A nineteen years old male patient showed a cystic lesion in left maxillary canine to premolar area (#23-#25). This lesion was asymptomatic, and found during his routine radiological check in local clinic. In the radiological observation the cystic lesion showed round radiolucent image containing many calcified bodies which were usually small but irregular in shape, expanding tumorously and resulted in the displacement of canine and second premolar in the absence of first premolar. The lesion was surgically enucleated, and a cystic fibrous tissue containing abnormal teeth was removed and examined pathologically. With the histological observation of tumorous odontogenic epithelium including many ghost cells, which were closely associated with abortive teeth, the lesion was finally diagnosed as CCOT associated with complex odontoma. The ghost cells of CCOT was strongly positive for β-catenin, GADD45, and LC3, and slightly positive for MMP-9, while they were rarely positive for BCL2, Wnt1, HSP-70, and p38. Therefore, it was presumed that the ghost cells of CCOT might undergo dormant cell state through altered cytodifferentiation stimulated by severe growth arrest, DNA damage signaling, and abundant autophage formation.

본 연구는 생물전환된 대두의 isoflavone 함량, 총 페놀함량, 항산화능(DPP radical 소거능, ORAC 지수) 및 β-Glucan 함량을 측정하였다. Isoflavone의 경우 추출용매에 상관없이 배당체가 모두 비배당체로 전환되는 것을 확인하였다. Total isoflavone 함량의 경우 hexane 탈지 대두박발효물에서 2577.96 μg/mL으로 가장 높은 값을 나타냈으며, ethanol 탈지 대두박 비발효물에서 428.27 μg/mL으로 가장 낮은 값을 나타내었다. 총 페놀 함량은 대두 원물에서 39.44 mg GAE/g으로 나타났으며, ethanol 탈지 대두박비발효물 및 hexane 탈지 대두박 비발효물은 27.07, 27.75 mg GAE/g으로 대두 원물보다 다소 낮은 값을 나타냈다. 생물전환된 대두의 총 페놀 함량은 hexane 탈지 대두박발효물 41.61 mg GAE/g, ethanol 탈지 대두박 발효물42.34 mg GAE/g으로 비발효물에 비해 약 1.5배 가량 증가된 함량을 보였다. DPPH radical 소거능의 경우 대두 원물에서 51.10%의 소거능을 나타내었고 hexane 탈지 대두박 비발효물에서 50.51%, ethanol 탈지 대두박 비발효물은 43.27%의 소거능을 나타냈다. 생물전환된 ethanol 탈지 대두박 발효물에서 59.92%로 radical 소거능이 증가되었지만 hexane 탈지 대두박 발효물은 31.30%로 비발효물에 비해 낮은 radical 소거활성을 보였다. ORAC 지수는 대두 원물이 384.47 μM TE/g을 보였으며, hexane 탈지 대두박 비발효물 및 ethanol 탈지 대두박 비발효물은 318.52, 247.48 μM TE/g으로 나타났다. 생물전환된 hexane 탈지 대두박 발효물은 786.36 μM TE/g, ethanol 탈지 대두박 발효물에서 721.96 μM TE/g으로 비발효물에 비해 ORAC 지수가 증가하는 것으로 나타났다. β-Glucan 함량은 0.09~0.11%의 범위로 나타났으며 대두 원물과 ethanol 탈지 대두박 발효물에서 가장 높은 0.11%를 보였고 hexane 탈지 대두박 비발효물과 hexane 탈지 대두박 발효물에서 0.09%로 가장 낮은 β-glucan 함량을 보였지만, 추출용매 및 생물전환에 따른 β-glucan 함량의 큰 차이는 나타나지 않았다.

Many transgenic domestic animals have been developed to produce therapeutic proteins in the mammary gland. However, purification of therapeutic proteins from transgenic milk are very important for productivity of recombinant protein. Development of a knock-in vector system is needed to improve production of therapeutic proteins. In this study, we are develop Knock-in vector to express human Erythropoietin protein (hEPO) using Gluthathione S-transferase (GST) fusion system on mouse β-casein exon 3 locus. The knock-in vector consisted of the 5 homologous arm (1.02 kb), GST, PreScission protease site, hEPO cDNA, BGH polyA signal, CMV-EGFP, and 3homologous arm(1.81 kb). The analysis of nucleotide and amino acid sequence revealed that GST-hEPO mRNA is probably translated with the mouse β-casein sequence and the β-casein-GST-hEPO fusion protein is probably secreted by ER-Golgi pathway. After that, the hEPO protein can be cleaved to remove the GST from the fusion protein by PreScission protease during purification of recombinant protein. This knock-in vector may help to create transgenic mouse expressing human Erythropoietin protein via the endogenous expression system of the mouse β-casein gene in the mammary gland.

The production of therapeutic proteins from transgenic animals is one of the most important successes of animal biotechnology. Endostatin is 20 KDa C-terminal fragment derived from type XVIII collagen and an endogenous inhibitor of tumor growth by inhibition of angiogenesis. In this study, we are developed knock-in vector consists of 5’ arm region (1.02 kb), human Endostatin cDNA, CMV-EGFP, and 3’ arm region (1.83 kb). To express Endostatin gene as transgene, the F2A sequence was fused to the 5’ terminal of Endostatin gene and inserted into exon 3 of the β -casein gene. If this knock-in vector is inserted into the porcine β-casein gene locus by homologous recombination, human Endostatin mRNA are expressed using the gene regulatory region of the β-casein. Also, the β-casein and Endostatin fusion protein is translated and Endostatin protein is separated by F2A self cleavage during translation. In conclusion, our knock-in vector may help to create transgenic pig expressing human Endostatin protein via the endogenous expression system of the porcine β-casein gene in the mammary gland.

Although in vitro production (IVP) techniques of porcine follicular oocytes have progressed and are well studied, the developmental potential of porcine oocytes matured in vitro remains low compared with those matured in vivo. It is well known that one of the reason occurred impair in vitro maturation (IVM) of porcine oocytes is the oxidative stress. Oxidative stress is mainly caused by reactive oxygen species (ROS) generation formed during cellular metabolism. β-cryptoxanthin (BCX) is one of the carotenoid pigment and possesses strong anti-oxidative and free radical scavenging activities and suppresses lipid peroxidation and nitrogen oxide production. The objective of this study was to examine the effects of BCX treatment on porcine oocyte during IVM and their in vitro developmental potential. The follicular oocytes were cultured in IVM medium supplemented with 0, 0.1, 1, 10 and 100 μM BCX (control, 0.1 B, 1 B, 10 B and 100 B). In analysis of intracellular ROS expression level after IVM, 1 B group was the lowest among all groups (p<0.05), while other BCX treated groups are similar to control group. Also, 1 B group was significantly decreased during the classified oocyte maturation stage (GVBD, MⅠ and MⅡ) than control (p<0.05). In addition, the relative mRNA expression level of antioxidant gene (superoxide dismutase-2 and peroxiredoxin-5) was significantly higher in 1 B group than control (p<0.05). After parthenogenetic activation, there was no different in the cleavage rate between two groups, however, the blastocyst formation rate was significantly higher in 1 B group than in control (p<0.05). In embryo quality, the total cell number and DNA fragmentation of blastocysts were no different between two groups. These results demonstrate that BCX is helpful for decreasing ROS level of porcine follicular oocytes and improves their developmental potential.

To increase the stability of anthocyanin from black rice, β-cyclodextrin (β-CD) formed a complex with anthocyanin, and thermal and UVB stability of the complex was investigated at various pH levels. The formation of β-CD complex with anthocyanin was confirmed by the decreased UV-Vis spectrophotometer absorbance (at 511nm) with increasing the concentration of β-CD at pH 2, 2.5, 3, 4, and 5. ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid)assay was conducted to measure radical scavenging capacity (RSC) of the complex at pH 2, 4, 6, and 8, after which the RSCs were recorded over the 24 h heating(95°C) and 24 h UVB irradiating periods. The RSC values of the complexes increased as the β-CD concentration increased (0.1-2%) at pH 6 and 8. Upon thermal treatment, the RSC readings of the complexes tended to decrease to a lesser extent compared to the anthocyanin control (without β-CD) at pH 2. This result suggests that the thermal stability of anthocyanin was more retained in the presence of β-CD at low pH (pH 2). However, in the case of UVB irradiation, the effect of β-CD complex on the anthocyanin RSC was not significant, though the RSC values for both the anthocyanin control and complexes trended downward as the UVB irradiation time increased at all pH ranges except for pH 8 (no downward trend). In short, this study suggests that the effect of β-CD complex on the stability of anthocyanin antioxidant capacity depends on pH and the susceptibility to the degradation process.

The glucosidase inhibitors are currently interested owing to their promising therapeutic potential in the treatment of disorders such as diabetes, human immunodeficiency virus (HIV) infection, metastatic cancer, and lysosomal storage diseases. Among them, β-d-glucosidase inhibitors can be used to elucidate the mechanisms of various biochemical reactions and to treat the Gaucher disease as potential therapeutics. In this study, the natural β-glucosidase inhibitor was isolated from jujube leaf extract. The isolation and purification are vital for the structure analysis. The jujube leaf was extracted with hot water (95°C, 15 min). Then the natural β-glucosidase inhibitor was isolated using size exclusion chromatography and semi-preparative HPLC. In the first purification process, the 35 fractions were collected according to molecular size using PD-10 column packed with Sephadex G-25 medium. The strongest inhibition on the activity of β -glucosidase was observed at 6 and 7 fractions. The chromatogram of these samples showed that 3 peaks were observed comparing with baseline. For further purification, the strongest inhibitory activity peak was isolated using semi-preparative HPLC with VDS C18 column. Highly purified inhibitor, which can be used for structure analysis and characterization, was obtained. Further study is necessary to identify the natural β-glucosidase inhibitor in the jujube leaf extract.

The aim of the present study was to develop an animal model for evaluation of temporomandibular (TMJ) nociception under TMJ inflammation. We also investigated the participation of IL-1β in inflammation-induced TMJ nociception. Experiments were carried out using male Sprague-Dawley rats. Intra-articular injection of 3% formalin was administered to evaluate hyperalgesia 3 days after CFA injection. Intra-articular injection of 3% formalin did not produce nociceptive behavior in normal rats. Although intra-articular injection of 3 doses of CFA produced TMJ inflammation, only 1:3 diluted CFA produced hyperalgesia when formalin was injected intra-articularly 3 days after CFA injection. Co-administration of IL-1 receptor inhibitor with formalin into the TMJ cavity 3 days after CFA injection was performed. Co-administration of IL-1 receptor inhibitor significantly inhibited formalin-induced hyperalgesia in rats with CFA-induced TMJ inflammation. These results suggested that intra-articular injection of formalin produced hyperalgesia under chronic TMJ inflammation. Moreover, IL-1β plays an important role in TMJ hyperalgesia under chronic inflammation and blockade of IL-1β is a potential therapeutic target for inflammatory TMJ pain.

β-carotene is present in carrots, pumpkins, and sweet potatoes. It suppresses many types of cancers by regulating cellular proliferation and apoptosis through a variety of mechanisms. However, the effects of β -carotene on oral cancer cells have not been clearly established. The main goal of this study was to investigate the effects of β-carotene on cell growth and apoptosis in oral cancer cells. Our results demonstrate that treatment with β-carotene induced inhibition of cell growth, and that the effect was dependent on β-carotene treatment time and concentration in KB cells. Furthermore, treatment with β-carotene induced nuclear condensation and fragmentation in KB cells. β-carotene promoted proteolytic cleavage of procaspase-3, -7, -8 and –9 with associated increases in the concentration of cleaved caspase-3, -7, -8 and –9. In addition, the level of cleaved PARP was increased by β-carotene treatment in KB cells. These results suggest that β-carotene can suppress cell growth and induce apoptosis in KB human oral cancer cells, and that it may have potential usefulness in anti-cancer drug discovery efforts.

HA (hydroxyapatite)/β-TCP (tricalcium phosphate) biomaterial (BCP; biphasic calcium phosphate) is widely used as bone cement or scaffolds material due to its superior biocompatibility. Furthermore, NH4HCO3 as a space holder (SH) has been used to evaluate feasibility assessment of porous structured BCP as bone scaffolds. In this study, using a spark plasma sintering (SPS) process at 393K and 1373K under 20MPa load, porous HA/β-TCP biomaterials were successfully fabricated using HA/β-TCP powders with 10~30 wt% SH, TiH2 as a foaming agent, and MgO powder as a binder. The effect of SH content on the pore size and distribution of the BCP biomaterial was observed by scanning electron microscopy (SEM) and a microfocus X-ray computer tomography system (SMX-225CT). The microstructure observations revealed that the volume fraction of the pores increased with increasing SH content and that rough pores were successfully fabricated by adding SH. Accordingly, the cell viabilities of BCP biomaterials were improved with increasing SH content. And, good biological properties were shown after assessment using Hanks balanced salt solution (HBSS).

국내외에서 수집한 표고 품종을 대상으로 영양성분 및 β-glucan 함량을 분석한 결과 수집한 표고의 유리당 함량 을 분석한 결과 총 4종의 유리당이 검출되었으며, 그 중 trehalose 함량은 시료별로 0.83 ~9%까지 큰 차이를 보였 다. 구성아미노산 함량을 분석한 결과 총 아미노산함량은 중국 톱밥배지 표고 JMI10050에서 17,672 mg%로 가장 높게 나타났으며, 필수아미노산의 함량은 국내 원목재배 표고 JMI10059에서 6,211 mg% 가장 높게 나타났다. 유 리아미노산 함량을 분석한 결과, 총 16종의 아미노산이 검출되었으며, 주요 유리아미노산으로는 histidine, glutamic acid 및 arginine으로 나타났다. 필수 유리아미노산인 threonine, valine, methionine, isoleucine, leucine, phenylalanine, histidine 및 lysine의 함량은 중국 톱밥배 지 표고 JMI10052에서 521.06 mg%로 가장 높게 나타 났다. 중국에서 수집한 표고에 비하여 국내에서 재배된 표고 의 β-glucan이 약 10% 가량 높게 검출되었고, 그 중 국내 원목재배 표고 JMI10066이 31.74%로 가장 높은 함량을 나타냈다.

The effects of dietary β-glucan, obtained from bacterial fermentation, on the intestinal mass, short chain fatty acids, lactate production and pH in Sprague-Dawley (SD) rats were evaluated. SD rats fed with 0% (control group), 1% or 5% β-glucan supplemented diets (w/w) for 3 weeks. The presence of β-glucan in the diets resulted in a significant increase in colonic contents in a dose dependent manner. The amount of short chain fatty acids increased in rats fed β-glucan diets. Rats fed the 5% β-glucan diets had higher levels of acetate, propionate and butyrate by 1.8, 1.7 and 3.0 fold of the control group in the cecum, and 2.2, 2.9 and 3.1 fold of the control group in the colon, respectively. The β-glucan diets also significantly increased the levels of cecal and colonic lactate by 1.4~3.4 fold, when compared to the control diet, indicating that dietary β-glucan stimulated the growth of lactic acid bacteria within the intestine. These results suggest that dietary β-glucan, by providing short chain fatty acids and reducing the cecal and colonic pH, may be beneficial in improving gut health, and provide evidence for the use of β-glucan as a dietary supplement for human consumption.

Integrin is a heterodimer protein that locates on cell membrane to interact with neighboring cells or extracellular matrix. A transcriptome analysis of the brassica leaf beetle, Phaedon brassicae, midgut identified both α and β subunits of integrin. RNA interference of β subunit genes significantly impaired survival of both larvae and adults of P. brassicae. A recombinant bacteria expressing double-stranded RNA specific to β integrin of P. brassicae were constructed and showed significant oral toxicities.

A response surface methodology (RSM) was used to evaluate how pH and ionic strength (IS) affect the fate (i.e. size and colloidal stability) of an SC formulation containing the pyrethroid β-cyfluthrin. The response surfaces determined under a range of environmentally relevant conditions were then used to assess the toxicity of the SC formulation of β-cyfluthrin to D. magna. The changes in hydrodynamic diameter (HDD) and colloidal stability as determined by zeta potential measurement were closely related to either or both of the change in pH and IS with the linear factor of IS being the most significant factor affecting those changes. Thus, the concentration of SC formulation of β-cyfluthrin remaining in the water column was dependent on the pH and IS conditions and highest when the colloidal suspension contained small particles or a lack of agglomeration leading to sedimentation of the particles. The toxicity results show correspondingly higher toxicity to D. magna when exposed to the SC formulation of β-cyfluthrin when pH and IS conditions favor formation of either the smallest HDD or most stable colloidal suspensions.

무질서 매질에서 형광, 산란과 응집의 영향은 파장과 산란된 형광세기로 나타내는데, laser induced fluorescence(LIF) 분광학에 의한 분자특성으로 나타난다. 산란매질에서 광학적 효과는 광학적 파라미터들(μs, μa, μt)에 의해 표현되고 응집은 고-액상 분리공정과 Photodynamic therapy에서 중요 하게 활용되고 있다. 따라서 입자가 서로 접근될 때 콜로이드 입자들의 상호작용을 LIF와 응집효과로 분석하였다. 우리는 레이저 광원에서 검출기까지 거리의 함수에 의해 in vitro 시료의 산란과 형광 스펙 트라를 측정하였다. 산란계수 μs는 산란체의 입자가 증가함에 크게 나타났다. 그리하여 purple membrane vesicle과 β-carotene의 혼합물의 매질에서 광원에서 검출기에 의한 거리에 대한 측정된 값 (I, δ)이 거리가 가까워짐에 따라 크게 나타났다.

The objective of the present study was to examine the effects of β-glucan originating from Aureobasidium on full-thickness skin wound healing in diabetic C57BL/KsJ-db/db mouse models. In the diabetic C57BL/KsJ-db/db model, test articles were topically applied twice a day for 20 days starting from 1 day after wounding. The results were compared to that of MadecassolTM ointment (madecassol; 1% Centella asiatica extracts) topically applied at a concentration of 100 mg/kg. Treatment with β-glucan resulted in significant (p<0.01 or p<0.05) and dose-dependent decreases in wound size compared with that of vehicle control showing increased wound size (WS, %). In addition, 50% contraction time (CT50) was dramatically and dose-dependently reduced, and inflammatory cells in granulation tissues of the wound area were significantly (p<0.01 or p<0.05) and dose-dependently reduced compared with that of vehicle control showing increased numbers of micro-vessels and fibroblasts as well as re-epithelialization. In the madecassol group, similar changes in inflammatory cells and fibroblasts with re-epithelialization were also observed, but madecassol did not influence angiogenesis. No meaningful changes in body weight were detected in all tested groups compared with the vehicle control. Therefore, these data suggest that β-glucan has a beneficial effect on diabetic delayed skin wound healing and may be useful to manage incurable skin wounds in diabetic animals.

국내에서 생산되어 시판되고 있는 보리쌀 제품을 수집하여아밀로오스와 β-glucan 함량을 분석하고 아밀로오스 함량에 따른 찰성 및 메성 보리쌀의 호화 및 취반 특성을 비교하였다.보리쌀 제품은 제품에 표기된 바에 따라 찰보리쌀과 메보리쌀로 분류하여 이들의 아밀로오스 함량을 측정하였다. 분석결과보리쌀 제품의 아밀로오스 함량은 4.46 ~ 30.68% (평균 16.33%)로 다양하게 나타났으며, 찰보리쌀 제품은 대부분 아밀로오스함량이 5 ~ 10% 정도의 분포를 보였다. 보리쌀 제품의 β-glucan 함량은 2.49 ~ 6.79% 범위(평균 4.57%)로 분석되었다.시판 보리쌀 제품을 찰보리쌀(아밀로오스 10% 이하)과 메보리쌀(아밀로오스 20% 이상) 그룹으로 구분하여 각각 선발한보리쌀 제품을 비교하였으며, 찰보리쌀이 메보리쌀에 비해 β-glucan 함량이 약 2% 높게 나타났다. 찰보리쌀은 메보리쌀에비해 신속점도측정기(RVA)에 의한 호화개시온도, 최고점도,trough, breakdown, 최종점도, setback이 낮게 나타났다. 보리쌀의 취반특성에서 시판 찰보리쌀은 메보리쌀에 비해 수분흡수율과 퍼짐성은 높은 반면 용출고형분이 낮았으며, 텍스쳐(경도)는 찰보리쌀이 낮아 보리밥의 취반특성 및 식감이 보다 좋은 것으로 평가되었다.