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Development of High Intensity Focused Ultrasound (HIFU) Mediated AuNP-liposomal Nanomedicine and Evaluation with PET Imaging

  • Ji Yoon Kim;Un Chul Shin;Ji Yong Park;Ran Ji Yoo;Soeku Bae;Tae Hyeon Choi;Kyuwan Kim;Young Chan Ann;Jin Sil Kim;Yu Jin Shin;Hokyu Lee;Yong Jin Lee;Kyo Chul Lee;Suhng Wook Kim;Yun-Sang Lee
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.9 no.1
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    • pp.9-16
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    • 2023
  • Liposomes as drug delivery system have proved useful carrier for various disease, including cancer. In addition, perfluorocarbon cored microbubbles are utilized in conjunction with high-intensity focused-ultrasound (HIFU) to enable simultaneous diagnosis and treatment. However, microbubbles generally exhibit lower drug loading efficiency, so the need for the development of a novel liposome-based drug delivery material that can efficiently load and deliver drugs to targeted areas via HIFU. This study aims to develop a liposome-based drug delivery material by introducing a substance that can burst liposomes using ultrasound energy and confirm the ability to target tumors using PET imaging. Liposomes (Lipo-DOX, Lipo-DOX-Au, Lipo-DOX-Au-RGD) were synthesized with gold nanoparticles using an avidin-biotin bond, and doxorubicin was mounted inside by pH gradient method. The size distribution was measured by DLS, and encapsulation efficiency of doxorubicin was analyzed by UV-vis spectrometer. The target specificity and cytotoxicity of liposomes were assessed in vitro by glioblastoma U87mg cells to HIFU treatment and analyzed using CCK-8 assay, and fluorescence microscopy at 6-hour intervals for up to 24 hours. For the in vivo study, U87mg model mouse were injected intravenously with 1.48 MBq of 64Cu-labeled Lipo-DOX-Au and Lipo-DOX-Au-RGD, and PET images were taken at 0, 2, 4, 8, and 24 hours. As a result, the size of liposomes was 108.3 ± 5.0 nm at Lipo-DOX-Au and 94.1 ± 12.2 nm at Lipo-DOX-Au-RGD, and it was observed that doxorubicin was mounted inside the liposome up to 52%. After 6 hours of HIFU treatment, the viability of U87mg cells treated with Lipo-DOX-Au decreased by around 20% compared to Lipo-DOX, and Lipo-DOX-Au-RGD had a higher uptake rate than Lipo-DOX. In vivo study using PET images, it was confirmed that 64Cu-Lipo-DOX-Au-RGD was taken up into the tumor immediately after injection and maintained for up to 4 hours. In this study, drugs released from liposomes-gold nanoparticles via ultrasound and RGD targeting were confirmed by non-invasive imaging. In cell-level experiments, HIFU treatment of gold nanoparticle-coupled liposomes significantly decreased tumor survival, while RGD-liposomes exhibited high tumor targeting and rapid release in vivo imaging. It is expected that the combination of these models with ultrasound is served as an effective drug delivery material with therapeutic outcomes.

Evaluation of Anti-wrinkle Effect of Peanut Shell Extract and Stability of the Extract in Cosmetic Products (땅콩 겉껍질 추출물의 주름개선 효능 및 화장품 제형에서 추출물의 안정성 평가)

  • Narae Han;Jin Young Lee;Mihyang Kim;Eun Young Choi;Bong-Jeun An;Yu-Young Lee;Moon Seok Kang;Hyun-Joo Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.203-212
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    • 2023
  • This study was conducted to investigate the possibility of peanut shell, a by-product of peanut, as a functional cosmetic ingredient. Peanut shell extract showed high antioxidant activity with IC50 values of 75.00, 46.33, and 472.83 ㎍/mL for DPPH and ABTS radical scavenging and SOD-like activity, respectively. Furthermore, peanut shell extract was efficiently decreased the MMP-1 and MMP-3 protein level in the UVB treated-HaCaT cell and maintained procollagen protein level similar to normal control. Similar to anti-wrinkle related protein expression assay, the IC50 value of elastase and collagnease inhibition in peanut shell extract was lower as 0.30 and 0.09 mg/mL, respectively, than that of the positive control. Additionally, eriodictyol and luteolin, which are isolated from peanut shell extract, showed 53.8 and 98.0% elastase inhibition rate, respectively, and 60.1 and 72.5% collagenase inhibition rate, respectively, at a concentration of 0.1 mg/mL. Thus, luteolin was assumed to be the effective ingredient for wrinkle inhibition in peanut shell extract. As a result of stability evaluation of lotion and cream formulations containing peanut shell extract, it was confirmed to be a stable formulation with no significant changes. Therefore, it is considered that peanut shell extract can be applied as a cosmetic ingredient for wrinkle inhibition.

The Protective Role of Gleditsiae fructus against Streptococcus pneumoniae (폐렴 구균에 대한 조협의 보호 역할 연구)

  • Jun-ki Lee;Se-Hui Lee;Dong Ju Seo;Kang-Hee Lee;Sojung Park;Sun Park;Taekyung Kim;Jin-Young Yang
    • Journal of Life Science
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    • v.33 no.2
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    • pp.158-168
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    • 2023
  • Natural products have been used to mitigate the effects of cancer and infectious diseases, as they feature diverse bioactivities, such as antioxidant, antibacterial, anti-inflammatory, and immunomodulatory effects. Here, we chose 10 natural products that are well-known as pulmonary enhancers and investigated their bactericidal effects on Streptococcus pneumoniae. In the disk diffusion assay, the growth of S. pneumoniae was significantly regulated by G. fructus treatment regardless of extraction method used. We first adopted spraying as a novel delivery method for G. fructus. Interestingly, mice exposed to G. fructus three times a day for 2 weeks were resistant to S. pneumoniae intranasal infection (shown both through body weight loss and survival rates compared to the control group). Moreover, we confirmed that exposure to G. fructus regulated the colonization of the bacteria despite the sustained inflammation in the lung after exposure to S. pneumoniae, indicating that migrated inflammatory immune cells may involve a host defense mechanism against pulmonary infectious diseases. While a similar number of granulocytes (CD11b+Ly6C+Ly6G+), neutrophils (CD11b+Ly6CintLy6G+), and monocytes (CD11b+Ly6CintLy6G-) were found between groups, a significantly increased number of alveolar macrophages (CD11b+CD11chiF4/80+) was detected in BAL fluids of mice pre-exposed to G. fructus at 5 days after S. pneumonia infection. Taken together, our data suggest that this usage of G. fructus can induce protective immunity against bacterial infection, indicating that facial spray may be helpful in enhancing the defense mechanism against pulmonary inflammation and in evaluating the efficacy of natural products as immune enhancers against respiratory diseases.

Brain-expressed X-linked 2 Binds to Kinesin Superfamily Protein 3A (Brain-expressed X-linked (Bex) 2와 heterotrimeric kinesin-2의 KIF3A와의 결합)

  • Kim, Mooseong;Jeong, Young Joo;Park, Sung Woo;Seo, Mi Kyoung;Kim, Sang Jin;Lee, Won Hee;Urm, Sang-Hwa;Lee, Jung Goo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.32 no.3
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    • pp.189-195
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    • 2022
  • Kinesin-2 comprises two subfamilies of the heterotrimeric or homodimeric motors found in mammalian cells. Heterotrimeric kinesin-2 consists of kinesin superfamily proteins (KIFs) 3A and 3B and kinesin-associated protein 3 (KAP3), which is a molecular motor protein that moves along microtubules. It plays diverse roles in cargo transport, including anterograde trafficking in cilia, and interacts with many different cargoes and proteins, but their binding proteins have not yet been fully identified. In this study, the yeast two-hybrid assay was used to identify the proteins that interact with the cargo-binding domain (CBD) of KIF3A, and an interaction between KIF3A and brain expressed X-linked 2 (Bex2) was found. Bex2 bound to the CBD-containing C-terminal tail region of KIF3A but did not interact with the same region of KIF3B or KIF5A (a motor protein of kinesin-1). KIF3A interacted with another isoform, Bex1, but did not interact with Bex3. In addition, glutathione S-transferase (GST) pull-downs showed that KIF3A specifically interacts with GST-Bex1 and GST-Bex2 but not with GST alone. When co-expressed in HEK-293T cells, Bex2 co-localized with KIF3A and co-immunoprecipitated with KIF3A and KIF3B but not KIF5B. In combination, these results suggest that Bex2 is capable of binding to heterotrimeric kinesin-2 and may serve as an adaptor protein that links heterotrimeric kinesin-2 with cargo.

Protective Effect of Mixed Extract including Hovenia dulcis Thunberg against Chronic Ethanol Treatment-induced Cytotoxicity in a Brain and Liver Tissue (만성 알코올 섭취로부터 유도되는 뇌 및 간 조직 독성에 대한 지구자 혼합 추출물의 보호 효과)

  • Kim, Jong-Min;Park, Seon-Kyeong;Guoa, Tian-Jiao;Kang, Jin-Yong;Ha, Jeong-Su;Lee, Du-Sang;Kwon, O-Jun;Lee, Uk;Heo, Ho Jin
    • Journal of agriculture & life science
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    • v.50 no.2
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    • pp.125-138
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    • 2016
  • To assess the industrial possibility of mixed-extracts containing Hovenia dulcis Thunberg and 12 different botanical ingredients, a protective effect was confirmed in the chronic ethanol-induced the liver, brain, and blood injury in mouse. Blood glucose levels of the normal control group(NG) and ethanol administration group(EG) were respectively 119.43mg/dL and 305.25mg/dL, and the mixed-extracts administration group(100, 200mg/kg body weight + 25% ethanol 5g/kg body weight respectively; ME100 & ME200) were decreased to 272.76mg/dL and 234.60mg/dL. Blood ethanol contents were decreased in ME100 and ME200(3.85mg/dL, 3.08mg/dL) compared to EG(4.08mg/dL), and blood acetaldehyde contents were also decreased in ME(15.76mg/dL, 15.16mg/dL) compared to EG(18.72mg/dL). The contents of hepatotoxic indicators such as glutamine pyruvic transaminase(GPT) and glutamic oxaloacetic transaminase (GOT), nephrotoxic indicators such as blood urea nitrogen(BUN), and creatine(CRE), and total cholestero(TCHO), and triglyceride(TG) in mouse blood serum were significantly decreased in the ME compared to EG. The acetylcholinesterase(AChE) activity of ME(109.00% and 108.47%, respectively) in mouse brain tissues was decreased in ME compared to EG(116.10%). Finally, ME was remarkable in vivo antioxidant activities in the mouse liver and brain tissues by superoxide dismutase(SOD), oxidized glutathione(GSH)/total GSH ratio and the malondialdehyde (MDA) assay. Therefore, the mixed-extracts was considered to be effective a high value food with protective effect against chronic ethanol traetment-induced cytotoxicity in liver and brain tissues.

Anti-tumor Effect of a Combination of Hongyoung Ethanol Extract and Cisplatinin YD-10B Oral Cancer Cells (YD-10B 구강암세포에서 홍영 에탄올 추출물과 시스플라틴 병용에 의한 항암 효과)

  • Eun-Jung Kim;Sung-Hee Hwang;Sangwook Park
    • Journal of Life Science
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    • v.33 no.6
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    • pp.498-505
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    • 2023
  • Solanum tuberosum Linnaeus cv Hongyoung, which represents red potato, was developed in Korea. Hongyoung is known to have anti-oxidant, anti-inflammatory, anti-viral, and anti-tumor properties, but no research has been conducted on the growth inhibition and apoptosis effects of hongyoung in YD-10B oral cancer cells. In this study, the combined treatment of hongyoung ethanol extract (HEE) and cisplatin were examined to determine its ability to inhibit cancer cell growth, induce apoptosis, and inhibit matrix metalloproteinases (MMP)-2 and MMP-9 cancer metastasis. The cell viability was investigated using a 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H- tetrazolium monosodium salt (MTS) assay, and the ability to induce apoptosis was analyzed using an FACS analyzer. The mRNA expression and protein activity of MMP-2 and MMP-9 were measured via RT-PCR and zymography. The YD-10B oral cancer cells showed an increase in growth inhibition as the concentration of HEE increased. The combination of 200 µM cisplatin and 500 ㎍/ml HEE reduced the growth of the YD-10B oral cancer cells by more than 50% compared to cisplatin alone. When phorbol 12-myristate 13-acetate (PMA)-treated YD-10B oral cancer cells were co-treated with 200 µM cisplatin and 500 ㎍/ml HEE, both the mRNA expression and protein activity of the MMP-2 and MMP-9 decreased. In addition, the percentage of the sub-G1 phase, which indicates apoptosis ability, more than doubled when treated in combination with 200 µM cisplatin and 500 ㎍/ml HEE than when cisplatin alone was used. The results of this study therefore suggest the possibility of using a combination of HEE and cisplatin in the development of effective drugs to treat oral cancer.

Antioxidant Activity of Tea Made from Korean Mountain-Cultivated Ginseng Leaves and its Influence on Lipid Metabolism (장뇌삼 엽차의 항산화활성 및 지질대사에 미치는 영향)

  • Bae, Man-Jong;Kim, Soo-Jung;Ye, Eun-Ju;Nam, Hak-Sik;Park, Eun-Mi
    • Journal of the Korean Society of Food Culture
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    • v.24 no.1
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    • pp.77-83
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    • 2009
  • This study was conducted to investigate the chemical composition and biological function of tea made from mountain-cultivated ginseng leaves. The antioxidant activities of tea made from mountain-cultivated ginseng leaves were determined by measuring their electron-donating ability based on their DPPH and nitrite-scavenging ability. The electron-donating abilities of tea made from mountain-cultivated ginseng leaves (500 and 1,000 ppm) as determined by DPPH assay were 45.6 and 85.1%, respectively. The nitrite scavenging ability of tea made from mountain-cultivated ginseng leaves (500 and 1,000 ppm) at pH 6.0 were 32.8 and 51.4%, respectively. Furthermore, the nitrite scavenging activity increased in a dose-dependent manner at all pH values. The effects of tea made from mountain-cultivated ginseng leaves on Male Sprague-Dawley rats were also evaluated. To accomplish this, the rats were divided into three groups (A: normal diet group, B: high fat diet group and C: high fat diet supplemented with tea made from mountain-cultivated ginseng leaves group). The anti-obesity effects of tea made from mountain-cultivated ginseng leaves were then evaluated. The serum total lipid, total cholesterol and triglyceride contents in C group were lower than those of B group; however, these differences were not statistically significant. The HDL-cholesterol content was significantly higher in the C group than in the other groups. Taken together the results of this study suggest that tea made from mountain-cultivated ginseng leaves possesses antioxidant activity and improves lipid metabolism.

Biochemical Assessment of Deer Velvet Antler Extract and its Cytotoxic Effect including Acute Oral Toxicity using an ICR Mice Model (ICR 마우스 모델을 이용한 녹용 추출물의 생화학적 평가 및 급성 경구 독성을 포함한 세포 독성 효과)

  • Ramakrishna Chilakala;Hyeon Jeong Moon;Hwan Lee;Dong-Sung Lee;Sun Hee Cheong
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.430-441
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    • 2023
  • Velvet antler is widely used as a traditional medicine, and numerous studies have demonstrated its tremendous nutritional and medicinal values including immunity-enhancing effects. This study aimed to investigate different deer velvet extracts (Sample 1: raw extract, Sample 2: dried extract, and Sample 3: freeze-dried extract) for proximate composition, uronic acid, sulfated glycosaminoglycan, sialic acid, collagen levels, and chemical components using ultra-performance liquid chromatography-quadrupole-time-of-light mass spectrometry. In addition, we evaluated the cytotoxic effect of the deer velvet extracts on BV2 microglia, HT22 hippocampal cells, HaCaT keratinocytes, and RAW264.7 macrophages using the cell viability MTT assay. Furthermore, we evaluated acute toxicity of the deer velvet extracts at different doses (0, 500, 1000, and 2000 mg/kg) administered orally to both male and female ICR mice for 14 d (five mice per group). After treatment, we evaluated general toxicity, survival rate, body weight changes, mortality, clinical signs, and necropsy findings in the experimental mice based on OECD guidelines. The results suggested that in vitro treatment with the evaluated extracts had no cytotoxic effect in HaCaT keratinocytes cells, whereas Sample-2 had a cytotoxic effect at 500 and 1000 ㎍/mL on HT22 hippocampal cells and RAW264.7 macrophages. Sample 3 was also cytotoxic at concentrations of 500 and 1000 ㎍/mL to RAW264.7 and BV2 microglial cells. However, the mice treated in vivo with the velvet extracts at doses of 500-2000 mg/kg BW showed no clinical signs, mortality, or necropsy findings, indicating that the LD50 is higher than this dosage. These findings indicate that there were no toxicological abnormalities connected with the deer velvet extract treatment in mice. However, further human and animal studies are needed before sufficient safety information is available to justify its use in humans.

Application of the Extract of Zanthoxylum piperitum DC to Manufacturing Eco-friendly Nosocomial Infection Control Protective Materials (초피의 항균 활성을 이용한 원내 감염 제어 친환경 방호 소재 개발)

  • Shin Young Park;Ki Yun Kim;Do Youn Jun;Sung Chul Kim;Hyo-Il Jung;Young Ho Kim
    • Journal of Life Science
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    • v.33 no.10
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    • pp.820-827
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    • 2023
  • Since COVID-19 began at the end of 2019, the wearing time of protective clothing used to prevent pathogenic bacteria and virus infection has increased, and the development of safe protective materials that are human-friendly and have antibacterial and antiviral functions has been required. In this study, we investigated the possibility of developing natural antibacterial protection materials using ethanol extract of the medicinal plant Zanthoxylum Piperitum DC. The antibacterial activity assay of the 80% ethanol extract of Z. piperitum DC leaves against various nosocomial infectious bacteria, using the disk diffusion method, showed that Staphylococcus aureus ATCC 25923, Klebsiella pneumoniae ATCC 13883, Salmonella typhimurium, and Aeromonas hydrophila are sensitive to the inhibitory action of the extract. The IC50 values of the ethanol extract against S. aureus, K. pneumoniae, P. vulgaris and A. hydrophila were about 0.59 mg/ml, 0.50 mg/ml, 1.06 mg/ml, and 0.06 mg/ml, respectively. To determine whether the ethanol extract of Z. piperitum DC leaves can be applied to the development of antibacterial protective fabric, the ethanol extract was tested using a protective fabric from the KM Health Care Corp. using the JIS L1902-Absorption method. As a result, the bacteriostatic and bactericidal activity values of S. aureus ATCC 25923 and K. pneumoniae ATCC 13883 appeared to be more than 2.0 when treated with the ethanol extract at a concentration of 1% (w/v). Together, these results suggest that Z. piperitum DC leaves can be applied to develop natural antibacterial functional protective fabrics.

Effects of Microbial Fermentation on the Antioxidant Activities of Protaetia brevitarsis Larvae (미생물 발효가 흰점박이꽃무지(Protaetia brevitarsis) 유충의 항산화 활성에 미치는 영향)

  • Han Bi Kim;Hye Soo Kim;Soo Jeong Cho
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1052-1061
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    • 2023
  • This study was carried out to evaluate the effect of fermentation by B. subtilis (BPLE), L. brevis (LPLE), S. cerevisiae (SPLE) and C. militaris (CPLE) on the antioxidant activity of Protaetia brevitarsis larvae fed with mushroom substrates (king oyster mushroom). The total polyphenol content of Protaetia brevitarsis larvae (PLE), BPLE, LPLE, SPLE and CPLE were 58.07±0.67, 83.33±0.98, 79.21±1.32, 61.02±0.87 and 57.90±1.02 mg GAEs/extract g, respectively. The flavonoid contents of the PLE, BPLE, LPLE, SPLE and CPLE were 17.35±1.57, 19.49±0.95, 16.90±1.57, 18.12±0.95 and 16.99±0.95 mg QEs/extract g, respectively. The DPPH radical scavenging activity showed no significant difference between the PLE, BPLE, LPLE, SPLE and CPLE at a concentration of 0.2 mg/ml. However, at a concentration of 0.4 mg/ml or more, the DPPH radical scavenging activity of the BPLE and LPLE was higher than that of the PLE. The reducing power of the BPLE and LPLE was also higher than that of the PLE, and more than twice as high at a concentration of 0.8 mg/ml or more. The ORAC value of the BPLE (79.77±0.82 uM TEs/extract g) was higher than that of the PLE (61.34±0.97 uM TEs/extract g). A WST-1 assay of the RAW 264.7 cells indicated that the PLE, BPLE, LPLE, SPLE and CPLE showed no cytotoxicity.