The aim was to determine whether ultrasound targeted microbubble destruction (UTMD) promotes dual targeting of HSP72 and HSC70 for therapy of castration-resistant prostate cancer (CRPC), to improve the specific and efficient delivery of siRNA, to induce tumor cell specific apoptosis, and to find new therapeutic targets specific of CRPC.VCaP cells were transfected with siRNA oligonucleotides. HSP70, HSP90 and cleaved caspase-3 expression were determined by real-time quantitative polymerase chain reaction and Western blotting. Apoptosis and transfection efficiency were assessed by flow cytometry. Cell viability assays were used to evaluate safety. We found HSP72, HSC70 and HSP90 expression to be absent or weak in normal prostate epithelial cells (RWPE-1), but uniformly strong in prostate cancerous cells (VCaP). UTMD combined with dual targeting of HSP72 and HSC70 siRNA improve the efficiency of transfection, cell uptake of siRNA, downregulation of HSP70 and HSP90 expression in VCaP cells at the mRNA and protein level, and induction of extensive tumor-specific apoptosis. Cell counting kit-8 assays showed decreased cellular viability in the HSP72/HSC70-siRNA silenced group. These results suggest that the combination of UTMD with dual targeting HSP70 therapy for PCa may be most efficacious, providng a novel, reliable, non-invasive, safe targeted approach to improve the specific and efficient delivery of siRNA, and achieve maximal effects.
Pasqualotto, M.;Neres, M.A.;Guimaraes, V.F.;Klein, J.;Inagaki, A.M.;Ducati, C.
Asian-Australasian Journal of Animal Sciences
/
v.28
no.6
/
pp.807-815
/
2015
The present study aimed at evaluating the intensity of Tifton 85 conditioning using a mower conditioner with free-swinging flail fingers and storage times on dehydration curve, fungi presence, nutritional value and in vitro digestibility of Tifton 85 bermudagrass hay dry matter (DM). The dehydration curve was determined in the whole plant for ten times until the baling. The zero time corresponded to the plant before cutting, which occurred at 11:00 and the other collections were carried out at 8:00, 10:00, 14:00, and 16:00. The experimental design was randomised blocks with two intensities of conditioning (high and low) and ten sampling times, with five replications. The high and low intensities related to adjusting the deflector plate of the free iron fingers (8 and 18 cm). In order to determine gas exchanges during Tifton 85 bermudagrass dehydration, there were evaluations of mature leaves, which were placed in the upper middle third of each branch before the cutting, at every hour for 4 hours. A portable gas analyser was used by an infrared IRGA (6400xt). The analysed variables were photosynthesis (A), stomatal conductance (gs), internal $CO_2$ concentration (Ci), transpiration (T), water use efficiency (WUE), and intrinsic water use efficiency (WUEi). In the second part of this study, the nutritional value of Tifton 85 hay was evaluated, so randomised blocks were designed in a split plot through time, with two treatments placed in the following plots: high and low intensity of cutting and five different time points as subplots: cutting (additional treatment), baling and after 30, 60, and 90 days of storage. Subsequently, fungi that were in green plants as well as hay were determined and samples were collected from the grass at the cutting period, during baling, and after 30, 60, and 90 days of storage. It was observed that Tifton 85 bermudagrass dehydration occurred within 49 hours, so this was considered the best time for drying hay. Gas exchanges were more intense before cutting, although after cutting they decreased until ceasing within 4 hours. The lowest values of acid detergent insoluble nitrogen were obtained with low conditioning intensity after 30 days of storage, 64.8 g/kg DM. The in vitro dry matter of Tifton 85 bermudagrass did not differ among the storage times or the conditioning intensities. There was no fungi present in the samples collected during the storage period up to 90 days after dehydration, with less than 30 colony forming units found on plate counting. The use of mower conditioners in different intensities of injury did not speed up the dehydration time of Tifton 85.
It is important to find out a new material having high removal efficiency for the harmful algal blooms because the dispersion of Hwangto in a large amount to the sea water may bring some ecologically unfavorable problems. For this purpose, the efficiency of several natural and synthetic mineral species for the mitigation of algal blooms was measured. The mixing ratio of monominerals and the sea water with 3,000∼5,000 cells/$m\ell$ of Cochlodinium polykrikoides was 10 g/${\ell}$ and the removal ratio was measured by counting the living cells after the dispersion time of 10, 30 and 60 min., respectively. According to the experimental results, the removal ratio by illite, kaolinite, montmonmorillonite, red mud, Na-A type of zeolite ranged 84-92% after 1hr of contact time, which is comparable to that of Hwangto. The size of above monominerals ranged 3∼50${\mu}m$. Meanwhile, the amorphose material and hematite with the size of 50∼100 nm showed excellent removal ratio of more than 99% after 30min. of dispersion. The results of the study showed that the removal ratio was not related to the chemical composition and pH of the minerals applied but to the grain size. The experimental results strongly suggest that the main mitigation mechanism would be the contact and coagulation.
Jae-Ki Shin;Nan-Young Kim;Yongeun Park;Kyung-Lak Lee;Baik-Ho Kim;Yong-Jae Kim;Han-Soon Kim;Jung Ho Lee;Hak Young Lee;Soon-Jin Hwang
Korean Journal of Ecology and Environment
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v.56
no.3
/
pp.196-206
/
2023
Benthic attached diatoms (BADs), a major primary producer in lotic stream and river ecosystems are micro-sized organisms and require a highly magnified microscopic technique in the observation work. Thus, it is often not easy to ensure accuracy and precision in both qualitative and quantitative analyses. This study proposed a new technique applicable to improve quality control of aquatic ecosystem monitoring and assessment using BADs. In order to meet the purpose of quality control, we developed a permanent mounting slide technique which can be used for both qualitative and quantitative analyses simultaneously. We designed specimens with the combination of grid on both cover and slide glasses and compared their efficiency. As a result of observation and counting of BADs, the slide glass designed with the color-lined grid showed the highest efficiency compared to other test conditions. We expect that the method developed in this study could be effectively used to analyze BADs and contributed to improve the quality control in aquatic ecosystem health monitoring and assessment.
Journal of Korean Society for Atmospheric Environment
/
v.3
no.1
/
pp.27-33
/
1987
The environment and biological studies of tritium have been carried out in the advanced countries since the mid 1950's. In the case of a potential tritium exposure, the usual procedure is trifium bioassay (as HTO) in human urine in order to determine the amount of tritium deposited in the body called tritium body burden. The maximum permissible body burden(MPBB) of tritium in total body is about $30{\mu}Ci/{\ell}$ for body tissue. In the bioassay, the most common investigation level for detection of tritium in urine is 1/10th of MPBB. For this bioassay project, the first priority is given to obtaining a quench correction curve. This consideration is necessary because of the variability in color of human urine specimens. Quenching effect in this case mainly is caused by the absorption of scintillation light flashes by the urine sample. By the least squares method on the statistical basis, an estimated formula for quench correction curve was determined to be Y = 0.771 + 1.836 ${\tmes}10^{-4}$X, where the efficiency(Y) was ranged from about 12% to 31% in the liquid scientillation counting. In this paper, a brief theory concerning the biological half-life of tritium and the retention formula to apply to systematically distributed tritium are described.
Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
/
v.9
no.3
/
pp.141-148
/
2011
It is clarified in the radioactive waste transfer regulation that the concentration of radioactive waste for the major radio nuclide has to be examined when radioactive waste is guided to the radioactive waste stores. In case of the low level radioactive waste sample, the analytical results of radioactive waste concentration frequently show a value lower than minimum detectable activity (MDA). Since the MDA value basically depends on the amount of a sample, background value, measurement time, counting efficiency, and etc, it would be necessary to increase a sample amount with a intention of minimizing MDA. In order to measure a concentration of $^{129}I$ in low and medium level radioactive waste, $^{129}I$ was collected by using a distillation technique after leaching the simulated radioactive waste sample with a non-volatile acid. The recovery of $^{129}I$ measured was compared with that measured with column elution technique which is a conventional method using an anion-exchange resin. The recovery of inactive iodide by using the distillation method and column elution were found as $86.5{\pm}0.9%$ and $87.3{\pm}2.7%$, respectively. The recovery and MDA value calculated for distillation technique when 100 g of extracted solution of $^{129}I$ was taken, were found to be $84.6{\pm}1.6%$ and $1.2{\times}10^{-4}Bq/g$, respectively. Consequently, the proposed technique with simplified process lowered the MDA value more than 10 times compared to the column elution technique that has a disadvantage of limited sampling amount.
The purposes of this study were to produce cloned rabbit embryos and offsprings by nuclear transplantation(NT) using in vitro matured oocytes as nuclear recipient cytoplasm and to determine the effect of frozen nuclei donor embryos on the production efficiency of cloned embryos. The 8cell embryos were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline containing 10% fetal calf serum(FCS) at 40 hours after hGG injection. A portion of collected embryos were preserved at 4$^{\circ}C$ for 24 hours and a portion of them were frozen by vitrification method. The embryos used for donor nuclei were synchronized in the phase of Gi /S transition. The in vitro matured oocytes were used as recipient cytoplasm following removing the nucleus and the first polar body. The synchronized blastomeres from fresh, cooled or frozen embryos were injected into the enucleated oocytes by micromanipulation and were electrofused by electrical stimulation of three pulses for 60 $\mu$sec at 1.0 W /cm in 0.28 M mannitol solution. The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$incubator. Following in vitro culture of the NT embryos to blastocyst stage, they were stained with Hoechst 33342 dye for counting the number of blastomeres by fluorescence microscopy. The nuclear transplant embryos developed in vitro to 2- to 4-cell stage were transferred into the oviducts of synchronized recipient does. The results obtained were summarized as follows: 1. The fusion rates of the blastomeres from fresh, cooled and frozen embryos with the in vitro matured and enucleated oocytes were 100, 95.8 and 64, 3%, respectively. 2. Development in vitro to blastocyst was significantly(p<0.05) different between the cloned embryos with the blastomeres from fresh, cooled or frozen embryos as 39.0, 20. 9 and 15.7%, respectively. 3. The mean numbers of cell cycle per day during in vitro culture of cloned embryos blastomeres from fresh, cooled or frozen embryos was 1.31, 1.29 and 1.16, respectively. 4. A total of 77 nuclear transplant embryos were transferred into 6 recipient does, of which two offsprings were produced from a foster mother 31 days after embryo transfer.
Control of white rot which is one of the most serious apple diseases in Korea has mainly relied on periodical spray of protective fungicides. As the main inoculum source of the disease is pycnidiospores produced in the warts formed on affected stems of apple tree, it can be conceivable that inhibition of spore production might be an effective means for controlling the disease. Inhibitory efficacy of eight selected fungicides against sporulation of the fungus was assessed by counting the number of spores produced at detached warts treated with the fungicides of recommended dilution. They showed diverse effect on sporulation. Carbendazim and azoxystrobin suppressed sporulation almost completely, the former irreversively. Thiram and folpet promoted sporulation as producing much more number of spores than untreated control. Others showed almost no effect on sporulation. Effects of suppression and promotion in the sporulation shown by the fungicides on the control of white rot were examined by incidences of disease and infection at the plots adopted the spray programs of which the fungicide at late May was substituted by carbendazim, azoxystrobin, folpet and thiram, respectively. Disease incidence and infection frequency at the plots sprayed former two chemicals which suppressed sporulation were much lower than those of the plots adopted latter two chemicals and untreated plot at which the fungicide spray was skipped at that time. These facts were reconfirmed in the experiments conducted with carbendazim and thiram, in which 100 fruits were bagged just prior to each spray from late May to late July for elucidating the effect of the two fungicides on the action of subsequent ones. Disease incidence and infection frequency on the fruit bagged just prior to each spray were gradually increased as the seasons going on. The increase rate at the carbendazim plot was much lower than that of thiram. Especially, the fruit infected till late July at the carbendazim plot were almost completely cured by the three fungicides, iminoctadine-triacetate, tebuconazole and samzinwang, a combined formular of iminoctadine-triacetate and difenoconazole, sprayed at late July and hence. In thiram plot, infected fruit were also cured by the 3 fungicides but not remarkable. From these results, it can be concluded that control efficiency of white rot can be greatly enhanced by selecting the fungicide capable of suppress the sporulation of white rot fungus at the season when the mass dispersal of spores is not initiated.
This paper discusses a method for optimizing the semi-actuated traffic signal control system by adjusting the initial interval according to the number of vehicles waiting for the green light in the actuated phase. We also present a Left-Turn actuated traffic signal control strategy that examines the vehicular noise in the detection area and determines the phase extension and the gap-out. In order to detect the vehicles in real-time, an image detector's Video Image Tracking technology was adopted. A 'Zone in Zone'method was implemented, and the image detection area is segmented into three zones: 1) Zone1 for verifying a vehicles obligatory presence, 2) Zone2 for counting the standby vehicles, and 3) Zone3 for examining the number of vehicles that have passed. The on-site assessment of the Left Turn Actuated Control is carried out using CORSIM, and the results show that the Control Delay decreased by 23.10%, 15.06%, and 4.34% compared to the delays resulted from pre-timed control, semi-actuated control-1 and semi-actuated control-2 traffic signal control systems respectively. The Queue Time also decreased by 36.24%, 20.10% and the Total Time by 14.36%, 7.02% for the same scenario. Which clearly demonstrates the operational efficiency. A sensitivity analysis reveals that the improvement from the propose traffic control strategy tends to increase as the through traffic volume reaches a saturated condition and the left-turn traffic volume decreases.
In order to assess antifungal activity of a wooden storage box, which was made of Paulownia tomentosa and used for keeping ancient documents, antifungal activity of volatile organic compounds emitted from the box was investigated along with qualitative analysis on major substances of the compounds. After collecting floating microorganisms inside air tester, the fungal activity was assessed by counting the number of colonies growing on TSA media. Compared to the control which collected 85 colonies from outdoor, 72 colonies were observed showing reduction rate of 14.82%. Through GC/MS and TDS system analysis, limonene was detected from the volatile organic compounds as characteristic features. When the fungal activity was assessed through fumigation by adding natural biocide BI and BII containing eugenol and anethole as major substances, both biocides showed a strong fungal activity with respectively 92.6%(inside the box) and 99.9%(outdoor) of reduction rate. Although these results didn't clarify antifungal activity of the volatile organic compounds emitted from the Paulownia-wood storage box and their functional components, it was at least confirmed that there is application possibility of natural biocide to use for preservation of ancient documents with increased efficiency in controlling pests of wooden storage boxes.
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