T3-L1 Adipocytes

Oleic acid (C18:1) play an important role in proliferation of fat cells. In this study, the effect of oleate on cells viability in 3T3-L1 cells (fat cells) was investigated. The 3T3-L1 cells were treated with various concentrations of oleate in the presence of 23 mM glucose. Oleate was added to adipogenic media (day 0) to investigate the influence of oleate on proliferation of postconfluent preadipocytes after 24 h induction. 0.1 mM oleate promoted cell division by increasing 33.9% number of cells from basal control in postconfluent preadipocytes. However, there were no significantly different in cells viability with control cells when oleate concentrations were increased up to 0.5 mM. When added to differentiated adipocytes (day 12) for 48 h, the number of cells decreased as oleate concentrations increased. 92.7% of cells lost demonstrated apoptosis and necrosis after 48 h with 0.5 mM oleate. The fluorochrome staining was examined under fluorescence microscopy using acridine orange and ethidium bromide double staining. Furthermore, the presence of high lactate (60.6% increased from basal control) released into plasma has shown the direct cytotoxicity of 0.5 mM oleate on adipocytes.