• Title/Summary/Keyword: cell morphology

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Novel Isoquinolinamine and Isoindoloquinazolinone Compounds Exhibit Antiproliferative Activity in Acute Lymphoblastic Leukemia Cells

  • Roolf, Catrin;Saleweski, Jan-Niklas;Stein, Arno;Richter, Anna;Maletzki, Claudia;Sekora, Anett;Escobar, Hugo Murua;Wu, Xiao-Feng;Beller, Matthias;Junghanss, Christian
    • Biomolecules & Therapeutics
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    • v.27 no.5
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    • pp.492-501
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    • 2019
  • Nitrogen-containing heterocycles such as quinoline, quinazolinones and indole are scaffolds of natural products and have broad biological effects. During the last years those structures have been intensively synthesized and modified to yield new synthetic molecules that can specifically inhibit the activity of dysregulated protein kinases in cancer cells. Herein, a series of newly synthesized isoquinolinamine (FX-1 to 8) and isoindoloquinazolinone (FX-9, FX-42, FX-43) compounds were evaluated in regards to their anti-leukemic potential on human B- and T- acute lymphoblastic leukemia (ALL) cells. Several biological effects were observed. B-ALL cells (SEM, RS4;11) were more sensitive against isoquinolinamine compounds than T-ALL cells (Jurkat, CEM). In SEM cells, metabolic activity decreased with $10{\mu}M$ up to 26.7% (FX-3), 25.2% (FX-7) and 14.5% (FX-8). The 3-(p-Tolyl) isoquinolin-1-amine FX-9 was the most effective agent against B- and T-ALL cells with IC50 values ranging from 0.54 to $1.94{\mu}M$. None of the tested compounds displayed hemolysis on erythrocytes or cytotoxicity against healthy leukocytes. Anti-proliferative effect of FX-9 was associated with changes in cell morphology and apoptosis induction. Further, influence of FX-9 on PI3K/AKT, MAPK and JAK/STAT signaling was detected but was heterogeneous. Functional inhibition testing of 58 kinases revealed no specific inhibitory activity among cancer-related kinases. In conclusion, FX-9 displays significant antileukemic activity in B- and T-ALL cells and should be further evaluated in regards to the mechanisms of action. Further compounds of the current series might serve as templates for the design of new compounds and as basic structures for modification approaches.

Understanding of Intrauterine Environment Changes based on Proteomics and Bioinformatics during Estrous Cycle (단백체학과 생물정보학을 이용한 자궁 내 환경의 이해)

  • Lee, Sang-Hee;Lee, Seunghyung
    • Journal of Life Science
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    • v.29 no.5
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    • pp.621-630
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    • 2019
  • Fertilization is the beginning of a new life that occurs in the female uterine. The female reproductive tract is composed ovary, oviduct, uterine, vagina and cervix, their physiological features are regulated by estrous cycle. Of these, uterine is a main point to establish embryo development and implantation, and intercommunication between embryo and uterine environment is necessary for suitable pregnancy. Endometrium is part of the uterine, its morphology is repetitively changed by hormones, and characteristic of uterine fluid from endometrium is also changed. Recently, massive proteins of endometrium and uterine fluid can be detected according to develop proteomics and bioinformatics and have been accelerated the understanding of the reproductive biology fields. Moreover, the massive protein information is actively studying with deeply studied theory such as sex hormone signal pathway and angiogenesis in mammals. In this paper, we review understanding of endometrium remodeling, uterine gland and fluid during estrous cycle, additionally studies on endometrium and uterine fluid based on proteomics techniques. Lastly, we introduced methods of the protein-protein correlation using bioinformatics tool that interaction with hormone receptors, representative angiogenetic factors and detected proteins using proteomics in endometrium and uterine fluid. This review will be useful to understanding the study on search of new cell mechanism in endometrium and uterine fluid.

The antioxidant icariin protects porcine oocytes from age-related damage in vitro

  • Yoon, Jae-Wook;Lee, Seung-Eun;Park, Yun-Gwi;Kim, Won-Jae;Park, Hyo-Jin;Park, Chan-Oh;Kim, So-Hee;Oh, Seung-Hwan;Lee, Do-Geon;Pyeon, Da-Bin;Kim, Eun-Young;Park, Se-Pill
    • Animal Bioscience
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    • v.34 no.4
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    • pp.546-557
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    • 2021
  • Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

Practical Approach for Quantitative and Qualitative Analyses of Marine Ciliate Plankton (해양 섬모충플랑크톤 정량과 정성분석의 현실적 접근)

  • KIM, YOUNG OK;KIM, SUN YOUNG;CHOI, JUNGMIN;KIM, JAESEONG
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.26 no.3
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    • pp.248-262
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    • 2021
  • Marine planktonic ciliates include two major groups, loricated tintinnids and naked oligotrichs. The study of marine ciliate plankton in Korea began with taxonomic efforts on tintinnids based on the morphology of lorica, a vase-shaped shell. Despite polymorphism in the lorica, it is utilized as a key characteristic in identification of tintinnid species. However, oligotrichs have been studied only recently in Korea due to challenges associated with the observation of ciliary arrangements and the technical development for cell staining. Species diversity and phylogenetic classification of the ciliates have been informed by recent advances in morphological and molecular analyses. Illustrations of the planktonic ciliate in Korea have been published on the basis of taxonomic data of tintinnids and oligotrichs. Planktonic ciliates acting as the major consumers of pico- and nanoplankton as well as the prey of mesozooplankton, has been monitored by spatial and temporal investigations in Korean coastal waters. A practical approach addressing the limitations and potential of marine ciliate studies in Korea is proposed here to improve the data quality of planktonic ciliates, providing an enhanced basis for quality control of ciliate monitoring.

Direct-fed Enterococcus faecium plus bacteriophages as substitutes for pharmacological zinc oxide in weanling pigs: effects on diarrheal score and growth

  • Oh, Sang-Hyon;Jang, Jae-Cheol;Lee, Chul Young;Han, Jeong Hee;Park, Byung-Chul
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1752-1759
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    • 2022
  • Objective: Effects of direct-fed Enterococcus faecium plus bacteriophages (EF-BP) were investigated as potential substitutes for pharmacological ZnO for weanling pigs. Methods: Dietary treatments were supplementations to a basal diet with none (NC), 3,000-ppm ZnO (PC), 1×1010 colony-forming units of E. faecium plus 1×108 plaque-forming units (PFU) of anti-Salmonella typhimurium bacteriophages (ST) or 1×106 PFU of each of anti-enterotoxigenic Escherichia coli K88 (F4)-, K99 (F5)-, and F18-type bacteriophages (EC) per kg diet. In Exp 1, twenty-eight 21-day-old crossbred weanling pigs were individually fed one of the experimental diets for 14 days and euthanized for histological examination on intestinal mucosal morphology. In Exp 2, 128 crossbred weanling pigs aged 24 days were group-fed the same experimental diets in 16 pens of 8 piglets on a farm with a high incidence of post-weaning diarrhea. Results: None of the diarrheal score or fecal consistency score (FCS), average daily gain (ADG), gain: feed ratio, structural variables of the intestinal villus, and goblet cell density, differed between the EF-BP (ST+EC) and NC groups, between EF-BP and PC, or between ST and EC, with the exception of greater gain: feed for EF-BP than for PC (p<0.05) during days 7 to 14 (Exp 1). In Exp 2, ADG was less for EF-BP vs PC during days 0 to 7 and greater for EF-BP vs NC during days 7 to 14. FCS peaked on day 7 and declined by day 14. Moreover, FCS was less for EF-BP vs NC, did not differ between EF-BP and PC, and tended to be greater for ST vs EC (p = 0.099). Collectively, EF-BP was comparable to or slightly less effective than PC in alleviating diarrhea and growth check of the weanling pigs, with ST almost as effective as PC, when they were group-fed. Conclusion: The E. faecium-bacteriophage recipe, especially E. faecium-anti-S. typhimurium, is promising as a potential substitute for pharmacological ZnO.

Lectin histochemistry of the olfactory mucosa of Korean native cattle, Bos taurus coreanae

  • Sungwoong, Jang;Bohye, Kim;Jeongmin, Lee;Sohi, Kang;Joong-Sun, Kim;Jong-Choon, Kim;Sung-Ho, Kim;Taekyun, Shin;Changjong, Moon
    • Journal of Veterinary Science
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    • v.23 no.6
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    • pp.88.01-88.14
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    • 2022
  • Background: The olfactory mucosa (OM) is crucial for odorant perception in the main olfactory system. The terminal carbohydrates of glycoconjugates influence chemoreception in the olfactory epithelium (OE). Objectives: The histological characteristics and glycoconjugate composition of the OM of Korean native cattle (Hanwoo, Bos taurus coreae) were examined to characterize their morphology and possible functions during postnatal development. Methods: The OM of neonate and adult Korean native cattle was evaluated using histological, immunohistochemical, and lectin histochemical methods. Results: Histologically, the OM in both neonates and adults consists of the olfactory epithelium and the lamina propria. Additionally, using periodic acid Schiff and Alcian blue (pH 2.5), the mucus specificity of the Bowman's gland duct and acini in the lamina propria was determined. Immunohistochemistry demonstrated that mature and immature olfactory sensory neurons of OEs express the olfactory marker protein and growth associated protein-43, respectively. Lectin histochemistry indicated that numerous glycoconjugates, including as N-acetylglucosamine, mannose, galactose, N-acetylgalactosamine, complex type N-glycan, and fucose groups, were expressed at varied levels in the different cell types in the OMs of neonates and adults at varying levels. According to our observations, the cattle possessed a well-developed olfactory system, and the expression patterns of glycoconjugates in neonatal and adult OMs varied considerably. Conclusions: This is the first study to describe the morphological assessment of the OM of Korean native cattle with a focus on lectin histochemistry. The findings suggest that glycoconjugates may play a role in olfactory chemoreception, and that their labeling properties may be closely related to OM development and maturity.

Effect of Stewartia koreana Nakai Branch Extracts on the Biofilm Formation of Porphyromonas gingivalis (노각나무 가지 추출물이 Porphyromonas gingivalis의 바이오필름 형성에 미치는 영향)

  • Park, Min Jeong;Kim, Hye Soo;Kim, Han Bi;Park, JunHo;Yu, Chan Yeol;Cho, Soo Jeong
    • Journal of Life Science
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    • v.32 no.1
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    • pp.56-62
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    • 2022
  • This study was investigated to evaluate the potential of Stewartia koreana Nakai as an oral healthcare material. The inhibitory effects of extracts on the biofilm formation and fimbriae genes expression of Porphyromonas gingivalis were determined by minimal inhibitory concentrations (MIC), biofilm biomass staining, SEM, and qRT-PCR analysis. The S. koreana Nakai branch was extracted into 70% ethanol, and bacteriostatic MIC of extracts against P. gingivalis were 0.6 mg/ml. In P. gingivalis cultures treated with 0.2-2.0 mg/ml of extract, biofilm production rate was significantly decreased in a concentration-dependent manner. The morphology of treated and untreated samples was observed by SEM, and cell aggregation and biofilm were only observed in those treated with extract. Subsequently, qRT-PCR analysis showed that the mRNA expression on fimbriae genes fimA and fimB was suppressed in a concentration-dependent manner. Based on these results, it can be suggested that S. koreana branch extract has the potential to be used as naturally derived oral healthcare material because of its bacteriostatic action and inhibition of P. gingivalis biofilm formation.

Induction of Apoptosis by β-Lapachone in Hep3B Human Hepatocellular Carcinoma Cells Is Caspase-Dependent and Associated with Inactivation of PI3K/Akt Signaling (Hep3B 인간 간암세포에서 caspase 의존적이며 PI3K/Akt 신호전달의 불활성화와 관련된 β-lapachone의 세포사멸 유도)

  • Jae Im Kwon;Yung Hyun Choi;Hyun Hwangbo
    • Journal of Life Science
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    • v.34 no.2
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    • pp.94-104
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    • 2024
  • β-Lapachone is a natural quinone compound originally obtained from the bark of the lapacho tree (Tabebuia vellanedae), which has been used in traditional medicine in several South and Central American countries for treating various diseases. Although β-lapachone has been reported to have potent anticancer activity in many types of cancer cells, its effect on the proliferation of hepatocellular carcinoma (HCC) cells is still unclear. Therefore, in this study, we investigated the effect of β-lapachone on the proliferation of human HCC Hep3B cells. According to our results, the decrease in cell viability of Hep3B cells caused by β-lapachone was closely related to the induction of apoptosis, which was confirmed through changes in nuclear morphology and flow cytometry. In addition, in Hep3B cells treated with β-lapachone, the expression of Bcl-2, an anti-apoptotic factor, was decreased, while the expression of Bax, an apoptosis inducer, was increased, and the activity of the caspase cascade was also increased. However, in the presence of a pan-caspase inhibitor, β-lapachone-induced apoptosis was weakened, indicating that the induction of apoptosis by β-lapachone was caspase-dependent. Moreover, β-lapachone treatment activated extracellular-regulated kinase (ERK) signaling while inhibiting activation of the phosphoinositide 3 kinase (PI3K)/Akt pathway. Furthermore, the effect of the ERK inhibitor on suppressing the induction of apoptosis by β-lapachone was minimal, and the PI3K inhibitor significantly increased β-lapachone-induced apoptosis. The findings from this study will contribute to a better understanding of the anticancer activity of β-lapachone in HCC cells.

A STUDY ON THE EXTRACELLULAR MATRIX IN THE ARTIFICIALLY CHEATED CLEFT LIP WOUND HEALING OF RABBIT FETUSES (토끼 태자에 형성시킨 구순열상의 치유과정에서 세포외기질 분포에 관한 연구)

  • Yang, Won-Sik;Baek, Seung-Hak
    • The korean journal of orthodontics
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    • v.28 no.1 s.66
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    • pp.1-15
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    • 1998
  • Adult wound healing is accompanied with inflammation and eventual scar formation, whereas fetal wounds heal rapidly by mesenchymal proliferation without significant inflammatory cell participation and with minimal or no scar formation. The cellular mechanisms underlying these differing forms of wound healing are unknown but the extracellualr matrix through its effects on cell function, may play a key role. Therefore the purpose of this study is to investigate the spatial and temporal deposition of several component of extracellular matrix, which are known to be involved with scar formation, in the artificially created cleft lip wound healing of fetuses. The author had undergone hysterotomy and created cleft lip-like defects on fetuses of New Zealand White Rabbit in mid-third trimester(24 days). Fetuses were divided into the repaired group, the unrepaired group and the sham-operated control group. At 1, 2, 3, 5, 7 days after procedure, fetuses were obtained by Caeserem section. After documenting the viability of fetuses, they were photographed to compare size and facial morphology and sectioned for histological examination by H & E stain and spatial and temporal deposition of collagen typeI, III, IV, V and fibronectit laminin by immunohistochemical method. The findings are summarized as follows 1. There were lack of inflammation in the repaired and the unrepaired group during experimental periods. 2. The reepithelialization of the unrepaired group was slower than that of the repaired group. 3. Collagen I, III, V were found from post-op. third day. There were no difference of distribution in the control, the repaired and the unrepaired group. Collagen types I, III, V were present in all groups with restoration of the normal collagen pattern in the fetus. This implies that lack of scarring in fetal wounds is due to the difference of collagen organization pattern within wound and not simply lack of collagen formation. 4. Collagen IV was slightly increased at post-op. third day and decreased after post-op. fifth day. Eventually there were no differences in the control, the repaired and the unrepaired group. Lminin was found at post-op. fifth day and maintained staining density until post-op. seventh day. There were no differences in the control, the repaired and the unrepaired group. According to staining of laminin and collagen type IV in epithelial basement membrane, formation of epithelial basement membrane was not completed until reepithelialization was finished. 5. According to staining of laminin and collagen type IV, there were no increase of neovascularity in the repaired and the unrepaired group. 6. Fibronectin was increased until post-op. third day at fibrin clot, wound base and margin and decreased after post-op. fifth day. Eventually, there were no differences in the control, the repaired and the unrepaired group. So it implies fibronectin plays a role as provisional matrix for fetal wound healing.

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Differentiation of Dopaminergic and Cholinergic Neurons from Mesenchymal-like Stem Cells Derived from the Adipose Tissue (사람 지방 유래 중간엽 줄기세포의 도파민성 및 콜린성 신경세포분화)

  • Hong, In-Kyung;Jeong, Na-Hee;Kim, Ju-Ran;Do, Byung-Rok;Kim, Hea-Kwon;Kang, Sung-Goo
    • Development and Reproduction
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    • v.12 no.1
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    • pp.31-39
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    • 2008
  • Neural tissue has limited intrinsic capacity of repair after injury, and the identification of alternate sources of neural stem cells has broad clinical potential. We isolated mesechymal-like stem cells from human adipose tissues (AT-MSCs), and studied on transdifferentiation-promoting conditions in neural cells. Dopaminergic and cholinergic neuron induction of AT-MSCs was also studied. Neural differentiation was induced by adding bFGF, EGF, dimethyl sulphoxide (DMSO) and butylated hydroxyanisole(BHA) in N2 Medium and N2 supplement. The immunoreactive cells for $\beta$-tubulin III, a neuron-specific marker, GFAP, an astrocyte marker, or Gal-C, an oligodendrocyte marker, were found. AT-MSCs treated with bFGF, SHH and FGF8 were differentiatied into dopaminergic neurons that were immunopositive for TH antibody. Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor (bFGF), retinoic acid (RA) and sonic hedgehog (Shh). AT-MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including neuro D1, $\beta$-tubulin III, GFAP and nestinwas markedly elevated during this acute differentiation. While the stem cell markers such as SCF, C-kit, and Stat-3 were not expressed after preinduction medium culture, we confirmed the differentiation of dopaminergic and cholinergic neurons by TH/$\beta$-tubulin III or ChAT/ $\beta$-tubulin III positive cells. Conclusively, AT-MSCs can be differentiated into dopaminergic and cholinergic neuronsand these findings suggest that AT-MSCs are alternative cell source of treatment for neurodegenerative diseases.

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