• BMB Reports
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• v.48 no.8
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• pp.438-444
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• 2015

Lysosomal storage diseases (LSDs) are a group of inherent diseases characterized by massive accumulation of undigested compounds in lysosomes, which is caused by genetic defects resulting in the deficiency of a lysosomal hydrolase. Currently, enzyme replacement therapy has been successfully used for treatment of 7 LSDs with 10 approved therapeutic enzymes whereas new approaches such as pharmacological chaperones and gene therapy still await evaluation in clinical trials. While therapeutic enzymes for Gaucher disease have N-glycans with terminal mannose residues for targeting to macrophages, the others require N-glycans containing mannose-6-phosphates that are recognized by mannose-6-phosphate receptors on the plasma membrane for cellular uptake and targeting to lysosomes. Due to the fact that efficient lysosomal delivery of therapeutic enzymes is essential for the clearance of accumulated compounds, the suitable glycan structure and its high content are key factors for efficient therapeutic efficacy. Therefore, glycan remodeling strategies to improve lysosomal targeting and tissue distribution have been highlighted. This review describes the glycan structures that are important for lysosomal targeting and provides information on recent glyco-engineering technologies for the development of therapeutic enzymes with improved efficacy. [BMB Reports 2015; 48(8): 438-444]

• Fisheries and Aquatic Sciences
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• v.18 no.1
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• pp.65-71
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• 2015

The functional utility of a transgenic marine medaka Oryzias dancena strain carrying the red fluorescent protein (RFP) gene driven by an endogenous choriogenin H (chgH) promoter was evaluated for its ability to detect waterborne $17{\alpha}$-ethinylestradiol (EE2), a synthetic estrogen derivative. The chgH:rfp transgenic marine medaka larvae showed an age-dependent tendency in the efficiency of EE2-mediated transgene expression, in which transgenic larvae older than 6 days post-hatching displayed a more effective response in their transgene expression to EE2 than did younger hatchlings. During experimental exposures to high concentrations of EE2 (200 to 1,000 ng/L), the transgenic responses in the hatchlings were broadly dose- and duration-dependent. With exposures using lower doses of EE2 (25, 50 and 100 ng/L), EE2-induced transgenic RFP was also observed in the transgenic larvae, although the lower doses required exposure of longer duration. Under the EE2 exposure and microscope assay conditions used in our study, transgenic marine medaka larvae exhibited a similar degree of EE2-mediated RFP phenotype expression at various salinity levels (0, 15 and 30 ppt).

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1776-1783
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• 2002

A population segregating for the naked neck (Na) gene was used to evaluate its effect on fast growing broilers at heat stress. An experimental stock comparable to those of modern broilers was established by backcrossing to colour synthetic male and female lines. Matings between heterozygous (Na/na) males and females produced normally feathered (na/na), heterozygous (Na/na) and homozygous (Na/Na) chicks for the present study. Day old to seven week old coloured broilers of three genotypes viz. normally feathered (na/na), heterozygous naked neck (Na/na) and homozygous naked neck (Na/Na) were compared for heat dissipation, growth performance, body conformation traits, blood biochemical parameters and carcass traits in tropical climate. In hot climate, naked neck broilers had significantly less body temperature and better heat dissipation capabilities as compared to normal broilers. The naked neck broilers had significantly higher body weight and better feed conversion ratio than na/na broilers. The Na/Na or Na/na broilers exhibited higher giblet yield, blood loss and lower feather mass compared to na/na broilers. The results indicated that the reduction in feather coverage in Na/Na and Na/na broilers facilitates better heat dissipation with lower body temperature, more body weight gain, better FCR and carcass traits compared to normal broilers.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.2305-2309
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• 2012

Crocin, the main pigment of Crocus sativus L., has been shown to have antiproliferative effects on cancer cells, but the involved mechanisms are only poor understood. This study focused on probable effect of crocin on the immortality of hepatic cancer cells. Cytotoxicity of crocin ($IC_{50}$ 3 mg/ml) in hepatocarcinoma HepG2 cells was determined after 48 h by neutral red uptake assay and MTT test. Immortality was investigated through quantification of relative telomerase activity with a quantitative real-time PCR-based telomerase repeat amplification protocol (qTRAP). Telomerase activity in 0.5 ${\mu}g$ protein extract of HepG2 cells treated with 3 mg/ml crocin was reduced to about 51% as compared to untreated control cells. Two mechanisms of inhibition, i.e. interaction of crocin with telomeric quadruplex sequences and down regulation of hTERT expression, were examined using FRET analysis to measure melting temperature of a synthetic telomeric oligonucleotide in the presence of crocin and quantitative real-time RT-PCR, respectively. No significant changes were observed in the $T_m$ telomeric oligonucleotides, while the relative expression level of the catalytic subunit of telomerase (hTERT) gene showed a 60% decrease as compared to untreated control cells. In conclusion, telomerase activity of HepG2 cells decreases after treatment with crocin, which is probably caused by down-regulation of the expression of the catalytic subunit of the enzyme.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.804-811
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• 2001

In many Gram-negative bacteria, autoinducers, such as N-acyl-L-homoserine lactone(acyl-HSL) and its derivative molecules, mediate the cell-density-dependnet expression of certain operons. The current study identified the autoinducers produced by Burkholderia cepacia G4, a trichloroethylene-degrading lagoon isolate, using TLC bioassays with Agrobacterium tumefaciens NT1(pDCI141E33) and Chromobacterium violaceum CVO26, and a GC-MS analysis. The ${R_f}\;and\;{R_t}$ values and mass spectra were compared with those of synthetic compounds. Based on the analyses, it was confirmed that G4 produces N-hexanoyl (C6)-, N-octanoyl (C8)-, N-decanoyl (C10)-, N-dodecanoyl (C12)-HSL, and an unknown active species. The integration of the GC peak areas exhibited a ratio of C8-HSL:C10-HSL:C12-HSL at 3:17:1 with C6-HSL and C10-HSL production at trace and micromolar levels, respectively, in the culture supernatants. Nutants partially defective in producing acyl-HSLs were also partially defective in the biosynthesis of an antibiotic substance. These results indicate that the autoinducer-dependent gene regulation in G4 is dissimilar to the clinical B. cepacia strains isolated from patients with cystic fibrosis.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.445-448
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• 2002

The Pseudomonas fluorescens KCTC 1767, a selected and identified as potential candidate for stereo-specific resolution of rac-ketoprofen ethyl ester, was systematically investigated in order to induce the high level expression and detailed characterization of the expressing enzyme esterase. We cloned the esterase gene from chromosomal DNA of Pseudomonas fluorescens KCTC 1767 by PCR with two synthetic primers that desinged for simple purification. The recombinant esterase from Pseudomonas fluorescens KCTC 1767 exibited a high conversion rate and enantioselectivity to the (S)-ketoprofen ethyl ester as expected. The enzyme was easily purified to homogeniety by using a metal chelating affinity chromatography as a protein with poly histidine taq, and thus obtained 0.6 mg of protein from a 100 mL culture broth in a single step. The purified enzyme was steadily stable at the pH range from 7.0 to 10. The activity was also retained to be about 70% after the preincubation at $40^{\circ}C$ but over $50^{\circ}C$ lost the activity completely. The molecular mass of the esterase was estimated to be about 43 kDa on SDS-PAGE, and an identical result was also shown in gel filteration chromatography. The specific activity was calculated 27 mM/mg-protein/min by using the rac-ketoprofen ethly ester as a substrate.

• Toxicological Research
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• v.23 no.3
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• pp.207-214
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• 2007

Chronic oxidative stress produced by exposure to environmental chemicals or pathophysiological states can lead animals to aging, carcinogenesis and degenerative diseases. Indirect antioxidative mechanisms, in which natural or synthetic agents are used to coordinately induce the expression of cellular antioxidant capacity, have been shown to protect cells and organisms from oxidative damages. Electrophile and free radical detoxifying enzymes, which were originally identified as the products of genes induced by cancer chemopreventive agents, are members of this protective system. The NFE2 family transcription factor Nrf2 was found to govern expression of these detoxifying enzymes, and screening for Nrf2-regulated genes has identified many gene categories involved in maintaining cellular redox potential and protection from oxidative damage as Nrf2 downstream genes. Further, studies using Nrf2-deficient mice revealed that these mutant mice showed more susceptible phenotypes towards exposure to environmental chemicals/carcinogens and in oxidative stress related disease models. With the finding that cancer chemopreventive efficacy of indirect antioxidants (enzyme inducers) is lost in the absence of Nrf2, a central role of Nrf2 in the antioxidative protective system has been firmly established. Promising results from cancer prevention clinical trials using enzyme inducers propose that pharmacological interventions that modulate Nrf2 can be an effective strategy to protect tissues from oxidative damage.

• KSII Transactions on Internet and Information Systems (TIIS)
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• v.12 no.8
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• pp.3606-3629
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• 2018

The discovered high potential utility itemsets (HPUIs) have significant influence on a variety of areas, such as retail marketing, web click analysis, and biological gene analysis. Thus, in this paper, we propose an algorithm called HPUIM-QGA (Mining high potential utility itemsets based on a quantum genetic algorithm) to mine HPUIs over uncertain datasets based on a quantum genetic algorithm (QGA). The proposed algorithm not only can handle the problem of the non-downward closure property by developing an upper bound of the potential utility (UBPU) (which prunes the unpromising itemsets in the early stage) but can also handle the problem of combinatorial explosion by introducing a QGA, which finds optimal solutions quickly and needs to set only very few parameters. Furthermore, a pruning strategy has been designed to avoid the meaningless and redundant itemsets that are generated in the evolution process of the QGA. As proof of the HPUIM-QGA, a substantial number of experiments are performed on the runtime, memory usage, analysis of the discovered itemsets and the convergence on real-life and synthetic datasets. The results show that our proposed algorithm is reasonable and acceptable for mining meaningful HPUIs from uncertain datasets.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.3
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• pp.269-274
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• 2015

Chronic inflammation has been proposed as one of the main molecular mechanisms of aging and age-related diseases. Although evidence in humans is limited, short-term calorie restriction (CR) has been shown to have anti-inflammatory effects in aged experimental animals. We reported on the long-term treatment of daumone, a synthetic pheromone secreted by Caenorhabditis elegans in an energy deficient environment, extends the life-span and attenuates liver injury in aged mice. The present study examined whether late onset short-term treatment of daumone exerts anti-inflammatory effects in the livers of aged mice. Daumone was administered orally at doses of 2 or 20 mg/kg/day for 5 weeks to 24-month-old male C57BL/6J mice. Increased liver macrophage infiltration and gene expression of proinflammatory cytokines in aged mice were significantly attenuated by daumone treatment, suggesting that short-term oral administration of daumone may have hepatoprotective effects. Daumone also dose-dependently suppressed tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$ )-induced nuclear factor-${\kappa}B$ (NF-${\kappa}B$) phosphorylation in HepG2 cells. The present data demonstrated that short-term treatment of daumone has anti-inflammatory effects in aged mouse livers possibly through suppression of NF-${\kappa}B$ signaling and suggest that daumone may become a lead compound targeting aging and age-associated diseases.

• Biomolecules & Therapeutics
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• v.2 no.2
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• pp.155-160
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• 1994

Human immunodeficiency virus type 1 (HIV-1) protease is essential for the replication of the virus and it is therefore an attractive target for antiviral drugs of HIV-1. Several dipeptide isosteres containing 2-isoxazoline or $\alpha$-hydroxy ketomethylene have been synthesized and their inhibitory effects on the HIV-1 protease examined. The enzymatically active HIV-1 protease was purified to homogeniety from E. coli transformed with a recombinant plasmid (pMAL-pro) containing the entire gene encoding the protease. The purified protease had the substrate specificity with Km value of 9.8$\mu$M when an undecapeptide His-Lys-Ala-Arg-Val-Leu-(p-nitro)Phe-Glu-Ala-Nle-Ser-amide was used as a substrate, and the products from the substrate after specific cleavage by HIV-1 protease were analyzed by HPLC. The synthetic compounds containing dipeptide isosteres showed specific inhibitory effects while a dipeptide isostere containing an isoxazoline ring inhibited the HIV-1 protease competitively with Ki value of 500 $\mu$M. Even if the inhibition effects of HIV-1 protease were not very high, these novel dipeptide isosteres can be used as key structural moieties for developing specific inhibitors of HIV-1 protease.