Many transcription factors are involved in directing the growth of porcine oocytes. The localization and expression level of a given transcription factor often differ at each stage of early embryonic growth, which spans from fertilization to the formation of the blastocyst. A hallmark of the blastocyst stage is the separation of the endodermal and mesodermal ectoderm. The embryo's medium and its effects are known to be crucial during early development compared to the other developmental stages, and thus require a lot of caution. Therefore, in many experiments, early development is divided into the quality of oocyte and cumulus cells and used in experiments. We thought that we were also heavily influenced by genetic reasons. Here, we examined the expression patterns of five key transcription factors (CDX2, OCT4, SOX2, NANOG, and E-CADHERIN) during porcine oocyte development whose expression patterns are controversial in the pig to the literature. Antibodies against these transcription factors were used to determine the expression and localization of them during the early development of pig embryos. These results indicate that the expressions of key transcription factors are generally similar in mouse and pig early developing embryos, but NANOG and SOX2 expression appears to show species-specific differences between pig and mouse developing embryos. This work helps us better understand how the expression patterns of transcription factors translate into developmental effects and processes, and how the expression and localization of different transcription factors can crucially impact oocyte growth and downstream developmental processes.

Clematis patens는 미나리아재비과 으아리속에 속하는 다년 생 숙근초로 꽃잎화한 꽃받침을 가지고 있는 점이 특징이며 흰색, 분홍색, 자주색, 보라색, 등 다양한 화색이 존재한다. 본 연구에서는 C. patens ‘The President’에서 flavonoid 생합성 경로에 관여하는 유전자 중 delphinidin 계열 anthocyanin 색 소 합성을 유도하는 F3'5'H(ClF3'5'H)를 분리 동정하였으며, 화색 및 발달 단계별 ClF3'5'H 유전자의 발현 패턴을 분석하 였다. 또한 클레마티스 화색별 anthocyanin의 함량을 알아보 았다. 그 결과, 기존 보고된 Aconitum carmichaelii의 F3'5'H 유전자의 아미노산 서열과 79% 일치하여 높은 상동성을 갖는 것을 확인하였으며, ClF3'5'H 유전자의 발현 패턴은 흰색을 띠 는 클레마티스에서는 ClF3'5'H 유전자의 발현량이 많지는 않 았으나 다른 발달 단계에 비해 완전 개화한 단계에서 발현이 강했다. 분홍색을 띠는 클레마티스에서는 모든 발달 단계에서 ClF3'5'H 유전자의 발현이 검출되지 않았으며, 자주색과 보라 색을 띠는 클레마티스에서는 개화 직전 및 완전 개화 단계에 서 발현하였다. 이 결과로부터 클레마티스의 다양한 화색과 클 레마티스의 F3'5'H 유전자 발현과의 상관관계가 시사되었다.

Identification of specific marker proteins in cells is useful for isolating cells and determining their cellular characteristics and functions. Based on our previous study showing that matrix metalloproteinase 9 (MMP-9) can be used as a marker for porcine spermatogonia, the expression pattern of MMP-9 was determined in both pre- (5-month old) and post-pubertal (11–month old) bovine testes. Histological analysis revealed that spermatogonia were located near the basement membrane in both testes, while spermatozoa were not detected in the 5-month old pre-pubertal bovine testes and epididymides. Mature spermatozoa were observed in the 11-month bovine testes and epididymides, and MMP-9 expression in 11-month old bovine testes was lower than 5-month old testes, according to reverse transcription-PCR and real-time-PCR data. To determine the specific expression sites of MMP-9 in the bovine testes, immunohistochemistry was performed. Expression of MMP-9 was observed in cells near the basement membrane of seminiferous tubules in both 5- and 11-month old testes. Furthermore, MMP-9 positive cells expressed protein gene product 9.5 (PGP9.5) and deleted in azoospermia (DAZL) that are already known as bovine spermatogonial stem cells markers. In the present study, MMP-9 expression was identified in both pre- and post-pubertal bovine spermatogonia expressing PGP9.5 and DAZL, and located near the basement membrane of seminiferous tubules. Thus, MMP-9 can be used as a marker for bovine spermatogonia, and may provide useful platforms for understanding the interaction between germ cells and extracellular matrix during spermatogenesis in the seminiferous tubules.

Pheromone biosynthesis activating neuropeptide (PBAN) produced in the subesophageal ganglion is known to stimulate pheromone production in the pheromone gland. A cDNA isolated from female adult heads of Maruca vitrata encodes 197 amino acids including PBAN, designated as Mvi-PBAN, and four other neuropeptides (NPs): diapause hormone (DH) homologue, α-NP, β-NP and γ-NP. All of the peptides are amidated in their C-termini and shared a conserved motif, FXPR(or K)L-NH2 structure. Mvi-PBAN consists of 35 amino acids as previously reported (Chang and Ramasamy, 2014). RT-PCR analysis revealed that Mvi-PBAN cDNA was expressed in all examined body parts. Nucleotide sequence analysis of RT-PCR products indicated the Mvi-PBAN sequence was identical in all examined body parts of both sexes. These results suggest that Mvi-PBAN expression is maintained in examined stages or tissues.

전자코와 GC/MS 기기를 이용하여 나도풍란의 꽃에서발현되는 주요 향기성분 및 향기발현패턴을 분석하였다.전자코와 GC/MS 의 분석을 통해 전자코에서는 약 9개의 chromatogram peak 를 얻어냈고, GC/MS 기기분석에서는 약 13개 정도의 chromatogram peak를 얻어냈다. 전자코 peak 중에서 6가지 종류가 GC/MS chromatogrampeak와 공통적으로 나타났으며 즉 retention time 3.2초,4.2초, 5.4초, 5.8초, 6.3초, 6.9초의 peak였으며 그 peak에 해당되는 향기성분들은 각각 2-furanmethanol, linalool,citronellol, neral, nerodidol, benzoic acid, hexadecanoicacid, 1,2-benzenedicarboxylic acid로 추정되었다. 나도풍란군집에서 개체별로 향기발현량과 향기패턴을 비교분석한결과 주요성분으로 예측되는 6개 peak는 모든 개체에서발현되는 것으로 나타났지만 개체간 발현량에 차이가 있는 것으로 나타났다. 꽃의 발달단계별 향기발현정도를 분석하기 위한 실험에서 꽃의 발달 단계중 꽃봉오리 상태와 노화된 꽃에서는 향기발현량이 적었으며 꽃이 완전개화한 화서 중앙부위에 있는 꽃들에서 가장 많은 양의 향기가 발생되는 것으로 나타났다. 꽃의 기관별 향기분석에서는 나도풍란 꽃의 주요향기성분이 주로 sepal과 petal조직에서 가장 많이 발현되는 것으로 확인되었으며 column과 spur에서는 발생량이 매우 적은 것으로 나타났다. 일중 시간대별 주요향기성분의 발현량과 패턴을 분석한 결과 오전11시에 가장 높은 향기발현량을 보였으며 오후 5시 이후부터 향기발현량이 현저히 줄기 시작하여 저녁 8시 이후에는 향기성분이 발생되지 않았다. 빛 조사가 향기발현에 미치는 영향을 알아보기 위해 암처리와 광처리후 향기양과 패턴을 분석한 결과 암처리에서는 40시간 이후부터는 대체로 향기성분이 계속 줄어들었으며, 40시간연속적으로 빛을 조사한 후 20시에 전자코 분석을 한 결과 오전 시간대와 동일한 향기발현량과 발현패턴을 보여주었다. 이 결과를 통해 빛의 조사시간 및 생체리듬주기가 향기발현에 큰 영향을 줄 수 있는 요인임을 확인할수 있었다.

염 또는 건조 스트레스 처리에 의한 톨 페스큐 종자의 발아율 변화와 유식물체 수준에서의 유전자 발현을 조사하기 위하여 in vitro 조건에서 NaCl과 PEG를 처리하여 분석하였다. NaCl 처리시 톨 페스큐 품종별 발아율은 50 mM 농도에서 발아율이 서서히 감소하기 시작하였으며 350 mM의 농도에서는 모든 품종에서 발아가 되지 않는 경향을 보였다. NaCl 처리 농도에 따른 발아율 감소율은 Fawn 품종이 가장 큰 변화를 보였으며 Kentucky-31(E-) 품종이 가장 강한 내성을 보였다. 또한, PEG 처리시 톨 페스큐 품종별 발아율의 변화도 NaCl 처리시와 유사한 경향을 보였으며 고농도인 30% PEG 처리구에서는 모든 품종에서 발아가 되지 않는 경향을 보였으며 Kentucky-31(E-) 품종이 가장 강한 내성을 보였다. 톨 페스큐 유식물체 수준에서 염해와 건조 스트레스에 의한 유전자 발현양상을 조사하기 위하여 DEGs(differentially expressed genes) 탐색을 위한 ACP-based GeneFishingTM PCR 분석을 통해 NaCl 또는 PEG 처리에 따른 발현량의 차이를 보이는 총 4개의 DEG를 선발하여 클로닝하고 염기서열을 분석하였다. 무처리구에 비해 NaCl 처리시 4개의 DEG가 증가하였고 감소하는 DEG는 확인 되지 않았으나, PEG 처리에서는 3개의 DEG (DEG 1, 3, 및 4)가 증가하였고 1개의 DEG가 감소하는 경향을 나타내었다. 발굴된 DEG들을 blastx 검색에 의하여 rubisco large subunit (DEG1), microsomal glutathion S-transferase (GST) 3-like isoform 1 (DEG2) 유전자로 동정되었다.

Procymidone is a fungicide with anti-androgenic properties widely used to protect fruits from fungal infection, which induces an excessive reactive oxygen species production in male reproductive organs. In this study, to clarify whether procymidone affect the cellular antioxidant system of prostate at onset of puberty, gene expression patterns of the representative antioxidant enzymes such as cytoplasmic glutathione peroxidase (GPx1), phospholipid hydroperoxide GPx (PHGPx), selenoprotein P (SePP), cytoplasmic copper/zinc superoxide dismutase (SOD1), and manganese SOD (SOD2) were investigated in the rat ventral prostates exposed to procymidone using real-time RT-PCR analyses. Seven-week-old Sprague-Dawley rats castrated at 6 weeks old were treated with procymidone (25, 50, or 100 mg/kg per day) orally for 7 consecutive days after testosterone propionate (0.4 mg/kg per day) administration by subcutaneous injection. As compared to normal control animals, GPx1 mRNA expression in prostates significantly increased by the administration with TP and/or procymidone. However, PHGPx and SOD1 mRNA levels significanatly decreased by over 25 mg/kg of procymidone treatment and SePP and SOD2 mRNA levels was significanatly reduced by over 50 mg/kg of procymidone treatment. These findings indicate that procymidone may affect the antioxidant system of prostatic cells in up-regulation mode of GPx1, but in down-regulation modes of PHGPx, SePP, SOD1, and SOD2, suggesting that procymidone may affect differently the cellular antioxidant system of prostate according to the exposure doses.

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a unique antioxidant enzyme involved in reduction of peroxidized phospholipids within biomembranes. To investigate the expression pattern of the PHGPx gene during fetal development, in situ hybridization analyses were performed using mouse FITC-labeled PHGPx cRNA probes in fetal tissues on embryonic days (Ed) 13.5-18.5. During these periods, PHGPx mRNA appeared in the developing telencephalon, diencephalon, spinal cord, and spinal ganglion. In particular, PHGPx mRNA was strongly expressed in pyramidal cells of the cerebral cortex. On Eds 17.5-18.5, PHGPx mRNA was detected in various tissues including liver, intestinal villi and crypt, pancreas, lung, and olfactory epithelium of the nasal cavity. In addition, PHGPx mRNA was highly expressed in the inner ear on Eds 14.5-18.5, brown fat on Ed 17.5, and adrenal gland on Ed 18.5. It is conceivable that PHGPx may act as an important antioxidant against fetal oxidative stress during mouse organogenesis.

The black soldier fly (BSF), Hermetia illucens, is known as a beneficial insect and feeds on organic materials derived from animals and human, resulting in reduction of food waste and conversion of organic materials. Despite of many studies on the BSF, there have been no reports of cloned genes encoding serine proteases in the BSF. Thus, the primary objective of this study is to clone and to investigate expression pattern of genes encoding serine proteases released from the midgut of the BSF larvae in order to gain a better understanding of expression mechanism of serine proteases. We cloned two serine proteases from the BSF larva. Based on phylogenetic tree analysis, one was chymotrypsin, the other was trypsin. The open reading frame (ORF) of chymotrypsin was 804bp, which encoded a polypeptide of 267 amino acids. In case of trypsin, the ORF was 744bp, which encoded a polypeptide of 247 amino acids. To investigate expression pattern of two serine proteases, we conducted semi-quantitative RT-PCR at different tissues and different developmental stages. A chymotrypsin and trypsin transcripts were revealed strongly in mid gut. Especially, a chymotrypsin was detected largely at feeding stage more than molting stage, while trypsin was expressed similarly between feeding stage and molting stage

The current study aims to link Korean tradition to modern culture; to re-produce patterns of lotus depicted in Minwha, The study also aims to discover the beauty of Korean tradition and to modernize it. The current study is based upon document searches(including research papers) and the Internet searches. Through these searches, it investigates the concept of Minhwa, the origin of lotus depiction and its symbolic meanings, the traits of such pattern. Based upon this investigation, the study attempts to modernize the patterns of lotus and apply the modernized patterns to designing shirts. The process which employs the lotus patterns illustrated in Minhwa to shirt design includes four sub-processes: selecting, allocating, coloring, and selecting production techniques. The subprocess of selecting patterns is two folded: the first stage covers carbon-copying the distinctive features of lotus, lotus leaf, lotus bud, lotus pip, and lotus stem; the second stage is making these features suitable to shirt sizes. For the process of coloring those shirts, Piccaso’s work(Pablo Picasso, 1881～1973) has been selected and the colors in his work have been adopted to dye the rest of the shirts as well as the lotus features. The process of selecting production techniques includes ornament tail in order to modernize the patterns allocated in the shirts. Once these processes are completed, the shirts are made on a scale of real-life size. These processes of creating shirt design by modernizing traditional patterns will hopefully contribute to researchers expanding the domain of shirt design.

Innate immune response is initiated by the recognition of unique microbial molecular patterns through pattern recognition receptors (PRRs). The purpose of this study is to dissect the expression of various PRRs in gingival epithelial cells of differentiated versus undifferentiated states. Differentiation of immortalized human gingival epithelial HOK-16B cells was induced by culture in the presence of high Cα²+ at increased cell density. The expression levels of various PRRs in HOK-16B cells were examined by realtime reverse transcription polymerase chain reaction (RTPCR) and flow cytometry. In addition, the expression of human beta defensins (HBDs) was examined by real time RT-PCR and the amounts of secreted cytokines were measured by enzyme linked immunosorbent assay. In undifferentiated HOK-16B cells, NACHT-LRR-PYDcontaining protein (NALP) 2 was expressed most abundantly, and toll like receptor (TLR) 2, TLR4, nucleotide-binding oligomerization domain (NOD) 1, and NOD2 were expressed in substantial levels. However, TLR3, TLR7, TLR8, TLR9, ICE protease-activating factor (IPAF), and NALP6 were hardly expressed. In differentiated cells, the levels of NOD2, NALP2, and TLR4 were different from those in undifferentiated cells at RNA but not at protein levels. Interestingly, differentiated cells expressed the increased levels of HBD-1 and -3 but secreted reduced amount of IL-8. In conclusion, the repertoire of PRRs expressed by gingival epithelial cells is limited, and undifferentiated and differentiated cells express similar levels of PRRs.

This study was conducted to examine the mRNA expression of apoptosis-related and imprinted genes and methylation pattern of the differentially methylated region (DMR) of H19 gene in day 35 of SCNT pig fetuses. The day 35 of natural mating (control) or cloned (clone) pig fetuses were recovered from uterus. Endometrium from dam and liver from fetus were obtained, respectively. mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. The Bcl-2 mRNA expression in clone was significantly lower than that of control (p<0.05). The mRNA expression of H19 gene in both endometrium and liver was significantly higher in clone than that of control, respectively (p<0.05). The level of IGF-2 mRNA in liver of clone was significantly lower than that of control (p<0.05), whereas the mRNA expression of IGF2-R gene in liver of clone was significantly higher than that of control (p<0.05). The DMR of H19 was lower methylation pattern in clone than that of control. These results suggest that the aberrant mRNA expression of apoptosis-related and imprinted genes and the lower DMR methylation pattern of imprinted gene may be closely related to the inadequate fetal development of cloned fetus.