Two experiments were conducted to study the nature of protein degradation in fish muscle postmortem, first one with English sole (Paraphyrus vetulus) followed by another with rockfish (Sebastodes spp.). In the first one, proteolysis was measured by the increase of amino-N in gutted fish during storage in ice and in the homogenates prepared from fish of different ice storage during $20^{\circ}C-incubation$. In order to test the possible involvement of fish muscle a cathepsin, a portion of each homogenate sample was exposed to 0.5 Mrad of gamma radiation to destroy viable microorganisms prior to the incubation. Proteolysis was not detected until viable count reached a level above $10^7$ cells per gm fish flesh, corresponding to 31 days of ice storage. Even if fish flesh were mechanically disrupted by means of homogenization and subsequently incubated at $20^{\circ}C$, proteloysis attributable to muscle cathepsin was not detected. In the second with rockfish muscle aseptically prepared from freshly killed fish, the samples were inoculated with a proteolytic strain of fish spoilage Pseudomonad or irradiated at 0, 0.5 and 3.0 Mrad. The four samle groups were stored at $0-2^{\circ}C$ to compare the spoilage pattern of sterile and non-sterile muscle. In sterile muscle both total-N (extracted in 0.5M KCl) and amino-N $(soluble\;in\;70\%\;ethanol)$ declined slightly while the inoculated muscle showing increase in parallel with the increase of number of inoculated bacterium. The results indicate that proteolysis is a part of normal fish spoilage and the onset of proteolysis is delayed until viable count reaches its maximum level. Contribution of fish muscle cathepsin to protein degradation in white flesh fish muscle post-mortem is nil.
Kim, Soo-Kil;Jeung, Tae-Sig;Lim, Sang-Wook;Park, Yeong-Mouk;Park, Dahl
Progress in Medical Physics
/
v.21
no.1
/
pp.86-92
/
2010
The purpose of this study was to measure curvature contour skin dose using radiochromic film and TLD for a conventional open field. We also attempted to quantify the degradation of skin sparing associated with use of immobilization devices for high energy photon beams and to calculate the skin dose with a help of Monte Carlo (MC) simulation. To simulate head-and-neck and shoulder treatment, a cylindrical solid water phantom 11 cm in diameter was irradiated with 6 MV x-rays using $40{\times}40\;cm^2$ field at 100 cm source axis distance (SAD) to the center of the phantom. Aquaplastic mesh mask was placed on the surface of the cylindrical phantom that mimicked relevant clinical situations. The skin dose profile was obtained by taking measurements from $0^{\circ}$ to $360^{\circ}$ around the circumference of the cylindrical phantom. The skin doses obtained from radiochromic film were found to be 47% of the maximum dose of $D_{max}$ at the $0^{\circ}$ beam entry position and 61% at the $90^{\circ}$ oblique beam position without the mask. Using the mask (1.5 mm), the skin dose received was 59% at $0^{\circ}$ incidence and 78% at $80^{\circ}$ incidence. Skin dose results were also gathered using thin thermoluminescent dosimeters (TLD). With the mask, the skin dose was 66% at $0^{\circ}$ incidence and 80% at $80^{\circ}$ incidence. This method with the mask revealed the similar pattern as film measurement. For the treatments of the head-and-neck and shoulder regions in which immobilization mask was used, skin doses at around tangential angle were nearly the same as the prescription dose. When a sloping skin contour is encountered, skin doses may be abated using thinner and more perforated immoblization devices which should still maintain immoblization.
Purpose: Adaptive statistical iterative reconstruction (ASIR) technique is a reconstruction method of CT image using statistical noise modeling which is known to reduce image noise and to preserve image quality despite reducing radiation dose. The aim of this study is to evaluate images using ASIR on bone SPECT/CT which is primarily performed in our hospital. Materials and Methods: We compared the images of applied ASIR (ASIR level: 20-80%) and none ASIR by changing the mA based on 120 kVp, 100 mA using Discovery NM/CT 670 (GE, U.S.A). First, we evaluated attenuation correction in SPECT image by changing the ASIR level using Anthropomorphic phantom. Second, we compared the contrast to noise ratio (CNR), image noise and spatial resolution in CT image using ACR phantom. Third, after selecting the ASIR level applicable patient using lower torso phantom, we examined 2 patients who followed up bone SPECT/CT and we performed blind test. Results: The degree of attenuation correction in SPECT image showed no significant difference between applied ASIR and none ASIR (P>0.05). When applied ASIR, the noise of CT image were reduced at least 17 up to 52% by changing the mA. The CNR of image with ASIR was maintained more than 0.8 at 40 mA (ASIR 60%) while those without ASIR showed 0.42 at standard 40 mA. In comparison of the high contrast object, we distinguished 12 line pairs/cm at 40 mA regardless of appling ASIR. Comparison of the patients image applied ASIR level 60% (40 mA) which found out by spine image of lower torso phantom showed no signigicant difference between applied ASIR and none ASIR in blind test. The CTDIvol and DLP for applied ASIR 60% showed decreased by 60%, 60% on average than using standard mA. Conclusion: The study show that the radiation dose in SPECT/CT using ASIR can be reduced despite degradation of SPECT and CT images. In addition, higher ASIR level could be possibly applied characteristics of SPECT/CT that region of interest is limited to bone.
Recently, many landfills are constructed by using geosynthetics with earth materials. Geotextiles in geosynthetics are applied for the various purposes such as filters of the leachate collection and removal system and protectors of geomembrane liner. However geotextiles can be exposed to direct sunlight during the construction of landfill for several months. As you know the exposure of polymers to sunlight can be a major source of degradation of them. This study is to suggest a criteria on the installation of heavy weight geotextiles at the landfill. For this purpose, several different geotextiles were evaluated by outdoor exposure at two different locations and under the different seasons for one and half years. As the results of outdoor exposure, polypropylene geotextiles with 500g/$m^2$ and 700g/$\textrm{m}^2$were maintained around 40% of the initial strength during for one and half years. However, the polypropylene geotextile of 1000g/$\textrm{m}^2$showed 65% of the retained strength for the same duration of exposure. The retained strength of all polypropylene geotextile reduced dramatically with the cumulative sunshine radiation on a horizontal surface of around 1500MJ/$\textrm{m}^2$.
The purpose of this study was to investigate the degree of image degradation and the improvement of image quality caused by the density difference between the orthodontic filling material and the surrounding anatomical structure during the examination of the facial CT by quantitative and qualitative analysis. The teeth were scanned using 64-MDCT (Discovery 750 HD, GE HEALTH CARE, Milwaukee, USA). The teeth were scanned and compared according to tube voltage, silicone application, and MAR application. As a result, 10.36% CT value decreased at 140 kVp and 5.81% decrease at the application of silicon material. As a result of the qualitative evaluation, it was evaluated that 7 of the 10 observers and 3 of the acceptors were applied to the MAR algorithm. Therefore, it is possible to reduce the unnecessary burden on the radiation exposure dose as well as to reduce the loss of image data by reducing the high density artifacts, as well as the inspection parameters used in the current clinical application and various algorithms that can reduce the high density artifacts. It can be expected to provide a lot of image information.
Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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v.13
no.1
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pp.73-86
/
2015
The use of complimentary indicators, other than radiation dose and risk, to assess the safety of radioactive waste disposal has been discussed in a number of publications for providing the reasonable assurance of disposal safety and convincing the public audience. In this study, the radionuclide flux was selected as performance indicator to appraise the performance of engineered barriers and natural barrier in the Wolsong low- and intermediate-level waste disposal facility. Radionuclide flux showing the retention capability by each compartment of the disposal system is independent of assumptions in biosphere model and exposure pathways. The scenario considered as the normal scenario of disposal facility has been divided into intact or degraded silo concrete conditions. In the intact silo concrete, the radionuclide flux has been assessed with respect to the radionuclide retardation performance of each engineered barrier. In the degraded silo concrete, the radionuclide flux has been explored based on the performance degradation of engineered barriers and the relative significance of natural barrier quantitatively. The results can be used to optimally design the near-surface disposal facility being planned as the second project phase. In the future, additional complimentary indicators will be employed for strengthening the safety case for improving the public acceptance of low- and intermediate-level waste disposal facility.
Colussi, Francieli;Serpa, Viviane;Da Silva Delabona, Priscila;Manzine, Livia Regina;Voltatodio, Maria Luiza;Alves, Renata;Mello, Bruno Luan;Nei, Pereira Jr.;Farinas, Cristiane Sanches;Golubev, Alexander M.;Santos, Maria Auxiliadora Morim;Polikarpov, Igor
Journal of Microbiology and Biotechnology
/
v.21
no.8
/
pp.808-817
/
2011
Because of its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum has a considerable potential in biomass hydrolysis applications. Trichoderma harzianum cellobiohydrolase I (ThCBHI), an exoglucanase, is an important enzyme in the process of cellulose degradation. Here, we report an easy single-step ion-exchange chromatographic method for purification of ThCBHI and its initial biophysical and biochemical characterization. The ThCBHI produced by induction with microcrystalline cellulose under submerged fermentation was purified on DEAE-Sephadex A-50 media and its identity was confirmed by mass spectrometry. The ThCBHI biochemical characterization showed that the protein has a molecular mass of 66 kDa and pI of 5.23. As confirmed by smallangle X-ray scattering (SAXS), both full-length ThCBHI and its catalytic core domain (CCD) obtained by digestion with papain are monomeric in solution. Secondary structure analysis of ThCBHI by circular dichroism revealed ${\alpha}$- helices and ${\beta}$-strands contents in the 28% and 38% range, respectively. The intrinsic fluorescence emission maximum of 337 nm was accounted for as different degrees of exposure of ThCBHI tryptophan residues to water. Moreover, ThCBHI displayed maximum activity at pH 5.0 and temperature of $50^{\circ}C$ with specific activities against Avicel and p-nitrophenyl-${\beta}$-D-cellobioside of 1.25 U/mg and 1.53 U/mg, respectively.
Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.
Joo, Ji-Hye;Seok, Ji Hyun;Hong, In-kee;Kim, Nam Kyoung;Choi, Eunmi
Journal of the Society of Cosmetic Scientists of Korea
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v.42
no.1
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pp.57-64
/
2016
Just like UV radiation, heat increases collagen degradation and accumulation of abnormal elastin fiber and this is termed thermal skin aging. Dunaliella salina (DS), a green alga, is known for its beta-carotene accumulation, having various applications in the health and nutritional products. However, the effects of DS on heat-induced skin aging remain unexplored. In this study, we performed anti-thermal aging tests of the ethanol extract of DS (DSE). We measured the cellular levels of type I procollagen and MMP-1 using ELISA in human dermal fibroblast cells after heat shock. DSE reduced the expression of MMP-1 protein and increased the expression of type I procollagen. In addition, DSE upregulated the mRNA expression of HSP47 reduced by heat shock, which is involved in collagen synthesis. Also, DSE reduced the expression of inflammation mediator (TGF-${\beta}$, IL-12, etc). We demonstrate that DSE regulates the heat-induced solar elastosis through the regulation of tropoelastin and fibrillin-1, two major proteins of elastic fibers, and MMP-12 expression. These results suggest that DSE may be effective for preventing thermally induced skin aging.
Mustafa, Ebtihal H;Mahmoud, Huda T;Al-Hudhud, Mariam Y;Abdalla, Maher Y;Ahmad, Iman M;Yasin, Salem R;Elkarmi, Ali Z;Tahtamouni, Lubna H
Asian Pacific Journal of Cancer Prevention
/
v.16
no.8
/
pp.3213-3222
/
2015
Background: Cancer metastasis depends on cell motility which is driven by cycles of actin polymerization and depolymerization. Reactive oxygen species (ROS) and metabolic oxidative stress have long been associated with cancer. ROS play a vital role in regulating actin dynamics that are sensitive to oxidative modification. The current work aimed at studying the effects of sub-lethal metabolic oxidative stress on actin cytoskeleton, focal adhesion and cell migration. Materials and Methods: T47D human breast cancer cells were treated with 2-deoxy-D-glucose (2DG), L-buthionine sulfoximine (BSO), or doxorubicin (DOX), individually or in combination, and changes in intracellular total glutathione and malondialdehyde (MDA) levels were measured. The expression of three major antioxidant enzymes was studied by immunoblotting, and cells were stained with fluorescent-phalloidin to evaluate changes in F-actin organization. In addition, cell adhesion and degradation ability were measured. Cell migration was studied using wound healing and transwell migration assays. Results: Our results show that treating T47D human breast cancer cells with drug combinations (2DG/BSO, 2DG/DOX, or BSO/DOX) decreased intracellular total glutathione and increased oxidized glutathione, lipid peroxidation, and cytotoxicity. In addition, the drug combinations caused a reduction in cell area and mitotic index, prophase arrest and a decreased ability to form invadopodia. The formation of F-actin aggregates was increased in treated T47D cells. Moreover, combination therapy reduced cell adhesion and the rate of cell migration. Conclusions: Our results suggest that exposure of T47D breast cancer cells to combination therapy reduces cell migration via effects on metabolic oxidative stress.
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