Although rational genetic engineering is nowadays the favored method for microbial strain improvement, building up mutant libraries based on directed evolution for improvement is still in many cases the better option. In this regard, the demand for precise and efficient screening methods for mutants with high performance has stimulated the development of biosensor-based high-throughput screening strategies. Genetically encoded biosensors provide powerful tools to couple the desired phenotype to a detectable signal, such as fluorescence and growth rate. Herein, we review recent advances in engineering several classes of biosensors and their applications in directed evolution. Furthermore, we compare and discuss the screening advantages and limitations of two-component biosensors, transcription-factor-based biosensors, and RNA-based biosensors. Engineering these biosensors has focused mainly on modifying the expression level or structure of the biosensor components to optimize the dynamic range, specificity, and detection range. Finally, the applications of biosensors in the evolution of proteins, metabolic pathways, and genome-scale metabolic networks are described. This review provides potential guidance in the design of biosensors and their applications in improving the bioproduction of microbial cell factories through directed evolution.
Woo Young Jeon;Jung Ho Park;Jeong-Kui Ku;Jin-A Baek;Seung-O Ko
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.49
no.5
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pp.287-291
/
2023
Odontogenic keratocysts (OKCs) located in the maxillae have rarely been reported in the literature. Standard treatment modalities for OKC range from marsupialization to marginal resection. However, most of the studies on OKC treatment have been related to mandibular OKCs. The anatomical structure and loose bone density of the maxillae and the empty space of the maxillary sinus could allow rapid growth of a lesion and the ability to tolerate tumor occupancy in the entire maxilla within a short period of time. Therefore, OKCs of the maxillae require more aggressive surgery, suchas resection. As an alternative, this report introduces a modified Carnoy's solution, a strong acid, as an adjuvant chemotherapy after cyst enucleation. This report describes the clinical outcomes of enucleation using a modified Carnoy's solution in patients with large OKCs on the posterior maxillae. In three cases, application of a modified Carnoy's solution had few side effects or morbidity. Each patient was followed for four to six years, and none showed any signs of recurrence. In conclusion, adjuvant treatment with a modified Carnoy's solution can be considered a treatment option capable of reducing the recurrence rate of OKC in the maxillae.
The potential of tivozanib as a treatment for oral squamous cell carcinoma (OSCC) was explored in this study. We investigated the effects of tivozanib on OSCC using the Ca9-22 and CAL27 cell lines. OSCC is a highly prevalent cancer type with a significant risk of lymphatic metastasis and recurrence, which necessitates the development of innovative treatment approaches. Tivozanib, a vascular endothelial growth factor receptor inhibitor, has shown efficacy in inhibiting neovascularization in various cancer types but has not been thoroughly studied in OSCC. Our comprehensive assessment revealed that tivozanib effectively inhibited OSCC cells. This was accompanied by the suppression of Bcl-2, a reduction in matrix metalloproteinase levels, and the induction of intrinsic pathway-mediated apoptosis. Furthermore, tivozanib contributed to epithelial-to-mesenchymal transition (EMT) inhibition by increasing E-cadherin levels while decreasing N-cadherin levels. These findings highlight the substantial anticancer potential of tivozanib in OSCC and thus its promise as a therapeutic option. Beyond reducing cell viability and inducing apoptosis, the capacity of tivozanib to inhibit EMT and modulate key proteins presents the possibility of a paradigm shift in OSCC treatment.
Objective : In order to develop a novel nerve guidance channel using porcine small intestinal submucosa (SIS) for nerve regeneration, we investigated the possibility of SIS, a tissue consisting of acellular collagen material without cellular immunogenicity, and containing many kinds of growth factors, as a natural material with a new bioactive functionality. Methods : Left sciatic nerves were cut 5 mm in length, in 14 Sprague-Dawley rats. Grafts between the cut nerve ends were performed with a silicone tube (Silicon group, n=7) and rolled porcine SIS (SIS group, n=7). All rats underwent a motor function test and an electromyography (EMG) study on 4 and 10 weeks after grafting. After last EMG studies, the grafts, including proximal and distal nerve segments, were retrieved for histological analysis. Results : Foot ulcers, due to hypesthesia, were fewer in SIS group than in Silicon group. The run time tests for motor function study were 2.67 seconds in Silicon group and 5.92 seconds in SIS group. Rats in SIS group showed a better EMG response for distal motor latency and amplitude than in Silicon group. Histologically, all grafts contained some axons and myelination. However, the number of axons and the degree of myelination were significantly higher in SIS group than Silicon group. Conclusion : These results show that the porcine SIS was an excellent option as a natural biomaterial for peripheral nerve regeneration since this material contains many kinds of nerve growth factors. Furthermore, it could be used as a biocompatible barrier covering neural tissue.
Journal of The Korean Society of Agricultural Engineers
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v.56
no.1
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pp.89-99
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2014
The cultivation of biomass crops is now global demand for decreasing emissions of carbon dioxide ($CO_2$) from fossil fuel. Miscanthus species have been studied as a suitable crop for biomass production, due to its characteristics of fast growth and high biomass. In Korea, Miscanthus species have gained wide attention as an option for biomass production alternative to fossil fuels, recently. New strain of giant Miscanthus has been developed and two large trial sites for the giant Miscanthus production were built in the lower reaches of the Geum River. To evaluate the ecological influence of the giant Miscanthus as an bioenergy crop for the future, we investigated the impact of the construction of the giant Miscanthus production fields on the biota and also compared it with biota in paddy fields near the study sites. The biota including plants, amphibians, reptiles, mammals, avifauna, insects, and bugs was investigated. The plant diversity of the giant Miscanthus production fields was poorer than the paddy fields because the high height of the giant Miscanthus might hinder the growth of other plants. However, the giant Miscanthus production fields serves habitat to animals, leading to rich diversity of animals including avifauna, insects, and bugs. The rich diversity of the animals in the giant Miscanthus production fields coincides with the fact that the giant Miscanthus was grown without any pesticide, herbicide, and fertilizer. This study showed that the giant Miscanthus can influence on biota and further long term study is needed to elucidate the interaction between the diversity of biota and the giant Miscanthus.
Journal of the Korean Institute of Landscape Architecture
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v.33
no.2
s.109
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pp.32-47
/
2005
This study was carried out to investigate and propose a green-wall system with evergreen plants for urban greening of Tongyeong City. To achieve these goals, the requirements and possibilities for wall greening were investigated and evaluated considering the location, topography, and climate of Tongyeong City. Existing walls were analyzed and then a suitable green wall system is proposed. Tongyeong City and its 151 islands covers the central and the southern parts of the Goseong peninsula. Most of the land is covered with hills and mountains; $43.9\%$ of the land area has a slope greater than $15\%$ and most hills and mountains near the urban area have a slope of more than $30\%$. As a result of the topographical properties, concrete retaining walls can often be seen along the streets in urbanized areas. These retaining walls are not only unattractive, but they also create environmental problems, and thus should be replaced with native evergreen plants. Options for replacing the retaining walls include evergreen vine-plants such as Hedaa spp. and Euonymus radicans, but native evergreen shrubs such as Pittosporum tobira, Nandina domestica, Raphiolepis umbellata, Ilex cornuta, flex crenata, Fatsia japonic, and Aucuba japonica may be a more attractive option. Current wall conditions are unsuitable for planting vines, therefore, a reservoir-drainage-type plant box filled with a light artificial substrate is required for greening these concrete retaining walls. These might be irrigated in the dry season and fertilized annually by an appropriate system. These plant boxes could be attached along the entire walls. An experiment investigating effects of substrates and bark-chip mulching on the growth of Hedera spp. showed that the mixture of cerasoil and field soil(v/v, 4:6) was superior to field soil alone and to the mixture of perlite small grain, large grain, and field soil(v/v/v, 2:2:6). Bark-chip mulching tended to increase the growth of Hedera spp..
Background: Breast cancer evolution and tumor progression are controlled by complex interactions between steroid receptors and growth factor receptor signaling. Aberrant growth factor receptor signaling can augment or suppress estrogen receptor function in hormone-dependent breast cancer cells. Thus, we aimed to investigate antitumor effects of sorafenib and lapatinib alone and in combination on MCF-7 breast cancer cells. Materials and Methods: Cytotoxicity of the sorafenib and lapatinib was tested in MCF-7 cells by XTT assays. 50, 25, 12.5 and $6.25{\mu}M$ concentrations of sorafenib and 200, 100, 50 and $25{\mu}M$ concentrations of lapatinib were administered alone and in combination. Results were evaluated as absorbance at 450nM and $IC_{50}$ values are calculated according to the absorbance data Results: Both sorafenib and lapatinib showed concentration dependent cytotoxic effects on MCF-7 cells. Sorafenib exerted cytotoxic effects with an $IC_{50}$ value of $32.0{\mu}M$; in contrast with lapatinib the $IC_{50}$ was $136.6{\mu}M$. When sorafenib and lapatinib combined, lapatinib increased cytotoxic effects of sorafenib at its ineffective concentrations. Also at the concentrations where both drugs had cytotoxic effects, combination show strong anticancer effects and killed approximately 70 percent of breast cancer cells. Conclusions: Combinations of tyrosine kinase inhibitors and cytotoxic agents or molecular targeted therapy has been successful for many types of cancer. The present study shows that both sorafenib and lapatinib alone are effective in the treatment of breast cancer. Also a combination of these two agents may be a promising therapeutic option in treatment of breast cancer.
Background: Saponins are a major active component for the traditional Chinese medicine, Rubus parvifolius L., which has shown clear antitumor activities. However, the specific effects and mechanisms of saponins of Rubus parvifolius L. (SRP) remain unclear with regard to human chronic myeloid leukemia cells. The aim of this study was to investigate inhibition of proliferation and apoptosis induction effects of SRP in K562 cells and further elucidate its regulatory mechanisms. Materials and Methods: K562 cells were treated with different concentrations of SRP and MTT assays were performed to determine cell viability. Apoptosis induction by SRP was determined with FACS and DAPI staining analysis. Western blotting was used to detect expression of apoptosis and survival related genes. Specific inhibitors were added to confirm roles of STAT3 and AMPK pathways in SRP induction of apoptosis. Results: Our results indicated that SRP exhibited obvious inhibitory effects on the growth of K562 cells, and significantly induced apoptosis. Cleavage of pro-apoptotic proteins was dramatically increased after SRP exposure. SRP treatment also increased the activities of AMPK and JNK pathways, and inhibited the phosphorylation expression level of STAT3 in K562 cells. Inhibition of the AMPK pathway blocked the activation of JNK by SRP, indicating that SRP regulated the expression of JNK dependent oon the AMPK pathway. Furthermore, inhibition of the latter significantly conferred resistance to SRP pro-apoptotic activity, suggesting involvement of the AMPK pathway in induction of apoptosis. Pretreatment with a STAT3 inhibitor also augmented SRP induced growth inhibition and cell apoptosis, further confirming roles of the STAT3 pathway after SRP treatment. Conclusions: Our results demonstrated that SRP induce cell apoptosis through AMPK activation and STAT3 inhibition in K562 cells. This suggests the possibility of further developing SRP as an alternative treatment option, or perhaps using it as adjuvant chemotherapeutic agent for chronic myeloid leukemia therapy.
A study was conducted to compare the effect of halal slaughter without stunning and gas stun killing followed by bleeding on residual blood content and storage stability of rabbit meat. Eighty male New Zealand white rabbits were divided into two groups of 40 animals each and subjected to either halal slaughter without stunning (HS) or gas stun-kill (GK). The volume of blood lost during exsanguination was measured. Residual blood was further quantified by determination of haemoglobin content in Longissimus lumborum (LL) muscle. Storage stability of the meat was evaluated by microbiological analysis and measuring lipid oxidation in terms of thiobarbituric acid reactive substances (TBARS). HS resulted in significantly higher blood loss than GK. HS had significantly lower residual haemoglobin in LL muscle compared to GK. Slaughter method had no effect on rabbit meat lipid oxidation at 0, 1, and 3 d postmortem. However, at 5 and 8 days of storage at $4^{\circ}C$, significant differences (p<0.05) were found, with meat from the GK group exhibiting significantly higher levels of MDA than that from HS. At day 3, greater growth of Pseudomonas aeroginosa and E. coli were observed in the GK group (p<0.05) with B. thermosphacta and total aerobic counts remained unaffected by slaughter method. At days 5 and 7 postmortem, bacterial counts for all tested microbes were affected by slaughter method, with GK exhibiting significantly higher growth than HS. It can be concluded that slaughter method can affect keeping quality of rabbit meat, and HS may be a favourable option compared to GK due to high bleed out.
Journal of the Korean Society of Environmental Restoration Technology
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v.21
no.3
/
pp.99-110
/
2018
Compared to yellow dust coming from China or particulate matter created naturally in spring due to Total Suspended Particulate(TSP), particulate matter in winter season have much more serious effect on human body as they penetrate cell membranes. Although such particulate matter are becoming a social issue, there are no concrete plans on how to reduce them. Air-purifying plants are limited in maintaining the indoor air quality of large area because it is usually difficult to quantify their performance. In order to improve this, a bio-filter that can be connected to air conditioner is suggested as an option. This study seeks to improve air conditioning model-based monitoring method for bio-filters from prior studies and objectify correlations between applied vegetation and growing environment into quantitative indicators. By doing so, this study seeks to provide criteria on plants applied to artificial soil-based vegetation bio-filters and basic information to set air-conditioning features. The study results confirmed significant tendency on the growing stability of each purifying plant in mechanical air-conditioning environment. Among three models selected for bio-filter vegetation models, epipremnum aureum showed high performance in quantitative indicators, including soil moisture, EC, and leaf temperature, etc., indicating that it would assure the highest growing stability in this test air-conditioning environment.
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