• Journal of the Korean Society of Tobacco Science
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• v.2 no.1
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• pp.1-7
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• 1980

Gel filtration column chromatography (Sephadex G-75), dialysis an d Brushite column chromatography were carried out to separate the brown pigmented macromolecule from water extracts of the cured leaf tobaccos. The two distinct macromolecules having different molecular weight were separated by the Sephadex column chromatography. Brushite also separated two different species of macromolecules which might have different electronic structures. According to the enzymatic degradation of protein in Burley and Hicks, chymotrypsin showed the best degradation ratio, ie., 16-30% in Burley and 38-57% in Hicks. Similar effect was observed with pepsin. However, very low effect of degradation was revealed with trypsin. The sample treated with the proteolytic enzymes revealed the disappearance of the first peak and the slight decrease of the 2nd peak height in the separation profile of Sephadex. After dialysis, the brown pigmented macromolecule was pyrolyzed at $300^{\circ}C$ and the strongly fluorescent components not identified before pyrolysis were detected with TLC separation. Absorption spectrum of these fluorescent compounds was monitored in benzene and the absorption maxima at 265nm and 275 nm were obtained. Considering absorption maxima and shape of the spectrum, those fluorescent compounds seem to be PAH derivatives.

• Parasites, Hosts and Diseases
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• v.36 no.4
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• pp.261-268
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• 1998

The present study was undertaken to investigate the role of cysteine proteinase of Trichomonas vaginalis in escaping from host defense mechanism. A cysteine proteinase of T. vaginalis was purified by affinity chromatography and gel filtration. Optimum pH for the purified proteinase activity was 6.0. The proteinase was inhibited by cysteine and serine proteinase inhibitors such as E-64, NEM, IAA, leupeptin. TPCK and TLCK, and also by $Hg^{2+}$, but not affected by serine-, metallo-, and aspartic proteinase inhibitors such as PMSF, EDTA and pepstatin A. However, it was activated by the cysteine proteinase activator, DTT. The molecular weight of a purified proteinase was 62 kDa on gel filtration and 60 kDa on SDS-PAGE. Interestingly, the purified proteinase was able to degrade serum IgA, secretory IgA, and serum IgG in time- and dose-dependent manners. In addition, the enzyme also degraded hemoglobin in a dose-dependent manner. These results suggest that the acidic cysteine proteinase of T. vaginalis may play a dual role for parasite survival in conferring escape from host humoral defense by degradation of immunoglobulins, and in supplying nutrients to parasites by degradation of hemoglobin.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.182-191
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• 2007

Bioabsorbable devices have been utilized and experimented in many aspects of orthopaedic surgery. Depending upon their constituent polymers, these materials can be tailored to provide sufficient rigidity to allow bone healing, retain mechanical strength for certain period of time, and then eventually begin to undergo degradation. The objective of this study was to estimate extent in which Poly-L-latic acid (PLLA) implants had bioabsorbability and biocompatibility with bone and soft tissue in dogs and also to develop bioabsorbable, biocompatible materials with the appropriate strength and degradation characteristics to allow for regular clinical use for treating orthopedic problems in humans as well as animals. Eighteen dogs were used as experimental animals and were inserted two types of PLLA implants. PLLA rods were inserted into subcutaneous tissue of back or the abdomen wall. And the rods were tested for material properties including viscosity, molecular weight, melting point, melting temperature, crystallinity, flexural strength, and flexural modulus over time. PLLA screws were inserted through cortical bone into bone marrow in the femur of the dogs and stainless steel screw was inserted in the same femur. Radiographs were taken after surgery to observe locations of screw. Histological variations including cortical bone response, muscular response, bone marrow response were analyzed over the time for 62weeks. The physical properties of PLLA rods had delicate balances between mechanical, thermal and viscoelastic factors. PLLA screws did not induce any harmful effects and clinical complications on bone and soft tissue for degradation period. These results suggest that PLLA implants could be suitable for clinical use.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.12
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• pp.23-32
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• 2020

This study investigated the thermal degradation and fading behavior of PTT dyed with vat dye and its dyeing ability. The PTT sample was dyed with vat dye using an acid treatment and pad-steam method to improve the dyeing performance. This method made dye particle nanosize and allowed it to penetrate the polymer material easily. The sample dyed using the pad-steam method showed level dyeing and enhanced dyeing affinity, compared to the batch-dyeing method. The degradation behavior of PTT dyed with vat dye after each heat and UV treatment was examined with the change in tensile strength or K/S value on the sample. The tensile strength and K/S values of the sample dyed with vat dye after the heat and UV treatment decreased with increasing temperature and exposure time. Although they showed high degradation under severe conditions, its rate constant was improved compared to the samples dyed with disperse dye. Consequently, acid treatment and the pad-steam method resulted in the introduction of vat dye on PTT. In addition, the PTT dyed with vat dye showed enhanced thermal and UV resistance because of their high molecular weight and chemical structure for stable adsorption behavior.

• Journal of Life Science
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• v.26 no.3
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• pp.296-301
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• 2016

An antibacterial peptide from skin extract of the catfish Silurus asotus was purified and characterized. The acidified skin extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into flow-through (F.T.), 10% methanol-elute (RM10), 60% methanolelute (RM60), and 100% methanol-elute (RM100) fractions. RM10, RM60, and RM 100 showed antimicrobial activity against Escherichia coli D31. On the other hand, the F.T. fraction did not show antimicrobial activity. Among the various fractions, RM 60 had the highest activity. RM 60 was partially purified on a cation exchange column (CM52) by a stepwise gradient. The ammonium acetate (pH 5.15) 0.02 M – 0.8 M fraction showed antimicrobial activity. Then an antimicrobial peptide was purified using a 0.6M fraction with strong antibacterial activity through a series of five C18 reversed-phase HPLC columns. For the characterization of the purified peptide, the molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of this peptide was about 4182.1 [M+H]⁺. The amino acid sequence of this peptide was partially determined as follows: PALXXKARREAKVKF. These findings suggest that this peptide plays a significant role in the innate defense system of catfish skin.

• Journal of Life Science
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• v.27 no.1
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• pp.50-56
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• 2017

An antibacterial peptide was purified from an acidified gill extract of the pufferfish Takifugu pardalis. The acidified gill extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into a flow-through (F.T.) and 60% methanol fraction (RM 60). Among the eluents, RM 60 had potent antibacterial activity against Bacillus subtilis KCTC 1021. RM 60 was partially purified on a cationic-exchange column (SP-5PW) by a linear gradient, and the antibacterial peptide was then further purified, using a series of cationic-exchange and $C_{18}$ reversed-phase HPLC columns. For characterization of the purified peptide, its molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of the peptide was about 1171.6 Da. The amino acid sequence of the peptide was partially determined as: STKEKAPRKQ. A comparison of the N-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the N-terminus of the histone H3 protein, which belongs to the histone H3 family. Thus, this peptide was designated as a puffer fish gill (PFG)-related antimicrobial peptide. This is the report to describe an antimicrobial function for the N-terminus of histone H3 of an animal species. The findings suggest that this peptide plays a significant role in the innate defense system of the pufferfish.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.433-444
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• 1996

We describe a novel approach to evaluate quantitatively the amounts of denatured proteins in cells upon heat exposure. A thiol compound, diamide [azodicarboxylic acid bis (dimethylamide)] causes protein cross-linking with exposed sulfyhydryl residues of denatured proteins. Since denatured proteins expose normally well-hidden sulfhydryl groups, these will be preferentially cross-linked by diamide. Thus diamide acts to 'trap' denatured proteins. We observed that protein aggregates (high molecular weight protein aggregates, HMA) appeared on SDS-polyacrylamide gels run under non-reducing conditions and that the amount of HMA can be quantified by scanning the gels using a gas flow counter. Heating cells followed by a fixed dose of diamide exposure resulted in HMA increases in a heat-dose dependent manner, demonstrating that the quantitation of HMA could serve as a measure of heat-denatured proteins. We compared thermotolerant and nontolerant cells and found decreased HMA in tolerant cells upon heat treatment. As an attempt to examine the kinetics of protein renaturation (or 'repair'), we measured the amounts of aggregates formed by the addition of diamide at various times after heat shock. Such experiments demonstrate an equally rapid disappearance of HMA in previously unheated and in thermotolerant cells. Levels of HMA in tolerant cells increased significantly after electroporation of HSP70 specific mAbs, suggesting an involvement of HSP70 in reducing HMA levels in thermotolerant cells upon heat exposure. Immunoprecipitation studies using anti-HSP70 antibody indicated an association of HSP70 with heat-denatured proteins. Our results suggest that heat induces protein denaturation, and that elevated level of HSP70 present in thermotolerant cells protects them by reducing the level of protein denaturation rather than by facilitating the 'repair' (or degradation) process.

• 보존과학연구
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• s.34
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• pp.20-29
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• 2013

Paper, textile and wood materials are mainly consisted of cellulose. Cellulose is high molecule and make up the strong crystalline structure by hydrogen bonds. In particular, the polymerization degree of cellulose are closely related to the strength of fiber, and the permanence. the useful life of fiber, also depends on the degradation of this substance. The viscosity of cellulose is considered to be an important indicator of fiber damage in high molecule polymers. The viscosity measurements with CED solution is used to measure the molecular weight and the degree of polymerization of cellulose. Cellulose viscosity of wood fibers is measured with TAPPI standard method T230. However, TAPPI standard method T230 is difficult to completely dissolving the cellulose of high molecular weight and large degree of polymerization, such as Korea traditional papers and fabrics made with mulberry, ramie, cotton fibers. In this study, The high viscosity of hanji and fabric was measured with TAPPI standard method T254. T254 method is that the cellulose specimen with the proper amount of weaker (0.167M CED) solution, and completely dissolved with the stronger (1.0M CED) solution. It was found that cellulose with high degree of polymerization was dissolved more easily in general CED method.

• KSBB Journal
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• v.21 no.2
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• pp.133-139
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• 2006

In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

• Korean Journal of Food Science and Technology
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• v.18 no.2
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• pp.158-162
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• 1986

Various cell wall polysaccharides such as ionically associated pectin (IAP), covalently bounded pectin (CBP),4N potassium hydroxide soluble hemicellulosic fraction (HF,) and 0-3N soluble hemicellulosic fraction (HF,) were fractionated from crude cell wall of the fresh and soft persimmon by chemical method. The changesin cell wall polysaccharides were studied by gel filteration chromatography . The content of crude cell wall remarkably decreased in the soft persimmon. The decreasing rates of IAP, CBP and $HF_2$ were 59, 60 and 74%, respectively, while $HF_1$ and cellulose changed only a little during softening. Sugar compositions of IAP and CBP were 72-84% uronic acid, 5-1% hexose and 11-16% pentose, and also the hemicellulose was composed of uronic acid besides hexose and pentose that was hemicellulosic components. The loss rate of pentose in IAP, of hexose in CBP, of hexose and uronic acid in $HF_2$, of pentose in $HF_1$ increased during softening. Though apparent average molecular freight of all polysaccharides shifted from high molecular freight to low molecular weight polymer, the shifting degree of CBP and $HF_2$ was especially remarkable during softening. It is suggested that the severe softening phenomenon of persimmon involved the degradation and dissolution of wall bound-CBP and $HF_2$ which were associated with each other.