• Title/Summary/Keyword: in vitro/in vivo studies

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Involvement of a LiCl-Induced Phosphoprotein in Pigmentation of the Embryonic Zebrafish (Danio rerio) (LiCl에 의해 유도되는 phosphoprotein이 embryonic zebrafish (Danio rerio)의 pigmentation에 미치는 영향)

  • Jin, Eun-Jung;Thibaudeau, Giselle
    • Journal of Life Science
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    • v.18 no.9
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    • pp.1219-1224
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    • 2008
  • The embryonic zebrafish (Danio rerio) is rapidly becoming an important model organism for studies of early events in vertebrate development. Neural crest-derived pigment cell precursors of the embryonic zebrafish give rise to melanophores, xanthophores, and/or iridophores. Cell-signaling mechanisms related to the development of pigmentation and pigment pattern formation remain obscure. In this study, zebrafish embryos were treated with various signaling-related molecules - LiCl (an inositol-phosphatase inhibitor), forskolin (a protein kinase-A activator), a combination of LiCl/forskolin, and LiCl/heparin (an IP3 inhibitor) in order to identify the mechanisms involved in pigmentation. LiCl treatment resulted in ultrastructural and morphological alterations of melanophores. To identify the possible proteins responsible for this ultrastructural and morphological change, phosphorylation patterns in vitro and in vivo were analyzed. LiCl and LiCl/forskolin treatment elicited dramatic increases in the phosphorylation of a 55-kDa protein which was inhibited by heparin treatment. LiCl treatment also induced phosphorylation of a 55-kDa protein in melanophores purified from adult zebrafish. Collectively these results suggest that a LiCl-induced 55-kDa phosphoprotein plays a role in melanophore morphology and ultrastructure and ultimately effects gross pigmentation.

Studies on the prevention of tuberculosis in pet dogs 1. The effects of BCG pretreatment in pet dogs inoculated experimentally with Mycobacterium bovis (애완견의 결핵예방에 관한 연구 1. Mycobacterium bovis를 실험적으로 접종한 애완견에 있어서 BCG의 전처치 효과)

  • Kang, Jong-koo;Kim, Chang-ki
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.117-125
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    • 1992
  • Dogs were divided into 3 groups of two each; Bacillie Calmette-Guerin(BCG) pretreatment, M bovis only treatment and uninfected control group. BCG were vaccinated intradermally with 0.2ml before 3weeks of M bovis intraperitoneal infection. Infection at necropsy 4months later was readily in the both treated dogs. Histopathologically, the BCG pretreated dogs produce the moderate accumulation of macrophages and focal granuloma formation in the lung, whereas the M bovis only treared dogs produce the accumulation of predominantly macrophages, occasionaly polymorphonuclear cells and the more larger granuloma Bronchoalveolar lavage(BAL) was obtained and total and differential cell counts were examined. Total number of BAL cells harvested from uninfected dogs is lower compared with those of the both treated groups. The total cell number of M bovis only treated dogs were singificantly higher 1.8 times than that of the BCG pretreated dogs. The Fe receptor activity and the growth of organism in alveolar macrophages obtained from BCG pretreated dogs were compared with that in macrophages from M bovis only treated dogs. BCG vaccination resulted in substantial macrophage activation, measured as increased Fc receptor mediated phagocytosis and rosette formation, as wells as the inhibition of intracellular mycobacteria multiplication. However, actibated macrophages taken from BCG pretreated dogs are incapable of killing the M bovis. Thus, these results suggest that BCG pretrearment in the dog may produce a protective effect against tuberculosis because active alveolar macrophages have acquired antituberculous immunity, although few mycobacteria within the lung remain in a metabolically active state.

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Function of 27-Hydroxycholesterol in Various Tissues and Diseases (다양한 조직 및 질병에서 27-하이드록시콜레스테롤의 역할 및 기전 고찰)

  • Shim, Wan-Seog;Lee, Chanhee;Azamov, Bakhovuddin;Kim, Koanhoi;Lee, Dongjun;Song, Parkyong
    • Journal of Life Science
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    • v.32 no.3
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    • pp.256-262
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    • 2022
  • Oxysterols are oxygenated metabolites of cholesterol generated by serial enzymatic reactions during bile acid synthesis. Similar to cholesterol, oxysterols move rapidly to the intracellular region and modulate various cellular processes, such as immune cell responses, lipid metabolism, and cholesterol homeostasis. Different nuclear transcription factors, such as glucocorticoid, estrogen, and liver X receptors, can be modulated by oxysterols in multiple tissues. The most abundant oxysterol, 27-hydroxycholesterol (27-OHC), is a well-known selective modulator that can either activate or suppress estrogen receptor activity in a tissue-specific manner. The contribution of 27-OHC in atherosclerosis development is apparent because a large amount of it is found in atherosclerotic plaques, accelerating the transformation of macrophages into foam cells that uptake extracellular modified lipids. According to previous studies, however, there are opposing opinions about how 27-OHC affects lipid and cholesterol metabolism in metabolic organs, including the liver and adipose tissue. In particular, the effects of 27-OHC on lipid metabolism are entirely different between in vitro and in vivo conditions, suggesting that understanding the physiology of this oxysterol requires a sophisticated approach. This review summarizes the potential effects of 27-OHC in atherosclerosis and metabolic syndromes with a special discussion of its role in metabolic tissues.

Antitumor Activity of CBT-AK5 Purified from Lactobacillus casei against Sarcoma-180 Infected ICR Mice (유산균의 배양물로부터 분리된 물질에 대한 암 유발 ICR Mice의 항암 효과)

  • Yeo, Moon-Hwan;Kim, Dong-Myung;Kim, Yong-Hwan;Kim, Jung-Hwa;Baek, Hong;Chung, Myung-June
    • Journal of Dairy Science and Biotechnology
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    • v.26 no.2
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    • pp.23-30
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    • 2008
  • This study implicated that the CBT-AK5 purified from Lactobacillus casei (LAFTI L26) which showed antitumor activity in ICR mice. Hence, ICR mice were inoculated intraperitoneally Sarcoma-180 as well as CBT-AK5. Then we observed the life span and tumor increment of those ICR mice. Here our studies showed effect on two different way of treatment as intraperitoneally and orally treated in Sarcoma-180 infected ICR mice. We found that intraperitoneally treatment of Sarcoma-180 and CBT-AK5 is more effective than orally fed. The life span of the ICR mice were highly reduced after the inoculation of Sarcoma-180. Those effects like increment of body weight, the growth of ascites and solid were inhibited significantly after the treatment of CBT-AK5 in Sarcoma-180 infected ICR mice. Finally these studies suggested that CBT-AK5 isolated form Lactobacillus casei showed excellent antitumor activity against Sarcoma-180 infected ICR mice.

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Odorant Receptors Containing Conserved Amino Acid Sequences in Transmembrane Domain 7 Display Distinct Expression Patterns in Mammalian Tissues

  • Ryu, Sang Eun;Shim, Tammy;Yi, Ju-Yeon;Kim, So Yeun;Park, Sun Hwa;Kim, Sung Won;Ronnett, Gabriele V.;Moon, Cheil
    • Molecules and Cells
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    • v.40 no.12
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    • pp.954-965
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    • 2017
  • Mammalian genomes are well established, and highly conserved regions within odorant receptors that are unique from other G-protein coupled receptors have been identified. Numerous functional studies have focused on specific conserved amino acids motifs; however, not all conserved motifs have been sufficiently characterized. Here, we identified a highly conserved 18 amino acid sequence motif within transmembrane domain seven (CAS-TM7) which was identified by aligning odorant receptor sequences. Next, we investigated the expression pattern and distribution of this conserved amino acid motif among a broad range of odorant receptors. To examine the localization of odorant receptor proteins, we used a sequence-specific peptide antibody against CAS-TM7 which is specific to odorant receptors across species. The specificity of this peptide antibody in recognizing odorant receptors has been confirmed in a heterologous in vitro system and a rat-based in vivo system. The CAS-TM7 odorant receptors localized with distinct patterns at each region of the olfactory epithelium; septum, endoturbinate and ectoturbinate. To our great interests, we found that the CAS-TM7 odorant receptors are primarily localized to the dorsal region of the olfactory bulb, coinciding with olfactory epithelium-based patterns. Also, these odorant receptors were ectopically expressed in the various non-olfactory tissues in an evolutionary constrained manner between human and rats. This study has characterized the expression patterns of odorant receptors containing particular amino acid motif in transmembrane domain 7, and which led to an intriguing possibility that the conserved motif of odorant receptors can play critical roles in other physiological functions as well as olfaction.

The Effects of Simvastatin on Bone Healing in Mandible Fractured Rats. (백서의 하악골 골절 치유에서 Simvastatin이 미치는 영향)

  • Jeong, Jae-Oo;Kwon, Yong-Seok;Kim, Seok-Kwun;Lee, Keun-Cheol
    • Archives of Plastic Surgery
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    • v.36 no.5
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    • pp.525-530
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    • 2009
  • Purpose: The hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) are widely used in the treatment of dyslipidemia for the lowering of cholesterol. And studies about simvastatins have been shown to enhance bone formation in vitro and in vivo in rodents. But some other researchers have reported that there was no anabolic effect abouts simvastatins on bone. The peripheral distribution beyond the liver represents a small fraction of an orally administered dose. We hypothesize that this poor peripheral distribution is the likely reason that simvastatins, yield ambiguous results as anabolic agents. We therefore investigated whether the effects of simvastatins on bone may be enhanced by subcutaneous administration, providing better peripheral delivery of these drugs. Methods: 36 rat unilaterally mandible fractured models were prepared and divided into two groups. The simvastatin treated group where 1 mg/kg of simvastatin was daily injected subcutaneously. The same dose of normal saline was injected on the control group. And 3 rats in each group were sacrificed and taken bone samples in each week. Bone sample was evaluated with tensile strength and histological morphology after 1, 2, 3, 4, 5 and 6 weeks. Results: In simvastatin treated group, the fracture healing process, chondrocyte aggregation, collagen formation and trabecular bone formation was rapidly proceeded than the control group in histologically. The tensile strength of the simvastatin treated group was 1.02, 2.25, 3.95, 4.42, 5.49 and $6.00N/mm^2$ by weeks. The control group data was 0.60, 1.05, 2.17, 3.75, 4.15 and $5.17N/mm^2$ by weeks. The average tensile strength was higher by $1.04N/mm^2$ in simvastatin treated group. Conclusion: The currently available data on the effects of simvastatin on bone has done to confirm the finding that simvastatin helps fracture healing. And the potential for simvastatin to be used as anabolic agents for bone when delivered by the subcutaneous route.

Induction of Apoptosis by Pectenotoxin-2 Isolated from Marine Sponges in U937 Human Leukemic Cells (인체 혈구암세포 U937에서 해양해면동물에서 추출된 Pectenotoxin-2에 의한 Apoptosis의 유발에 관한 연구)

  • Shin, Dong Yeok;Kang, Ho Sung;Bae, Song-Ja;Jung, Jee H.;Choi, Yung Hyun
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.2
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    • pp.63-70
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    • 2006
  • Natural product compounds are the source of numerous therapeutic agents. The marine environment produces natural products from a variety of structural classes exhibiting activity against numerous disease targets including anticancer agents. Among these, pectenotoxin-2 (PTX-2), which was first identified as a cytotoxic entity in marine sponges, which depolymerizes actin filaments, was found to be highly effective and more potent to activate an intrinsic pathway of apoptosis in p53-deficient tumor cells compared to those with functional p53 both in vitro and in vivo. However, the anti-proliferative mechanism of the compound at non-cytotoxic concentrations has not yet been explored. In the current study, we sought to investigate anti-proliferation and apoptosis of PTX-2 against U937 human leukemic cells and its underlying molecular mechanism. Exposure of U937 cells to PTX-2 resulted in growth inhibition and induction of apoptosis in dose- and time-dependent manner as measured by MTT assay, fluorescent microscopy and flow cytometric analysis. The anti-proliferative effect of PTX-2 was associated with a marked increase in the expression of cyclin-dependent kinase p21 (WAF1/CIP1) mRNA which was tumor suppressor p53-independent. The increase in apoptosis was connected with a time-dependent down-regulation of anti-apoptotic Bcl-XL and inhibitor of apoptosis proteins (IAPs) family such as XIAP and cIAP-2. Though additional studies are needed, these findings suggested that PTX-2-induced inhibition of U937 cells was associated with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of PTX-2.

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Possibility of Cancer Treatment by Cellular Differentiation into Adipocytes (지방세포로의 분화를 통한 악성 종양의 치료 가능성)

  • Byeong-Gyun Jeon;Sung-Ho Lee
    • Journal of Life Science
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    • v.33 no.6
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    • pp.512-522
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    • 2023
  • Cancer with unlimited cell growth is a leading cause of death globally. Various cancer treatments, including surgery, chemotherapy, radiation therapy, immunotherapy, and targeted therapy, can be applied alone or in combination depending on the cancer type and stage. New treatments with fewer side effects than previous cancer treatments are continually under development and in demand. Undifferentiated stem cells with unlimited cell growth are gradually changed via cellular differentiation to arrest cell growth. In this study, we reviewed the possibility of treating cancer by using cellular differentiation into the adipocytes in cancer cells. In previous in vitro studies, oral antidiabetic drugs of the thiazolidinedione (TDZ) class, such as rosiglitazone and pioglitazone, were induced into the adipocytes in various cancer cell lines via increased peroxisome proliferator-activated receptor-γ (PPAR γ) expression and glucose uptake, which is the key regulator of adipogenesis and the energy metabolism pathway. The differentiated adipogenic cancer cells treated with TDZ inhibited cell growth and had a less cellulotoxic effect. This adipogenic differentiation treatment suggests a possible chemotherapy option in cancer cells with high and abnormal glucose metabolism levels. However, the effects of the in vivo adipogenic differentiation treatment need to be thoroughly investigated in different types of stem and normal cells with other side effects.

Modulacon of Cell Cycle Control by Histone Deacetylase Inhibitor Trichostatin A in A549 Human Non-small Cell Lung Cancer Cells (인체폐암세포 A549의 세포주기 조절인자에 미치는 histone deacetylase inhibitor trichostatin A의 영향)

  • Hwang Ji Won;Kim Young Min;Hong Su Hyun;Choi Byung Tae;Lee Won Ho;Choi Yung Hyun
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.726-733
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    • 2005
  • Histone deacetylase (HDAC) inhibitors target key steps of tumor development. They inhibit proliferation, induce differentiation and/or apoptotic cell death, and exhibit potent antimetastatic and antiangiogenic properties in cancer cells in vitro and in vivo. Although they are emerging as a promising new treatment strategy in malignancy, how they exert their effect on human non-small cell lung cancer cells is as yet unclear. The present study was undertaken to investiate the underlying mechanism of a HDAC inhibitor trichostatin A (TSA)-induced growth arrest and its effect on the cell cycle control gene products in a human lung carcinoma cell line A549. TSA treaoent induced the growth inhibition and morphological changes in a concentration-dependent manner. Treatment of A549 cells with TSA resulted in a concentration-dependent increased G1 (under 100 ng/ml) and/or G2/M (200 ng/ml) cell population of the cell cycle as determined by flow cytometry Moreover, 200 ng/ml TSA treatment significantly induced the population of sub-G1 cells (23.0 fold of control). This anti-proliferative effect of TSA was accompanied by a marked inhibition of cyclins, positive regulators of cell cycle progression, and cyclin-dependent kinases (Cdks) expression and concomitant induction of tumor suppressor p53 and Cdk inhibitors such as p21 and p27 Although further studies are needed, these findings provide important insights into the possible molecular mechanisms of the anti-cancer activity of TSA in human lung carcinoma cells.

Anti-inflammatory activities of Sagunja-tang(Sijunzi-tang) (사군자탕(四君子湯)의 항염증 효능 연구)

  • Lee, Jin-Ah;Ha, Hye-Kyung;Jung, Da-Young;Lee, Ho-Young;Lee, Nam-Hun;Lee, Jun-Kyoung;Seo, Chang-Seob;Kim, Jung-Hoon;Lee, Mee-Young;Shin, Hyeun-Kyoo
    • Herbal Formula Science
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    • v.18 no.1
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    • pp.87-94
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    • 2010
  • Objectives : To provide the information of efficacy for Sagunja-tang (Sijunzi-tang; SG), it was evaluated the anti-inflammatory effect. SG, a widely used herbal formula in tranditional Korean medicine, has been used to treat for the Boki-invigorating. In many studies, plant-derived anti-inflammatory efficacies have been investigated for their potential inhibitory effects on lipopolysaccharide (LPS)-stimulated macrophages. This study was performed to examine the anti-inflammatory effects of SG extract on LPS-stimulated RAW 264.7 cells. Methods : The productions of nitric oxide (NO), prostaglandin (PG)$E_2$, interleukin (IL)-6 and tumor necrosis factor (TNF)-$\alpha$ were examined in a macrophage cell line, RAW 264.7 cells, in the presence of the SG extract. RAW 264.7 cells were incubated with LPS $1\;{\mu}g/mL$ and SG extract for 18 hours. The anti-inflammatory activity of SG was investigated by carrageenin-induced paw edema in rats. The paw volume was measured at 0, 2 and 4 hours following carrageenin-induced paw edema in rats. Results : SG extract showed inhibitory effect on $PGE_2$, IL-6 and TNF-$\alpha$ by LPS-stimulated RAW 264.7 cells. But SG extract was not inhibitory effect on NO by LPS-stimulated RAW 264.7 cells. And administration of SG extract (1 g/kg) showed a reduction in carrageenin-induced paw edema on rats. Conclusions : These results suggest that SG extract has anti-inflammatory activities in vitro and in vivo models.