• Journal of The Korean Society of Inherited Metabolic disease
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• v.12 no.1
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• pp.5-13
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• 2012

I-cell disease (mucolipidosis type II; MIM 252500) and pseudo-Hurler polydystrophy (mucolipidosis type III; MIM 252600) are disorders caused by abnormal lysosomal transport in cells. The presence of numerous inclusion bodies in the cytoplasm of fibroblasts, a lack of mucopolysacchariduria, increased lysosomal enzyme activity in serum, and decreased GlcNAc-phosphotransferase activity are hallmark. Here, we attempted to investigate phenotypical and biochemical characteristics of the knockoutmouse of GlcNAc-phosphotransferase ${\alpha}/{\beta}$ subunits; in addition, we also attempted to determine whether the lysosome enriched fraction derived from placenta can be beneficial to phenotype and biochemistry of the knockout mouse.We found that the knockout mouse failed to thrive and had low bone density, as is the case in human. In addition, skin fibroblasts from the animal had the same biochemical characteristics, including increased lysosomal enzyme activity in the culture media, in contrast to the relatively low enzyme activity within the cells. Intravenous injection of the lysosome rich fraction derived from placenta into the tail vein of the animal resulted in a gain of weight, while saline injected animals didn't.In conclusion, our study demonstrated the phenotypical and biochemical similarities of the knockout mouse to a mucolipidosis type II patient and showed the therapeutic potential of the lysosome enriched fraction. We admit that a larger scale animal study will be needed; however, the disease model and the therapeutic potential of the lysosome enriched fraction will highlight the hope for a novel treatment approach to mucopolipidosis type II, for which no therapeutic modality is available.

• Journal of Life Science
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• v.24 no.11
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• pp.1244-1251
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• 2014

Platycodon grandiflorum (PG) has been known to possess many biological effects, including anti-inflammatory and anti-allergy activity and anti-obesity and hyperlipidemia effects. However, little research has been conducted regarding its anticancer effects, with the exception of its ability to stimulate apoptosis in skin cells. There has also been no study regarding PG-induced autophagy. The modulation of autophagy is recognized as one of the hallmarks of cancer cells. Depending on the type of cancer and the context, autophagy can suppress or help cancer cells to overcome metabolic stress and the cytotoxicity of chemotherapy. Therefore, the present study was designed to investigate whether or not extracts from PG-induced cell death were connected with autophagy and apoptosis in HCT-116 human colon cancer cells. PG stimulation decreased cell proliferation in a dose- and time-dependent manner and induced apoptosis, which was partially dependent on the activation of caspases. PG treatment also resulted in the formation of autophagic vacuoles simultaneously with regulation of autophagy-related genes. Interestingly, a PG-mediated apoptotic effect was further triggered by pretreatment with the autophagy inhibitors 3-methyladenin and bafilomycin A1. However, cell viability recovered quite well with bafilomycin A1 treatment. These findings show that PG treatment promotes both autophagy and apoptosis and that PG-induced autophagic response might play a role in the autophagic cell death of HCT-116 cells.

• Biomolecules & Therapeutics
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• v.27 no.5
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• pp.492-501
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• 2019

Nitrogen-containing heterocycles such as quinoline, quinazolinones and indole are scaffolds of natural products and have broad biological effects. During the last years those structures have been intensively synthesized and modified to yield new synthetic molecules that can specifically inhibit the activity of dysregulated protein kinases in cancer cells. Herein, a series of newly synthesized isoquinolinamine (FX-1 to 8) and isoindoloquinazolinone (FX-9, FX-42, FX-43) compounds were evaluated in regards to their anti-leukemic potential on human B- and T- acute lymphoblastic leukemia (ALL) cells. Several biological effects were observed. B-ALL cells (SEM, RS4;11) were more sensitive against isoquinolinamine compounds than T-ALL cells (Jurkat, CEM). In SEM cells, metabolic activity decreased with $10{\mu}M$ up to 26.7% (FX-3), 25.2% (FX-7) and 14.5% (FX-8). The 3-(p-Tolyl) isoquinolin-1-amine FX-9 was the most effective agent against B- and T-ALL cells with IC50 values ranging from 0.54 to $1.94{\mu}M$. None of the tested compounds displayed hemolysis on erythrocytes or cytotoxicity against healthy leukocytes. Anti-proliferative effect of FX-9 was associated with changes in cell morphology and apoptosis induction. Further, influence of FX-9 on PI3K/AKT, MAPK and JAK/STAT signaling was detected but was heterogeneous. Functional inhibition testing of 58 kinases revealed no specific inhibitory activity among cancer-related kinases. In conclusion, FX-9 displays significant antileukemic activity in B- and T-ALL cells and should be further evaluated in regards to the mechanisms of action. Further compounds of the current series might serve as templates for the design of new compounds and as basic structures for modification approaches.

• Journal of Life Science
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• v.17 no.11
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• pp.1497-1504
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• 2007

To investigate the function and secretion of human thrombopoietin (TPO) in mammalian cells, hTPO cDNA was cloned using human liver cDNA, and recombinant hTPO (rec-hTPO) was produced in CHO cell lines. In addition, six N-linked glycosylation sites were substituted for Ala to elucidate the role of each carbohydrate chain. To analyze the biological activity, rec-hTPO protein was injected subcutaneously. Blood was withdrawn for platelet determination. The metabolic clearance rate (MCR) was also analyzed at the 1, 4, 10 and 24 hr after tail vein injection. Wild-type TPO (WT) was efficiently secreted into the medium. However, a hTPO mutant with 116 deleted nucleotides detected by PCR cloning was not secreted. The N-linked glycosylation sites had nearly the same expression quantity as rec-hTPO WT apart from mutants 3 and 4. The glycosylation site of mutant 4 appeared to be an indispensable site for hTPO secretion. Also characterized was the biological activity through an injection with rec-hTPO (10 ng) to ICR mice (7 weeks). The result of the blood analysis showed a considerable increase in the platelet number six days after He injection. To analyze the pharmacokinetics, rec-hTPO was injected into the tail vein (5 ng). The result was 200 pg/ml 1hr after this injection. Following this, it dramatically decreased and virtually disappeared 10 hours after the injection. Thus, rec-hTPO may be a treatment for thrombopenia by the production of the high active rec-hTPO. In addition, hTPO can permit the development of potent new analogues that stimulate the platelet value.

• Journal of Nutrition and Health
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• v.48 no.1
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• pp.1-8
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• 2015

Purpose: Anthocyanins from purple sweet potato (PSP) have been investigated in vitro and in animals and found to have a protective effect against oxidative hepatic damage. In this study, we investigated that aqueous extract of PSP can ameliorate the dysfunction of lipid metabolism in mice fed a high fat/cholesterol diet. Methods: Forty C57BL/6J mice were randomly divided into 5 groups (n = 8) and fed one of the following diets for 8 weeks; normal fat (NF) diet; high fat/cholesterol (HFC) diet; HFC with 1.25% PSP (HFPL) diet; HFC with 2.5% PSP (HFPM) diet; HFC with 5% PSP (HFPH) diet. Results: Non-alcoholic fatty liver was manifested in the HFC group by showing increased levels in plasma alanine aminotransferase (ALT) activity, total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C), increased level of TC and presence of many large lipid droplets in the liver, and increased fat cell size in the HFC group compared with the NF group. However, administration of HFC induced a significant decrease in food intake, resulting in decrease in fat mass. Co-administration of PSP did not lead to reversal of body weight changes, ALT activity, and lipid levels in plasma and the liver, but suppressed excess enlargement of the fat cell size through increasing carnitine palmitoyltransferase-1 (CPT-1) gene expression in the liver. Accordingly, the number of fat droplets in the liver was reduced in PSP administered groups. Conclusion: Taken together, these results suggest that PSP may have a protective effect on the dysfunction of lipid metabolism. Conduct of further studies on the coordinated regulation of PSP for lipid metabolic homeostasis at the liver-adipose tissue axis is needed.

• Journal of Animal Science and Technology
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• v.58 no.1
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• pp.2.1-2.10
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• 2016

Background: Heat stress adversely affects the physiological and metabolic status, and the productive performance of buffalo. Methods: The present study was conducted to explicate the effect of misting and wallowing cooling strategies during heat stress in lactating Murrah buffalo. The study was conducted for three months (May-July) of which first two months were hot dry and last month was hot humid. Eighteen lactating buffaloes, offered the same basal diet, were blocked by days in milk, milk yield and parity, and then randomly allocated to three treatments: negative control (no cooling), cooling by misting, and cooling by wallowing. Results: The results showed higher (P < 0.05) milk yield in buffaloes of misting and wallowing group compared to control during the experimental period however wallowing was found more (P < 0.05) effective during July (hot humid period). Both the treatments resulted into significant (P < 0.05) reduction in rectal temperature (RT) and respiratory rate (RR) compared to control animals during study period whereas wallowing was found to be effective on pulse rate (PR) only during July. Both treatments were resulted in mitigating the heat stress mediated decrease in packed cell volume (PCV), lymphocytopnoea and neutrophilia whereas decrease in total erythrocyte count (TEC) and monocytes was only mitigated by wallowing. Heat load induced alteration in serum creatinine and sodium concentration was significantly (P < 0.05) ameliorated by misting and wallowing whereas aspartate aminotransferase, alkaline phosphatase and superoxide dismutase activity, and reactive oxygen species concentration could be normalized neither by misting nor by wallowing. The significant (P < 0.05) increment in serum cortisol and prolactin levels observed in June and July period in control animals was significantly (P < 0.05) prevented by misting and wallowing. Conclusions: It can be concluded that misting and wallowing were equally effective in May and June (hot dry period) whereas wallowing was more effective during hot humid period in preventing a decline in milk production and maintaining physiological, metabolic, endocrine and redox homeostasis.

• Journal of Korean Society of Water and Wastewater
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• v.35 no.6
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• pp.405-415
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• 2021

MMBR system has been suggested as a promising system to resolve harvesting problems induced from low settling efficiency of microalgae. And recently, a lot of research on reducing fouling at the MMBR system has investigated focused on EPS in many cases. EPS of microalgae mainly consists of polysaccharides and protein components, and is produced through photosynthesis and nitrogen-carbon metabolic pathways. Especially, P-EPS is one of major compounds which occur membrane fouling phenomenon, as its hydrophobic protein components cause floc formation and cake layer accumulation. And it is already known that almost every microalgae can metabolize P-EPS or Chl-a when nitrogen sources as a substrate is insufficient or exhausted situation. With the above backgrounds, uptake rates of P-EPS or Chl-a by Scenedesmus quadricauda according to the type of carbon source and nitrogen concentration were evaluated in order to verify correlation between carbon source vs P-EPS production, and indeed Scenedesmus quadricauda uses P-EPS or Chl-a when the amounts of nitrogen sourc es in the feed is not satisfied. As a result, it was shown that P-EPS and Chl-a production were increased proportional to nitrogen concentration under organic carbon condition. And especially, the amo unts of P-EPS and Chl-a in the cell were diminished with the nitrogen source becomes insufficient or exhausted. Because P-EPS accelerates fouling at the MMBR system, P-EPS degradation by Scenedesmus quadricauda in order to get nitrogen source may contribute to reducing fouling. About a affects of N-consumed Chl-a to the MMBR fouling, more survey is needed. On the contrary, considering the purpose of MMBR system of this study, i.e. harvesting useful high value microalgae efficiently feeding adequate industrial process wastewater, it seems like difficult to maintain satisfied metabolic activity and to harvest with high yield rate using nitrogen-poor MMBR feed.

• Korean Journal of Weed Science
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• v.12 no.3
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• pp.261-270
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• 1992

Many herbicides that are applied at the soil before weed emergence inhibit plant growth soon after weed germination occurs. Plant growth has been known as an irreversible increase in size as a result of the processes of cell divison and cell enlargement. Herbicides can influence primary growth in which most new plant tissues emerges from meristmatic region by affecting either or both of these processes. Herbicides which have sites of action during interphase($G_1$, S, $G_2$) of cell cycle and cause a subsequent reduction in the observed frequency of mitotic figures can be classified as an inhibitor of mitotic entry. Those herbicides that affect the mitotic sequence(mitosis) by influencing the development of the spindle apparatus or by influencing new cell plate formation should be classified as causing disruption of the mitotic sequence. Sulfonylureas, imidazolinones, chloroacetamides and some others inhibit plant growth by inhibiting the entry of cell into mitosis. The carbamate herbicides asulam, carbetamide, chlorpropham and propham etc. reported to disrupt the mitotic sequence, especially affecting on spindle function, and the dinitroaniline herbicides trifluralin, nitralin, pendimethalin, dinitramine and oryzalin etc. reported to disrupt the mitotic sequence, particularly causing disappearence of microtubles from treated cells due to inhibition of polymerization process. An inhibition of cell enlargement can be made by membrane demage, metabolic changes within cells, or changes in processes necessary for cell yielding. Several herbicides such as diallate, triallate, alachlor, metolachlor and EPTC etc. reported to inhibit cell enlargement, while 2, 4-D has been known to disrupt cell enlargement. One potential danger inherent in the use of soil acting herbicides is that build-up of residues could occur from year to year. In practice, the sort of build-up that would be disastrous is unikely to occur for substances applied at the correct soil concentration. Crop injury caused by soil applied herbicides can be minimized by (1) following the guidance of safe use of herbicides, particularly correct dose at correct time in right crop, (2) by use of safeners which protect crops against injury without protecting any weed ; interactions between herbicides and safeners(antagonists) at target sites do occur probably from the following mechanisms (1) competition for binding site, (2) circumvention of the target site, and (3) compensation of target site, and another mechanism of safener action can be explained by enhancement of glutathione and glutathione related enzyme activity as shown in the protection of rice from pretilachlor injury by safener fenclorim, (3) development of herbicide resistant crops ; development of herbicide-resistant weed biotypes can be explained by either gene pool theory or selection theory which are two most accepted explanations, and on this basis it is likely to develop herbicide-resistant crops of commercial use. Carry-over problems do occur following repeated use of the same herbicide in an extended period of monocropping, and by errors in initial application which lead to accidental and irregular overdosing, and by climatic influence on rates of loss. These problems are usually related to the marked sensitivity of the particular crops to the specific herbicide residues, e.g. wheat/pronamide, barley/napropamid, sugarbeet/ chlorsulfuron, quinclorac/tomato. Relatively-short-residual product, succeeding culture of insensitive crop to specific herbicide, and greater reliance on postemergence herbicide treatments should be alternatives for farmer practices to prevent these problems.

• Journal of Life Science
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• v.21 no.12
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• pp.1752-1760
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• 2011

In this study, the effect of the Opuntiahumifusa extracts on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and ROS level of a cell was investigated using an osteoblast. Opuntiahumifusawas separated intoOpuntiahumifusapeel (OH-P), seed (OH-Se) and stem (OH-St).These were subjected to extraction by using hot water and ethanol. The proliferation of the MC3T3-E1 osteoblastic cells that were treated with OH-Se water extract were increased by approximately 120%. Regarding the effects of OH-Se on ALP activity, the $50{\mu}g/ml$ ethanol extract group showed the highest activity. The synthesis of collagen increased significantly in response to treatment with OH-Se water extract. The ROS scavenging effects of Opuntiahumifusawere investigated for involvement of oxidativedamage, cell culture and staining. Also, when OH-Se water extract $100{\mu}g/ml$ was added, the ROS level decreased by 54%. These results indicate that Opuntiahumifusa extracts have an anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

• Journal of Microbiology and Biotechnology
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• v.21 no.6
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• pp.637-645
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• 2011

Production of recombinant proteins by excretory expression has many advantages over intracellular expression in Escherichia coli. Hyperexpression of a secretory exoglucanase, Exg, of Cellulomonas fimi was previously shown to saturate the SecYEG pathway and result in dramatic cell death of E. coli. In this study, we demonstrated that overexpression of the PspA in the JM101(pM1VegGcexL-pspA) strain enhanced excretion of Exg to 1.65 U/ml using shake-flask cultivation, which was 80% higher than the highest yield previously obtained from the optimized JM101(pM1VegGcexL) strain. A much higher excreted Exg activity of 4.5 U/ml was further achieved with high cell density cultivation using rich media. Furthermore, we showed that the PspA overexpression strain enjoyed an elevated critical value (CV), which was defined as the largest quotient between the intracellular unprocessed precursor and its secreted mature counterpart that was still tolerable by the host cells prior to the onset of cell death, improving from the previously determined CV of 20/80 to the currently achieved CV of 45/55 for Exg. The results suggested that the PspA overexpression strain might tolerate a higher level of precursor Exg making use of the SecYEG pathway for secretion. The reduced lethal effect might be attributable to the overexpressed PspA, which was postulated to be able to reduce membrane depolarization and damage. Our findings introduce a novel strategy of the combined application of metabolic engineering and construct optimization to the attainment of the best possible E. coli producers for secretory/excretory production of recombinant proteins, using Exg as the model protein.