• Title/Summary/Keyword: Insect cells

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Disseminated Histiocytic Sarcoma with Ureteric Involvement in a Jindo Dog

  • Cho, Hee-Soo;Cheong, Jong-Tae;Park, Hyun-Jung;Kim, Jae-Hoon;Jung, Ji-Youl;Yun, Youngmin;Song, Woo-Jin
    • Journal of Veterinary Clinics
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    • v.38 no.5
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    • pp.249-253
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    • 2021
  • We describe the case of a Jindo dog that presented with a 2-month history of an abscess-like mass on the skin of the left hip. The patient was initially diagnosed with a suspected insect bite and was treated with a topical dressing. After observing no improvements, surgical debridement was done to remove the abscess. Nine days after surgery, the patient started showing severe anorexia, vomiting, lethargy, and oliguria. Azotemia was detected, and the patient was diagnosed with acute kidney injury. Ultrasonography depicted an abdominal mass (4.6 × 7.5 cm) in the right ureter, and severe hydronephrosis in the right kidney. The patient was euthanatized. Results of the necropsy, histopathologic examination, and immunohistochemistry for tumor cells, confirmed that the patient had developed hydronephrosis and disseminated histiocytic sarcoma which had metastasized to the lungs. To the best of our knowledge, this is the first report of acute renal failure produced by ureter invasion and urinary tract obstruction resulting from the abdominal mass of disseminated histiocytic sarcoma in dogs.

Gene functional analysis of Harmonia axyridis by in vitro transcription

  • Park, Sang-Eun;Youn, Young-Nam
    • Korean Journal of Agricultural Science
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    • v.46 no.3
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    • pp.471-488
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    • 2019
  • Random genes were screened in two transforming ways to investigate the new genes of a ladybug using the Harmonia axyridis cDNA library stock cell cloned in the LITMUS 28i vector in a previous study. Phenotypic variation was observed after injection of the synthesized double-stranded RNA through the in vitro transcription process. The cDNA library of H. axyridis was transformed into E. coli $DH5{\alpha}$ and 10B competent cells by heat shock. Analysis of the nucleotide sequences of the 42 clones with the insert DNAs revealed that 21 clones were homologous with the genes of insects, and only one clone had a gene from H. axyridis. Thirteen of the 21 insect genes were homologous with genes from coleopteran insects. Fourteen genes were selected, which were identified by the gene screening results, and were synthesized as double-stranded RNA through in vitro transcription. One microgram of the synthesized double-stranded RNA between segments T1 and T2 were injected using a syringe into each anesthetized fourth larvae which were under 2 days old. As a result, a phenotypic variation appeared in the larva injected with the two genes. While the eggs of H. axyridis injected with distilled water hatched out three days after oviposition, the eggs of H. axyridis injected with dsHma 06 did not hatch but become shrivel a week after oviposition. Most of the H. axyridis injected with dsHma 08 died and were unable to complete the pupation or eclosion during ecdysis.

The Anti-inflammatory and Antiallergic Effects of Allomyrina dichotoma Larva Hot-water Extract (장수풍뎅이 유충 열수 추출물에 의한 항알레르기와 항염증 효과)

  • Lee, Hwa Jeong;Seo, Minchul;Kim, In-Woo;Lee, Joon Ha;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1130-1136
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    • 2017
  • Traditionally, the larvae of Allomyrina dichotoma (AD), a species of the rhinoceros beetle, have been widely used for their antidiabetic, antihepatofibrotic, antineoplastic, and antiobesity effects. The United Nations' Food and Agriculture Organization has reported on the possibility of using edible insects in human dietary supplements in the future. However, despite the growing interest in insect-based bio-active products, the biological activities of these products have rarely been studied. Previously, we reported that AD larvae inhibit the in vitro differentiation of adipocytes via transcription factor downregulation. In this study, our objective was to evaluate the effects of a hot-water extract of AD larvae on allergy and inflammation. To investigate the inhibitory effect of the extract on allergic reactions, we measured the levels of ${\beta}-hexosaminidase$, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin-4 (IL-4), and cyclooxygenase-2 (COX-2) after activation of RBL-2H3 cells using Compound 48/80. In addition, the inhibitory effect of the extract on inflammation was determined using Raw 264.7 cells after lipopolysaccharide (LPS) stimulation. The extract significantly inhibited the ${\beta}-hexosaminidase$, $TNF-{\alpha}$, IL-4, and COX-2 levels in RBL-2H3 cells. Furthermore, it effectively inhibited the inflammatory cytokine IL-6, nitric oxide, and inducible nitric oxide synthase expression in LPS-stimulated Raw 264.7 cells. These results suggest that AD larval extract can be potentially developed as an antiallergic and anti-inflammatory therapeutic agent.

Optimal Cultur Conditions for the Production of Insecticidal Toxin by Xenorhabdus nematophilus Isolated from Steinernema carpocapsae (Steinernema carpocapsae로부터 분리된 Xenorhabdus nematophilus에 의한 살충물질 생산을 위한 최적 배양조건)

  • 유연수;박선호
    • KSBB Journal
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    • v.15 no.1
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    • pp.100-105
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    • 2000
  • Optimal medium composition, culture conditions, characteristics of phase variation and activity of insecticidal toxin by Xenorhabdus nematophilus isolated and identified from Korean entomopathogenic nematode Steinernema carpocapsae were examined. Optimal medium composition of this strain was 50-70 g/L yeast extract, 3 g/L $K_{2}HPO_{4}$, 1g/L $NH_{4}H_{2}PO_{4}$, 2g/L ${MgSO}_4$$\cdot$${7H}_{2}O$, 10g/L NaCl and, these, yeast extract was found as a limiting nutrient for cell growth. When Monod equation was applied, maxmum specific growth rate and Monod constant were estimated as 0.13 $hr^{-1}$ and 20g/L, respectively. The pH of culture medium increased up to 8.5-9.5 regardless of initial pH 6-7 as the cells continued to grow. The specific growth rate in a 7 L fermentor was 0.18 $hr^{-1}$, which was enhancement 1.4 fold compared to a flask culture. In case of phase variation, phase I fraction was maintained above 90% at the stationary phase for both flask and fermentor cultures. According to oral toxicity test of Gallena mellonella by Xenorhabdus nematophilus, the addition of cell pellets into feed inhibited normal growth of insect larvae and killed completely then after 20 days cultivation. When culture supernatant of this strain was injected into hemolymph of insect larva, the toxicity was strongest at 24hr cultivation in the early exponential phase and gradually decreased as the culture time proceeded.

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Comparative Analysis of Host Insect Immunodepression Induced by Two Entomopathogenic Bacteria, Xenorhabdus nematophilus and Staphylococcus gallinarum, with Differential Pathogenicities (병원력 차이를 보이는 두 곤충병원세균(Xenorhabdus nematophilus와 Staphylococcus gallinarum)의 면역저하 능력 비교 분석)

  • 박영진;김길호;김용균
    • Korean journal of applied entomology
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    • v.42 no.4
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    • pp.353-360
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    • 2003
  • Immunodepression can be required for entomopathogenic bacteria to induce their potent pathogenicities to the target insects. Here, we raise a hypothesis that the capacity of a pathogenic bacterium to induce the target insect immunodepression has positive relationship with the degree of pathogenicity. X. nematophilus had 1,200 times as potent as another entomopathogenic bacterium, Staphylococcus gallinarum against the fifth instar larvae of silkworm, Bombyx mori, when they were Injected into the hemocoel. Although both bacteria had significant cytotokic effect on the hemocytes of B. mori, X. nematophilus gave faster and greater cytotoxicity than did S. gallinarum. In cellular immune reactions, B. mori could form 20 hemocyte nodules against the bacterial injection with 5${\times}$10$\^$5/ cells. The number of the hemocyte nodules was significantly depressed when live X. nematophilus was inject-ed, but not in S. gallinarum. Activation of prophenoloxidase (proPO) was depressed in the bacterial injection. The depression of PO activation was significantly greater in X. nematophilus infection than in S. gallinarum injection. Lysozyme activity was induced by the injection of S. gallinarum at 4 h after the treatment, but not induced in X. nematophilus at all the time. These results showed that X. nemato-philus induced greater immunodepression against B. mori and resulted in higher pathogenicity than did S. gallinarum. Therefore, this study suggests that the immunodepression induced by entomopathogenic bacteria has positive relationship with their pathogenicity.

Occurrence of Powdery Mildew Caused by Erysiphe abeliicola on Glossy Abelia in Korea (Erysiphe abeliicola에 의한 꽃댕강나무 흰가루병 발생)

  • Cho, Sung-Eun;Park, Ji-Hyun;Lee, Seung-Kyu;Lee, Sang-Hyun;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.18 no.2
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    • pp.133-138
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    • 2012
  • In November 2009, a powdery mildew on glossy abelia (Abelia ${\times}$ grandiflora) was found in Seogwipo, Jeju Island, Korea. Further survey in the southern part of Korea, e.g., Jeju, Busan, and Tongyeong confirmed occurrence of the disease. White colonies were present on leaves, young stems, and flowers, detracting from their beauty in landscape plantings. Severely infected lesions were discolored to red-purplish. Based on the morphological characteristics and analysis of rDNA, the fungus associated with the symptoms was identified as Erysiphe abeliicola U. Braun & S. Takam. This work provides the morphological feature of its anamorph for the first time, which is characterized by having multi-lobed hyphal appressoria and short foot-cells of conidiophores. Morphological characteristics of mature chasmothecia were consistent with the previous Japanese record of this species. The sequence of internal transcribed spacer region of ribosomal DNA obtained from a Korean sample showed that this species places in the section Microsphaera of the genus Erysiphe in phylogenetic position, corresponding with the classical taxonomy. This is the first report of E. abeliicola and its host plant in Korea. The host plant A. ${\times}$ grandiflora is newly listed in the host range of E. abeliicola.

Neuronal Mechanisms that Regulate Vitellogenesis in the Fruit Fly (노랑초파리 난황형성과정 제어 신경 메커니즘)

  • Kim, Young-Joon;Zhang, Chen
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.109-115
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    • 2022
  • Vitellogenesis is the process by which yolk accumulates in developing oocytes. The initiation of vitellogenesis represents an important control point in oogenesis. When females of the model insect Drosophila melanogaster molt to become adults, their ovaries lack mature vitellogenic oocytes, only producing them after reproductive maturation. After maturation, vitellogenesis stops until a mating signal re-activates it. Juvenile hormone (JH) from the endocrine organ known as the corpora allata (CA) is the major insect gonadotropin that stimulates vitellogenesis, and the seminal protein sex peptide (SP) has long been implicated as a mating signal that stimulates JH biosynthesis. In this review, we discuss our new findings that explain how the nervous system gates JH biosynthesis and vitellogenesis associated with reproductive maturation and the SP-induced post-mating response. Mated females exhibit diurnal rhythmicity in oogenesis. A subset of brain circadian pacemaker neurons produce Allatostatin C (AstC) to generate a circadian oogenesis rhythm by indirectly regulating JH and vitellogenesis through the brain insulin-producing cells. We also discuss genetic evidence that supports this model and future research directions.

Involvement of TLR4-JNK/NF-κB signaling pathway in RAW264.7 cell activation of Protaetia brevitarsis seulensis larvae extracts (흰점박이꽃무지 유충 추출물의 RAW264.7 세포 활성화에서 TLR4-JNK/NF-κB 신호전달 경로의 관여)

  • Ju-Hwi Park;Jongbeom Chae;Joon Ha Lee;Dongyup Hahn;Ju-Ock Nam
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.447-454
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    • 2023
  • In the environment in which humans live, there are various antigens that invade the human body and interfere with humans leading a healthy life, so the immune system recognizes the antigen then removes them through a complex mechanism. Macrophages are widely distributed immune cells involved in the innate immune system, and produce various immune modulators such as inducible nitric oxide synthase-induced nitric oxide, cyclooxygenase-2 induced prostaglandin E2 and proinflammatory cytokines such as tumor necrosis factor-alpha. On the other hand, Protaetia brevitarsis seulensis larvae are a type of edible insect that have emerged as an alternative to the future food supply problem. The immuno-modulatory effect through the activation of murine macrophage RAW264.7 cell via mitogen-activated protein kinases (MAPKs)/nuclear factor-kappa B (NF-κB) signaling pathways has been reported. Based on this report, in this study, we confirmed how the expression of immune modulators induced by Protaetia brevitarsis seulensis larvae extracts in RAW264.7 cells was changed by treatment with pharmacological inhibitors of toll-like receptor 4 (TLR4), MAPKs and NF-κB signaling pathways. As a result, reduction of immune modulators was confirmed in the c-Jun N-terminal kinase (JNK) inhibitor treatment group and NF-κB inhibitor treatment group among the Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell. Furthermore, in the TLR4 inhibitor-treated group, decreases in phosphorylation of JNK and NF-κB factors were confirmed in Protaetia brevitarsis seulensis larvae-treated RAW264.7 cell, as well as decreases in immune modulators. This results suggest that Protaetia brevitarsis seulensis larvae activates RAW264.7 cells by the engagement of TLR4-JNK/NF-κB signaling pathway.

Bombyx mori Protein Disulfide Isomerase (bPDI) Protects Sf9 Cells from Endoplasmic Reticulum (ER) Stress (소포체 스트레스에 대한 Protein Disulfide Isomerase의 세포보호효과)

  • Goo, Tae-Won;Yun, Eun-Young;Kim, Sung-Wan;Choi, Kwang-Ho;Kang, Seok-Woo;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.8 s.88
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    • pp.1129-1134
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    • 2007
  • In the previous our study, a cDNA that encodes protein disulfide isomerase from Bombyx mori (bPDI)was isolated and characterized. bPDI has an open reading frame of 494 amino acids contained two PDI-typical thioredoxin active site of WCGHCK and ER (endoplasmic reticulum) retention signal of the KDEL motif at its C-terminal. Recent studies have demonstrated that misfolded proteins are accumulated in many diseases including Alzheimer’s, goiter, emphysema, and prion infections. bPDI was over-expressed or knock-downed in Sf9 cells to study the relationship between bPDI expression and protections against protein misfolding. bPDI gene was cloned in insect expression vector pIZT/V5-His for over-expression and bPDI double-stranded RNA (dsRNA) was generated for knock-down. Over-expression of bPDI significantly improved survival rate, but bPDI dsRNA transfection significantly reduced survival rate after 48 hours exposure. In mock-transfected or wild-type cells had no significant effect. The results support the view that bPDI is one of the important intracellular components for cell protect mechanism, especially, against ER stress such as protein misfolding.

Technique to Generate Sterile Males of Striped Fruit Flies, Zeugodacus scutellata, using Electron Beam Irradiation and their Application to Genetic Control (전자빔 조사를 이용한 호박꽃과실파리의 불임 수컷 제조 기술과 유전방제 응용)

  • Kim, Yonggyun;Al Baki, Md. Abdullah;Kwon, Gimyeon
    • Korean journal of applied entomology
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    • v.59 no.1
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    • pp.25-35
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    • 2020
  • The striped fruit fly, Zeugodacus scutellata, is endemic in Korea, but it has been regarded as one of the serious quarantine pests throughout the world. Sterile insect release technique (SIT) has been used to eradicate quarantine fruit flies. This study developed a technique to generate sterile males and applied SIT to control Z. scutellata. First of all, the reproductive systems of Z. scutellata adults were examined with fluorescent microscope. Polytrophic ovaries comprises of around 100 follicles with developing oocytes. Each follicle contains an oocyte with several nurse cells and are surrounded with follicular epithelium. Oocyte development began at 10 days after adult emergence (DAE) and formed chorionated oocytes after 20 DAE. On the other hand, male testes were well developed just after adult emergence. The vas deferens was filled with motile sperms. To generate sterile males, different doses (0~1,000 Gy) doses of electron beam were irradiated to 3~5 days old pupae of Z. scutellata. When male pupae were irradiated with electron beam at 200 Gy, they developed and mated with females without any significant difference compared to untreated males. Although the untreated females mated with the 200 Gy-irradiated males laid eggs, no eggs did not hatch. The 200 Gy-irradiated males were then applied to untreated male and female flies in a density ratio of 1:9 (untreated males : treated males). The laid eggs suffered significant infertility. These results suggest that electron beam-irradiated pupae at 200 Gy resulted in male sterility and the resulting males would be applied to SIT.