• Title/Summary/Keyword: maceration

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Cloning and Sequencing of the pelCl Gene Encoding Pectate Lyase of Erwinia carotovora subsp. carotovora LY34 (Erwinia carotovora subsp. carotovora LY34에서 pelCI 유전자 클로닝)

  • Lim, Sun-Tech;Park, Yong-Woo;Yun, Han-Dae
    • Applied Biological Chemistry
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    • v.40 no.5
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    • pp.380-387
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    • 1997
  • Phytopathogenic Erwinia carotovora subsp. carotovora (Ecc) LY34 causes plant tissue maceration by secretion of pectinolytic enzymes such as pectate Iyase (PL) existed as multiple isoenzyme form. Genomic DNA from Ecc LY34 was digested with Sau3Al and ligated into the BamHI site of pBluescript ll $SK^+$. Among them, a clone hydrolyzing polypectate was selected and its DNA was digested with BamHI. Through the subsequent subcloning the resulting 3.1 kb fragment, corresponding to a peICI, was subcloned into pLYPA 100. The structural organization of a peICI gene encoding a 374 amino acid residues consists of an open reading frame (ORF) of 1,122 bp commencing with a ATG start codon and followed by a TAA stop codon. PeICI contained a typical prokaryotic signal peptide of 22-amino acid. Since the deduced amino acid sequences of PeICl protein was very similar to those of PelIII of Erwinia carotovora subsp. carotovora, and to those of Pel3 of Erwinia carotovora subsp. atroseptica, and to those of PeIC of Erwinia carotovora subsp. carotovora, it belong to the same family PLbc group. The 374-amino acld PeICI had a calculated Mr of 40,507 and pI of 7.60.

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Neuroprotective Effect of Root Extracts of Berberis Vulgaris (Barberry) on Oxidative Stress on SH-SY5Y Cells

  • Rad, Elham Shahriari;Eidi, Akram;Minai-Tehrani, Dariush;Bonakdar, Shahin;Shoeibi, Shahram
    • Journal of Pharmacopuncture
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    • v.25 no.3
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    • pp.216-223
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    • 2022
  • Objectives: Oxidative stress plays a key role in chronic and acute brain disorders and neuronal damage associated with Alzheimer disease (AD) and other neurodegeneration symptoms. The neuroprotective effects of berberine and Berberis vulgaris (barberry) root extract against apoptosis induced by hydrogen peroxide (H2O2) in the human SH-SY5Y cell line were studied. Methods: The methanolic extraction of barberry root was performed using a maceration procedure. Oxidative stress was induced in SH-SY5Y cells by H2O2, and an MTT assay was applied to evaluate the neuroprotective effects of berberine and barberry root extract. The cells were pretreated with the half maximal inhibitory concentration (IC50) of each compound (including berberine, barberry root extract, and H2O2), and the anti-apoptotic effects of all components were investigated using RT-PCR. Results: The SH-SY5Y cell viability increased in both groups exposed to 75 and 150 ppm barberry extract compared with that in the H2O2-treated group. The data showed that exposing SH-SY5Y cells to 30 ppm berberine significantly increased the cell viability compared with the H2O2-treated group; treatment with 150 and 300 ppm berberine and H2O2 significantly decreased the SH-SY5Y cell viability and was associated with berberine cytotoxicity. The mRNA levels of Bax decreased significantly under treatment with berberine at 30 ppm compared with the control group. A significant increase in Bcl-2 expression was observed only after treatment with the IC50 of berberine. The expression level of Bcl-2 in cells exposed to both berberine and barberry extracts was also significantly higher than that in cells exposed to H2O2. Conclusion: The outcomes of this study suggest that treatment of SH-SY5Y cells with barberry extract and berberine could suppress apoptosis by regulating the actions of Bcl-2 family members.

A New Screening Method for the Selection of Calla Lily Zantedeschia aethiopica Cultivars Resistant to Calla Lily Soft Rot (칼라의 무름병 저항성 품종 선발을 위한 검정 방법 개발)

  • Joung, Hyang Young;Choi, Mok Pil;Han, Kyung Sook;Kim, Su;Goo, Dae Hoe;Kang, Yun Im;Choi, Youn Jung;Park, Sang Kun
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.366-370
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    • 2013
  • This experiment was carried out to establish the screening methods for the selection of Zantedeschia aethiopica cultivars resistant to calla lily soft rot caused by Pectobacterium carotovorum subsp. carotovorum. A center of leaf and petiole tissues of four Z. aethiopica cultivars were wounded and inoculated with three different inoculum concentrations, $1{\times}10^7cfu/mL$, $1{\times}10^8cfu/mL$, and $1{\times}10^9cfu/mL$, of EccNHRI-21 isolate. And they were evaluated at 4, 18, and 26 hours after inoculation. The lesion of maceration was developed well in the leaf tissues inoculated with $1{\times}10^9cfu/mL$ concentration. And evaluation of resistance in 18 hours after inoculation was correlated with field resistance positively. Using this method, ten Z. aethiopica commercial cultivars and four wild types were screened. 'Crowbrough' and 'White Cutie' were selected as highly resistant genotypes and 'Mont Blanc' and 'Silky White' as resistant genotypes. 'Wedding March' and 'Kiwi Blush' were evaluated as moderate resistant and the others including 'Childsiana' were susceptible. And all of four wild types were evaluated as more than moderate resistant. In spite of control through cultural, biological and chemical methods, the use of resistant cultivars is most efficient to overcome calla lily soft rot. This newly developed screening method was helpful for breeding new varieties resistant to calla lily soft rot.

A Comparison of Illness Behavior among Patients with Somatoform Disorders, Depressive Disorders and Psychosomatic Disorders (신체형장애, 우울장애 및 정신신체장애 환자들간의 질병행동의 비교)

  • Koh, Kyung-Bong;Ki, Sun-Wan
    • Korean Journal of Psychosomatic Medicine
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    • v.5 no.2
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    • pp.185-194
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    • 1997
  • A comparison was made regarding illness behavior among patients with somatoform disorders, depressive disorders and psychosomatic disorders. The subjects consisted of out-patients with somatoform disorders(N=52), depressive disorders(N=52) and psychosomatic disorders(N=51). illness behavior was assessed by illness Behavior Assessment Schedule and the questionnaire about help-seeking behavior. The patients with somatoform disorders and psychosomatic disorders more often affirmed the presence of somatic disease, were more likely to have phobia of disease, had more preoccupation with ideas of disease and more frequently shopped around oriental clinics than the patients with depressive disorders. The patients with somatoform disorders more often attributed its cause to physical factors, less often attributed the origin of affective disturbance to psychological causes, showed Less depression and irritability, and were less likely to accept psychiatric treatment recommended by other physicians than depressive patients. The patients with somatoform disorders were more likely to report having been told that they suffered from a mild illness than those with psychosomatic disorders. The patients with somatoform disorders with psychological problems tended to inhibit expression of their emotion. Female patients with somatoform disorders more often affirmed the presence of psychological disorder and attributed its cause to psychological factors than male ones. These results suggest that in illness behavior, patients with somatoform disorders are different from depressive patients, whereas the former patients are similar to psychosomatic patients except the discrepancy between therapists and patients regarding evaluation of their symptoms. Thus, it is emphasized that first, therapists need to approach patients with somatoform disorders somatically with understanding of their underlying need to deny psychological problems, followed by either psychological or biopsychosocial approach.

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STUDIES ON THE EXTRACTION OF SEAWEED PROTEINS 1. Extraction of Water Soluble Proteins (해조단백질의 추출에 관한 연구 1. 수용성 단백질의 추출)

  • RYU Hong-Soo;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.10 no.3
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    • pp.151-162
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    • 1977
  • Distribution of marine algae is diverse in Korea and the resource of edible algae is abundant marking 239,037 tons of yearly production in 1976. They have been known as a protein source and used as a supplement in Korean diet. It is necessary to estimate the potentiality and properties of usable algal proteins especially as food resources and studies of extraction and separation of the proteins, therefore, are basically required for this purpose. In this study, the influence of various factors including the sample treatment, extraction time and temperature, sample us extraction solvent ratio and pH upon the extractability of the water soluble protein was determined. And the effect of precipitation treatment for isolation of the algal protein from the extracts was also tested. Nine species of algae, the major ones in consumption as food namely Porphyra suborbiculata, Undaria pinnatifida, Hizikia fusiforme, Sargassum fulvellu, Enteromorpha linza, Codium fragile, Sargassum kjellmanianum and Ulva pertusa were collected as fresh from Kijang, Yangsan Gun, in the vicinity of Busan city. The content of crude protein $(N\times6.25)$ of the algae ranged from $9.46\%\;to\;24.14\% showing the highest value in Porphyra suborbiculata and the minimum in Hizikia fusiforme. In the effort of maceration of blending methods on the extractability, immersion freezing in dry ice-methanol solution appeared most effective yielding 1.5 to 2.5 times extractability than that of the mortar grinding method. The effect of the ratio of sample vs solvent on extractability differed from species. It was enhanced at the ratio of 1:20 (w/v) in Ulva pertusa and Enteromorpha linza while the ratio was 1:30 (w/v) for Cedium fragile, Undaria pinnatifida, Hizikia fusiferme, Sargassum fulvellum and Porphyra suborbiculata and 1:40 for Sargassum kjellmanianum respectively. The effect of extraction time and temperature was revealed differently from species which might be caused by differences in the constitution of algal tissues resulting in that the extraction for 1 hour at $50^{\circ}C$ gave the maximum extractabilily in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Hikikia fusiforme, Undaria pinnatifida, Sargassum kjellmanianum and 3 hours in Codium fragile. And the extractability was higher at $50^{\circ}C$ to $60^{\circ}C$ for the most of the tested samples except Hizikia fusiforme. The optimum pH for the extraction was 9 to 12. The recovery of extractable nitrogen to the total nitrogen was $63\%$ in average with the first extracts and $8.6\%$ with the second extracts respectively. Both extracts were prepared by 2 hour extraction at $50{\pm}1^{\circ}C$ with dry ice-methanol frozen and seasand macerated materials. And these conditions assumed to be an optimum for the extraction of water soluble algal proteins since the nitrogen content after the first extraction covered $90\%$ of the total water extractable nitrogen. In the precipitation of the extracted proteins, Barnstein method and methanol treatment seemed to be more efficient than other precipitation methods.

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The Developmental Effects of Radiation on ICR Mouse Embryos in Preimplantation Stage (착상전기(着床前期)에 있어서 ICR Mouse의 태아(胎兒)에 대한 방사선(放射線) 개체(個體) Level 영향(影響)의 연구(硏究))

  • Gu, Yeun-Hwa
    • Journal of Radiation Protection and Research
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    • v.21 no.4
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    • pp.273-284
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    • 1996
  • Embryos and fetuses are more sensitive to various environmental agents than are adults or children. The biological effects such as intrauterine death and malformation are closely connected with prenatal exposure very various agents. The sensitivity of these embryonic/fetal effects depends on the stage of pregnancy. From the viewpoint of fetal development, embryonic and fetal stages can be divided into three stages : Preimplantation, organogenetic and fetal. Each stage corresponds to 0 to 4.5days, 4.5 to 13.5days, and 13.5days of gestation in mice, respectively. Many studies on the biologcal effects of mice irradiated by ${\gamma}-rays$ at various stages during organogenesis and fetal period have been performed. Based on these results, the dose-effect and dose-response relationships in malformations, intrauterine death, or retardation of the physical growth have been practically modeled by the ICRP(International Commission on Radiological Protection) and other international bodies for radiation protection. Many experimental studies on mice have made it clear that mice embryos in the preimplantation period have a higher sensitivity to radiation for lethal effects than the embryos/fetuses on other prenatal periods. However, no eratogenic effects of radiation at preimplantation stages of mice have been described in many textbooks. It has been believed that 'all or none action results' for radiation of mice during the preimplantation period were applied. The teratogenic and lethal effects during the preimplantation stage are one of the most important problems from the viewpoint of radiological protection, since the preimplantation stage is the period when the pregnancy itself is not noticed by a pregnant woman. There are many physical or chemical agents which affect embryos/fetuses in the environment. It is assumed that each agents indirectly effects a human. Then, a safety criterion on each agent is determined independently. The pregnant ICR mice on 2, 48, 72 or 96 hours post-conception (hpc), at which are preimplantation stage of embryos, were irradiated whole body Cesium-gamma radiation at doses of 0.1, 0.25, 0.5, 1.5, and 2.5 Gy with dose rate of 0.2 Gy/min. In the embryos from the fetuses from the mice irradiated at various period in preimplantation, embryonic/fetal mortalities, incidence of external gross malformation, fetal body weight and sex ratio were observed at day 18 of gestation. The sensitivity of embryonic mortalities in the mice irradiated at the stage of preimplantation were higher than those in the mice irradiated at the stage of organogenesis. And the more sensitive periods of preimplantation stage for embryonic death were 2 and 48 hpc, at which embryos were one cell and 4 to 7 cell stage, respectively. Many types of the external gross malformations such as exencephaly, cleft palate and anophthalmia were observed in the fetuses from the mice irradiated at 2, 72 and 96 hpc. However, no malformations were observed in the mice irradiated at 48 hpc, at which stage the embryos were about 6 cell stage precompacted embryos. So far, it is believed that the embryos on preimplantation stage are not susceptible to teratogens such as radiation and chemical agents. In this study, the sensitivity for external malformations in the fetuses from the mice irradiated at preimplantation were higher than those in the fetuses on stage of organogenesis.

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