• 제목/요약/키워드: in vivo tissue

검색결과 854건 처리시간 0.03초

Targeting of Nuclear Encoded Proteins to Chloroplasts: a New Insight into the Mechanism

  • Lee, Yong-Jik;Kim, Yong-Woo;Pih, Kyeong-Tae;Hwang, Inhwan
    • 식물조직배양학회지
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    • 제27권5호
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    • pp.407-409
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    • 2000
  • Outer envelope membrane proteins of chloroplasts encoded by the nuclear genome are transported without the N-terminal transit peptide. Here, we investigated the targeting mechanism of AtOEP7, an Arabidopsis homolog of small outer envelope membrane proteins in vivo. AtOEP7 was expressed transiently in protoplasts or stably in transgenic plants as fusion proteins with GFP. In both cases AtOEP7:GFP was targeted to the outer envelope membrane when assayed under a fluorescent microscope or by Western blot analysis. Except the transmembrane domain, deletions of the N- or C-terminal regions of AtOEP7 did not affect targeting although a region closed to the C-terminal side of the transmembrane domain affected the targeting efficiency. Targeting experiments with various hybrid transmembrane mutants revealed that the amino acid sequence of the transmembrane domain determines the targeting specificity The targeting mechanism was further studied using a fusion protein, AtOEP7:NLS:GFP, that had a nuclear localization signal. AtOEP7:NLS:GFP was efficiently targeted to the chloroplast envelope despite the presence of the nuclear localization signal. Taken together, these results suggest that the transmembrane domain of AtOEP7 functions as the sole determinant of targeting specificity and that AtOEP7 may be associated with a cytosolic component during translocation to the chloroplast envelope membrane.

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Benzochloroporphyrin Derivative Induced Cytotoxicity and Inhibition of Tumor Recurrence During Photodynamic Therapy for Osteosarcoma

  • Gong, Hai-Yang;Sun, Meng-Xiong;Hu, Shuo;Tao, Ying-Ying;Gao, Bo;Li, Guo-Dong;Cai, Zheng-Dong;Yao, Jian-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권5호
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    • pp.3351-3355
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    • 2013
  • Photodynamic therapy (PDT) is a promising cancer treatment modality that uses dye-sensitized photooxidation of biologic matter in target tissue. This study explored effects of the photosensitizer BCPD-17 during PDT for osteosarcoma. LM-8 osteosarcoma cells were treated with BCPD-17 and cell viability after laser irradiation was assessed in vitro with the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The effects of BCPD-17 during PDT recurrence were then examined on tumor-bearing mice in vivo. BCPD-17 had dosedependent cytotoxic effects on LM-8 osteosarcoma cells after laser irradiation which also had energy-dependent effects on the cells. The rate of local recurrence was reduced when marginal resection of mice tumors was followed by BCPD-17-mediated PDT. Our results indicated BCPD-17-mediated PDT in combination with marginal resection of tumors is a potentially new effective treatment for osteosarcoma.

고추 추출물의 경구 투여에 의한 피어스판 면역세포 활성화 작용 (Immunomodulatory Effects of Orally Administrated Capsicum Extract on Peyer's Patches)

  • 박민영;김동희;진미림
    • 동의생리병리학회지
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    • 제24권3호
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    • pp.446-451
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    • 2010
  • We investigated whether oral administration with capsicum extract (Capsicum annuum var. cheongyang) would affect the immune system by examining the immune cells of Peyer's patch (PP), a gut associated lymphoid tissue, ex vivo. The mice were orally administrated with capsicum extract (100 mg/kg/day), capsaicin (10 mg/Kg), and the vehicle for four consecutive days, and PPs were isolated from intestines 2 days later. When the PP cells were cultured in the presence of Concanavalin A for 72 hr, the levels of cytokines, including IL-2 and IFN-${\gamma]$, were dramatically increased, while the levels of IL-4 remained unchanged compared with the control. Data from the FACS analysis of PP cells indicated that capsicum extract significantly increased the number of CD3+ and CD4+ T cells as well as CD 19+ B cells compared with the control but not CD11b+ cells. Furthermore, the percentages of IL-2+ /CD4+ cells and IFN-${\gamma}+$/CD4+ were greatly increased. These data suggested that oraladministration with capsicum extract might activate the CD4+ T cells leading to cytokine production as well as CD19+ B cells in Peyer's patches. As such, capsicum extract might have potential as an immune modulating agent.

Antimicrobial Efficacy of Penicillium amestolkiae elv609 Extract Treated Cotton Fabric for Diabetic Wound Care

  • Rozman, Nur Amiera Syuhada Binti;Hamin, Nurhanis Syafiqah Binti Mohd Nor;Ring, Leong Chean;Nee, Tan Wen;Mustapha, Mahfuzah Binti;Yenn, Tong Woei
    • Mycobiology
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    • 제45권3호
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    • pp.178-183
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    • 2017
  • Diabetes mellitus is a chronic disorder which affects millions of population worldwide. Global estimates published in 2010 reported the world diabetic prevalence as 6.4%, affecting 285 million adults. Foot ulceration and wound infection are major forms of disabilities arising from diabetic diseases. This study was aimed to develop a natural antimicrobial finishing on medical grade textile that meets American Association of Textiles Chemists and Colorists (AATCC) standard. The textile samples were finished with the ethanolic extract of Penicillium amestolkiae elv609, an endophytic fungus isolated from Orthosiphon stamineus Benth (common name: cat's whiskers). Endophyte is defined as microorganism that reside in the living plant tissue, without causing apparent disease symptom to the host. The antimicrobial efficacy of the ethanolic extract of P. minioluteum was tested on clinical pathogens isolated from diabetic wound. The extract exhibited significant inhibitory activity against 4 bacteria and 1 yeast with the minimal inhibitory concentration ranged from 6.25 to 12.5 mg/mL. The results indicate different susceptibility levels of the test microorganism to the ethanolic extract. However, the killing activity of the extract was concentration-dependent. The finished medical textile showed excellent antimicrobial efficacy on AATCC test assays. All the microbial cultures treated with the textile sample displayed a growth reduction of 99.9% on Hoheinstein Challenge Test. The wash durability of the finished textile was found good even after 50 washes with commercial detergent. Besides, the gas chromatography mass spectrometry analysis showed that 6-octadecenoic acid and diethyl phthalate were the main bioactive constituents of the extract. In conclusion, the developed medical textile showed good antimicrobial efficacy on laboratory tests. This work can be extended to in vivo trials for developing healthcare textile products for antimicrobial applications.

Differential Regulation of Cytochrome P450 Isozyme mRNAs and Proteins by Femur Fracture Trauma

  • Lee, Woo-Young;Lee, Sun-Mee
    • Archives of Pharmacal Research
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    • 제26권12호
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    • pp.1079-1086
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    • 2003
  • The aim of this study was to investigate the effect of trauma on cytochrome P450 (CYP) gene expression and to determine the role of Kupffer cells in trauma-induced alteration of CYP isozymes. Rats underwent closed femur fracture (FFx) with associated soft-tissue injury under anesthesia. To deplete Kupffer cells in vivo, gadolinium chloride ($GdCl_3$) was intravenously injected at 7.5 mg/kg body wt., 1 and 2 days prior to FFx surgery. At 72 h of FFx, liver tissues were isolated to determine the mRNA and protein expression of CYP isozymes and NADPH-P450 reductase by reverse transcription-polymerase chain reaction and Western immunoblotting, respectively. In addition, the mRNA levels of tumor necrosis factor alpha (TNF-$\alpha$), inducible nitric oxide synthase (iNOS) and heme oxygenase-1 (HO-1) were evaluated. FFx increased the mRNA level of CYP1A1; an increase that was not prevented by $GdCl_3$. There were no significant differences in the mRNA expression of CYP1A2, 2B1 and 2E1 among any of the experimental groups. The protein levels of CYP2B1 and 2E1 were significantly decreased by FFx; a decrease that was not prevented by $GdCl_3$ treatment. The gene expression of NADPH-P450 reductase was unchanged by FFx. FFx significantly increased the expression of TNF-$\alpha$ mRNA; an increase that was attenuated by $GdCl_3$. The mRNA expression of HO-1 was increased by FFx, but not by $GdCl_3$ . Our findings suggest that FFx differentially regulates the expression of CYP isozyme through Kupffer cell-independent mechanisms.

Proliferation of Mouse Prostate Cancer Cells Inflamed by Trichomonas vaginalis

  • Kim, Sang-Su;Kim, Kyu-Shik;Han, Ik-Hwan;Kim, Yeseul;Bang, Seong Sik;Kim, Jung-Hyun;Kim, Yong-Suk;Choi, Soo-Yeon;Ryu, Jae-Sook
    • Parasites, Hosts and Diseases
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    • 제59권6호
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    • pp.547-556
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    • 2021
  • Our objective was to investigate whether inflammatory microenvironment induced by Trichomonas vaginalis infection can stimulate proliferation of prostate cancer (PCa) cells in vitro and in vivo mouse experiments. The production of CXCL1 and CCL2 increased when cells of the mouse PCa cells (TRAMP-C2 cell line) were infected with live T. vaginalis. T. vaginalis-conditioned medium (TCM) prepared from co-culture of PCa cells and T. vaginalis increased PCa cells migration, proliferation and invasion. The cytokine receptors (CXCR2, CCR2, gp130) were expressed higher on the PCa cells treated with TCM. Pretreatment of PCa cells with antibodies to these cytokine receptors significantly reduced the proliferation, mobility and invasiveness of PCa cells, indicating that TCM has its effect through cytokine-cytokine receptor signaling. In C57BL/6 mice, the prostates injected with T. vaginalis mixed PCa cells were larger than those injected with PCa cells alone after 4 weeks. Expression of epithelial-mesenchymal transition markers and cyclin D1 in the prostate tissue injected with T. vaginalis mixed PCa cells increased than those of PCa cells alone. Collectively, it was suggested that inflammatory reactions by T. vaginalis-stimulated PCa cells increase the proliferation and invasion of PCa cells through cytokine-cytokine receptor signaling pathways.

Dynamic Cardiac Magnetic Resonance Fingerprinting During Vasoactive Breathing Maneuvers: First Results

  • Luuk H.G.A. Hopman;Elizabeth Hillier;Yuchi Liu;Jesse Hamilton;Kady Fischer;Nicole Seiberlich;Matthias G. Friedrich
    • Journal of Cardiovascular Imaging
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    • 제31권2호
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    • pp.71-82
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    • 2023
  • BACKGROUND: Cardiac magnetic resonance fingerprinting (cMRF) enables simultaneous mapping of myocardial T1 and T2 with very short acquisition times. Breathing maneuvers have been utilized as a vasoactive stress test to dynamically characterize myocardial tissue in vivo. We tested the feasibility of sequential, rapid cMRF acquisitions during breathing maneuvers to quantify myocardial T1 and T2 changes. METHODS: We measured T1 and T2 values using conventional T1 and T2-mapping techniques (modified look locker inversion [MOLLI] and T2-prepared balanced-steady state free precession), and a 15 heartbeat (15-hb) and rapid 5-hb cMRF sequence in a phantom and in 9 healthy volunteers. The cMRF5-hb sequence was also used to dynamically assess T1 and T2 changes over the course of a vasoactive combined breathing maneuver. RESULTS: In healthy volunteers, the mean myocardial T1 of the different mapping methodologies were: MOLLI 1,224 ± 81 ms, cMRF15-hb 1,359 ± 97 ms, and cMRF5-hb 1,357 ± 76 ms. The mean myocardial T2 measured with the conventional mapping technique was 41.7 ± 6.7 ms, while for cMRF15-hb 29.6 ± 5.8 ms and cMRF5-hb 30.5 ± 5.8 ms. T2 was reduced with vasoconstriction (post-hyperventilation compared to a baseline resting state) (30.15 ± 1.53 ms vs. 27.99 ± 2.07 ms, p = 0.02), while T1 did not change with hyperventilation. During the vasodilatory breath-hold, no significant change of myocardial T1 and T2 was observed. CONCLUSIONS: cMRF5-hb enables simultaneous mapping of myocardial T1 and T2, and may be used to track dynamic changes of myocardial T1 and T2 during vasoactive combined breathing maneuvers.

백합 경단 및 인편배양으로부터 유식물체 분화 및 자구형성에 미치는 생장조절제의 영향 (Effects of Growth Regulators on Shoot Differentiation and Bulblet Formation in Shoot-Tip and Bulb-Scale Cultures of Lilium longiflorum)

  • 이은모;정해준;민병훈;이영복
    • 식물조직배양학회지
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    • 제22권2호
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    • pp.83-87
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    • 1995
  • 백합 경단 및 인편배양에 있어서 유식물체분화 및 자구형성에 미치는 생장조절제의 효과를 구명하기 위하여 실험을 수행하였다. 백합 경단배양 시 유식물체분화는 NAA 0.1 mg/L 또는 NAA 0.1 mg/L + BA 0.1 mg/L 복합처리구에서 비교적 양호하였으나, 뿌리분화는 생장조절제가 무첨가구가 양호하였다. 인편 조직절편 배양 시 생장조절제 첨가없이도 유식물체분화는 가능하였으나, NAA 0.2 mg/L 처리구에서 가장 양호하였며, BA 단독처리구에서는 양호하지 못하였다. NAA대신 IBA를 처리할 경우에는 분화된 유식물체의 자구 형성을 촉진시켰고, 특히 IBA 0.1 mg/L에서 효과적이었다. NAA 0.2 mg/L를 처리할 때 sucrose 3% 첨가보다 6% 첨가가 자구형성을 촉진시켰다.

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PLGA 종류와 담체의 형성 방법에 따른 인간의 조직공학적 연골형성 (Tissue Engineered Cartilage Formation on Various PLGA Scaffolds)

  • 김유미;임종옥;정호윤;박태인;백운이
    • 대한의용생체공학회:의공학회지
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    • 제23권2호
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    • pp.147-153
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    • 2002
  • 본 연구에서는 관상의 Poly(lactic-co-glycolic acid) (PLGA) 담체에 대한 인간의 초자연골과 탄성연골의 형성정도를 살펴보았다. 담체는 PLGA의 분자량에 따라서는 110,000 g/mol과 220,000 g/mol을 비교하였고 내경 유지를 위하여 내경측에는 220,000 g/mol, 외경측에는 110,000 g/mol 의 복합체를 만들거나, 비분해성 고분자 폴리에틸렌 튜브와 110,000 g/mol PLGA의 담체와의 결합도 시도하였다. PLGA 담체들은 주사전자현미경으로 단면 구조를 관찰하였다. 각각의 담체에 20세 미만의 환자들의 비중격에서 채취된 초자연골과 귀에서 채취된 탄성연골에서 분리한 연골세포를 심었다. 분리된 연골세포는 두 번의 계대배양을 거쳐 각각의 PLGA 담체에 심었고 일주일동안 생체 외 환경에서 배양하였다. 각각의 세포와 담체의 복합체를 nude mouse의 배부 좌, 우로 피하조직에 이식하고 8주 뒤 H&E 염색으로 조직 검사를 시행하였다. 110,000 g/mol의 PLGA담체의 연골조직은 잘 형성되어 있었지만 그 내경은 유지되지 못하였다. 반면 220,000 g/mol의 PLGA담체의 연골조직은 내경은 유지하였으나 연골조직이 부분적으로 형성되어 있고 성숙한 연골조직의 양이 많지 않았다. 초자연골 세포에 비교하여 탄성연골 세포가 같은 조건하에서 연골조직을 더 많이 형성한 것으로 나타났다. 관상의 유지를 위하여 220,000 9/mol PLGA 담체를 내경측에 110,000 g/mol PLGA 담체를 외경측으로 한 담체에서는 연골조직 형성이 잘 되지 않았으나 내경측에 폴리에틸렌 튜브를 끼운 110,000 g/mol PLGA 담체에서는 조직 형성과 내경유지가 잘 되었고 원래의 담체와 거의 유사한 형태로 유지되었다. 분화된 연골세포도 조직 소견으로 확인할 수 있었다. 이 1mm 내경의 관상 연골조직은 인공 기관지나 식도 등을 위한 동물 실험과 인공 합성 튜브의 대체 등 앞으로 많은 응용분야가 기대된다.

콘택트렌즈용 다목적용액(MPS)의 세포증식저해 및 실험용 토끼 각막에 미치는 영향 (The Cytotoxicity and Effect on the Experimental Rabbit Cornea of Soft Contact Lens Multi-Purpose Solution (MPS))

  • 고은경;채수철;박수경;김덕송;이종빈
    • Applied Microscopy
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    • 제36권3호
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    • pp.141-153
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    • 2006
  • 본 연구는 시중에서 유통되고 있는 콘택트렌즈 관리용액인 다목적용액 (multi-purpose solution, MPS)이 세포에 미치는 증식저해정도와 실험용 토끼눈의 각막 상피 및 내피조직에 미치는 손상정도를 비교 관찰하고자 시행하였다. MPS는 $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea)를 사용하였다. 세포증식 저해율은 L929세포주를 배양 후 MTT assay로 검정하였고, 형태학적으로는 광학 현미경과 Hematoxylin & Eosin staining 표본을 제작하여 관찰하였다. In vivo 실험은 백색 가토 9마리 (18안)를 3군으로 분류하여 실험군인 좌안(9안)에는 각 MPS제품을, 대조군인 우안(9안)에는 보존제가 포함되지 않은 멸균생리식염수를 점안하였다. 일정기간 점안 후, 실험용 토끼눈의 각막표면을 Rose bengal로 염색하여 관찰하였고 각막상피 및 내피조직의 변화는 주사전자현미경 (scanning electron microscopy, SEM)으로 관찰하였다. 세포증식 저해율은 각각 54, 73, 36%로 나타났으며, 형태학적 변화는 L929세포주의 일반적인 형태와 달리 타원형 또는 원형으로 위축되었다. Rose bengal 염색 결과, 각 실험군은 대조군과 달리 전반적으로 붉게 염색되었고 주사전자현미경 (SEM)관찰 결과, 각막상피는 다각형 모자이크(polygonal mosaic)형태의 균일도가 감소하였고, 각막내피 또한 형태가 불규칙하였으며 섬유아세포만 국소적으로 관찰되었다. 따라서 MPS의 항미생물의 효과와 더불어 생체세포의 안전성면에서 고려해 볼 때 합성보존제의 농도개선과 세포독성이 없는 새로운 보존제 개발의 지속적인 연구가 필요할 것으로 판단된다.