• Archives of Plastic Surgery
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• v.34 no.3
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• pp.291-297
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• 2007

Purpose: Platelet-rich plasma(PRP) contains protein growth factors, which are actively secreted by platelets to promote wound healing. However, it is not clear whether the injection of PRP into the autologous fat grafts increases the survival rate and the degree of angiogenesis. Methods: New Zealand White rabbit ears were injected fat with PRP, saline, insulin or isoproterenol (n=8/each group) for observation of the survival and degree of angiogenesis of the injected fat. The volume of the harvested fat and the degree of angiogenesis from dorsum of rabbit ears were evaluated 4, 8, and 12 weeks after the autologous fat graft. The degree of angiogenesis was measured with microvascular density (MVD) counts. Results: The volume of harvested fat decreased in a time-dependent manner after autologous fat grafts, but the decrease rate in volume of harvested fat was slower in PRP-injected group compared to that of other control groups. The difference in the volume of the harvested fat between PRP-injected group and other control groups became significant from 4 weeks after the autologous fat graft, and was maintained up to 12 weeks. However, there was no significant difference between PRP-injected group and insulin-injected group 8 and 12 weeks after the autologous fat graft. On the contrary, MVD counts increased in a time-dependent manner after autologous fat grafts. The MVD counts were significantly higher in PRP-and insulin-injected groups than in other control groups from 4 weeks after the autologous fat graft, and these differences were maintained up to 12 weeks. There was no correlation between mean platelet numbers and the volume of harvested fat. Conclusion: The present study demonstrates that PRP-injection into autologous fat grafts increases the survival rate and the degree of angiogenesis. Thus, PRP injection with autologous fat grafts would be a promising tool for maintaining the volume of the grafted fat.

• Korean Journal of Pharmacognosy
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• v.36 no.2 s.141
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• pp.129-135
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• 2005

Lectins from Allomyrina dichotoma (ADL) and Bombyx mori (BML) were partially purified by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. An assay for cytokine expression was carried out by using reverse transcription polymerase chain reaction(RT-PCR). mRNA isolated from PBMC(human peripheral blood mononuclear cells) were stimulated with ADL(O.D.=0.2) and BML(O.D.=0.1) for various times(1,4,8,24,48 and 72 h) and various cytokine mRNA assessed by RT-PCR were shown as follows: The patterns of bands for IL-1 mRNA of BML were very similar with those from ADL and these bands were decreased along the increasing reaction times after showing a strong band at 1 h. However mRNA expressions for IL-2, IL-6, $IFN{\gamma}$ and $TNF{\alpha}$ showed different patterns between ADL and BML. With the effect of ADL, the expression of IL-2 and IL-6 mRNA were continuously detected until 72 h with the strongest band of IL-2 mRNA at 24 h. The strong bands of $IFN{\gamma}$ mRNA were observed from 4 to 8 h but the strongest one of $TNF{\alpha}$ was just observed at 1 h. Meanwhile with BML, the bands for IL-2 and $IFN{\gamma}$ were increased along the increasing reaction times until 72 h. The strongest bands were showed from 4 to 8 h with IL-6 and at 8 h with $TNF[\alpha}$. To verify quantitatively ELISA was used for assay of protein secretions of the cytokine gene with IL-2 and $IFN{\gamma}$ expressed markedly different in RT-PCR. The highest cytokine secretion for IL-2 was demonstrated at 48 h. The production of $IFN{\gamma}$ was markedly increased at 24 h and secreted highest at 72 h. These result suggest that ADL and BML, as inducers of cytokines, can elicit detectable cytokine mRNA from PBMC within the first few hours of stimulation and maintain the production of cytokines for a few days by the methods of RT-PCR and ELISA.

• KSBB Journal
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• v.24 no.3
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• pp.246-252
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• 2009

Rice cells were transformed to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter. hCTLA4Ig was produced and secreted into culture media inducibly when sugar was depleted. The obstacles of this system are the cell death and release of proteases by sugar starvation. These problems resulted in the losses of stability and productivity of hCTLA4Ig. Therefore, the effects of proline as an inhibitor of cell death were investigated. When 4 mM proline was added in sugar-free media, the cell death and release of proteases were reduced. As a consequence, the production of hCTLA4Ig was enhanced. In addition, the effects of protein stabilizers such as gelling agents were studied. It was found that the application of 0.01 g/L gelatin led to an increase in hCTLA4Ig production. This increase might be originated from the stabilization of hCTLA4Ig. In conclusion, the production of hCTLA4Ig could be enhanced by the additions of proline and gelatin in transgenic rice cell cultures.

• Journal of Life Science
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• v.25 no.2
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• pp.121-129
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• 2015

Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

• Journal of Life Science
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• v.25 no.2
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• pp.214-222
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• 2015

Aeromonas hydrophila is the most common water fish pathogen and cause diseases such as hemorrhagic septicemia, dropsy, ulceration and asymptomatic septicemia. A. hydrophila secretes many extracellular products (ECPs) which contribute to effective infection, wide distribution and great adaptability to environmental changes. Crude ECPs of A. hydrophila CK257, a strain used in this study, exhibits a toxic activity to the animals including mouse, rabbit and fish. Toxic symptoms were indicated by tissue damage and skin injuries in animal. When ECPs were subcutaneously injected to animals, skin damages were observed, appearing like necrosis. Preliminary research demonstrated that the active factors are protein component. The crude ECPs were collected after ammonium sulfate precipitation of cell-free culture supernatant. ECPs were fractionated with the use gel filtration chromatography. Five ECP fractions were obtained, of which one fraction was found to be toxic to goldfish. MALDI-TOF analyses provided two interesting proteases called M35 and M28. Both M35 and M28 are known as metalloprotease. Accordingly, proteins in an active fraction exhibited caseinolytic activity. These proteins were difference of caseinolytic activity under different metallic ions. Also active fraction has elastolytic activity. These results suggested that peptidase M28 and M35 may be a candidate factor for tissue necrosis activity about infection with A. hydrophila.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.3
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• pp.480-487
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• 1997

Vitellogenin (Vg) was purified from plasma of $estradiol-17\beta(E_2)-treated$ spotted flounder, verasper variegatus, by preripitation with cold distilled water, followed by fractionation using a Sepharose CL-6B column chromatography. $E_2-induced$ protein was identified as Vg by SDS-PAGE and western blot analysis. The molecular weight of the purified Vg was estimated 175 kD as determined by SDS-PAGE. In order to measure the Vg level, monoclonal antibodies against Vg were produced by hybridoma technique. Purified Vg was immunized into Balb/c mice and then the spleen cells from mice were fused with NS-1 myeloma cells. The hybridoma cells were screened by enzyme-linked immunosorbent assay (WLISA) and subcloned by limiting dilution. The hybridoma clones which secreted antibodies highly reactive to the purified Vg were designated as 4D6. Its specificity was demonstrated by western blot from plasma of untreated, $E_2-treated$ male fish, and purified Vg.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.2
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• pp.137-142
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• 2008

Echinacea purpurea, an indian traditional plant medicine has been widely used as herbal remedy for the treatment of disease such as colds or other infections. However, Echinacea purpurea extracts recently have been applied as a cosmetic ingredient for skin care. We artificially cultured Echinacea purpurea by using the bioreactor culture system for this study. We induced callus from Echinacea purpurea and separated adventitious roots, harvested and extracted after cultured in bioreactors. Previously, several studies have been reported on anti-oxidant and immuno-enhancing effects of Echinacea purpurea extract but other efficacies were not well known. In this study, we investigated the whitening, anti-wrinkle and anti-oxidant effects to know applicable value of tissue-cultured Echinacea purpurea adventitious roots extract(TCEPARE) as a cosmetic ingredient. TCEPARE did not show cytotoxicity until a concentration of 2% and showed the anti-oxidative effect in DPPH and NBT tests. Also, the extract decreased tyrosinase expression in a dose-dependent manner and inhibited melanin synthesis in B16 melanoma cells. TCEPARE reduced protein levels of MMP-1, 2 secreted in culture medium or in cell lysates. From these results we suggest that TCEPARE has potential benefits applicable as to cosmetic ingredient for skin care products.

• The Korean Journal of Zoology
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• v.39 no.4
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• pp.337-351
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• 1996

Melatonin Is a multifunctional hormone secreted from the pineal gland in the middle of cerebrum and cerebellum. Its synthesis and release reflect photopedod;Photopedod is a yearly predictable ambient factor that most animals utilize as an environmental cue for maximum survival. Hamsters maintaln reproductive activity in summer during which day length exceeds night time. Upon the advent of autumnal equinox they undergo gonadal regression. The photoperiodic effects are prevented by removal of the pineal gland and restored by the timed repiacument of melatonin. The results suggest that melatonin constitutes part of control mechanism whereby environmental information is transduced to neuroendocrine signal responsIble for the functional integrity of the reproductive system. From the studies for the action site of melatonin following the treatment of photopedod or melatonin in the lesion of a spedflc portion of hypothalamus, suprachiasmatic nuclei and pars tuberalis are shown to be a consensus site for melatonIn. The action of melatonin. In the regulation of reproduction is largely unknown. It is mainly due to the lack of acute effect of melatonin on gonadotropin secretion. However, reduction of the gonadotropln release and augmentation of the hypothalamic gonadotropin-releasing hormone (GnRH) content by long-term treatment of melatonln Indicate that constant presence of melatonln may partidpate in the regulation of sexual activity via the GnRH neuronal system. The action mechanism by which melatonin exerts Its effect on GnRH neuron needs to be eluddated. The inability of opiold analogues to affect the reproductive hormones in sexually regressed animals by inhibftory photopedod and melatonin suggests that the opioldergic neuron may be a prime intervening mediator. Recent cloning of melatonin receptor will contribute to investigate its anatomical Identification and the action mechanism of melatonin on target tissues at the molecular level.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.229-246
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• 1997

Implantation is a most important biological process during pregnancy whereby conceptus establishes its survival as well as maintenance of pregnancy. During the periimplantation period, both uterine endometriurn and conceptus synthesize and secrete a host of growth factors and cytokines which mediate the actions of estrogen and /or progesterone and also exert their steroid-independent actions. Growth factors expressed by the materno-conceptal unit en masse have important roles in cell migration, stimulation or inhibition of cell proliferation, cellular differentiation, maintenance of pregnancy and materno-conceptal communications in an autorcrine /paracrine manner. The present review focuses on the role of the intrauterine IGF system during periimplantation conceptus development. The IGF system comprises of IGF- I and IGF- II ligands, types I and II IGF receptors and six or more IGF-binding proteins(IGFBPs). IGFs and IGFBPs are expressed and secreted by uterine endometrium with tissue, pregnancy stage and species specificities under the influence of estrogen, progesterone and other growth factor(s). Conceptus also synthesizes components of the IGF system beginning from a period between 2-cell and blastocyst stages. Maternal IGFs are utilized by both maternal and conceptal tissues; conceptus-derived growth factors are believed to be taken up primarily by conceptus. IGFs enhance the development of both maternal and conceptal compartments in a wide range of biological processes. They stimulate proliferation and differentiation of endometrial cells and placental precursor cells including decidual transformation from stromal cells, placental formation and the synthesis of some steroid and protein hormones by differentiated endometrial cells or placenta. It is also well-documented in a number of experimental settings that both IGFs stimulate preimplantation embryo development. In slight contrast to these, prenatal mice carrying a null mutation of IGF and /or IGF receptor gene do not exhibit any apparent growth retardation until after implantation. Reason (s) for this discrepancy between the knock-out result and the in vitro ones, however, is not known. IGFBPs, in general, are believed to inhibit IGF action within the materno-conceptal unit, thereby allowing endometrial stromal cell differentiation as well as dampening ex cessive placental invasion into maternal tissue. There is evidence, however, indicating that IGFBP can enhance IGF action depending on environrnental conditions perhaps by directioning IGF ligand to the target cell. There is also a third possibility that certain IGFBPs and their proteolytic fragments may have their own biological activities independent of the IGF. In addition to IGFBPs, IGFBP proteases including those found within the uterine tissue or lumen are thought to enhance IGF bioavailability by degrading their substrates without affecting their bound ligand. In this regard, preliminary results in early pregnant pigs suggest that a partially characterized IGFBP protease activity in uterine luminal fluid enhances intrauterine IGF bioavailability during conceptus morphological development. In summary, a number of in vitro results indicate that IGFs stimulates the development of the rnaterno-conceptal unit during the periimplantation period. IGFBPs appear to inhibit IGF action by sequestering their ligands, whereas IGFBP proteases are thought to enhance intrauterine bioavailability of IGFs. Much is remaining to be clarified, however, regarding the roles of the individual IGF system components. These include in vivo evidence for the role of IGFs in early conceptus development, identification of IGF-regulated genes and their functions, specific roles for individual IGFBPs, identification and characterization of IGFBP proteases. The intrauterine IGF club house thus will be paying a lot of attention to forthcoming results in above and other areas, with its door wide-open!

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.847-850
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• 2005

The aim of this study was to increase production of leucocin A, a kind of bacteriocin, in a transformed variety of S. cerevisiae. We investigated optical density, total secreted protein, protease activity, and antibacterial activity for the transformed S. cerevisiae in different carbon sources. The production of leucocin A growth-associated, and antibacterial activity, according to carbon source, was in the order of sucrose, glucose, glycerol, and fructose. Antibacterial activity was $10.6\%$ higher in the presence of sucrose than glucose. This is the first report regarding the effect of carbon sources on the production of leucocin A in transformed S. cerevisiae, as far as we ascertain. Our results could prove useful in the industrial production of natural preservatives.