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Effects of Sea Tangle (Laminaria japonica) Extract and Fucoidan Drinks on Oxygen Radicals and Their Scavenger Enzymes in Stressed Mouse (스트레스 부하 마우스의 활성산소 및 제거효소에 미치는 다시마(Laminaria japonica)와 후코이단 음료의 영향)

  • CHOI Jin-Ho;KIM Dae-Ik;PARK Soo-Hyun;KIM Dong-Woo;KIM Chang Mok;KOO Jae Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.6
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    • pp.764-769
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    • 1999
  • This study was designed to investigate the effects of sea tangle (Laminaria japonica) extract (Dasi-Ex group: dry base $4.0\%$) and fucoidan-added (Fuco-I, II, III group: fucoidan of $1,0\%,\;2.0\%,\;3.0\%$ added to Dasi-Ex) drinks on the formation of oxygen radicals and scavenger enzyme activities of stressed mice. ICR male mice (20 $\pm$2 g) were fed experimental diets and these drinks instead of water for 18 days including 4 days of sociopsychological stress. Dasi-Ex and Fuco-I, II and III groups resulted in a marked decreases $20\~40\%$ in basal oxygen radical (BOR) formation, and $15\~25\%$ in induced oxygen radical (IOR) formation compared with control group. Hydroxyl radical formations were significantly inhibited about $10\%$ in Dasi-Ex group, while remarkably inhibited $30\~40\%$ in Fuco-I, II and III groups. lipid peroxide (ISO) levels in Dasi-Ex group were not significantly different from those of control group, tut Fuco-I, II and III groups resulted in a significant decreases about $10\%$ in LPO levels compared with control group, Dasi-Ex, Fuco-I, II and III groups resulted in a marked decreases ($31\%,\;36\%,\;39\%$ and $42\%$, respectively) in oxidized protein levels through production of carbonyl group. Significant differences in nitric oxide (NO) levels in Dasi-Ex group were not obtained, but NO levels were slightly inhibited about $7\%$ in Fuco-I and II groups and $20\%$ in Fuco-III group compared with control group. Significant differences in superoxide dismutase (SOD) and catalase (CAT) activities in Dasi-Ex and Fuco-I groups were not obtained, but Fuco-II and III groups resulted in a significant increases $25\~40\%$ in SOD activities, and about $10\%$ in CAT activities compared with control group. These results suggest that the sociopsychological stress and aging process could be effectively inhibited by biological activity of sea tangle and fucoidan components.

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

  • 김유미;임종옥;정호윤;박태인;백운이
    • Journal of Biomedical Engineering Research
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    • v.23 no.2
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    • pp.147-153
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    • 2002
  • The purpose of this study was to evacuate the effect of different types of Poly(lactic-co-glycolic acid) (PLGA) scaffolds on the formation of human auricular and septal cartilages. All of the scaffolds were formed in a tubular shape for potential application for artificial trachea or esophagus with either 110,000 g/mol PLGA. 220,000 g/mol PLGA. or a combination of both. In order to maintain the tubular shape in vivo, two methods were used. One method was inserting polyethylene tube at the center of scaffolds made of 110,000 g/mol PLGA. The other method involved combination of the two different molecular weight PLGA's. The inner surface of tubular shaped scaffold made with 110,000 g/mol PLGA was coated with 220,000 9/mol PLGA to give more mechanical rigidity. Elastic cartilage was taken from the ear of a patient aged under 20 nears old and hyaline cartilage was taken from the nasal septum. The chondrocytes were then isolated. After second passage, the chondrocytes were seeded on the PLGA scaffolds followed by in vitro culture for one week. The cells-PLGA scaffold complex were implanted subcutaneously on the back of nude mice for 8 weeks. The tissue engineered cartilages were separated from nude mice and examined histologically after staining with the Hematoxylin Eosin. The morphology of the scaffolds were examined by scanning electron microscopy. The pores were well formed and uniformly distributed in the various PLGA scaffolds. After 8 weeks in vivo culture, cartilage was well formed with 110,000 g/mol PLGA. however lumen had collapsed. In contrast. a minimal amount of neocartilage was formed with 220,000 g/mol PLGA, while the architecture of scaffold and lumen were well preserved. Elastic cartilage formed more neocartilage than hyaline. Hyaline and elastic neocartilage were well formed on 110,000 g/mol PLGA with the polyethylene tube, exhibiting mature chondrocytes and preservation of the tubular shape. It was found that 110,000 g/mol PLGA was more appropriate for cartilage formation but higher molecular weight polymer was necessary to maintain the three dimensional shape of the scaffold.

Different Expressions of HIF-$1\alpha$, Bcl-2 and Baxin DU145 Prostate Cancer Cells Transplanted in Nude Mouse between X-Ray and Neutron Irradiation (누드마우스에 주입된 DU-145 전립샘암에서 엑스선과 중성자선에 의한 HIF-$1\alpha$, Bcl-2, Bax 발현의 차이)

  • Kong, Moon-Kyoo;Kang, Jin-Oh;Kim, Sang-Ki;Shin, Dong-Oh;Park, Seo-Hyun;Kim, Chang-Ju;Chang, Hyun-Kyung
    • Radiation Oncology Journal
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    • v.27 no.4
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    • pp.218-227
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    • 2009
  • Purpose: To investigate the radiobiologic effects of neutron and X-ray irradiation on DU-145 prostate carcinoma cells by identifying the differences of HIF-$1\alpha$ expression and apoptosis. Materials and Methods: Nude mice were injected with the human prostate cancer cell line, DU-145, and then irradiated with 2 Gy and 10 Gy X-rays, or 0.6 Gy and 3.3 Gy neutrons, respectively. The mice were sacrificed at 24 hours and 120 hours after irradiation. The expression levels of HIF-$1\alpha$, Bcl-2 and Bax were compared with immunohistochemical staining and western blotting. The apoptotic indexes were compared with the Terminal deoxynucleotidyl biotin-dUTP nick and labeling (TUNEL) assay. Results: At day 1, HIF-$1\alpha$ and Bcl-2 expression decreased, while Bax expression and the number of TUNEL positive cells increased in neutron irradiated groups for the control and X-ray irradiated groups. The Bcl-2/Bax ratio was significantly lower in the neutron irradiated groups regardless of dose (p=0.001). The same pattern of the differences in the expressions of the HIF-$1\alpha$, Bcl-2, Bax, Bcl-2/Bax ratio, and apoptotic indexes were indentified at day 5. HIF-$1\alpha$ expression was related with Bcl-2 (p=0.031), Bax (p=0.037) expressions and the apoptotic indexes (p=0.016) at day 5. Conclusion: Neutron irradiation showed a decrease in HIF-$1\alpha$, Bcl-2 expression, and Bcl-2/Bax ratio, but increased Bax expression regardless of dose. This study suggests that the differences radiobiological responses between photon and neutron irradiation may be related to different HIF-$1\alpha$ expression and subsequent apoptotic protein expressions.

The Clinical Significance of Cathepsin D and p53 Expression in Locally Advanced Rectal Cancer (국소진행된 직장암에서 Cathepsin D와 p53 발현의 임상적 의의)

  • Kim, Jun-Sang;Lee, Sheng-Jin;Kim, Jin-Man;Cho, Moon-June
    • Radiation Oncology Journal
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    • v.26 no.1
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    • pp.56-64
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    • 2008
  • Purpose: Cathepsin D(CD) is a lysosomal acid proteinase that is related to malignant progression, invasion, and a poor prognosis in several tumors. The aim of this study was to evaluate the prognostic clinical significance of CD and p53 expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer who were treated with preoperative chemoradiation. Materials and Methods: Eighty-nine patients with locally advanced rectal cancer(cT3/T4 or N+) were included in this study. Preoperative chemoradiation consisted of a dose of 50.4 Gy of pelvic radiation and two concurrent cycles of administration of 5-fluorouracil and leucovorin. Surgery was performed six weeks after chemoradiation. CD and p53 expression in pretreatment formalin-fixed paraffin-embedded tumor biopsy specimens were assessed by immunohistochemical staining using a CD and p53 monoclonal antibodies. The threshold value for a positive stain in tumor tissue and stromal cells was 1+ intensity in 10% of the tumors or stromal cells, respectively. Results: Positive CD expression was found in 57(64%) of the tumors and 32(35%) of the stromal cell specimens. There was no association with CD expression of the tumor or stromal cells and patient characteristics. There was a correlation between tumor CD expression with stromal cell CD expression(p=0.01). Overexpression of p53 was not a significant prognostic factor. The 5-year overall survival(OS) and disease-free survival(DFS) rates were not different between tumor CD-negative and positive patient biopsy samples(69% vs. 65%, 60% vs. 61%, respectively). The 5-year OS rates in the tumor-negative/stromal cell-negative, tumor-negative/stromal cell-positive, tumor-positive/stromal cell-negative and tumor-positive/stromal cell-positive biopsy samples were 75%, 28%, 62%, and 73%, respectively. Stromal cell staining only without positive tumor staining demonstrated the worst overall survival prognosis for patients(p=0.013). Conclusion: Overexpression of p53 in rectal biopy tissue was not associated with prognostic significance. In the pretreatment biopsy specimens, an exclusive increase in CD expression in stromal cells without tumor expression was related to poor overall survival in patients with locally advanced rectal cancer treated with preoperative chemoradiation.

$HgCl_2$ Dysregulates the Immune Response of Balb/c Mice (수은에 의한 마우스의 면역반응 조절장애)

  • Ki, No-Suk;Koh, Dai-Ha;Kim, Chong-Suh;Lee, Jung-Sang;Kim, Nam-Song;Lee, Hwang-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.1 s.45
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    • pp.11-24
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    • 1994
  • The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

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Facile Fabrication of Animal-Specific Positioning Molds For Multi-modality Molecular Imaging (다중 분자 영상을 위한 간편한 동물 특이적 자세 고정틀의 제작)

  • Park, Jeong-Chan;Oh, Ji-Eun;Woo, Seung-Tae;Kwak, Won-Jung;Lee, Jeong-Eun;Kim, Kyeong-Min;An, Gwang-Il;Choi, Tae-Hyun;Cheon, Gi-Jeong;Chang, Young-Min;Lee, Sang-Woo;Ahn, Byeong-Cheol;Lee, Jae-Tae;Yoo, Jeong-Soo
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.5
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    • pp.401-409
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    • 2008
  • Purpose: Recently multi-modal imaging system has become widely adopted in molecular imaging. We tried to fabricate animal-specific positioning molds for PET/MR fusion imaging using easily available molding clay and rapid foam. The animal-specific positioning molds provide immobilization and reproducible positioning of small animal. Herein, we have compared fiber-based molding clay with rapid foam in fabricating the molds of experimental animal. Materials and Methods: The round bottomed-acrylic frame, which fitted into microPET gantry, was prepared at first. The experimental mice was anesthetized and placed on the mold for positioning. Rapid foam and fiber-based clay were used to fabricate the mold. In case of both rapid foam and the clay, the experimental animal needs to be pushed down smoothly into the mold for positioning. However, after the mouse was removed, the fabricated clay needed to be dried completely at $60^{\circ}C$ in oven overnight for hardening. Four sealed pipet tips containing $[^{18}F]FDG$ solution were used as fiduciary markers. After injection of $[^{18}F]FDG$ via tail vein, microPET scanning was performed. Successively, MRI scanning was followed in the same animal. Results: Animal-specific positioning molds were fabricated using rapid foam and fiber-based molding clay for multimodality imaging. Functional and anatomical images were obtained with microPET and MRI, respectively. The fused PET/MR images were obtained using freely available AMIDE program. Conclusion: Animal-specific molds were successfully prepared using easily available rapid foam, molding clay and disposable pipet tips. Thanks to animal-specific molds, fusion images of PET and MR were co-registered with negligible misalignment.

Effects of Scutellaria radix Extract on Osteoblast Differentiation and Osteoclast Formation (황금 추출물이 조골세포와 파골세포의 활성에 미치는 영향)

  • Shin, Jeong-Min;Park, Chan-Kyung;Shin, Eun-Ju;Jo, Tae-Hyung;Hwang, In-Kyeong
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.674-679
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    • 2008
  • Scutellaria radix (SR) has been utilized as a traditional medicine for a variety of diseases including Rheumatoid arthritis and its major flavonoids - baicalein, baicalin, and wogonin - have been reported to exert beneficial health effects, including anti-bacterial, anti-viral, anti-inflammatory, and free-radical scavenging. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of SR on osteoblast and osteoclast cells. SR extract was prepared using 70% ethanol solvent. Osteoblastic MC3T3-E1 cells and osteoclast precursor Raw 264.7 macrophage cells were utilized. SR extract increased MC3T3-E1 cell proliferation and stimulated alkaline phosphatase activity dose-dependently, 152.0% of the control at concentration $1{\mu}g/mL$. Additionally, SR extract ($1{\mu}g/mL$) stimulated Bone nodule formation activity in MC3T3-E1 cells, approximately 223.3% of the control, 20 days after the exposure. In addition, SR extract significantly reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from Raw 264.7 cells. In conclusion, SR extract stimulates the proliferation and bioactivities of boneforming osteoblasts, and inhibits the activities of bone-resorbing osteoclasts to a certain degree.

Isolation and Characterization of vasa Gene of Triploid and Diploid Human Lung Flukes (Paragonimus westermani) (폐흡충의 이배체와 삼배체 vasa 유전자 분석 및 특징)

  • Lee, Keun-Hee;Yu, Hak-Sun;Hur, Jae-Won;Yu, Sung-Suk;Choi, Sun-Hee;Park, Sang-Kyun;Lee, Sun-Joo;Chung, Dong-Il;Kong, Hyun-Hee;Ock, Mee-Sun;Jeong, Hae-Jin
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.462-469
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    • 2007
  • In this study, we isolated, characterized, and compared the vasa homologous genes of diploid and triploid Paragonimus westermani and localized VASA homologous proteins in both lung fluke types. Open reading frames of Pw-vasa-2n and Pw-vasa-3n were of 1812 bp, and encoded deduced proteins of 622 amino acids with calculated molecular weights of 69.0 kDa and 68.9 kDa and pI's of 9.11 and 9.03, respectively. A comparison of these two VASA deduced protein sequences showed that only 6 of the 622 amino acids differed. The deduced sequences of Pw-VASA-2n and Pw-VASA-3n contained eight consensus sequences characteristic of the DEAD-box protein family and their N-terminal regions contained four arginine-glycine-glycine (RGG) motifs. These two lung fluke VASA-like proteins were more similar to those of other VASA proteins than to those of other DEAD-family proteins isolated from several organisms (planarian, zebra fish, mouse, and human). vasa homologous gene transcription and VASA protein expressions in triploid type lung flukes was slightly stronger than in the diploid type. Immunostaining showed that testes and a portion of the ovaries of both diploid and triploid lung flukes reacted strongly to anti-Pw-VASA antibody.

The Anticancer Effect and Mechanism of Photodynamic Therapy Using 9-Hydroxypheophorbide-a and 660 nm Diode Laser on Human Squamous Carcinoma Cell Line. (9-hydroxypheophorbide-a와 660 nm 다이오드 레이저를 이용한 광역학치료의 항암효과와 치료기전에 대한 연구)

  • Ahn, Jin-Chul
    • Journal of Life Science
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    • v.19 no.6
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    • pp.770-780
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    • 2009
  • A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

Cadms/SynCAMs/Necls/TSLCs Interact with Multi-PDZ Domain Protein MUPP1 (Cadms/SynCAMs/Necls/TSLCs와 multi-PDZ domain protein MUPP1 단백질의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1276-1283
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    • 2014
  • Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.