Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results: The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS (KISS1-hepatitis B surface antigen S) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction. Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.
Thermal acid hydrolysis pretreatment of Kappaphycus alvarezii was carried out with 12% (w/v) seaweed slurry and 180 mM H2SO4 at 140°C for 5 min. Utility of the thermotolerant yeast Kluyveromyces marxianus KCTC7150 was evaluated with respect to cell growth and ethanol fermentation at 40°C was close to optimal for enzymatic hydrolysis. This could lead to the integration of both the saccharification and fermentation processes. The levels of ethanol production by simultaneous saccharification and fermentation (SSF) with non-adapted and adapted K. marxianus KCTC7150 were 9.1 g/l with an ethanol yield (YEtOH) of 0.24 and 10.2 g/l with an ethanol yield (YEtOH) of 0.27 at 156 h, respectively. The two-phase SSF process was employed in this study to improve the efficiency of ethanol fermentation. Adapted K. marxianus KCTC7150 using the two-phase SSF process produced 13.5 g/l with an ethanol yield (YEtOH) of 0.35 at 96 h. Development of the two-phase SSF process could enhance the overall ethanol fermentation yields of the seaweed K. alvarezii.
Kim, Bae-Chul;Han, Rong-Xun;Kim, Myung-Yoon;Shin, Young-Min;Park, Chang-Sik;Jin, Dong-Il
Reproductive and Developmental Biology
/
v.33
no.2
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pp.107-111
/
2009
The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.
Journal of the Institute of Electronics Engineers of Korea TC
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v.46
no.4
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pp.29-39
/
2009
The need for radio spectrum is recently considered as a huge hurdle towards the rapid development of wireless networks. Large parts of the spectrum are allocated to licensed radio services in proprietary way. However, enormous success of the wireless services and technologies in the unlicensed bands has brought new ideas and innovations. In recent years cognitive radio has gained much attention for solving the spectrum scarcity problem. It changes the way spectrum is regulated so that more efficient spectrum utilization is possible. Multi-hop relay technology on the other hand has intensively been studied in the area of ad hoc and peer-to-peer networks. But in cellular network, only recently the integration of multi-hop capability is considered to enhance the performance significantly. Multi-hop relaying can extend the coverage of the cell to provide high data rate service to a greater distance and in the shadowed regions. Very few papers still exist that combine these methods to maximize the spectrum utilization. Thus we propose a network architecture combining these two technologies in a way to maximize the system throughput. We present the throughput capacity equations for the proposed system model considering various system parameters like utilization factor by the primary users and primary users' transmission radius and through extensive numerical simulations we analyze the significance of work.
Journal of the Institute of Electronics and Information Engineers
/
v.49
no.12
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pp.209-218
/
2012
Recently, the Field Programmable Stateful Logic Array (FPSLA) was proposed as one of the most promising system integration technologies which will extend the life of the Moore's law. This work is the first proposal of the FPSLA design automation flow, and the approaches to logic synthesis, synchronization, physical mapping, and automatic placement of the FPSLA designs. The synchronization at each gate for pipelining determines the x-coordinates of cells, and reduces the placement to 1-dimensional problems. The objective function and its gradients for the non-linear optimization of the net length and placement density have been remodeled for the reduced global placement problem. Also, a recursive algorithm has been proposed to legalize the placement by relaxing the density overflow of bipartite bin groups in a top-down hierarchical fashion. The proposed model and algorithm are implemented, and validated by applying them to the ACM/SIGDA benchmark designs. The output state of a gate in an FPSLA needs to be duplicated so that each fanout gate can be connected to a dedicated copy. This property has been taken into account by merging the duplicated nets into a hyperedge, and then, splitting the hyperedge into edges as the optimization progresses. This yields additional 18.4% of the cell count reduction in the most dense logic stage. The practicality of the FPSLA can be further enhanced primarily by incorporating into the logic synthesis the constraint to avoid the concentrated fains of gates on some logic stages. In addition, an efficient algorithm needs to be devised for the routing problem which is based on a complicated graph. The graph models the nanowire crossbar which is trimmed to be embedded into the FPSLA fabric, and therefore, asymmetric. These CAD tools can be used to evaluate the fabric efficiency during the architecture enhancement as well as automate the design.
B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi1) is a polycomb group protein and a core component of polycomb repressive complex 1. Initial research into Bmi1 has focused on its role in tumorigenesis, and it is generally accepted that it is important for the proliferation and survival of cancer cells. However, more recent studies have revealed that Bmi1 is downregulated in brains with neurodegenerative disease and that it regulates the function of mitochondria and reactive oxygen species levels. In this study, we tested the therapeutic potential of Bmi1 in pilocarpine-induced seizures in Bmi1-knockout mice. Bmi1 expression transiently increased in the hippocampal CA1 and CA3 and the dentate gyrus following pilocarpine-induced status epilepticus (SE). In terms of seizure behavior, SE induction was 43.14% and 53.57% for Bmi1+/+ and Bmi1+/- mice, respectively. However, there was no significant difference in mortality or hippocampal damage between the two groups. Two months after SE induction, the frequency of epileptic seizures in the Bmi1+/- mice was 50% lower than in the control group, although the difference was not statistically significant. In addition, mossy fiber outgrowth in the Bmi1+/- mice was significantly higher than in their wild-type littermates. Taken together, these data indicate that reduced Bmi1 activity increases pilocarpine-induced seizure probability and mossy fiber outgrowth.
To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. In situ hybridization demonstrated that the transfection rate of sperm cells with and without BSA was 41 and 68% respectively, when the cells were treated with liposome/DNA complexes and 13% for DNA alone. LSC analysis showed that the binding of exogenous DNA was greatly reduced by DNase I treatment which digests DNA bound onto spermatozoa, suggesting that some of the DNA was internalized into the sperm membrane. To find out whether transfected DNA was internalized into sperm intracytomembrane, sperm DNA was amplified by inverse PCR. No PCR products were detected from sperm cells, indicating that the foreign DNA was simply bound onto the sperm membrane. To investigate transfer rates of exogenous DNA into oocytes via sperm cells, we used immunofluorescene method to follow the distribution of foreign DNA via spermatozoa: a few exogenous DNA was located in the cytoplasm of early embryos (13/60, 21.7% for DNA+/liposome+/BSA) and was not located in the pronucleus and/or nucleus. These results suggest that most of the transfected sperm cells could carry the foreign DNA into the egg by in vitro fertilization, but that the transferred DNA is degraded in the developing embryos without stable integration into the zygote genome. Therefore, we have directly injected with transfected sperm cell into oocyte cytoplasm and observed that some of the exogenous DNA was detected in preimplantation embryonic cytoplasm and expressed at preimplantation stages, suggesting that exogenous DNA in early zygote has their integrity. In this study, we have not identified a noble mechanism that interfering transportation of foreign DNA into zygote genome via spermatozoa. Our data, however, demonstrated that inverse PCR and immunofluorescene methods would be used as a new tool for follow-up of gene distribution in oocyte via sperm cells.
The non infecting, plant associated bacteria have attracted increased attention for stimulating plant growth and as environmental friendly plant protecting agents. Pink-pigmented facultatively methylotrophic bacteria (PPFMs), classified as Methylobacterium spp., are persistent colonizers of plant leaf surfaces. As the leaves of most or all plants harbor PPFMs that utilize leaf methanol as their sole source of carbon and energy, which is a specific attribute of the genus Methylobacterium. Although they are not well known, these bacteria are co-evolved, interacting partners in plant metabolism. This claim is supported, for example, by the following observations: (1) PPFMs are seed-transmitted, (2) PPFMs are frequently found in putatively axenic cell cultures, (3) Low numbers of seed-borne PPFMs correlate with low germinability, (4) Plants with reduced numbers of PPFM show elevated shoot/root ratios, (5) Foliar application of PPFMs to soybean during pod fill enhances seed set and yield, (6) Liverwort tissue in culture requires PPFM-produced vitamin B12 for growth, (7) treated plants to suppress or decrease disease incidence of sheath blight caused by Rhizoctonia solani in rice, and (8) the PPFM inoculation induced number of stomata, chlorophyll concentration and malic acid content, they led to increased photosynthetic activity. Methylobacterium spp. are bacterial symbionts of plants, shown previously to participate in plant metabolism by consuming plant waste products and producing metabolites useful to the plant. There are reports that inform about the beneficial interactions between this group of bacteria and plants. Screening of such kind of bacteria having immense plant growth promoting activities like nitrogen fixation, phytohormone production, alleviating water stress to the plants can be successfully isolated and characterized and integration of such kind of organism in crop production will lead to increased productivity.
To understand the basic physics underlying large spatial fluctuations of intensity and Doppler shift, we have investigated the dynamical charctersitics of the transition region of the quiet sun by analyzing a raster scan of high resolution UV spectral band containing H Lyman lines and a S VI line. The spectra were taken from a quiet area of $100'\times100'$ located near the disk center by SUMER on board SOHO. The spectral band ranges from 906 A to 950 A with spatial and spectral resolution of 1v and $0.044 {\AA}$, respectively. The parameters of individual spectral lines were determined from a single Gaussian fit to each spectral line. Then, spatial correlation analyses have been made among the line parameters. Important findings emerged from the present analysis are as follows. (1) The integrated intensity maps of the observed area of H I 931 line $(1\times10^4 K)$ and S VI 933 line $(2\times10^5 K)$ look very smilar to each other with the same characterstic size of 5". An important difference, however, is that the intensity ratio of brighter network regions to darker cell regions is much larger in S VI 933 line than that in H I 931 line. (2) Dynamical features represented by Doppler shifts and line widths are smaller than those features seen in intensity maps. The features are found to be changing rapidly with time within a time scale shorter than the integration time, 110 seconds, while the intensity structure remains nearly unchanged during the same time interval. (3) The line intensity of S VI is quite strongly correlated with that of H I lines, but the Doppler shift correlation between the two lines is not as strong as the intensity correlation. The correlation length of the intensity structure is found to be about 5.7' (4100 km), which is at least 3 times larger than that of the velocity structure. These findings support the notion that the basic unit of the transition region of the quiet sun is a loop-like structure with a size of a few $10^3 km$, within which a number of unresolved smaller velocity structures are present.
The cryparin of Cryphonectria parasitica belongs to a cell wall associated fungal hydrophobin. The cryparin, though it is encoded by a single copy gene, is known for the high expression during the liquid culture of C. parasitica, and it turns out that 22% of total mRNA was transcribed for cryparin at 48hr after the liquid culture. In addition, it is also known as one of down-regulated fungal proteins by the presence of double stranded RNA virus, Cryphonectria hypovirus 1. In previous studies (Kim et al., 1999), we have constructed a cryparin-null mutant by replacing the cryparin gene with hygromycin B resistance gene due to site directed homologous recombination. In order for the promoter analysis of cryparin which seems to be very strong as well as mycoviral specific, it is preferable to have a strain with only a target promoter replaced and a discernable target site for incoming vectors. However, the cryparin-null mutant revealed the presence of an additional copy of transforming vector except the one which replaced the cryparin gene. In addition, the cryparin-null mutant did not contain any markers for targeted integration of incoming vectors. This prompts us to design an experiment to obtain a strain for promoter analysis of cryparin gene. A different mating type strain EP6(Mata, $met^-$) was mated with the cryparin-null mutant ${\triangle}$Crp194-7(MatA, Crp${\triangle}$::hph) to make the progenies with only a single replacement vector and $met^-$ characteristic remained. Nutritional assay as well as Southern blot analysis revealed that the progeny, ${\triangle}$Crp194-a6, was the methionine auxotroph with a single replacing vector in genome. Northern blot analysis and PAGE showed that there was no cryparin produced in this bred strain either.
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