• Title/Summary/Keyword: intact cells

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Effect of simulated Acid rain on Foliar Structural of Changes of Ginkgo biloba and Pinus thunbergii (은행나무와 곰솔에 처리된 인공산성비에 의한 잎의 형태변화)

  • 소웅영
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.79-86
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    • 1995
  • Visible injury symptoms such as necrosis, chlorosis and premature senscence in the leaves of Ginkgo bloba and Pinus thunbergii treated with acid rain of pH 3.2 or below were observed. The epicuticular wax erosions were observed by SEM after exposure to acid rain of pH 2.4 and 3.2 in G. biloba and pH 4.0 below in P. thunbergii. The adaxial epidennal cells and sponge parenchyma cells were compressed, and those were distorted in the leaves of G. biloba treated with simulated acid rain of pH 3.2 or below. However, vascular tissue was intact. With increase of acidity, mesophyll cells were smaller than those of control while intercellular space in mesophyU was increased. In P. thunbergii, sponge parenchyma cells and vascular tissue except epidennis were distorted after exposure to acid rain of pH 2.4. The size change of stomata in foliar injury was not observed, but the stomatal index and size of stomatal aperture in leaves treated with acid rain increased. The stomata of injured leaf were opened in both species examined.amined.

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Continuous Fermentationof L-Lysine by Immobilized Corynebacterium glutamicum (Corynebacterium glutamicum 고정화균체에 의한 L-라이신 연속발효)

  • 이인선;조정일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.2
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    • pp.322-327
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    • 1994
  • For the improvement of L-lysine productivity, development of the continuous fermentation system by a bioreactor assembly was attempted. Primarily, optimal conditions on the whole cell immobilization of Corynebacterium glutamicum ATCC21514 were studied and 76.2% of immobilization ratio was obtained when the cells were entrapped with 4% k-carrageenan showing 4.0kg gel strength. A bioreactor system was set up using the immobilized cells was applied for the continuous production of L-lysine. The results obtained under the optimum conditions were compared with those of the batchwise fermentation. Experimental results obtained from 14 day continuous fermentation showed 36.7% of sugar conversion to L-lysine while the productivity of L-lysine was disclosed as 4.96mg/ml mg-dry cell weight /hr which is 2.5times and 4.1 times higher than those of the batchwise fermentation by the intact cells and by the immobilized cells, respectively.

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Electron Microscopic Observations on the tissues infected with Nuclear Polyhedosis Virus of Bombyx mori during eary stage (가잠 핵다각체병 바이러스 초기감염에 관한 전자현미경적 관찰)

  • 유강선;강석권
    • Journal of Sericultural and Entomological Science
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    • v.32 no.1
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    • pp.49-57
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    • 1990
  • To investigate the pathway of Nuclear Polyhedrosis Virus(NPV) of Bombyx mori in early stage infection of 2nd instar larva, the larval tissues were observed under electron microscope at interval of 6, 12, 24, and 48 hours after virus inoculation. The results are as follows. 1. The intact and enveloped nucleocapsids released from the polyhedra protein in the gut lumen apparently entered with the microvilli. 2. Virus progenies were observed in columnar cell nuclei 24 hours after inoculation, but polyhedra was not seen. The enveloped virus was observed in some of the intercellular spaces between mid-gut cells. 3. Many enveloped virus particles appeared in the basement membrane. These enveloped virus particles passed the basement membrane and gathered at blood cells in heomocoeal. 4. The NPV muliplicates in nuclei of the blood cells and the tracheal cells normally.

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An Optimized Method for the Construction of a DNA Methylome from Small Quantities of Tissue or Purified DNA from Arabidopsis Embryo

  • Yoo, Hyunjin;Park, Kyunghyuk;Lee, Jaehoon;Lee, Seunga;Choi, Yeonhee
    • Molecules and Cells
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    • v.44 no.8
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    • pp.602-612
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    • 2021
  • DNA methylation is an important epigenetic mechanism affecting genome structure, gene regulation, and the silencing of transposable elements. Cell- and tissue-specific methylation patterns are critical for differentiation and development in eukaryotes. Dynamic spatiotemporal methylation data in these cells or tissues is, therefore, of great interest. However, the construction of bisulfite sequencing libraries can be challenging if the starting material is limited or the genome size is small, such as in Arabidopsis. Here, we describe detailed methods for the purification of Arabidopsis embryos at all stages, and the construction of comprehensive bisulfite libraries from small quantities of input. We constructed bisulfite libraries by releasing embryos from intact seeds, using a different approach for each developmental stage, and manually picking single-embryo with microcapillaries. From these libraries, reliable Arabidopsis methylome data were collected allowing, on average, 11-fold coverage of the genome using as few as five globular, heart, and torpedo embryos as raw input material without the need for DNA purification step. On the other hand, purified DNA from as few as eight bending torpedo embryos or a single mature embryo is sufficient for library construction when RNase A is treated before DNA extraction. This method can be broadly applied to cells from different tissues or cells from other model organisms. Methylome construction can be achieved using a minimal amount of input material using our method; thereby, it has the potential to increase our understanding of dynamic spatiotemporal methylation patterns in model organisms.

Chemoquiescence with Molecular Targeted Ablation of Cancer Stem Cells in Gastrointestinal Cancers

  • Jong-Min Park;Young-Min Han;Migyeong Jeong;Eun Jin Go;Napapan Kangwan;Woo Sung Kim;Ki Baik Hahm
    • Journal of Digestive Cancer Research
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    • v.4 no.1
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    • pp.1-9
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    • 2016
  • The abundance of multi-drug resistance ATPase binding cassette and deranged self-renewal pathways shown in cancer stem cells (CSCs) played a crucial role in tumorigenesis, tumor resistance, tumor recurrence, and tumor metastasis. Therefore, elucidation of CSCs biology can improve diagnosis, enable targeted treatment, and guide the follow up of GI cancer patients. In order to achieve chemoquiescence, seizing cancer through complete ablation of CSCs, CSCs are rational targets for the design of interventions that will enhance responsiveness to traditional therapeutic strategies and contribute in the prevention of local recurrence as well as metastasis. However, current cancer treatment strategies fail to either detect or differentiate the CSCs from their non-tumorigenic progenies mostly due to the absence of specific biomarkers and potent agents to kill CSCs. Recent advances in knowledge of CSCs enable to produce several candidates to ablate CSCs in gastrointestinal (GI) cancers, especially cancers originated from inflammation-driven mutagenesis such as Barrett's esophagus (BE), Helicobacter pylori-associated gastric cancer, and colitis-associated cancer (CAC). Our research teams elucidated through revisiting old drugs that proton pump inhibitor (PPI) and potassium competitive acid blocker (p-CAB) beyond authentic acid suppression, chloroquine for autophage inhibition, sonic hedgehog (SHH) inhibitors, and Wnt/β-catenin/NOTCH inhibitor can ablate CSCs specifically and efficiently. Furthermore, nanoformulations of these molecules could provide an additional advantage for more selective targeting of the pathways existing in CSCs just like current molecular targeted therapeutics and sustained action, while normal stem cells intact. In this review article, the novel approach specifically to ablate CSCs existing in GI cancers will be introduced with the introduction of explored mode of action.

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Morphogenetic Alterations of Alternaria alternata Exposed to Dicarboximide Fungicide, Iprodione

  • Kim, Eunji;Lee, Hye Min;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.33 no.1
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    • pp.95-100
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    • 2017
  • Fungicide-resistant Alternaria alternata impede the practical control of the Alternaria diseases in crop fields. This study aimed to investigate cytological fungicide resistance mechanisms of A. alternata against dicarboximide fungicide iprodione. A. alternata isolated from cactus brown spot was cultured on potato-dextrose agar (PDA) with or without iprodione, and the fungal cultures with different growth characteristics from no, initial and full growth were observed by light and electron microscopy. Mycelia began to grow from one day after incubation (DAI) and continued to be in full growth (control-growth, Con-G) on PDA without fungicide, while on PDA with iprodione, no fungal growth (iprodione-no growth, Ipr-N) occurred for the first 3 DAI, but once the initial growth (iprodione-initial growth, Ipr-I) began at 4-5 DAI, the colonies grew and expanded continuously to be in full growth (iprodione-growth, Ipr-G), suggesting Ipr-I may be a turning moment of the morphogenetic changes resisting fungicidal toxicity. Con-G formed multicellular conidia with cell walls and septa and intact dense cytoplasm. In Ipr-N, fungal sporulation was inhibited by forming mostly undeveloped unicellular conidia with degraded and necrotic cytoplasm. However, in Ipr-I, conspicuous cellular changes occurred during sporulation by forming multicellular conidia with double layered (thickened) cell walls and accumulation of proliferated lipid bodies in the conidial cytoplasm, which may inhibit the penetration of the fungicide into conidial cells, reducing fungicide-associated toxicity, and may be utilized as energy and nutritional sources, respectively, for the further fungal growth to form mature colonies as in Ipr-G that formed multicellular conidia with cell walls and intact cytoplasm with lipid bodies as in Con-G.

Therapeutic Effect of Cyclosporine Administration in a Dog with Pemphigus Foliaceus (풍산견에서 발생한 낙엽성 천포창의 사이클로스포린 적용의 치료적 효과 증례)

  • Kim, Jung-Hyun;Sur, Jung-Hyang;Park, Chul;Yoo, Jong-Hyun;Kim, Ha-Jung;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.26 no.4
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    • pp.340-343
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    • 2009
  • A 3-year-old, intact female Poong-san dog was presented with a 4-month history of erosive, erythematous dermatitis unresponsive to systemic antibiotics and glucocorticoids. Crust, erosion, and alopecia were noted on the ear pinnae, bridge of nose, and forelimb. Cytological evaluation of intact pustules showed isolated free-floating rounded acantholytic keratinocytes admixed with non-degenerated neutrophils and eosinophils. Results of histopathologic examination revealed the intra-epidermal pustules with predominant neutrophils, less eosinophils, and isolated and clustered acantholytic cells. A diagnosis of pemphigus foliaceus (PF) was made based on the history, clinical, cytological and histopathological results. The skin lesions had improved after systemic cyclosporine therapy. This case report demonstrates that cyclosporine, an immunosuppressive agent, can be used in the management of PF in dogs.

Studies on Sexing of Bisected Mouse Embryos by Rat H-Y Antibody (Rat H-Y 항체에 의한 생쥐 분할란의 성 조절에 관한 연구)

  • 정장용;박희성;박충생
    • Korean Journal of Animal Reproduction
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    • v.15 no.3
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    • pp.179-187
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    • 1991
  • This experiment was carried out to develop a new technique of identifying XX of XY-bearing bisected embryos prior to implantation by immunological method. H-Y antiserum prepared in inbred Wastar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the concentraton or affected H-Y antibody was also investigated by culturing embryos under the concentration or affected H-Y antibody and culture rate. However, production of live young or sex rates of male and female from embryos transferred with psudopregnant. The biological test with the morula stage embryos showed that H-Y antibody was formed in all female rats immunized with spleen cell, but it was formed only in 80% female rats immunized with the antigen. When the bisected mouse embryos were cultured in vitro for 5~6 hours in morula stage, of 457 bisected embryos 81.4% of then were developed to the blastocyst stage. When the concentration rate of complement to H-Y antiserum varied from 1.0~5.0${mu}ell$, the lysis-rate of embryo was 19.5 to 67.3%. The concentration rate of complement did not influence the lysis-rate of embryos(P<0.05). The morphology embryos of bisected, zona-free and intact embryos showed the embryos lysis rate of 58.6, 42.7 and 48.5% respectively(P<0.05). Pregnancy rate were 50.0, 45.5 and 57.1% in psudopregnant recipient transferred with bisected, zona-free and intact blastocyst embryos. However, production of live youngs, sexual rate of male or female was 24(50.0:50.0), 22(45.5:55.5) and 36(58.3:41.7)mice, but affected and non affected half embryos with H-Y antiserum treatment was 23.1 and 26.7%. Also production of live youngs and sexual rate was 14(92.9:7.1) and 17(17.6:82.4)mice in affected and non affected half embryos in H-Y antiserum treatment(P<0.05).

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Lysophosphatidylcholine Attenuates Endothelium-dependent Relaxation Responses through Inhibition of ACh-induced Endothelial $[Ca^{2+}]_i$ Increase

  • Kwon, Seong-Chun;Lee, Yong-Ho;Nam, Taick-Sang;Ahn, Duck-Sun
    • The Korean Journal of Physiology and Pharmacology
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    • v.10 no.1
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    • pp.25-30
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    • 2006
  • Lysophosphatidylcholine (LPC), which accumulates in atherosclerotic arteries, has been reported to inhibit endothelium-dependent relaxation (EDR) in many different species. However, the underlying mechanism of LPC-induced inhibition of EDR is still uncertain. In the present study, we measured simultaneously both isometric tension and cytosolic free $Ca^{2+}$ ($[Ca^{2+}]_i$) in rabbit carotid strips, and examined the effect of LPC on tension and $[Ca^{2+}]_i$. In carotid strips with intact-endothelium, high $K^+$ (70 mM) increased both tension and $[Ca^{2+}]_i$, and cumulative addition of acetylcholine (ACh) from 0.1 to $10{\mu}M$ induced dose dependent increase of $[Ca^{2+}]_i$ with concomitant relaxation. In the presence of L-NAME (0.1 mM), ACh increased $[Ca^{2+}]_i$ without affecting the amplitude of high $K^+-induced$ tension. These ACh-induced change of $[Ca^{2+}]_i$ and tension was abolished by removal of endothelium or 10 nM 4-DAMP (muscarinic receptor antagonist) pretreatment. Pretreatment of LPC ($10{\mu}M$) inhibited ACh ($10{\mu}M$)-induced change of tension and $[Ca^{2+}]_i$ in endothelium-intact carotid artery. On the other hand, LPC had no effect on ACh-induced change of tension and $[Ca^{2+}]_i$ in endothelium denuded artery. In $Ca^{2+}$-free external solution, ACh transiently increased $[Ca^{2+}]_i$, and pretreatment of LPC significantly inhibited ACh-induced transient $[Ca^{2+}]_i$ change. Based on the above results, it may be concluded that LPC inhibits the ACh-induced $[Ca^{2+}]_i$ change through inhibition of $Ca^{2+}$ mobilization in vascular endothelial cells, resulting in decreased production of NO and concomitant inhibition of endotheliumdependent vascular relaxation.

Expression of the VP2 protein of feline panleukopenia virus in insect cells and use thereof in a hemagglutination inhibition assay

  • Yang, Dong-Kun;Park, Yeseul;Park, Yu-Ri;Yoo, Jae Young;An, Sungjun;Park, Jungwon;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.19.1-19.7
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    • 2021
  • Feline panleukopenia virus (FPV) causes leukopenia and severe hemorrhagic diarrhea, killing 50% of naturally infected cats. Although intact FPV can serve as an antigen in the hemagglutination inhibition (HI) test, an accidental laboratory-mediated infection is concern. A non-infectious diagnostic reagent is required for the HI test. Here, we expressed the viral protein 2 (VP2) gene of the FPV strain currently prevalent in South Korea in a baculovirus expression system; VP2 protein was identified by an indirect immunofluorescence assay, electron microscopy (EM), Western blotting (WB), and a hemagglutination assay (HA). EM showed that the recombinant VP2 protein self-assembled to form virus-like particles. WB revealed that the recombinant VP2 was 65 kDa in size. The HA activity of the recombinant VP2 protein was very high at 1:215. A total of 143 cat serum samples were tested using FPV (HI-FPV test) and the recombinant VP2 protein (HI-VP2 test) as HI antigens. The sensitivity, specificity, and accuracy of the HI-VP2 test were 99.3%, 88.9%, and 99.3%, respectively, compared to the HI-FPV test. The HI-VP2 and HI-FPV results correlated significantly (r = 0.978). Thus, recombinant VP2 can substitute for intact FPV as the serological diagnostic reagent of the HI test for FPV.