• Plant Disease and Agriculture
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• v.5 no.1
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• pp.27-33
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• 1999

Frost injury of crops is closely related to the epiphytic population dynamics of ice nucleation-active (INA) bacteria, and the injury can be reduced by decreasing the INA bacterial population. In order to predict the epiphytic population of INA bacteria on crops, a rapid and accurate detection method has to be developed. In the previous report, we produced some antibodies against INA proteins purified from the outer membrane of INA bacteria. However it was difficult to produce the antibodies because the purification procedures of the INA proteins were complicated, and the final yield was too low. We designed a specific peptide from the N-terminal region of INA protein by computer analysis and synthesized the peptide in vitro in this experiment. The peptide sequence was Asp-Ser-Por-Leu-Ser-Leu-His-Ala-Asp, that is corresponding to the highly conserved region in several INA proteins, with predicted beta turn, coiling, and hydrophilic region. A polyclonal anti-INA peptide antiserum produced specifically recognized INA bacteria as few as 10 colony-forming units (CFU) in the ELISA reactions and did not respond to other non-INA bacteria. Serological specificity of the anti-INA peptide antiserum will facilitate the forecasting of the INA bacterial population dynamics on crops.

• Molecules and Cells
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• v.39 no.4
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• pp.316-321
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• 2016

The receptor activator of nuclear factor ${\kappa}B$ (RANK) and its ligand RANKL are key regulators of osteoclastogenesis and well-recognized targets in developing treatments for bone disorders associated with excessive bone resorption, such as osteoporosis. Our previous work on the structure of the RANK-RANKL complex revealed that Loop3 of RANK, specifically the non-canonical disulfide bond at the tip, performs a crucial role in specific recognition of RANKL. It also demonstrated that peptide mimics of Loop3 were capable of interfering with the function of RANKL in osteoclastogenesis. Here, we reported the structure-based design of a smaller peptide with enhanced inhibitory efficiency. The kinetic analysis and osteoclast differentiation assay showed that in addition to the sharp turn induced by the disulfide bond, two consecutive arginine residues were also important for binding to RANKL and inhibiting osteoclastogenesis. Docking and molecular dynamics simulations proposed the binding mode of the peptide to the RANKL trimer, showing that the arginine residues provide electrostatic interactions with RANKL and contribute to stabilizing the complex. These findings provided useful information for the rational design of therapeutics for bone diseases associated with RANK/RANKL function.

• Journal of Life Science
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• v.31 no.1
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• pp.83-89
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• 2021

Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.

• Journal of Animal Science and Technology
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• v.48 no.4
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• pp.541-554
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• 2006

Present review is to introduce an omasal sampling technique in rumen proteolysis and to consider some information on the omasal sampling technique with particular emphasis on methodological aspects. Use of the omasal sampling technique provides a new opportunity for accurate estimation of rumen metabolism with overcoming limitations of previous in vivo, in vitro and/or in situ methods. The potential advantages of the present technique compared with post-ruminal sampling techniques include following points; 1) only rumen cannulated animals are required, 2) less endogenous nitrogen (N) is contaminated in omasal digesta and 3) omasal digesta are devoid of exposure to acid peptide hydrolysis occurring in the abomasum. Estimates of soluble non-ammonia N (SNAN) in omasal digesta indicate that the assumptions underlying the in situ method that rapidly degradable N fraction can be degraded at an infinite rate and only insoluble dietary N escapes the rumen may be not valid. Quatitatively higher peptide concentration rather than free amino acid and soluble protein in escapable SNAN suggests that hydrolysis of peptide to amino acid may be the rate-limiting step in rumen proteolysis.

• The Journal of Korean Medicine
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• v.24 no.4
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• pp.120-126
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• 2003

Object : This study was done to evaluate the inhibitory effect of Samul-tang (Siwu-tang) on collagen production by cultured murine hepatic non-parenchymal cells. Methods : Hepatic non-parenchymal cells were cultured from normal Sprague-Dawley rats and established in a primary cell culture on uncoated plastic culture plates. The Samul-tang (Siwu-tang) was treated into the cell culture media for 72 hours and the cells were harvested for analysis. Analyses were done on cell proliferation, [3H]thymidine incorporation assay and procollagen type IC-peptide. Results : The cultured cells resembled fibroblasts in shape and produced procollagen which is consistent to fibrogenesis in vivo. Proliferation of the non-parenchymal cells was inhibited slightly and the [3H]thymidine incorporation assay showed a dose-dependent decrease by Samul-tang (Siwu-tang) treatment. Production of procollagen type I C-peptide was decreased by low-concentration treatment of the Samul-tang (Siwu-tang), but increased by high-concentration treatment. Conclusion : It seemed that the cells were responding to the Samul-tang (Siwu-tang) in low-concentration, thus producing less collagen. However, when the drug was administered with high enough concentration to cause excessive stimulation of cells, it seemed that the activated cells might overly produce procollagen, the precursor of collagen, thus aggravating fibrosis of the liver. So, it is considered that the proper concentration of Samul-tang (Siwu-tang) is important when treating patients with liver cirrhosis based on the patients' status.

• Proceedings of the PSK Conference
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• 2000.04a
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• pp.168.3-169
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• 2000

• International Journal of Industrial Entomology and Biomaterials
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• v.26 no.1
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• pp.31-40
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• 2013

Paralytic peptide binding proteins (PP-BP) are 30KP proteins that show similarity to ENF binding proteins. The ENF-BP act as active regulators of ENF peptides. ENF peptides are multifunctional insect cytokines. The comparison of gene expression in diapause induced and non-diapause eggs at different time intervals after oviposition showed an upregulation of PP at 18h as well as PP-BP at 12 and 18h after oviposition along with few other genes. The current study has been taken up to investigate the role of PP as well as PP-BP in diapause induction in polyvoltine silkworms and to study the multigene organization of PP-BP in the Bombyx mori genome. The tissue specific expression analysis revealed that, PP-BP is highly expressed in fat body followed by egg and brain while no expression was observed in midgut. The expression levels of PP and PP-BP in diapause and non-diapause eggs from 0h to 48h after oviposition, validated through realtime PCR revealed that PP is highly expressed at 18 and 24h while PP-BP expression is higher at 12 and 18h time intervals suggesting their possible role in diapause induction. The whole genome survey of the PP-BP paralogous sequences revealed a total of 46 B. mori PP-BP homologs that are classified into 3 categories viz., ENF-BP, Typical 30KPs and serine/threonine rich 30KPs. These paralogous sequences are distributed on chromosomes 7, 20, 22 and 24, all 30KP and S/T rich 30KP proteins are present in the same locus of chromosome 20.

• Journal of Korean Academy of Nursing
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• v.31 no.6
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• pp.1034-1043
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• 2001

To determine the frequency of past and present obesity among patients with NIDDM and to identify the differences of body fat, blood pressure and C-peptide/glucose ratio according to obese diabetic patients (BMI$\geq$25 kg/$m^2$) and nonobese (BMI<25 kg/$m^2$). Also the final factor is to observe the anthrometric change patterns in the study. Method: The weight at 20 years-old, previous maximal body weight, and acute weight loss were queried. Current height, body weight, BMI, waist & hip circumferences, waist-hip ratio, skinfold thicknesses, blood pressure, fasting blood glucose, and fasting C-peptide were measured in one hundred sixty-seven NIDDM patients. The differences of the parameters ccording to obese and nonobese, and three anthropometric change patterns were analyzed. Result: Results were as follows: 1. 66.5 % of the NIDDM patients had a history of past obesity as assessed by their maximum weight, while only 33.2% of them were currently obese (p's < 0.001). 2. The waist & hip circumferences, skinfold thicknesses, systolic, diastolic & mean arterial blood pressure in obese patients were greater than those of nonobese patients (all p's < 0.001). 3. The waist and the hip circumferences, and skinfold thicknesses (subscapula & triceps) were highest among the obese-obese group. WHR and abdominal skinfold thickness in the obese-obese and obese-nonobese groups were higher than those in the nonobese- nonobese group. Systolic & diastolic and mean arterial blood pressures in the obese-obese group were higher than those of obese-nonobese and nonobese-nonobese groups(all p's < 0.005). 4. The abdominal and subscapular skinfold thicknesses in female diabetic patients were greater than those of male patients (all p's <0.0001). Conclusion: Although most Korean NIDDM patients were previously obese, many of them were not obese during the course of the study. Greater central and upper body adiposicity and higher blood pressure was shown in obese diabetic patients. Also, greater central and upper body adiposicity was demonstrated in female diabetic patients.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.1
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• pp.1-8
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• 2014

Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

• Journal of Animal Reproduction and Biotechnology
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• v.37 no.4
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• pp.239-245
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• 2022

Protein and peptide candidates are screened to apply therapeutic application as a drug. Ensuring that these candidates are delivered and maximized effectiveness is still challenging and a variety of studies are ongoing. As drug delivery system vehicles, cell-penetrating peptide (CPP) can deliver various kinds of cargo into the cell cytosol. In a previous study, we developed Ara27 CPP, which are a zinc knuckle family protein of Arabidopsis, and confirmed internalization in human dermal fibroblasts and human dental pulp stem cells at low concentration with short time treatment condition without any toxicity. Ara27, an amphipathic CPP, could be modified and utilized in the biomedical field excluding the risk of toxicity. Therefore, we would like to confirm the non-toxic induced penetrating ability of Ara27 in various cell lines. The purpose of this study was to screen the cell internalization ability of Ara27 in various cell lines and to confirm Ara27 as a promising core CPP structure. First, Ara27 was screened to confirm non-toxicity concentration. Then, fluorescence-labeled Ara27 was treated on human normal cell lines, cancer cell lines and animal cell lines to identify the cellular internalization of Ara27. Ara27 was well intracellular localized in all cell lines and the intensity of fluorescence was remarkably increased in time pass manner. These results indicate that Ara27 has the potential as a core structure for applications in various drug delivery systems.