• Title/Summary/Keyword: "The cat"

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Identification of a Bacillus thuringiensis Surface Layer Protein with Cytotoxic Activity against MDA-MB-231 Breast Cancer Cells

  • Rubio, Viviana P.;Bravo, Alejandra;Olmos, Jorge
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.36-42
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    • 2017
  • In this work, we isolated a surface layer protein (SLP) from a Bacillus thuringiensis (Bt) strain to evaluate it cytotoxic effects against MDA-MB-231 human breast cancer cells. AP11 was selected from a g roup of Bt strains using SLP olig onucleotides developed from Bacillus conserved regions. The AP11 strain was grown in Luria Bertani medium until the late exponential phase; an 86 kDa protein was extracted using 5 M LiCl and identified by liquid chromatography-tandem mass spectrometry. It corresponded to a multispecies SLP highly similar to previously described SLPs in Bt. The MDA-MB-231 breast cancer cells $LC_{50}$ was obtained using $0.25{\mu}g/ml$ of the isolated SLP. HaCat non-cancerous cells presented 90% survival using the same protein concentration. Our data suggest that SLP cytotoxicity against MDA-MB-231 could be induced by an interaction with the CDH11 cell membrane receptor.

Dysmyelopoiesis in a cat with immune-mediated hemolytic anemia

  • Yu, Do-Hyeon;Lee, Jong-Hyun;Yoon, Ji-Seon;Li, Ying-Hua;Lee, Mi-Jin;Park, Jinho
    • Korean Journal of Veterinary Research
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    • v.48 no.2
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    • pp.203-207
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    • 2008
  • A 3-year-old spayed female Persian feline with non regenerative anemia showed persistent autoagglutination in EDTA anticoagulated blood. Primary immune-mediated hemolytic anemia (IMHA) was suspected and the underlying causes for IMHA were excluded by radiologic, sonographic, serologic and molecular studies. Cytologic examination of the bone marrow revealed that dysmyelopoiesis and dysplastic changes were prominent in the erythroid cells. These changes included asynchronous maturation of the nucleus and cytoplasm, binucleation, trinucleation, fragmented or lobulated nuclei and multilineages. Mild dysgranulopoiesis and dysmegakaryocytopoiesis were also detected including pseudo Pelger-Huet anomalies, giant band neutrophils, asynchronous maturation of the nucleus and cytoplasm in granulopoiesis and large hypolobulated forms as well as dwarf megakaryocytes in megakaryocytopoiesis. Myelodysplastic syndrome and congenital dysmyelopoiesis was ruled out by the low number of blast cells. Finally, secondary dysmyelopoiesis associated with IMHA was diagnosed and immunosuppressive treatment was successfully responsive.

Effects of Uncooked Korean Food on Blood Glucose and Antioxidant Fnzyme Activities of Streptozotocin-induced Diabetic Rats (당뇨환자용 식사대용식이 Streptozotocin으로 유발된 당뇨쥐의 혈당 및 항산화 효소 활성에 미치는 영향)

  • Bae, Han-Ho;Song, See-Won;Nam, Tae-Heung;Cho, Chung-Sik
    • The Korea Journal of Herbology
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    • v.23 no.1
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    • pp.63-74
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    • 2008
  • Objectives : This study has been carried out to understand the effect of Uncooked Korean Food(F-DM) on blood glucose and antioxidant enzyme activities in streptozotocin-induced Diabetic Rats. Methods : SD rats were separated into four groups(each with 20 rats). Except normal two group, the other two groups were injected into intra-peritoneal with streptozotocin 60 mg/kg. Experimental group was eated Feed with 25% F-DM for 4 weeks. The change of plasma glucose level, body weight were observed. After 4 weeks, liver and kidney weight, antioxidant enzyme activities, survival rate were observed with histological changes on liver, kidney and pancreas. Results : In experimental group, body weight and survival rate increased, plasma glucose level were decreased significantly. Liver and kidney weight, XOD activity were decreased in experimental group compared to control group. GSH-px and CAT activities, insulin- immunoreactive granules in p-cells were increased significantly in experimental group compared to control group. Conclusions : This study shows that the F-DM might be effective for treatment of diabetes and its complications, as well as reduction of the oxidative stress.

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Development of Leuconostoc sp. Host Vector System

  • Eom, Hyun-Ju;Park, Myeong-Soo;Ji, Geun-Eog;Han, Nam-Soo
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2004.06a
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    • pp.323-327
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    • 2004
  • Leuconostoc citreum CBUE isolated from kimchi proved to harbor a small cryptic plasmid, pNS75. The complete nucleotide sequence of pNS75 was 1,821 bp and had a low G+C content of 39.2%. Computer analysis using DNASIS revealed one open reading frame (ORF), having ATG as putatitive start condon and potentially encoding proteins with molecular mass of 38 kDa. The chimeric plasmid pLeuCM was first constructed wih pNS75, pUC19 and chroamphenicol acetyltransferase (CAT) from Staphylococcus sp.. pLeuCM replicated and expressed chroamphenicol acetyltransferase in Leuconostoc citerum CBNF after transformation. To test the availability of shuttle vector as cloning vehicle of foreign gene, $\alpha$-amylase gene of Streptococcus bovis was cloned and all transformants secreated the $\alpha$-amylase successfully. The result indicates that pLeuCM is a potential shuttle vector for Leuconostoc spp. and lactic acid bacteria.

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Purification and Characterization of a Thermostable Cellobiohydrolase from Fomitopsis pinicola

  • Shin, Keum;Kim, Yoon-Hee;Jeya, Marimuthu;Lee, Jung-Kul;Kim, Yeong-Suk
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1681-1688
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    • 2010
  • A screening for cellobiohydrolase (CBH) activity was performed and Fomitopsis pinicola KMJ812 was selected for further characterization as it produced a high level of CBH activity. An extracellular CBH was purified to homogeneity by sequential chromatography of F. pinicola culture supernatants. The molecular mass of the F. pinicola CBH was determined to be 64 kDa by SDS-PAGE and by size-exclusion chromatography, indicating that the enzyme is a monomer. The F. pinicola CBH showed a $t_{1/2}$ value of 42 h at $70^{\circ}C$ and catalytic efficiency of $15.8mM^{-1}s^{-1}(k_{cat}/K_m)$ for p-nitrophenyl-${\beta}$-D-cellobioside, one of the highest levels seen for CBH-producing microorganisms. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, the F. pinicola CBH is distinguished from other CBHs by its high catalytic efficiency and thermostability.

A Continuous Spectrophotometric Assay for NADPH-cytochrome P450 Reductase Activity Using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide

  • Yim, Sung-Kun;Yun, Chul-Ho;Ahn, Tae-Ho;Jung, Heung-Chae;Pan, Jae-Gu
    • BMB Reports
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    • v.38 no.3
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    • pp.366-369
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    • 2005
  • NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450 and also catalyzes the one-electron reduction of many drugs and foreign compounds. Various spectrophotometric assays have been performed to examine electron-accepting properties of CPR and its ability to reduce cytochrome $b_5$, cytochrome c, and ferricyanide. In this report, reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by CPR has been assessed as a method for monitoring CPR activity. The principle advantage of this substance is that the reduction of MTT can be assayed directly in the reaction medium by a continuous spectrophotometric method. The electrons released from NADPH by CPR were transferred to MTT. MTT reduction activity was then assessed spectrophotometrically by measuring the increase of $A_{610}$. MTT reduction followed classical Michaelis-Menten kinetics ($K_m\;=\;20\;{\mu}M$, $k_{cat}\;=\;1,910\;min^{-1}$). This method offers the advantages of a commercially available substrate and short analysis time by a simple measurement of enzymatic activity of CPR.

Effect of Electrical Hypothalamic Stimulation on Blood Glucose and Circulating Eosinophil Levels in Cat (시상하부 전기자극이 혈당량 및 혈중 Eosinophil 변동에 미치는 영향)

  • Lee, Kae-Yol;Lee, Myung-Hee;Lee, Bong-Koo;Yoo, Eun-Hak;Kim, Bong-Ju
    • The Korean Journal of Physiology
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    • v.1 no.2
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    • pp.181-184
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    • 1967
  • Hypothalamus is one of the most important center regulating voluntary and involuntary function of the body. We studied on the blood glucose and eosinophilic responses induced by electrical hypothalamic stimulation using stereotaxic apparatus. Blood glucose was measured according to Somogyi Nelson method and circulating eosinophil was counted with Hinkleman Stain. For the histological confirmation of electrode placement frozen sections were cut along electrode tract and stained with Hematoxylin-Eosin. The results obtained were as follows: 1. Anterior hypothalamic stimulation mostly leads to hypoglycemia but in some cases lead to hyperglycemia. 2. Hypothalamic stimulation leads to eosinopenia in majority cases. 3. There is no relationship between eosinopenia and hyperglycemia. 4. It appears that sympathetic and pararympathetic areas are not separate part but it coincides each other.

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Antioxidant and Anti-Inflammatory Effects of NCW Peptide from Clam Worm (Marphysa sanguinea)

  • Park, Young Ran;Park, Chan-Il;Soh, Yunjo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1387-1394
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    • 2020
  • Clam worms (Marphysa sanguinea) are a rich source of bioactive components such as the antibacterial peptide, perinerin. In the present study, we explored the physiological activities of a novel NCWPFQGVPLGFQAPP peptide (NCW peptide), which was purified from clam worm extract through high-performance liquid chromatography. Tandem mass spectrometry (MS/MS) revealed that NCW was a new peptide with a molecular weight of 1757.86 kDa. Moreover, NCW peptide exhibited significant antioxidant effects, causing a 50% inhibition of DPPH radical at a concentration of 20 μM without showing any cytotoxicity. These were associated with a reduction in the activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) in LPS-stimulated RAW264. 7 cells. Furthermore, NCW peptide exhibited anti-inflammatory effects in LPS-stimulated RAW264.7 macrophages via inhibition of the abnormal production of pro-inflammatory cytokines including nitric oxide (NO), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). These anti-inflammatory effects of NCW peptide were associated with the inhibition of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). Our results therefore suggest that this novel NCW peptide with antioxidant and anti-inflammatory effects could be a good therapeutic agent against inflammation-related diseases.

Deletion of Superoxide Dismutase Gene of Bombyx mori Nuclear Polyhedrosis Virus Affects Viral DNA Replication

  • Wang, Wenbing;Song, Zhixiu;Ji, Ping;Wu, Jun;Zhang, Zhifang;He, Jialu;Wu, Xiangfu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.2
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    • pp.225-228
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    • 2004
  • Superoxide dismutase (SOD) is an important enzyme which catalyzes superoxide radicals to hydrogen peroxide. A Cu, Zn sod-like gene was found in Bombyx mori nuclear polyhedrosis virus encoding 151 amino acids. To demonstrate its function, a recombinant virus named dsBmNPV with deleted sod gene was constructed. It was discovered that the sod gene was not essential for viral replication. Studies on growth of budded virus in BmN cells and superoxide dismutase and catalase activities in vivo after dsBmNPV infection showed that the titer of dsBmNPV decreased obviously comparing to wild type BmNPV, the sod gene was effective on genomic DNA replication of baculovirus, the peak of SOD activity of silkworm infected with wt-BmNPV appeared between 36 and 48 hrs post infection, and with dsBmNPV, it did not appear. And the changes of CAT activity after infection were similar to SOD activity.

Positive Regulator, a Rice C3H2C3-type RING Finger Protein H2-3(OsRFPH2-3), in Response to Salt Stress

  • Min Seok Choi;Cheol Seong Jang
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2023.04a
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    • pp.156-156
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    • 2023
  • Salinity is a major abiotic stress that limits rice productivity in many regions of the world. In order to develop salt stress tolerant rice plants, genetic engineering is a promising approach. We characterized the molecular function of rice C3H2C3 as a really interesting new gene (RING). Oryza sativa RING finger protein H2-3 (OsRFPH2-3) was highly expressed in 100 mM NaCl. To identify the localization of OsRFPH2-3, we fused vectors that include C-terminal GFP protein (35S;;OsRFPH2-3-GFP). OsRFPH2-3 was expressed in the nucleus in rice protoplasts. An in vitro ubiquitin assay demonstrated that OsRFPH2-3 possessed E3-ubiquitin ligase activity. However, the mutated OsRFPH2-3 were not possessed any E3-ubiquitin ligase activity. Under salinity conditions, OsRFPH2-3-overexpressing plants exhibited higher chlorophyll, proline, SOD, POD, CAT, and soluble sugar contents and lower H2O2 accumulation than wild-type plants, supporting transgenic plants with enhanced salinity tolerance phenotypes. OsRFPH2-3-overexpressing plants exhibited low Na+ accumulation and Na+/K+ ratios in their roots. Theses results suggest that overexpression of OsRFPH2-3 can make plant insensitivity about salinity conditions.

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