Green pesticides, derived from natural sources, have gained wider attention as an alternative approach to synthetic pesticides in managing polyphagous pests like Spodoptera litura. In this study, the methanolic flower extract of Nyctanthes arbor-tristis (Mx-Na-t) was subjected to chemical screening, and major peak area derivatives 3-Hydroxy-1,2-dimethyl-4(1H)-pyridone (3H-dp) and Tyrosol (Ty-ol) were identified. The toxicity against S. litura larvae of Mx-Na-t (at 500 ppm) was highest in third instars (96.4%), and for 3H-dp and Ty-ol (at 5 ppm) in II instars (76.5% and 81.4%, respectively). The growth and development of S. litura larvae and pupae were significantly reduced by all three extract and phytochemical treatments. Fecundity rates also declined with the treatments, from 1,020 eggs (control) to 540 eggs (Mx-Na-t), 741 eggs (3H-dp), and 721 eggs (Ty-ol). The extract and its active constituents decreased adult emergence and slowed total larval development in a dosedependent manner. The major gut enzymes of S. litura decreased in young larvae (II instar) exposed to Mx-Na-t, 3H-dp, and Ty-ol. Fourth instar midgut tissues were severely damaged by Mx-Na-t (250 ppm), 3H-dp, and Ty-ol (2.5 ppm) treatments, which induced structural damage to the epithelial cells and gut lumen. The earthworm Eisenia foetida was used to test for non-target toxicity. Crude Mx-Na-t at 500 ppm (13% and 3%) and 3H-dp (9.3% and 2.1%) and Ty-ol (10.2% and 1.5%) at 5 ppm produced lower mortality than the synthetic chemical cypermethrin at 1.0 ppm (27% and 18%) in filter paper and artificial soil assays, respectively. In addition, there was no significant change in earthworm weights under all three phytochemical treatments compared to controls. Additionally, the in-silico predictions of BeeTox and ProTox II indicated little or no toxicity toward honey bees and other nontargets associated with 3H-dp and Ty-ol. Overall these phyto-chemicals offer an effective pest management strategy.

This study explores the profound impact of varying oxygen content on microstructural and mechanical properties in specimens HO and LO. The higher oxygen concentration in specimen HO is found to significantly influence alpha lath sizes, resulting in a size of 0.5-1 μm, contrasting with the 1-1.5 μm size observed in specimen LO. Pore fraction, governed by oxygen concentration, is high in specimen HO, registering a value of 0.11%, whereas specimen LO exhibits a lower pore fraction (0.02%). Varied pore types in each specimen further underscore the role of oxygen concentration in shaping microstructural morphology. Despite these microstructural variations, the average hardness remains consistent at ~370 HV. This study emphasizes the pivotal role of oxygen content in influencing microstructural features, contributing to a comprehensive understanding of the intricate interplay between elemental composition and material properties.

최근 살진균제는 세계 식량 안보에 없어서는 안될 필수 요소이며, 그 사용량은 증가하고 있다. 살진균제는 직접적 또는 간접적으로 곤충에 영 향을 미쳐 유전자 및 분자 수준의 변화를 일으킨다. 곤충은 다양한 해독 매커니즘을 통해 살진균제를 포함한 농약으로부터 유발되는 활성산소 (ROS) 독성을 제거한다. 본 연구는 살진균제 캡탄의 비치명적 투여량(0.2, 2, and 20 μg/μL)을 주입 후 갈색거저리의 유충에서 해독효소의 mRNA 발현량을 분석했다. 갈색거저리의 전사체 분석을 통해 해독 매커니즘 관련 유전자인 퍼옥시다제(POX), 카탈라제(CAT), 슈퍼옥사이드 디스뮤타제(SOD) 및 글루타티온-S-트랜스퍼라제(GST)를 발굴하였다. 처리 24시간 후 TmPOX5 mRNA가 유의하게 증가한 것으로 나타났다. 처리 3 시간 후 TmSOD4의 mRNA가 유사하게 증가하였다. 또한 2 μg/μL 처리 24시간 후 TmCAT2의 mRNA 가 유의하게 증가하였다. 캡탄 노출 후 TmGST1 및 TmGST3의 mRNA 발현량도 증가하였다. 결론적으로, TmPOX5 및 TmSOD4 유전자는 갈색거저리에서 캡탄 노출에 대한 바이오마커 또는 생체이물 센서로 작용할 수 있음을 시사한다.

온도, pH, 빛 및 힘 등의 외부 자극에 반응하여 그 구조나 물리 화학적 특성이 변화 가능한 자극 감응형 하이드로젤에 대한 연구가 활발하게 진행되고 있다. 본 연구에서는 응력 감응형 분자인 스피로 피란을 사용하여 응력 및 pH 감응형 하이드로젤을 제조하였다. 먼저, 폴리에틸렌 다이아크릴레이트 (PEGDA)를 스피로피란 분자 양 끝에 접목시켜, 수용액에 쉽게 용해될 뿐만 아니라 하이드로젤 가교제 역할이 가능한 아령모양(PEG-spiropyran-PEG)의 SP-PEGDA 분자를 합성하였다. 이렇게 합성한 SP-PEGDA로 가교된 하이드로젤은 팽윤에 의해 발생하는 내부 응력에 의해 노란색의 스피로피란(SP) 분자를 보라색의 메로사인(MC) 형태로 변환시켰다. 또한 pH에 따라 양성화된 메로사인(MCH) 형태로 변환 하여 팽윤과 수축을 시각화 하였다.

본 연구에서는 강도를 조절한 마이크로겔을 사용하여 다양한 점탄성을 갖는 콜로이드 마이크로겔을 제조하였다. 하이드로겔의 화학적 가교제의 함량이 증가할수록 팽윤비는 2.0×104%에서 6.0×103%까지 감소하였고, 강도는 22.2 kPa에서 99.7 kPa까지 증가하였다. 이를 100 μm 크기로 분쇄하여 마이크로겔을 제작하였고 이온성 가교결합을 유도하는 분산액과 혼합하여 콜로이드 마이크로겔을 제작하였다. 그 결과, 가교제의 가교도와 분산액에 따라 10-1rad/s의 진동수에서 1.679 kPa.s에서 86.485 kPa.s까지 점도를 세밀하게 조절할 수 있었다. 본 연구에서는 콜로이드 마이크로겔의 물성을 제어하기 위해 하이드로겔의 가교도를 조절 또는 분산액의 종류와 함량을 조절하여 다양한 유변학적 거동을 갖는 콜로이드 마이크로겔을 제조하였다. 물성을 제어할 수 있는 콜로이드 마이크로겔을 사용하여 향후 콜로이드 현탁액 및 유화를 제조하는 화장품, 제약, 페인트 및 식품 산업에서 목적에 따라 적합한 물성을 갖는 콜로이드 마이크로겔을 제조할 수 있다.

Nanoemulsions are actively used in several applications for pharmaceutical, cosmetic and chemical industries. In this study, we propose the use of microfluidizer known as high pressure homogenizer to prepare lipid nanoemulsion as a potent cosmetic delivery carrier. The lipid nanoemulsions were prepared by O/W emulsion with hydrogenated lecithin and different type of oils. Effects of oil type on the stability of the lipid nanoemulsion were investigated with Dynamic Light Scattering (DLS) and Zeta-potential. Arbutin was used as model drug for transdermal administration through hairless mouse skin. Transdermal arbutin delivery using the lipid nanoemulsions was studied with HPLC method.

We developed a polymerase chain reaction (PCR)-based molecular method for sexing and identification using sexual dimorphism between the Zinc Finger-X and -Y (ZFX-ZFY) gene and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for mitochondrial DNA (mtDNA) cytochrome B (CYTB) gene in meat pieces and commercial sausages from animals of different origins. Sexual dimorphism based on the presence or absence of SINE-like sequence between ZFX and ZFY genes showed distinguishable band patterns between male and female DNA samples and were easily detected by PCR analyses. Male DNA had two PCR products appearing as distinct two bands (ZFX and ZFY), and female DNA had a single band (ZFX). Molecular identification was carried out using PCR-RFLP of CYTB gene, and showed clear species classification results. The results yielded identical information on the sexes and the species of the meat samples collected from providers without any records. The analyses for DNA isolated from commercial sausage showed that pig was the major source but several sausages originated from chicken and Atlantic cod. Applying this PCR-based molecular method was useful and yielded clear sex information and identified the species of various tissue samples originating from livestock.

Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

We have identified novel ricin-type (R-type) lectin by sequencing of random clones from cDNA library of the coleopteran beetle, T.molitor. The cDNA sequence is comprised of 495 bp encoding a protein of 164 amino acid residues and shows 49% identity with galectin of Tribolium castaneum. Bioinformatics analysis shows that the amino acid residues from 35 to 162 belong to ricin-type β-trefoil structure. The transcript was significantly upregulated after early hours of injection with peptidoglycans derived from Gram (+) and Gram (-) bacteria, beta-1, 3 glucan from fungi and an intracellular pathogen, L. monocytogenes suggesting putative function in innate immunity.

The objective of this study was to determine the effect of macrophages on growth of human colon cancer cells. The results showed that co-culture of colon cancer cells with macrophages inhibited the growth of colon cancer cells (HCT116 and SW620) depending on the number of macrophages, RAW 264.7 cells, and activated THP-1 cells accompanied by down regulation of pSTAT3 in cancer cells. We also found that expression and release of cancer cell growth inhibitory cytokines, IL-1 receptor antagonist (IL-1ra) and IL-10, was increased in macrophages. Blocking of the STAT3 pathway with specific inhibitor and siRNA of STAT3 abolished the growth of colon cancer cells and expression of IL-1ra and IL-10. In addition, neutralization of IL-1ra and IL-10 with antibodies resulted in reversal of macrophage-induced inhibition of cancer cell growth. These data showed that IL-1ra and IL-10 released from macrophages inhibit growth of colon cancer cells through inhibition of the STAT3 pathway.

The discovery of antibiotics has helped to save the lives of an uncountable number of people. Antibiotics have been grouped in different classes based on their origin, structure, and mechanism of action. An intrinsic and acquired mechanism of antimicrobial resistance has been identified in many bacterial strains that are of high clinical importance. This has seriously jeopardized the use of antibiotics and has also caused the spread of microbes that are resistant to effective first-choice, or “first-line” drugs. Thus, sensible use of antibiotics and the search for effective alternative measures are of high importance in order to minimize the effect due to existing and emerging antimicrobial resistant microbes.

본 연구에서는 가교 분자를 사용하여 0.7 kPa에서 17.7 kPa까지 다양한 강도를 갖는 콜라겐 겔을 성공적으로 제조하였다. 가교된 콜라겐 겔에 다공성 기공을 도입하고 진피세포를 내부에 담지하여, 겔 강도에 따른 세포 성장 및 거동을 확인하였다. 상대적으로 강도가 높은 겔에서 진피세포의 프로콜라겐 생합성이 47 ng에서 32 ng까지 감소하는 것을 확인하였다. 이렇게 제조된 인공피부 진피에 아데노신을 처리하였을 때, 특정 강도를 갖는 콜라겐 겔에서 프로콜라겐 생합성이 감소하는 것을 확인하였다. 반면에 기능성 펩타이드를 처리하였을 때 는 프로콜라겐 생합성이 콜라겐 겔의 강도에 크게 영향을 받지 않는 것을 확인할 수 있었다. 이러한 결과는 강도가 제어된 인공피부 제조 및 응용, 나아가 다양한 조직공학 분야의 기반 기술로 활용 가능하리라 기대된다.

본 연구에서는 생체재료인 콜라겐과 합성 단량체인 아크릴아마이드를 연속가교 하여, 하이드로젤 기반의 콜라겐 겔을 제조하였다. 아크릴아마이드의 함량 및 가교 정도에 따라, 1.5 kPa에서 3.0 kPa까지 다양한 강도 (E)를 갖는 콜라겐 겔을 제조할 수 있었다. 또한, 콜라겐 겔에 다공성 기공을 도입하고 진피세포를 내부에 담지하여, 겔 강도에 따른 세포 성장 및 거동을 확인하였다. 상대적으로 강도가 높은 겔에서 세포의 성장은 느렸지만 GAG 합성 및 분비는 활성화되는 것을 확인하였다. 콜라겐 겔의 기계적 물성에 따라 세포의 성장 및 활성이 영향을 받는 것을 알 수 있었으며, 이는 향후 인공피부 제조 및 응용, 나아가 다양한 조직공학 분야의 기반 기술로 활용 가능하리라 기대된다.

본 연구에서는 수용액상에서 자기조립 나노입자를 형성할 수 있는 레티노익산이 접목된 양친성 폴리아미 노산 유도체를 합성하였다. 합성한 양친성 폴리아미노산은 레티노익산의 접목도가 각각 5, 10, 30 mol%가 되도 록 조절하였다. 수용액 상에서 양친성 폴리아미노산은 소수성 레티노익산의 분자 결합에 의해 안정한 자기조립 나노입자를 형성하였다. 자기조립 나노입자는 레티노익산의 접목도가 증가할수록 크기는 작아지고 형태는 구형 에서 이중층 구조로 전이되었다. 또한 접목도가 10%일 때, 자기조립 입자의 구조 붕괴 없이 레티놀의 포집 및 전달이 가장 효과적인 것을 확인하였다. 접목도가 제어된 자기조립입자는 레티놀을 안정적으로 포집할 수 있기 때문에 주름개선제 및 다양한 기능성 화장품 전달체로 활용될 수 있다