• Journal of Life Science
• /
• v.25 no.10
• /
• pp.1164-1168
• /
• 2015

Previous research showed that chlorphenesin galactoside (CPN-Gal), a preservative in cosmetics, was safer than CPN against human skin cells [9]. To establish a stable and long-term process for CPN-Gal production, we investigated the repeated-batch process. In this process, β-gal-producing recombinant Escherichia coli cells were immobilized in calcium alginate beads, and CPN was converted to CPN-Gal by the transgalactosylation reaction. The process was conducted in a 300 ml flask, which contained E. coli cell-immobilized alginate beads, 33.8 mM of CPN, and 400 g/l of lactose. The pH and temperature were 7.0 and 40℃, respectively. During the repeated-batch operation, four consecutive batch operations were conducted successfully until 192 hr. The conversion yield of CPN to CPN-Gal was 64% during 192 hr, which was higher than the values in previous reports [3, 13]. Thereafter, however, the conversion yield gradually decreased until the operation was finished at 336 hr. Western blotting of immobilized E. coli cells revealed that β-gal gradually decreased after 192 hr. In addition, alginate beads were cracked when the operation was finished. It is probable that, including this loss of E. coli cells by cracks, deactivation, and product inhibition of E. coli β-gal might lead to a gradual decrease in the production of CPN-Gal after 192 hr. However, as the purification of β-gal is not necessary with β-gal-producing recombinant E. coli cells, β-gal-producing E. coli cells might be a practical and cost-effective approach for enzymatically synthesizing CPN-Gal. It is expected that this process will be extended to long-term production process of CPN-Gal for commercialization.

• Korean Chemical Engineering Research
• /
• v.48 no.3
• /
• pp.327-331
• /
• 2010

Porcine placenta was treated with alkali, acid and enzyme treatment to obtain extracts. Heavy metal contents such as Pb, As, and Hg were low enough to satisfy cosmetic agent standard. As a result of safety test(MTT assay), porcine placenta extracts showed over 80% of cell viability at $50{\mu}g/ml$, and cell toxicity was relatively lower. From antioxidation test using DPPH free radical scavenging assay, antioxidation effect was highest as 63% at $50{\mu}g/ml$ when porcine placenta was treated with alkali in pH 9. From whitening effect test using tyrosinase inhibition assay, tyrosinase inhibition effect was 30% at $50{\mu}g/ml$ concentration in alkali treated procine placenta, however, the efficiency was lower compared with arbutin or vitamin C. In anti-wrinkle effect test from elastase inhibition assay, elastase inhibition effects were 20~30% at $50{\mu}g/ml$ for 5 kinds of porcine placenta treatments, which was superior to standard, and especially, protease treated extracts showed best results. Skin formulation including 1% porcine placenta was made and the formulation was very stable for temperature and storage period. From this research, porcine placenta extract showed high potential for anti-wrinkle functional cosmetic agent.

• Journal of Life Science
• /
• v.19 no.10
• /
• pp.1389-1395
• /
• 2009

Generally, it takes a long time to purify wood vinegar, and it contains toxic compounds such as tar, methanol, phenol and benzopyrene. To reduce the toxicity of wood vinegar itself, we have developed a new purification method of wood vinegar using an oxidation-cohesion reaction and distillation with an active carbon. We have investigated the physico-chemical change (pH, specific gravity, refractive index and dissolved tar), the change of amount of toxic compounds (carbonyl group, phenol, benzopyrene and residual solvents) and organic acids (formic acid (FA), acetic acid (AA), propionic acid (PA)) of the purified wood vinegar under the long term and accelerated storage conditions. Also, we have evaluated the effect of the purified wood vinegar on hair growth using an alopecia model of C57BL/6 mice. As a result, we could find out that the purified wood vinegar was stable and remained without decay under the storage conditions and benzopyrene, a carcinogenic agent, was not detected in the purified wood vinegar. After topical treatment of the purified wood vinegar solution or minoxidil (MXD) for 2 weeks to dorsal skin, the hair regrowth of the mice accelerated faster than that of the control, with no clinical signs. In conclusion, we could suggest a guideline for quality control of process to reduce the toxic compounds in wood vinegar and it might be a useful hair growth promoter in the treatment of baldness or alopecia.

• Journal of Embryo Transfer
• /
• v.28 no.3
• /
• pp.243-250
• /
• 2013

Pyracantha is a genus of thorny evergreen large shrubs in the family of Rosaceae, with common names Firethorn or Pyracantha. It's extract has also been used in cosmetics as a skin-whitening agent and functioning through tyrosinase inhibition. Recent studies have shown that pyracantha extract possesses antioxidant activities and may significantly improve lipoprotein metabolism in rats. Although the mode of action of Pyracantha extract is not fully understood, a strong relationship was observed between antioxidant and apoptosis in some types of cells. Thus, the aim of this study was to evaluated the effect of pyracantha extract on blastocysts formation and their quality of the porcine parthenogenetic embryos. After parthenogenetic activation by chemicals, presumptive porcine parthenogenetic embryos were cultured in PZM-3 medium supplemented with extracts of pyracantha leaf, stalk and root for 6 day (1, 5 and $10{\mu}g/ml$, respectively). In our results, the frequency of blastocyst formation in pyracantha root extract ($5{\mu}g/ml$) treated group had increased that of other groups. Furthermore, blastocysts derived from pyracantha root extract ($5{\mu}g/ml$) treated group had increased the total cell numbers and reduced apoptotic index. Blastocyst development was significantly improved in the pyracantha root extract ($5{\mu}g/ml$) treated group when compared with the $H_2O_2$ treated group (p<0.05). Subsequent evaluation of the intracellular levels of ROS in pyracantha root extract ($5{\mu}g/ml$) treated groups under $H_2O_2$ induced oxidative stress were decreased (p<0.05). In conclusion, our results indicate that treatment of pyracantha root extract may improve in vitro development of porcine parthenogenetic embryos through its antioxidative and antiapoptotic effects.

• Applied Microscopy
• /
• v.22 no.2
• /
• pp.1-14
• /
• 1992

This experiment was performed to study the morphological responses of the parafollicular cells of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The head and neck region of the rat, under sodium thiopental anesthesia, was exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80 cm, and the dose rate was 200 rads/min. The rate of experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Pieces of the tissue taken from the thyroid gland were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH 7.3), and in 1% osmium tetroxide (0.1M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. Two types of the parafollicular cells, according to their electron densities, were found, i. e., light cells and dark cells. 2. Three types of the parafollicular cells, according to their sizes of secretory granules were found, i.e., small granule cells ($85nm{\pm}20.1;64{\sim}102nm$), medium granule cells ($120nm{\pm}26.5;77{\sim}179nm$), and large granule cells ($165nm{\pm}25.7;128{\sim}189nm$). 3. The differential ultrastructural changes of the cells according to their cell types, i.e., dark and light cell, or small, medium and large granule cells, were hardly observed in the time and dose range covered by this study. 4. The morphological changes of the parafollicular cells were not pronounced after exposure to 3,000 rads of X-ray. 5. Swollen cisternae of the granular endoplasmic reticulum and partial cytolysis were observed after exposure to 6,000 rads of X-ray. 6. Above results suggest that the parafollicular cells showed the alterations of mitochondrial and granular endoplasmic reticular swelling, and partial cytolysis, but only in doses of 6,000 rads.

• Korean Journal of Food Science and Technology
• /
• v.35 no.5
• /
• pp.865-870
• /
• 2003

The effects of storage atmosphere on the physicochemical and organoleptic properties of Asian pears (Pyrus pyrifolia Nakai cv. Niitaka) were investigated. The pears were stored at $0^{\circ}C$ for up to 8 months under CA conditions (3 kPa $O_2$＋1 kPa $CO_2$, 1 kPa $O_2$+5 kPa $CO_2$, 1 kPa $O_2$+1 kPa $CO_2$) controlled by a static system or air (21 kPa $O_2$+0 kPa $CO_2$). CA conditions retarded the loss of weight, flesh firmness, organoleptic properties (appearance, aroma, sweetness, texture), and the development of injuries in the fruits during storage. Low $O_2$ plus high $CO_2$ conditions maintained the highest flesh firmness. In addition, low $O_2$ reduced injuries more than the other conditions. Skin color (lightness, chroma, hue), soluble solids, titratable acidity, pH and vitamin C were unaffected by storage atmosphere.

• Journal of the Korean Chemical Society
• /
• v.62 no.3
• /
• pp.181-186
• /
• 2018

Nitrosamines are the nitrosocompounds which are produced by nitrosation reactions of the secondary amine and nitrite, and has been found to be produced through the vulcanization process during the production of rubber products Recently, nitrosamines have been detected in rubber products and become a major topic. Condoms are disposable medical devices, so safety is important because they come into direct contact with the skin and mucous membranes. In this study, we developed an analytical method for nitrosamines in condoms by applying ISO 29941 method. The samples were eluted by distilled water, and target compounds were extracted by liquid-liquid extraction with dichloromethane. And then after concentrated, and quantitatively analyzed by LC-MS/MS. The accuracies of the analytical method were ranged from 85.8 to 108.7%, precisions were lower than 11.5%, and the detection limits were from 0.11 (NDPA and NDBA) to 0.48 (NPYR) ng/mL. Among the 31 condom samples, NDBA was detected from 2 cases by extraction of distilled water, and NDMA were detected from 1 case, NDEA from 4 cases and NDBA from 26 cases by extraction of artificial saliva (pH 4.5). The total amount of nitrosamines in all samples were less than $500{\mu}g/kg$.

• Korean Journal of Microbiology
• /
• v.47 no.4
• /
• pp.348-355
• /
• 2011

Staphylococcus aureus is a major human pathogen that is associated with various types of local and systemic infection. Staphylococcal protin A (SPA), a highly expressed surface component of S. aureus, may have a role in virulence such as activating inflammation and interfering with immune clearance. We examined the effect of recombinant SPA on inflammatory response in human HaCaT keratinocytes. The recombinant SPA protein was prepared using the pET-28a Vector System in Escherichia coli. The expression of pro-inflammatory related adhesion molecules and cytokines in HaCaT cells incubated for 6, 12, and 24 h with SPA (2 ${\mu}g$/ml) was analyzed by comparative RT-PCR or ELISA. The expression of E-selectin, ICAM-1, MCP-1, IL-6 and IL-8 was significantly increased in HaCaT from 6 to 24 h after treatment with SPA. SPA showed the effect on the adhesion-promoting ability of U937 monocytes to HaCaT cells. Our data demonstrate that SPA stimulates inflammatory response of HaCaT cells, implicating an important factor for exacerbation of skin inflammation of immunologic disease.

• Journal of Microbiology
• /
• v.45 no.2
• /
• pp.158-167
• /
• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Journal of Environmental Science International
• /
• v.16 no.5
• /
• pp.665-670
• /
• 2007

In this study, we evaluated the property change and relationship between hematological and biochemical parameters in exposure group and control group by multiple inhalation exposure, of $NO_2$. In case of leukocyte, change of hematological property increased significantly in statistics (p<0.05) in mice experiment. in case of hematocrit of exposure group, it was of decreased significantly in statistics. the tendency of hemoglobin and hematocrit was similar to all for three case. in case of methmoglobin that of the exposure group in mice experiment was meaning increased(p<0.05). For the $NO_2$ influence in biochemical property, total protein and triglyceride was decreased but not significant in statistics, albumin was decreased in the exposure group in the animal experiments, blood urea nitrogen and creatinine increased significantly in statistics in the animal experiments. uric acid and lactate dehydrogenase in serum increased significantly in the exposure group. We have to evaluate risk assessment for the $NO_2$ of indoor air pollutant in low concentration and long time.