Humulus japonicus (HJ) is a widely used herbal medicine for pulmonary tuberculosis, hypertension, leprosy, and venomous wounds in Asia, particularly in China. Although HJ has certain physiological activities, such as longitudinal bone growth, antioxidation and alleviation of rheumatism, its anticancer activities, other than in colorectal and ovarian cancer, are yet to be studied. In this study, we investigated the anti-cancer activity and mechanism of methanol extracts of HJ (MeHJ) against human FaDu hypopharyngeal squamous carcinoma cells. MeHJ suppressed FaDu cell viability without affecting normal cells (L929), which was demonstrated using the MTT and Live & Dead assays. Furthermore, MeHJ effectively inhibited colony formation of FaDu cells, even at non-cytotoxic concentrations, and significantly induced apoptosis through the proteolytic cleavage of caspase-9, -3, -7, poly (ADP-ribose) polymerase and through the downregulation of BCL-2 and upregulation of BAX in FaDu cells, as determined by DAPI staining, flow cytometry, and western blot analyses. Collectively, these findings suggest that the inhibitory effects of MeHJ on the growth and colony formation of oral cancer cells may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeHJ has the potential to be used as a natural chemotherapeutic drug against human oral cancer.

벼메뚜기의 인공사료 개발을 위하여 일본의 Konno식 먹이를 대조구로 하고, 28개의 배합비율별 인공사료를 제작하여 급이시험을 하였다. 시험결과, Konno식 보다 6주 후 1마리당 체중은 22개 처리에서 높았고 우화성공율은 23개 처리에서 높았다. 폐사율은 부화입식 후 45일까지 Konno식 대비 24개 처리에서 높았다. 1마리당 체중, 우화성공율, 폐사율을 종합해 볼 때 탄수화물원으로는 옥수수 건조잎 분말 또는 통밀가루와 단백질원으로는 2~5%의 탈지대두분 또는 어분의 배합비율이 벼메뚜기의 인공사료로서 가능성이 높았다.

본 연구에서는 식물성 프로바이오틱스에 해조류인 미역을 첨가하여 프로바이오틱스의 체내 안정성 및 면역원성을 향상 시킬 수 있는 고농도 배양법을 개발하고자 하였다. 미역 첨가 배지를 생산 수준으로 유산균을 배양하였을 때 유산균수는 18시간째 1022, 24시간째 109으로 고농도 배양이 가능하였다. 미역첨가배지로 배양된 유산균의열 안정성, 위산 및 담즙 안정성의 효과도 기본배지와 비교 하여 유의적인 증가를 확인할 수 있었다. 미역첨가배지로 배양된 프로바이오틱스는 생균 및 사균 모두 면역증강효과를 나타내는 것을 알 수 있었다. 배양만으로 기능성이 향상된 프리바이오틱스를 개발함에 따라, 다양한 프로바이오틱스의 기능성 향상의 기반 기술로서 활용이 가능할 것으로 사료 된다.

2013년과 2014년에 경남 진주시 소재의 관행재배 단감원과 유기재배 단감원에서 쓸어잡기와 육안조사를 통하여 곤충상과 천적의 분포를 조사하였다. 2013년에 쓸어잡기로 조사한 결과 유기재배 단감원에서는 49종, 관행재배 단감원의 경우에는 18종의 곤충이 채집되었다. 2014년 유기재배 단감원에서는 총 7목 33과 66종, 관행재배 농가에서는 5목 15과 25종의 곤충이 채집되었다. 2013년과 2014년 모두 우점종은 노린재목으로 조사되었다. 두 단감원에서 우점종은 애긴노린재와 시골가시허리노린재였으며, 붉은잡초노린재와 톱다리개미허리노린재가 그 다음 우점종으로 조사되었다. 육안조사 결과 유기재배 단감원에서는 11목 72과 192종, 관행재배 단감원에서는 10목 52과 104종이 조사되었다. 유기재배 단감원의 우점종은 꽃매미와 양봉꿀벌, 지칭개수염진딧물로 조사되었고, 관행재배 단감원에서는 양봉꿀벌과 붉은잡초노린재로 조사되었다. 육안조사와 쓸어잡기 조사를 통해 천적 또한 채집되었다. 유기재배 단감원의 경우 쓸어잡기에서는 9종, 육안조사에서는 31종의 천적이 조사되었고, 관행재배 단감원의 경우 쓸어잡기에서는 5종, 육안조사에서는 18종의 천적이 조사되었다.

경남 창원과 진주의 농약 무살포 및 유기농 단감원에서 단감 잎을 가해하는 해충을 조사한 결과 국내에서 단감 해충으로 기록되지 않은 3종 을 채집 동정하였다. 동정된 종은 가을뒷노랑밤나방, 푸른빛집명나방, 감잎가는나방(신칭)이었다. 이들의 채집기록, 형태, 기주와 간단한 생태를 조사하여 보고한다.

Insect and natural enemy fauna were surveyed at conventional (CV) and organic-farming persimmon orchards (OF) at Jinju, Korea, using sweeping net and naked eyes in 2013 and 2014. Using sweeping net, 49 species at an OF and 18 species at a CV were observed in 2013. In 2014 too, more species were observed at OF (66 species, 33 families, 7 orders) than at a CV (25 species, 15 families, 5 orders). During both the years, dominant species were all hemipteran insects like Nysius plebejus (Lygaeidae) and Cletus punctiger (Coreidae) in both types of orchards, followed by Rhopalus maculatus (Rhopalidae), Riptortus pedestris (Alydidae). Similarly, naked eye inspection also found more species at OF (192 species, 72 families, 11 orders) than at CV (104 species, 52 families, 10 orders) in 2014. Dominant species in naked eye inspection were Lycorma emelianovi (Hemiptera: Fulgoridae), Apis mellifera (Hymenoptera: Apidae), Uroleucon cephalonopli (Hemiptera: Aphididae) at OF, and A. mellifera and R. maculatus at CV in 2014. Using naked eye inspection or sweeping net in 2014, more species of natural enemies were observed at OF (31 and 9 species, respectively) than at CV (18 and 5 species, respectively).

The role of cytochrome P-450 2E1 (P450 2E1) in the early phase of alcoholic fatty liver was examined. Female rats were pretreated with either allyl sulfide (200 mg/kg, po), disulfiram (500 mg/kg, po), YH 439 (250 mg/kg, po) or pyrazine (200 mg/kg/day × 2 days, ip). Marked changes in carbon tetrachloride-induced hepatotoxicity and carboxyhemoglobin (COHb) elevation due to dichloromethane administration were observed in rats treated with one of the P450 2E1 modulators. A single dose of ethanol (6 g/kg, po) increased the hepatic triglyceride contents approximately 2 fold, which was inhibited completely by YH 439 pretreatment. However, the other P450 2E1 modulators failed to alter the ethanol-induced hepatic triglyceride accumulation. In vitro hepatic microsomal enzyme activity was determined in 4 week old premature and 12 week old adult rats. Aminopyrine-N demethylation was not different, but p-nitrophenol hydroxylation and p-nitroanisole O-demethylation were significantly higher in premature rats compared to adult indicating that the P450 2E 1 activity decreases with the maturity of rats. However, no difference in the triglyceride accumulation induced by an intraperitoneal dose of ethanol (3 g/kg) was noted between premature and adult rats. The results suggest that the P450 2E1 activity dose not play an important role in the induction of acute alcoholic fatty liver.

본 연구는 4종의 별의별간(SS) 음료의 간보호 소재로서 의 이용 가능성을 조사하고자 항산화능 평가, t-BHP 와 CCl4로 산화적 손상 및 급성 간독성 유도한in vitro, in vivo 모델을 활용하여 간보호능을 평가하였다. 실험결과, 별의별간 01~04는 50 μM vitamin C 와 유사한 항산화 효과를 나타내었다. HepG2 세포에 t-BHP로 산화 스트레스를 유도한 뒤 나타나는 세포독성에 대해 별의별간 01 및 04에서 농도 의존적인 세포 보호효과를 보였으며, ROS 생성 억제에서 별의별간 01, 03, 04에서 농도의존적인 억제를 나타내었다. 미나리가 혼합된 별의별간 04에 대한 급성 간손상in vivo 모델을 활용하여 간보호능 검증 결과, 별의별간 04는 CCl4로 증가된 혈중 ALT, AST의 유의적 감소, 간 조직중 증가된 MDA 함량 감소 및 감소된 GSH의 유의적 증가를 나타내었다. 또한, 혈청 및 간 조직에서 증가 된 중성지방과 콜레스테롤을 유의적으로 감소시켰다. 이러 한 결과를 종합하며, 별의별간 04는in vitro 및in vivo 모델 에서 산화적 손상에 대해 간보호 효과를 나타내었다

산업적 유산균 배양의 가격대비 효율성을 높이기 위하여 질소원, 탄소원 및 무기염물을 기본량으로 기본배지를 제조하고, 효율적인 고농도 유산균체를 수득하기 위하여 pH조절제로서 따로 첨가되어지는 합성 화학물질을 대신하여 미역을 실험에 사용하였다. 미역의 최적의 첨가 방법을 찾고자 추출, 착즙, 분말의 형태로 배지를 제조하여 유산균증식능 평가한 결과, 착즙의 형태에서 유산균 증식능이 가장 큰 것으로 확인되었다. 이러한 실험 결과를 통해, 폐기되어 환경오염의 원인이 되는 가식부 외 미역을 유산균 배양배지에 착즙형태로 첨가함으로서 종래 pH 조절을 위해 완충액으로 첨가되는 합성 화학물질을 대체하여 활용될 수 있음을 확인하였다. 또한 미역첨가 기본배지 선정에 따른 배지 단가를 절감하면서 높은 균체 수율을 얻는 등 충분한 경제적 장점을 기대할 수 있다. 이러한 연구결과는 미역을 이용함

TCR subunits are members of membrane-bound receptors which allow the fast and efficient elimination of the specific fish pathogens have regulated function in adaptive immunity. Sequence structure of TCR subunits have been reported for various teleosts, but the information of each TCR subunit functional characterization through expression analysis in fish was unknown. In this study, we examined the gene expression of TCR subunits in the early developmental stages and observed transcript levels in various tissues from healthy adult olive flounder by RT-PCR. The mRNA expression of alpha subunit was already detected in the previous hatching step. But the transcripts of another TCR subunit were not observed during embryo development and increased after hatching and maintained until metamorphosis at the same level. It was found that all TCR subunits mRNAs are commonly expressed in the immune-related organ such as spleen, kidney and gill, also weak expressed in fin and eye. TCR alpha and beta subunit were expressed in brain, whereas gamma and delta were not expressed same tissue. The sequence alignment analysis shows that there are more than 80% sequence homology between TCR subunits. Because it has a high similarity of amino acid sequence to expect similar in function, but expression analysis show that will have may functional diversity due to different time and place of expression.

The innate immune system is the only defense weapon that invertebrates have, and it is the fundamental defense mechanism for fish. The innate immune response is important in newly hatched flounders because it is closely involved in the initial feeding phase, which is why it is essential for survival during the juvenile period. The expression analysis of genes involved in the innate immune response in the olive flounder (Paralichthys olivaceus) in the days after hatching is incomplete. Therefore, we have begun to examine the expression patterns of genes specifically induced during the development of the innate immune system in newly hatched flounders. Microscopic observation showed that pronephron formation corresponded with the expression of perforin-encoding gene. These results suggest that perforin plays a vital role in the innate immunity of the kidney during developmental stages. Perforin expression was strong at the start of the development of the innate immune response, and continued throughout all the development stages. Our findings have important implications with respect to perforin’s biological role and the evolution of the first defense mechanisms in olive flounder. Further studies are required to elucidate the perforin-mediated innate immunity response and to decipher the functional role of perforin in developmental stages.

Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.

Olive flounder (Paralichthys olivaceus) is one of the commercial important flatfish species in Korea. The ocular signal transduction pathway is important in newly hatched flounders because it is closely involved in the initial feeding phase thus essential for survival during the juvenile period. However, the study of gene expression during ocular development is incomplete in olive flounder. Therefore we examined the expression analysis of specifically induced genes during the development of the visual system in newly hatched flounders. We searched ocular development-involved gene in the database of expressed sequence tags (ESTs) from olive flounder eye and this gene similar to arrestin with a partial sequence homology. Microscopic observation of retinal formation corresponded with the time of expression of the arrestin gene in the developmental stage. These results suggest that arrestin plays a vital role in the visual signal transduction pathway of the retina during ocular development. The expression of arrestin was strong in the ocular system during the entirety of the development stages. Our findings regarding arrestin have important implications with respect to its biological role and evolution of G-protein coupled receptor (GPCR) signaling in olive flounder. Further studies are required on the GPCR-mediated signaling pathway and to decipher the functional role of arrestin.

Cathepsins are members of the multigene family of lysosomal cysteine proteinases and have regulated function in several life processes. The potential role of cathepsin F cysteine gene was expected as protease in the yolk processing mechanism during early developmental stage, but expression analysis was unknown after fertilization. The alignment analysis showed that amino acid sequence of cathepsin F from olive flounder liver expressed sequence tag (EST) homologous to cathepsin F of other known cathepsin F sequences with 87－98% identity. In this study, we examined the gene expression analysis of cathepsin F in various tissues at variety age flounder. Tissue distribution of the cathepsin F mRNA has been shown to be ubiquitous and constitutive pattern regardless of age in each group, although derived from cDNA library using liver sample. The mRNA level of cathepsin F more increased as developmental proceed during embryogenesis and early developmental stage, especially increased in the blastula, hatching stage and 3 days post hatching (dph). As a result, it may suggest that the proteolysis of yolk proteins (YPs) has been implicated as a mechanism for nutrient supply during early larval stages in olive flounder.

Olive flounder (Paralichthys olivaceus) is a most important aquaculture species in Korea. Like most marine fishes, olive flounders are stomachless at first feeding and aquired gastric function during the metamorphosis, so food was mainly digested by pancreatic enzyme from first feeding to premetamorphosis. However, comprehensive analysis of pancreatic and gastric digestive enzyme of olive flounder at early developmental period is still unclear. In the expression study of pancreatic and gastric digestive enzyme by real-time PCR at early developmental stage, pancreatic enzyme such as chymotrypsinogen 2, preproelastase 2 and 4, pancreatic protein somatomedin-B domain (PPSB) mRNA expression were initiated at first feeding and strongly expressed in the pancreas developmental stage, while gastric digestive enzyme signal was not at all detected during same period. Although, trypsinogens were secreted from pancreas and have similar amino acid sequence, trypsinogen 3 expression induction was detected both pancreas and stomach developmental stage, while trypsinogen 2 expression was significantly increased only post-metamorphosis period. Pepsinogen mRNA expression was only detected at metamorphosis according to stomach differentiation. Lipid digestive enzyme, lipase and intestine fatty acid binding protein 1 (I-FABP 1), were already reached a certain level at beginning of hatching and more increased during early developmental stage and then gradually decreased before metamorphosis. These results suggested that feed ingestion of olive flounder was exclusive charged by pancreatic digestive enyme, lipid digestive enzyme and trypsinogen 3 from first feeding and then fully swiched by gastric digestive enzyme and trypsinogen 2 from metamorphosis period.

For the study of population genetic structure with mtDNA, it is essential to measure genetic diversity at each mtDNA regions. Also, to evaluate the variation according to the each region should follow as well as to see if there are differences. In this study, we delved into the variations and dendrogram among samples of seven mtDNA regions (NDⅡ, NDⅤ, NDⅣ, NDⅣL, NDⅥ, NDⅠ, 12SrRNA) from wild Pacific abalone, Haliotis discus hannai collected in Yeosu, Korea. The region with the highest genetic variation was NDⅣ region (Haplotype diversity = 1.0000, Nucleotide diversity = 0.010823) with two to five times higher variation than the others. Furthermore, the study to see if there is a difference between the regions of samples showed that similar aspects of dendrogram in NDⅡ and NDⅠ(divergence of 90% and 87%), which forms a group with hd4, 7, 8 and 10 at bootstrap support, based on 1000 replications. Also, pair-wise FST between clusters within the regions showed high values; 0.4061 (P=0.0000), 0.4805 (P=0.0000) respectively. Therefore we can infer that it is the most efficient and accurate way to analyze the region of NDⅣ with the highest variation in addition to the regions of NDⅡ and NDⅠ, which formed clusters with high bootstrap value, for study of population genetic structure in this species.