The baculovirus expression system (BES) utilize the p10 or polyhedrin promoter, a very late promoter that exhibits strong transcriptional activity primarily at the end of viral infection, to produce useful recombinant proteins. The burst sequence of the very late promoter is essential for strong transcription, and VLF-1 is a transcription factor that binds specifically to the burst sequence, and it has been reported that it can regulate the amount and timing of expression of protein by the very late promoter. Recently, a VLF-1 constitutively expressing cell line was constructed to increase the production of the target protein, but the effect was minimal. In this study, to find the optimal VLF-1 expression conditions to increase target protein production efficiency, we controlled the expression of VLF-1 through various promoters and evaluated the target protein expression efficiency by the p10 promoter accordingly.

This work focuses on the fabrication of excellent magnetic structures for trapping breast cancer cells. Micromagnetic structures were patterned for trapping cancer cells by depositing 30 nm of permalloy on a silicon substrate. These structures were designed and fabricated using two fabrication techniques: electron beam lithography and laser direct writing. Two types of magnetic structures, rectangular wire and zig-zagged wire, were created on a silicon substrate. The length of each rectangular wire and each straight line of zig-zagged wire was 150 μm with a range of widths from 1 to 15 μm for rectangular and 1, 5, 10 and 15 μm for zigzag, respectively. The magnetic structures showed good responses to the applied magnetic field despite adding layers of silicon nitride and polyethylene glycol. The results showed that Si + Si3N4 + PEG exhibited the best adhesion of cells to the surface, followed by Si + Py + Si3N4 + PEG. concentration of 5-6 with permalloy indicates that this layer affected silicon nitride in the presence of Polyethylene glycolPEG.

Three different dogs who had immune-mediated hemolytic anemia (IMHA) were treated for more than two weeks with blood transfusion in an animal clinic. Despite this treatment and hospitalization, there was no clinical improvement in clinical signs as well as complete blood cell count (CBC) including hematocrit (HCT) and C-reactive protein (CRP). All cases were then injected two or three times with allogeneic stem cells through an intravenous route for treatment. Upon administrating stem cells to the IMHA dogs, clinical conditions and the indexes of HCT and CRP were clinically improved within or close to normal ranges.

가는잎향유[Elsholtzia angustifolia (Loes.) Kitag.]는 화형 이 아름답고, 정유 특유의 향기가 좋아서 분화용 및 지피용 관 상식물로 수요가 증가하고 있고, 전초에는 약효가 있다고 알 려져 있다. 본 연구는 가는잎향유의 육묘에 미치는 플러그 트 레이 셀 사이즈, 파종립수, 차광정도, 추비농도 등의 영향을 구명하기 위하여 수행되었다. 연구결과, 플러그 트레이 셀 사 이즈는 용량이 증가할수록 유묘의 초장, 엽수, 마디수, 근장, 지상부 생체중이 유의적으로 증가하였다. 파종 립수는 2립 파 종 시 가장 효율적이었고, 파종량이 증가할수록 생육이 감소 하였다. 차광정도가 높아질수록 초장은 증가하였고, 경직경, 엽수, 마디수는 55% 차광에서 가장 우수하였다. 추비 처리 시 공시비료 1000배 처리구에서 생육이 가장 양호하였다. 따라 서 가는잎향유의 육묘 시 162셀 트레이에 원예상토를 채운 다음 셀 당 2립 파종한 후 55% 차광막이 설치된 육묘상에서 공시비료 1000배로 엽면시비하는 것이 가장 효과적인 것으로 생각된다.

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

Despite advancements in therapeutic approaches, radiotherapy and cisplatin-based chemotherapy remain primary noninvasive treatments for patients with oral squamous cell carcinoma (OSCC). Moreover, the 5-year survival rate for patients with OSCC has remained almost unchanged for several decades, and many side effects of chemotherapy still exist. In this study, three-dimensional (3D) models of OSCC were established using fibroblasts, and the efficacy of various biological inhibitors was evaluated. A culture of epithelial cells with two types of fibroblasts (hTERT-hNOFs and cancer-associated fibroblasts) within a type I collagen matrix resulted in the formation of a continuous layer of tightly packed cells compared to models without fibroblasts. Furthermore, the effects of biological chemicals, including Y27632, latrunculin A, and verteporfin, on these models were investigated. The stratified formation of the epithelial layer and invasion in OSCC 3D-culture models were effectively inhibited by verteporfin, whereas invasion was weakly inhibited by Y27632 and latrunculin. Collectively, the developed OSCC 3D-culture models established with fibroblasts demonstrated the potential for drug screening, with verteporfin showing promising efficacy.

Oral bacterial infections substantially affect the development of various periodontal diseases and oral cancers. However, the molecular mechanisms underlying the association between Fusobacterium nucleatum (F. nucleatum ), a major periodontitis (PT)-associated pathogen, and these diseases require extensive research. Previously, our RNAsequencing analysis identified a few hundred differentially expressed genes in patients with PT and peri-implantitis (PI) than in healthy controls. Thus, in the present study using oral squamous cell carcinoma (OSCC) cells, we aimed to evaluate the effect of F. nucleatum infection on genes that are differentially regulated in patients with PT and PI. Human oral squamous cell carcinoma cell lines OSC-2O, HSC-4, and HN22 were used. These cells were infected with F. nucleatum at a multiplicity of infection of 100 for 3 hours at 37℃ in 5% CO2. Gene expression was then measured using reverse-transcription polymerase chain reaction. Among 18 genes tested, the expression of CSF3, an inflammation-related cytokine, was increased by F. nucleatum infection. Additionally, F. nucleatum infection increased the phosphorylation of AKT, p38 MAPK, and JNK in OSC-20 cells. Treatment with p38 MAPK (SB202190) and JNK (SP600125) inhibitors reduced the enhanced CSF3 expression induced by F. nucleatum infection. Overall, this study demonstrated that F. nucleatum promotes CSF3 expression in OSCC cells through p38 MAPK and JNK signaling pathways, suggesting that p38 MAPK and JNK inhibitors may help treat F. nucleatum-related periodontal diseases by suppressing CSF3 expression.

토복령은 우수한 항균, 항산화, 항염증 효능을 가진 소재로 알려져 있다. 이러한 토복령(Smilax china)의 추출물의 기능성을 화장품에 적용하기 위한 기초연구로써 토복령에서 발견되는 플라보노이드인 quercetin, catechin, naringenin의 농도별 경피 투과 특성을 조사할 필요성이 있다. Marzulli의 정의에 적 용한 결과 케르세틴의 Kp 값은 0.1 mg/mL에서 "빠름"으로 분류되었고, 0.2 및 0.4 mg/mL에서 "보통"으로 분류되었다. 특히, 농도가 증가함에 따라 투과 속도가 감소하는 경향이 있었다. 나린제닌의 경우 Flux 값은 각각 0.1, 0.2 및 0.4 mg/mL 농도에서 0.69, 1.07 및 1.42 μg/hr/cm²이었으며, 해당 Kp 값은 각각 6.95, 5.34 및 3.56이었다. 나린제닌의 Kp 값은 모든 농도에서 "보통" 범주에 속하며, 케르세틴과 관찰된 것과 같이 농도가 높아짐에 따라 투과 속도가 감소하였다. 카테킨의 경우 Flux 값은 각각 0.1, 0.2 및 0.4 mg/mL 농도에서 0.75, 1.09 및 1.66 μg/hr/cm²이었으며, 해당 Kp 값은 각각 7.55, 5.46 및 4.16이었다. 카테킨의 Kp 값은 모든 농도에서 일관되게 "보통"으로 분류되었다. 여드름 저해능 및 항염증 효능이 우수 한 토복령 추출물의 유효성분인 quercetin, catechin, naringenin의 경피 투과 특성이 보통 이상으로 나타나 기능성 화장품에 사용할 수 있는 우수한 천연물 소재인 것을 확인할 수 있었다.

The objectives of this paper are: (1) to conduct the thermal analyses of the disposal cell using COMSOL Multiphysics; (2) to determine whether the design of the disposal cell satisfies the thermal design requirement; and (3) to evaluate the effect of design modifications on the temperature of the disposal cell. Specifically, the analysis incorporated a heterogeneous model of 236 fuel rod heat sources of spent nuclear fuel (SNF) to improve the reality of the modeling. In the reference case, the design, featuring 8 m between deposition holes and 30 m between deposition tunnels for 40 years of the SNF cooling time, did not meet the design requirement. For the first modified case, the designs with 9 m and 10 m between the deposition holes for the cooling time of 40 years and five spacings for 50 and 60 years were found to meet the requirement. For the second modified case, the designs with 35 m and 40 m between the deposition tunnels for 40 years, 25 m to 40 m for 50 years and five spacings for 60 years also met the requirement. This study contributes to the advancement of the thermal analysis technique of a disposal cell.

자생 부추속 식물 중 강부추(Allium thunbergii for. rheophytum ined.)와 갯부추(A. pseudojaponicum Makino)는 관상용, 식 용 및 약용자원으로 가치가 있으나 육묘를 위한 생육환경조건 구명이 미비하여 연구할 필요성이 있다. 본 연구는 강부추와 갯부추의 육묘에 미치는 플러그 셀 크기, 차광률, 시비처리에 따른 영향을 구명하기 위하여 실험을 수행하였다. 강부추와 갯부추를 육묘한 결과, 플러그 셀 크기에서는 50, 72, 105, 128, 162, 200셀 처리 중 용적이 가장 큰 50셀에서 초장, 엽 수, 근수, 그리고 근장의 생육이 우수하였다. 그러나 생산비용 과 플러그 육묘의 효율성을 고려하여 105셀 이상의 플러그 트레이 중에서 선택하여 육묘하는 것이 효과적이라 판단된다. 차광률에 따른 유묘는 0, 30, 60, 90% 처리 중 30~60% 차 광처리에서 초장, 근수, 그리고 근장이 유의적으로 높게 측정 되어 생육이 양호하였다. 시비처리에서 생중량과 건중량을 제 외한 생육지표를 검토했을 때, 강부추의 적정 시비처리는 속 효성 고형비료(DO-PRO) 0.1g, 갯부추는 속효성 액체비료 (Peters) 주 1회 8mL 엽면시비처리였고 두 종 모두 속효성 시비처리가 효과적이었다. 강부추와 갯부추의 초기 생육에는 30~60% 차광처리가 된 재배플롯에서 원예상토가 충진된 128셀 플러그 트레이에 종자를 파종한후, DO-PRO 0.1g 또 는 Peters 8mL를 주 1회 엽면시비하면서 재배하는 것이 효 과적이라 판단된다.

수국은 수국과(Hydrangeaceae) 수국속(Hydrangea)의 낙엽관목 식물로 크고 화려한 화형을 가져 절화, 분화 및 조 경수로 전세계적 인기가 있는 식물이다. 나무수국은 수국 (H. macrophylla)과 비교하여 삽목율이 낮은 것으로 알려져 있지만 나무수국의 묘목생산을 위한 삽목 연구 및 두 종간 삽목율 차이 원인 규명에 관한 연구는 미미하다. 본 연구는 IBA(Indol-3-butyric acid) 500mg·L-1 처리시 삽수의 침지 시간에 따른 삽목율 조사를 통해 적정 호르몬 처리 시간을 제 시하고 나무수국과 수국의 해부학적 구조 관찰을 통한 삽목율 차이 발생의 원인을 규명하고자 실시하였다. 나무수국의 적정 호르몬 처리 시간을 규명하기 위해 IBA 500mg·L-1을 무처 리, 30분, 2시간, 4시간 침지처리를 하였다. 종간 삽목율 차이 발생의 원인 규명을 위해 나무수국과 수국의 줄기 단면과 삽 목 후 시간 경과에 따른 발근을 해부학적으로 관찰하였다. 연 구의 결과 나무수국의 삽목시 IBA 500mg·L-1에 2시간 이상 침지처리가 다른 처리구와 비교하여 발근율이 높고 발생 뿌리 수가 가장 많았다. 또, 나무수국의 삽목율이 수국과 비교하여 낮은 것은 줄기의 세포 구조상 방사조직의 형태, 섬유세포의 밀도, 도관의 발달, 전분 함유 세포의 수 등에 차이가 관찰되 었고 이러한 세포 구조적 차이들의 영향으로 나무수국이 수국 보다 삽목 후 뿌리 조직 세포분열이 7일 늦게 시작되는 것이 확인되었다. 본 연구의 결과로 나무수국의 삽목 번식의 기초 자료로 활용되어 묘목 생산 효율 증대에 활용되길 바란다.

Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2 , Gsr , HO-1, and catalase , were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2 , HO-1 , and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.

Uncoupling protein 1 (UCP1) is a unique mitochondrial membranous protein expressed in brown adipose tissue (BAT) in mammals. While its expression in response to cold temperatures and adipogenic inducers is well-characterized in mammals and human infants, the molecular characterization and expression of UCP1 in fish remain unexplored. To address this gap, we analyzed UCP1 expression in response to adipogenic inducers in a fish cell line, rainbow trout gonadal cells (RTG-2), and compared it with UCP1 expression in three mammalian preadipocytes, 3T3-L1, T37i, and WT1 exposed to the Peroxisome proliferator-activated receptor gamma (PPARγ) agonists, rosiglitazone (Rosi). In mammalian preadipocytes, UCP1 protein was highly expressed by Rosi, with an induction of adipogenesis observed in a time-dependent manner. This suggests that UCP1 plays a significant role in adipogenesis in mammals. However, RTG-2 cells showed no response to adipogenic inducers and exhibited only marginal expressions of UCP1. These results imply that RTG-2 cells may lack crucial responsive mechanisms to adipogenic signals or that the adipogenic response is regulated by other mechanisms. Further studies are needed to confirm these phenomena in fish preadipocytes when an appropriate cell line is established in future research.

Previous studies have shown that proline mutations in the heptad repeat region stabilize the coronavirus spike (S) protein in a pre-fusion state. To understand the impact of proline substitutions on the fusogenicity of the S protein, we engineered the swine acute diarrhea syndrome coronavirus (SADS-CoV) S protein with two proline substitutions (S-PP) and examined its fusogenicity using dual-split-protein based cell-cell fusion assay. Unlike the wild-type S (S-WT), S-PP rarely formed syncytia. Additionally, protein expression of S-PP was impaired compared to S-WT, as previously reported. Our results indicate that pre-fusion stabilized S protein is unable to induce membrane fusion and provide a better understanding of SADS-CoV S and vaccine antigen design.

In this review, the regulatory mechanisms of autophagy were described, and its interaction with apoptosis was identified. The role of autophagy in embryogenesis, tooth development, and cell differentiation were also investigated. Autophagy is regulated by various autophagy-related genes and those related to stress response. Highly active autophagy occurrences have been reported during cell differentiation before implantation after fertilization. Autophagy is involved in energy generation and supplies nutrients during early birth, essential to compensate for their deficient supply from the placenta. The contribution of autophagy during tooth development, such as the shape of the crown and root formation, ivory, and homeostasis in cells, was also observed. Genes control autophagy, and studying the role of autophagy in cell differentiation and development was useful for understanding human aging, illness, and health. In the future, the role of specific mechanisms in the development and differentiation of autophagy may increase the understanding of the pathological mechanisms of disease and development processes and is expected to reduce the treatment of various diseases by modulating the autophagic phenomenon.

헤스페리딘(Hesperidin, HD)은 다양한 식물체에 존재하는, 강한 항산화 기능을 가진 대표적인 flavonoid의 일종이다. 본 연구에서는 수용성 HD인 Hesperidin glucoside(HDG)가 가지는 세포손상 회복, 항염증 인자억제 및 melanin 생성억제 활성을 세포수준에서 비교하였다. HDG는 HD에 당전이 효소반응 으로 제조되었으며, HD에 비해 20,000배 이상 수용해도가 증가되었다. HaCaT 세포주에 대한 세포독성은 HDG가 HD에 비해 월등히 낮았다. HD와 HDG는 모두 자외선 조사된 HaCaT 세포에서 세포생존율 회 복효과를 나타내었다. 또한 HD와 HDG는 세포내 산화질소(NO), 종양괴사인자-α(TNF-α) 및 인터루킨 -6(IL-6)과 같은 염증 매개체 및 cytokine을 감소시켰으며, HD 보다는 HDG의 효과가 다소 우수하였다. Melanoma B16F10 세포주를 이용한 melanin 형성능과 tyrosinase 저해활성을 측정한 결과, HD와 HDG 모두 효과를 나타내었으며 HDG가 약간 우수한 결과를 보였다. 결론적으로, HD의 당전이체인 HDG는 HD에 비해 동등이상의 세포손상 회복, 염증성 매개체 및 cytokine 억제능과 melanin 형성억제능을 나타내 었으며, HDG의 높은 수용성과 낮은 세포독성 등의 특성은 다양한 분야에서의 용도를 확대시킬 수 있을 것으로 보인다.

The electrification of transportation is expected to greatly contribute to achieving the global climate change target. This study analyzed technological competitiveness in the fuel cell electric vehicle (FCEV) field based on patent family and citation index. Technology analysis was conducted by dividing FECV into six sub-technology areas based on IPC with fuel cell system, fuel cell technology, vehicle system, hydrogen storage and fueling, catalyst technology and etc. The largest number of patents are being filed in the fuel cell system technology field, and the fields with high growth rates over the past 10 years (2012-2022) were vehicle systems (12.4%) and hydrogen storage fuel field(11.5%). As of 2021, among global automakers, Toyota ranks the first in patent applications for FECV followed by Hyundai Motors in Korea, followed by Honda and Audi, with an average annual growth rate of 19.8%, the highest among competitors.

We investigated the anti-inflammatory effects of shiitake mushroom and kelp (SMK) mixture extracts in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 cells. Treatment of RAW 264.7 cells with LPS significantly increased NO (nitric oxide) production, pro-inflammatory cytokines (tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-6, and IL-1β), and inflammation-related genes (COX-2 and inducible nitric oxide synthase (iNOS)). In cytotoxicity testing using RAW 264.7 cells, SMK mixture extracts in the range of 1-16 μg/mL did not inhibit cell proliferation. However, SMK mixture extracts significantly inhibited NO production in a dose-dependent manner (p<0.05). SMK treatment significantly decreased TNF-α, IL-6, IFN-γ, and IL-1β levels compared to the LPS group, and similarly, pro-inflammatory cytokine mRNA levels also decreased. SMK mixture extracts reduced the mRNA expression of COX-2 and iNOS in RAW 264.7 cells compared to LPS (p<0.05). The above results show that SMK mixture extracts suppressed the inflammatory response induced by LPS. In particular, the extracts were shown to regulate the inflammatory response by suppressing the expression of inflammatory cytokines and inflammation-related enzymes.

본 연구에서는 산화 방지 특성이 있는 가리워진 아민기를 함유한 산화 그래핀(hindered amine grafted graphene oxide, HA-GO)을 합성하여 이를 도입한 나피온(Nafion) 기반의 복합 막을 제조한 후 고분자 전해질 막 연료전지 시스템에 응용하였다. HA-GO는 4-아미노-2, 2, 6, 6-테트라메틸-4-피페리딘(4-amino-2, 2, 6, 6-tetramethyl piperidine)에 존재하는 아민 기와 GO 표면에 존재하는 에폭시기의 개환 반응을 통해 제조하였으며, 합성된 HA-GO의 함량을 달리한 복합 막을 제조하여 순수 Nafion 막과 성능 특성을 비교하였다. HA-GO가 첨가된 복합 막은 Nafion 단일 막에 비해 기계적 물성, 화학적 안정성 및 수소이온 전도 특성이 향상되었다. 특히 HA-GO의 산화 방지 특성으로 인해 HA-GO가 첨가된 복합 막은 펜톤 평가 (Fenton’s test) 이후 수소이온 전도도의 유지 특성이 Nafion 단일 막에 비해 큰 폭으로 향상된 것을 확인할 수 있었다.

석유기반 플라스틱의 대체제인 폴리하드록시부틸레이트(polyhydroxybutyrate, PHB)의 기존 추출방법은 분자량 감 소 및 물성 변형을 일으킨다. 본 연구에서는 기능화 된 탄소나노튜브(carbon nanotube, CNT)를 부착한 돌기형 탄소나노튜브 분리막의 여과를 통해 물리적 파쇄를 발생시켜 미생물 내 축적된 PHB를 추출하고자 하였다. 돌기형 탄소나노튜브 분리막의 물리적 파쇄를 확인하기 위해 대장균 용액으로 여과 실험을 수행하여 불활성화를 관찰하였다. 또한 PHB를 축적한 미생물 용 액의 여과를 수행하여 PHB가 추출되었는지 확인하였더니 가장 대표적인 추출방법인 chloroform과 비교하여도 여과로 인한 추출이 4% 높은 성능을 가진 것을 관찰하였다. 본 결과를 통해 친환경적 바이오 플라스틱 회수를 위한 돌기형 탄소나노튜브 분리막의 적용 가능성을 확인하였다.