Periodontal disease is an inflammatory disease that affects the destruction of the bone supporting the tooth and connective tissues surrounding it. Periodontal ligament fibroblasts (PDLFs) induce overexpression of matrix metalloproteinase (MMP) involved in periodontal diseaseʼs inflammatory destruction. Osteoclasts take part in physiological bone remodeling, but they are also involved in bone destruction in many kinds of bone diseases, including osteoporosis and periodontal disease. This study examined the effect of baicalin on proteolytic enzymesʼ production and secretion of inflammatory cytokines in PDLFs and RAW 264.7 cells under the lipopolysaccharide (LPS)-induced inflammatory conditions. Baicalin inhibited the expression of the protein, MMP-1 and MMP-2, without affecting PDLFs’ cell viability, suggesting its possibility because of the inhibition of phosphorylation activation of mitogen-activated protein kinase’s p38, and the signal transduction process of nuclear factor κB (NFκB)-related protein. Also, baicalin reduced the expression of MMP-8 and MMP-9 in RAW 264.7 cells. This reduction is thought to be due to the inhibition of the signal transduction process of NFκB-related proteins affected by inhibiting p65RelA phosphorylation. Also, baicalin inhibited the secretion of nitric oxide and interleukin-6 induced by LPS in RAW 264.7 cells. These results suggest that baicalin inhibits connective tissue destruction in periodontal disease. The inhibition of periodontal tissue destruction may be a therapeutic strategy for treating inflammatory periodontal-diseased patients.

반추동물의 급여 단백질 효율성은 반추위 내 미생물 발효에 따른 우회단백질, 미생물체단백질, 사료 및 미생물체의 아미노산 구성 등에 의해 결정된다. 실험 1은 주요 단백질 사료원료 중 국내 한우 사육에 주로 이용되는 사료원료 15종[옥수수(남미), 옥수수(우크라이나), 단백피(국내), 단백피(중국1), 단백피(중국2), 대두박(국내), 대두박(미국), 대두박(브라질), DDGS(미국), 팜박(인도네시아), 채종박(캐나다), 루핀(미국), 미강(국 내), 주정박(국내), 타피오카(인도네시아)]의 반추위 내 단백질과 아미노산의 우회 수준을 측정하였다. 캐뉼라 장착 한우 암소를 이용하여 in situ 실험을 진행하였다. 우회 단백질 함량은 타피오카(인도네시아)에서 59.98%로 가장 높았다. 타피오카(인도네시아)는 총 우회 아미노산 함량이 73.73% 로 가장 높았으며, glutamic acid (Glu)와 alanine (Ala)이 72.17%, 75.23%로 가장 높았다. 실험 2에서는 실험 1과 같은 원료를 사용해 반추위 내 미생물 발효 특성에 따른 미생물체 아미노산 조성분석을 실시하였다. 실험 결과 채종박(캐나다)의 pH는 6.99 (p<0.05), 미생물 단백질 합성량은 326.53mg/100mL (p<0.05), NH3-N의 농도는 단백피(중국 1)가 48.20mg/100mL (p<0.05)로 높았다. 옥수수(우크라이나)의 총 VFA 농도는 97.92mM (p<0.05)이었다. 미생물의 총 아미노산 함량은 대두박(미국) 52.07%로 높았으며 아미노산 조성은 모든 시험 사료에서 Glu가 높았다. 따라서, 본 연구는 한우의 급여 단백질 사료원에 대한 반추위 내 우회단백질, 우회아미노산함량과 반추위 내 미생물 아미노산 조성에 대한 연구를 실시하였으며, 향후 한우 단백질 급여사료원에 대한 대사단백질(MP) 공급체계의 기초자료로 활용될 수 있을 것이다.

Autophagy is a cell survival mechanism that works for the survival of cells under various physiological and pathological conditions. ATG5 is a key protein in the process of autophagy propagation and is involved in tumor development and progression. Chemotherapeutic agents targeting ATG5 enhance the host's immune response in various human cancers and intensify the effectiveness of chemotherapy. However, the physiological role of ATG5 protein in oral squamous cell carcinoma (OSCC) has not been fully recognized. The purpose of this study was to examine the correlation between clinico-pathological factors of OSCC patients and ATG5 immunoexpression through immunohistochemistry (IHC) in the tissues of OSCC patients treated at our hospital, and to analyze the regarding influences and their mechanisms. The authors analyzed 20 OSCC patients from National University Dental Hospital, at Pusan National University from January 2002 to December 2007, which were eligible for the study. Data were obtained by reviewing the medical records of the OSCC patients, and ATG5 immunoexpression was obtained using IHC staining in the tissue samples of the OSCC patients. In the tissue sample of OSCC patients, the immunoexpression of ATG5 elevated, in comparison to the normal oral mucosa, and there was a significant correlation with Drinking, Pathological Stage. In regards to Cox regression analysis, Clinical stage, Tumor size, Histopathologic grade, Cervical nodal metastasis, Loco-regional recurrence, and ATG5 expression have statistically significant correlations. These results imply that the changes in the expression of ATG5 proteins in OSCC can be a prominent factor in the OSCC progression and the prognosis of OSCC patients.

To understand the role of small heat shock protein (sHSPs) in rice plant response to various stresses such as the heat and oxidative stresses, a cDNA encoding a 24.1 kDa mitochondrial small HSP (Oshsp24.1) was isolated from rice by rapid amplification of cDNA ends (RACE) PCR. The deduced amino acid sequence shows very high similarity with other plant small HSPs. DNA gel blot analysis suggests that the rice genome contains more than one copy of Oshsp24.1. High level of expression of Oshsp24.1 transcript was observed in rice seedlings in response to heat, methyl viologen, hydrogen peroxide, ozone, salt and heavy metal stresses. Recombinant OsHSP24.1 protein was produced in E. coli cells for biochemical assay. The protein formed oligomeric complex when incubated with Sulfo-EGS (ethylene glycol bis (succinimidyl succinate)). Our results shows that Oshsp24.1 has an important role in abiotic stress response and have potential for developing stress-tolerant plants.

L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl- 2 family and MAPK/Akt signaling pathways.

Heat shock proteins (HSPs) are highly conserved cellular proteins that contribute to adaptive responses of organisms to a variety of stressors. In response to stressors, cellular levels of HSPs are increased and play critical roles in protein stability, folding and molecular trafficking. The mRNA expression pattern of two well-known heat shock protein transcripts, HSP70 and HSP90 were studied in two tissues of nerve ganglia, cerebral ganglion and pleuropedal ganglion of Pacific abalone (Haliotis discus hannai). It was observed that both HSP70 and HSP90 transcripts were upregulated under heat stress in both ganglion tissues. Expression level of HSP70 was found higher than HSP90 in both ganglia whereas cerebral ganglion showed higher expression than pleuropedal ganglion. The HSP70 and HSP90 showed higher expression at Day-1 after exposed to heat stress, later decreased at Day-3 and Day-7 onwards. The present result suggested that HSP70 and HSP90 synthesize in nerve ganglion tissues and may provide efficient protection from stress.

오존은 수돗물 정수장에서 이용되는 소독 물질로 미세오염 물질들을 비롯해서 박테리아나 병원성 미생물체를 효과적으로 제거하는 것으로 많은 연구가 보고되어 있다. 본 연 구에서는 실내 사육 중인 붉은 체색을 지닌 Glyptotendipes tokunagai를 대상으로 서로 다른 농도의 오존 노출에 따른 영향을 파악하기 위해 치사율, 체색 변화와 heat shock protein 70 (HSP70) 유전자 발현을 측정하였다. 오존에 노출 된 G. tokunagai에서 농도-시간 의존적으로 치사율 증가가 관찰되었다. 또한 체색 변화는 오존 농도에 따라 붉은색의 체색이 체절마다 엷어지며 탈색되고 경직되는 현상이 보였다. HSP70 유전자 발현은 저농도인 0.2~0.5 ppm에서 노출 10분과 20분에 유의한 수준으로 높게 나타났으나 (P<0.05), 30 분 노출 후에는 발현량이 감소하는 경향을 보였다. 생리적으로 저산소층에 대해 적응능력이 뛰어난 깔따구 경우에도 오존은 매우 강력한 치사 효과를 유발하여 30분 노출 후 경직과 헤모글로빈 파괴로 인한 탈색이 유발되는 것을 보여주었다. 따라서 본 결과는 수돗물 정수장에서 병원성 미생물을 제거하는 데 사용되는 오존이 수생물에 주는 영향성을 파악하는 기초자료로서 활용될 수 있을 것이다.

Many parrots are considered endangered species due to threats from human activities. Gender determination is of great importance for biological studies and the conservation of endangered parrots. However, like other birds, gender determination in parrots is hindered due to the lack of external dimorphism between males and females. A molecular approach using the chromo-helicase-DNA binding protein 1 (CHD1) gene is commonly used for sexing birds. This study aimed to determine the gender of parrots from Korean zoos based on amplification and visualization of the partial CHD1 gene. The samples of 13 parrot species were collected from three different zoos in Korea and the extracted DNA templates were amplified using CHD1 gene primers. The gender of 27 samples of 13 species was determined by visualizing the PCR products on an agarose gel. While male parrots were indicated by a single band, female parrots were indicated by double bands. The findings provide additional information, which might be helpful for the management and care of parrots in Korean zoos.

Data on the crude protein requirements of elk doe are nonexistent and the data are essential for their management in Korea. Therefore, this study was conducted to evaluate the crude protein requirement for maintenance of elk doe. Three female elk deer were used in 3 × 3 Latin square design with three diets containing three levels of crude protein (CP) that contained low crude protein (approximately 12%), medium crude protein (15%), and high crude protein (18%). Each three elk doe trials included a 14-day preliminary period and a 5-day collection period. Crude protein intake was 4.83, 6.26, and 9.00 g/d for 12%, 15%, and 18% of CP level, respectively. Crude protein balances were 1.04, 1.41, and 4.14 for 12%, 15%, and 18% of CP level, respectively. The maintenance requirement for CP from the regression equation between CP intake and CP balance were 3.70 g/BW0.75.

Canine parvovirus (CPV) remains a leading infectious cause of death in canines, especially in young puppies. Though vaccination is being carried out regularly, immunization failures occur, and puppies may be exposed to infection. Virus-like particles (VLPs) act like a subunit vaccine, mimicking the structure of authentic viruses. Therefore, VLPs have the potential to be used as vaccine candidates. Since Viral Protein 2 (VP2), a major structural protein of CPV, is the crucial antigen for CPV, the purpose of this study was to produce a recombinant VP2 of new canine parvovirus-2a using the baculovirus expression system in SF9 insect cells. The results revealed that recombinant VP2 assembles to form VLPs with antigenic properties similar to those of natural CPV, the recombinant VLP can produce a hemagglutination assay (HA) titer (1:210) in SF9 cells. Expression of the recombinant 6-His-tagged VP2 in SF9 cells was confirmed by western blotting. These findings suggest that the recombinant VP2 expressed in this study could be used as an efficient subunit vaccine against CPV infection.

Runt related transcription factors (RUNX), a family of well-known transcription factors, play key regulatory roles in diverse biological processes, such as proliferation, differentiation, and DNA repair. Of RUNX family, RUNX3 is the least well characterized of the three family members. Nevertheless, the role of RUNX3 as a key regulator in essential biological pathways has been reported and inactivation of RUNX3 leads to a variety of disease, such as cancer, via regulation of Wnt signaling and K-ras mutations in many mammalian tissues. Recent studies using RUNX3-deficient cells and mice revealed an association with hematopoiesis and hypersensitivity to granulocytecolony stimulating factor. Nevertheless, protein dynamics associated with RUNX3 remain poorly understood. In the present study, we performed a large-scale protein study from Runx3 knockout (KO) mouse embryonic stem cells (mESC) using a stable isotope labeling by amino acids (SILAC)-based quantitative proteomics approach. The results showed that 67 proteins were significantly up and downregulated after Runx3 KO. Bioinformatic analyses that revealed that these proteins have diverse biological functions, such as substances transport and cellular structure. Thus, our results enhance our current understanding of the function of RUNX3 in mESCs and suggest potential roles for RUNX proteins in diverse diseases. Additionally, our results can be used as a database to help us understand the mechanism of action of RUNX3.

The fatty acid binding protein 4 (FABP4) gene plays an important role in lipid metabolism and homeostasis in adipocytes. The objective of this study was to analyze the association between a single nucleotide polymorphism (SNP), g.7516G>C, in the FABP4 gene and economic traits of Korean native cattle, Hanwoo. Primers were designed to target a region of the FABP4 gene between nucleotides 7417 and 7868 (AAFC01136716). The SNP, which was detected by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method using restriction enzyme MspA1I, was genotyped in 319 animals of Hanwoo steer population. Statistical analysis showed that the SNP genotype of the FABP4 gene significantly affected carcass weight (CW, p<0.01), longissimus muscle area (LMA, p<0.001), and marbling score (MS, p<0.001). GG allele of the SNP on 246 animals in a Wagyu × Limousin F2 reference population showed a higher MS (p<0.05) and subcutaneous fat depth (p<0.05) in previous report. But CC allele of the SNP showed greater values for MS, LMA, and CW in Hanwoo steers. These results suggest that the g.7516G>C SNP located in the FABP4 gene may affected differently depending on the cattle breed and can be used as a genetic selection marker in Korean native cattle.

노화는 말초 혈관 기능에 퇴행성 변화를 가져올 수 있으며 혈관의 기능은 혈관생성 지표의 영향을 받는 것으로 보고되고 있다. 따라서 이 연구는 젊은 흰쥐와 노화된 흰쥐를 대상으로 골격근(가자미근)에서 나타나는 혈관생성 지표인 Ang-1과 Ang-2의 단백질 발현의 차이를 비교하고 Ang-1 : Ang-2 ratio의 차이를 분석 및 관찰하기 위해 수행되었다. 연구의 목적을 위해 12주령 흰쥐(SD rats) 6마리와 20개월령 흰쥐 6마리를 Young 집단과 Old 집단으로 구분하였다. Old 집단의 경우 20개월 동안 직접 사육하여 노화 상태를 유도하였다. 골격근(가자미근)에서 관련 단백질의 발현 정도를 알아보기 위해 western blot 기법을 사용하였다. 연구결과, Ang-1 단백질은 젊은 집단에 비해 노화집단에서 낮게 나타나며 집단 간 차이를 보였다(p= .001). Ang-2 단백질의 경우, 젊은 집단에 비해 노화집단에서 유의하게 높게 나타났다(p= .007). 마지막으로 Ang-1:Ang-2 ratio는 젊은 집단에 비해 노화집단에서 유의하게 낮게 나타났다(p= .001). 따라서 골격근 내 혈관생성 관련 단백질들은 노화에 의해 변화되는 것으로 사료된다.

Viral protein 2 (VP2), which is the structural protein of parvovirus, can produce virus-like particles (VLPs) by a self-assembly process in vitro, making VLPs attractive vaccine candidates. VP2 of canine parvovirus (CPV) is responsible for neutralizing antibodies in immunized animals. In this study, VP2 protein of canine parvovirus-2c was expressed using a baculovirus expression system and assembled into parvovirus-like particles in insect cells. The results show that VP2 proteins assembled into virus-like particles (VLPs) with antigenic properties similar to those of natural CPV and a high hemagglutination (HA) titer (1:27). The recombinant 6-His-tagged VP2 protein with a molecular mass of about 65 kDa was detected by anti-His antibody and anti-PPV serum. This study provides a foundation for the application of VP2 protein in the clinical diagnosis of CPV and in the vaccination against CPV.

참다슬기 아가미 조직으로부터 heat shock protein 70 유전자를 분리 · 동정하였다. 참다슬기 HSP70 cDNA의 open reading frame (ORF)는 1,917 bp로 639개의 아미노산을 암호화하여 분자 량은 약 70 kDa으로 예측되었다. 생물정보학 배열분석에 의해 HSP 유전자 기능과 관여되어 있는 3가지 주요 signature motifs와 보존된 도메인을 확인하였다. 계통학적 분석을 통하여 참 다슬기 HSP70 유전자는 왕우렁이 Pomacea canaliculate와 같은 클러스트에 포함된다는 사실을 확인하였다. 수온 및 염분 변화에 따라, 참다슬기 HSP70 mRNA 유전자 레벨은 유의적으로 증 가하였으며(p < 0.05), 이는 외부자극요인을 파악할 있는 분자생물학적 마커로서 활용될 수 있 을 것으로 사료된다.

The objective of this study is to determine the effects of barrel temperature and moisture content on the physicochemical properties of texturized vegetable protein (TVP). The water absorption index, water solubility index, texture, integrity index, color, and scanning electron micrograph of the TVP were analyzed. The extrusion conditions consisted of barrel temperature (110, 130, and 150oC) and moisture content (40, 45, and 50%) at a fixed screw speed of 250 rpm. The TVP extruded at 150oC barrel temperature and bearing 50% moisture content had higher water absorption index and water solubility index. Elastic force, cohesiveness, and color differences were the highest in the TVP extruded at 150oC barrel temperature and possessing 40% moisture content. However, the TVP at 150oC barrel temperature and having 40% moisture content had a lower integrity index than the TVP carrying moisture contents of 45 and 50%. The structure of the TVP extruded at 150oC barrel temperature and having 40% moisture content was found similar to a chicken breast tissue’s structure. In conclusion, 150oC barrel temperature and 40% moisture content are optimal conditional characteristics for the texturization of soy protein isolate and gluten.