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Repair of UV-induced Cyclobutane Pyrimidine Dimers in Human Mitochonrial DNA-less Cells

  • Ikushima, Takaji;Gu, Ning;Tanizaki, Yuichi
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.479-481
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    • 2002
  • UV-induced DNA damage causes cell killing and mutations leading to carcinogenesis. In normal human cells, UV damage such as cyclobutane pyrimidine dimers (CPDs) and primidine-prymidone (6-4) photoproducts are mainly repaired by nucleotide excision repair mechanism. The molecular processes have been well characterized recently. To know the influence of mitochondrial genome on the nucleotide excision repair mechanism against CPDs, we comparatively examined the production of CPDs by UVC irradiation and their repair kinetics in human cells completely lacking mitochondrial DNA (mtDNA) and the parental HeLa S cells. Whole DNA extracted from the cells exposed to UVC was treated with T4-endonuclease V to break the phosphodiester bond adjacent to CPDs. The DNA was electrophoresed in a denaturing agarose gel, which was visualized by ethidium bromide staining. The relative amount of CPDs was determined by image analysis using NIH Image software. MtDNA- less (rho-O) cells were apparently more sensitive to UVC than HeLa S cells, while the level of induction of CPDs in rho-O and HeLa cells was comparable. The repair of CPDs was less efficient in rho-O cells compared with HeLa cells. The residual amount of CPDs after 24-h repair was larger in rho-O cells than in HeLa cells where more than 90 % of CPDs were repaired by then. The non-repaired CPDs would lead to apoptosis in rho-O cells. These results suggest that mitochondrial genome may contribute to some ATP-dependent steps in nucletide excision repair by supplying sufficient ATP which is generated through a respiratory chain in mitochondria.

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Antitumor and antioxidant activities of Bryonia laciniosa against Ehrlich's Ascites Carcinoma bearing Swiss albino mice

  • Sivakumar, T;Kumar, R Sambath;Perumal, P;Vamsi, MLM;Sivakumar, P;Kanagasabai, R;Baskaran, MV;Karki, Subhas S;Mazumder, UK;Gupta, M
    • Advances in Traditional Medicine
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    • v.5 no.4
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    • pp.322-330
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    • 2005
  • The plant Bryonia laciniosa (Family: Cucurbitaceae) has been indicated for the treatment of various diseases one among which is cancer. The purpose of this study was investigating experimentally the possible anti-tumor effect and antioxidant role of Bryonia laciniosa leaves in animal model. The methanol extract of Bryonia laciniosa (MEBL) administered at the doses of 62.5, 125 and 250 mg/kg in mice for 14 days after 24 h of tumor inoculation. The effect of MEBL on the growth of transplantable murine tumor, life span of EAC bearing mice, hematological profile and liver biochemical parameters (lipid peroxidation, antioxidant enzymes) were estimated. Treatment with MEBL decreased the tumor volume and viable cell count thereby increasing the life span of EAC bearing mice and brought back the hematological parameter more or less normal level. The effect of MEBL also decreases the levels of lipid peroxidation and increased the levels of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). The present work indicates that the methanol extract of Bryonia laciniosa exhibited significant antitumor and antioxidant activity in vivo.

Drying Characteristics of Barley and the Effect of Moisture Content and Drying Temperature on Milling Recovery (보리의 건조특성(乾燥特性)과 건조온도(乾燥温度) 및 함수율(含水率)이 정맥수율(精麥收率)에 미치는 영향(影響))

  • Lee, Yong Kook;Kim, Sam Do;Park, Seung Je
    • Journal of Biosystems Engineering
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    • v.8 no.2
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    • pp.62-68
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    • 1983
  • This study was to examine the drying characteristics of barley and the effect of moisture content of barley on milling performance. A barley variety, Jogang, having 35.0% of initial moisture content was used for this experiment. Thermo-hydrostatic dryer which consists of blower, condensor, heater, humidifier, drying chamber and control box, etc., was used for the drying experiment. The change in the weight of a barley sample was continuously measured by means of the ring type load cell installed inside the drying chamber. Milling test runs the samples having the predetermined moisture content were taken from each drying test run. A laboratory type barley miller was used for the milling test. The results of the study are summarized as follows: 1. The drying constants (k) applied for the thin layer drying model, (M-Me)/(Mo-Me) = $Ae^{-kt}$ were 0.155, 0.259 and 0.548, respectively, at the three levels of drying temperatures, $40^{\circ}C$, $50^{\circ}C$ and $60^{\circ}C$. The drying constants complied with the Arrhenius Equation, K = Ko exp (-C/T), were determined as $Ko=1.901455{\times}10^8$ and C = 6563. 2. The laboratory milling test indicated that the highest milled and head barley recovery was resulted from the sample which was dryed at $40^{\circ}C$. In general, the increase in the drying temperature from $40^{\circ}C$ to $60^{\circ}C$ indicated a negative effect on milling yields. 3. Also, the sample having 15% M.C. presented the highest milled and head barley recovery among the five moisture content levels (12, 15, 18,21 and 24%).

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Identification of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Golpayegani Cattle by DNA Test

  • Mosafer, J.;Nassiry, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1691-1695
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    • 2005
  • The bovine lymphocyte antigen (BoLA)-DRB3 gene encodes cell surface glycoproteins that initiate immune responses by presenting processed antigenic peptides to CD4 T helper cells. DRB3 is the most polymorphic bovine MHC class II gene which encodes the peptide-binding groove. Since different alleles favour the binding of different peptides, DRB3 has been extensively evaluated as a candidate marker for associations with various bovine diseases and immunological traits. For that reason, the genetic diversity of the bovine class II DRB3 locus was investigated by polymerase chain reaction-restriction fragment length polymorphism method (PCR-RFLP). This study describes genetic variability in the BoLA-DRB3 in Iranian Golpayegani Cattle. Iranian Golpayegani Cows (n = 50) were genotyped for bovine lymphocyte antigen (BoLA)-DRB3.2 allele by polymerase chain reaction and restriction fragment length polymorphism method. Bovine DNA was isolated from aliquots of whole blood. A two-step polymerase chain reaction followed by digestion with restriction endonucleases RsaI, HaeIII and BstYI was conducted on the DNA from Iranian Golpayegani Cattle. In the Iranian Golpayegani herd studied, we identified 19 alleles.DRB3.2${\times}$16 had the highest allelic frequency (14%), followed by DRB3.2${\times}$7 (11%). Six alleles (DRB3.2${\times}$25, ${\times}$24, ${\times}$22, ${\times}$20, ${\times}$15, ${\times}$3) had frequencies = 2%. Although additional studies are required to confirm the present findings, our results indicate that exon 2 of the BoLA-DRB3 gene is highly polymorphic in Iranian Golpayegani Cattle.

Prognostic Factors of Idiopathic Facial Palsy: A Retrospective Study

  • Park, Gi Nam;Jeong, Jeong Kyo;Kim, Eun Seok;Kim, Jung Ho;Kim, Young Il
    • Journal of Acupuncture Research
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    • v.34 no.3
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    • pp.23-38
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    • 2017
  • Objectives : The purpose of this study was to evaluate the clinical prognostic factors affecting facial palsy in 98 idiopathic facial palsy patients who were hospitalized and treated in 2015, using retrospective statistical analysis. Methods : We investigated patients with idiopathic facial nerve palsy, admitted to a Korean medical hospital in 2015, and examined patients' variables and therapeutic variables. For analysis of clinical data, an independent sample t-test, analysis of variance (ANOVA), and simple regression analysis were performed using IBM SPSS version 24.0. Results : 1. The initial degree of facial palsy showed statistical significance with age. The older the age, the more severe the initial palsy. 2. Following treatment degree of facial palsy was statistically significant with age, hypertension, and fasting blood sugar (FBS). The higher the value, the slower the recovery from facial palsy. There was a statistical significance with the number of treatments in a Korean medical hospital. The more frequent the treatment, the faster the facial palsy recovery. 3. Degree of facial palsy after 12 months was statistically significant with age, hypertension, diabetes, FBS, and the initial severity of facial palsy. The higher the value, the slower the facial palsy recovery. 4. Sex, left or right sided palsy, alcohol consumption, smoking, history of facial palsy, season of onset, total number of treatments and bio chemistry (BC), complete blood cell count (CBC), urinalysis (UA) factors had no statistical significance with prognosis of facial palsy. Conclusion : Age, season of onset, hypertension, diabetes, FBS, initial severity of facial palsy, and the number of treatments at a Korean medical hospital showed statistical significance. The number of treatments at the Korean medical hospital positively correlated with facial palsy prognosis, and the others variables showed a negative correlation with facial palsy prognosis.

Carbon Monoxide Dehydrogenase in Cell Extracts of an Acinetobacter Isolate (Acinetobacter sp.1의 일산화탄소 산화효소의 특성)

  • 조진원;김영민
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.133-140
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    • 1986
  • Extracts of CO-autotrophically grown cells of Acinetobacter sp. 1 were shown to use thionin, methylene blue, or 2,6-dichlorophenol-indophenol, but not NAD, NADP, FAD, or FMN, as electron acceptors for the oxidation of CO under strictly anaerobic conditions. The CO dehydrogenase (CO-DH) in the thes bacterium was found to be an inducible enzyme. The enzyme activity was determined by an assay based on the CO-dependent reduction of thionin. Maximal reaction rates were found at pH 7.5 and $60^{\circ}C$, and the Arrhenius plot revealed an activation energy of 6.1 kcal/mol(25.5kJ/mol). THe $K_m$ m/ for CO was $154{\mu}M$. Known metalchelating agents tested had no effects on the CO-DH activity. No divalent cations tested affect the enzyme activity significantly escept $Cu^{2+}$ which suppressed the activity completely. The enzyme was inhibited by glucose and succinate. The same extracts catalyzed oxidation of hydrogen gas and formate with thionin as electron acceptor. The CO-DH of Acinetobacter sp. 1 was to have no immunological relationship with that of Pseudomonas carboxydohydrogena.

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The Production of Chimeric Mice by Embryonic Stem Cell Carrying Human Luteinizing Hormone Gene (사람 LH 호르몬유전자를 도입한 배아주세포에 의한 카이미라 생쥐 생산)

  • ;Tokunaga, T.;Imai, H.
    • Journal of Embryo Transfer
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    • v.9 no.3
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    • pp.243-247
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    • 1994
  • 최근 의약적으로 유용한 단백질을 대량 생산키 위한 실현 가능한 방법이 유전자변환 가축의 이용과 관련되어 발전되어 왔다. 이러한 유전자 변환동물은 이종의 단백질을 유즙속으로 분비시키는 생체반응기로서 이용되고 있다. 이러한 전략적 목적을 위해 현재 유전자 변환동물의 생산을 위한 이용에 있어 여러 가지 방법들이 보고되고 있다. 그러나 ES 세포의 사용이 이러한 방법들 사이에서 가장 실질적인 것으로 추정되고 있다. 본 실험에서는 유전자 구축을 위해 사람 황체 호르몬(human luteinizing hormone; hLH)의 전사를 유도하기 위해 각각 2.2 및 0.5 kb의 토끼 $\beta$-casein pronoter 단편을 이용하여 생쥐의 유선에 hLH를 발현시키도록 조절하고 발현이 thynidine kinase(TK) pronoter에 의해 좌우되는 neo 유전자를 selectable marker로서 plasnid속에 삽입하였다. 그 결과 생긴 구축 유전자는 각각 pCas 2.2와 pCas 0.5로 명명하였다. 구축된 유전자로 2$\times$107의 TT-2 ES세포를 170V, 550$\mu$F로 100$\mu$g의 선상 plasmid에 의해 electroporation 시켰다. 감염된 colony들은 250$\mu$g/$m\ell$ G418을 함유하는 ESM 배양액에서 선별 7일 이후에 회수하여 성공적으로 감염된 ES세포는 PCR 및 Southern blot에 의해 확인되었고 그들 중 나머지는 trypsin 처리 후 각각 미세조작과 공배양 기술을 사용하여 ICR 생쥐의 8세포기 수정란 속에 도입하였다. 결국 24시간 동안 37$^{\circ}C$, 5% $CO_2$에서 배양된 배반포를 chimera의 생산을 위해 위임신 유기된 G418 선발처리 이후 400 및 275개의 ES 세포 colony가 생존하였으며, 3개의 ES 세포으 colony 의 genome 속에 임의적으로 plamid가 삽입된 것을 Southern blot에 의해 확인되었다. 총 13 chimera 생쥐가 3 colony로부터 생산되었으나 germ-line chimera는 현재 조사중이다. chimera 생산빈도는 공배양 기술보다 주입방법에서 현저히 높았다.

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Chest Compression Energy(kg) Measurement of 4 Types of Rescue Device on the Floor (4종류의 구급장비 및 마루(Floor) 위에서 시행한 가슴압박 에너지(kg)측정 실험에 관한 연구)

  • Shin, Dong-Min;Han, Yong-Taek;Kim, Seung-Yong;Park, Si-Eun
    • Journal of the Korean Society of Safety
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    • v.29 no.6
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    • pp.125-131
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    • 2014
  • The purpose of this study is to investigate the effect of high quality CPR using the 4 types of rescue device equipment and chest compressions energy measurement in pre-hospital settings. So, we used the mode to insert load cell in ALS Skill master Manikin to develop CFMM(Compression Force Measurement Manikin) on main stretcher, CPR board, long spine board, scoop stretcher and floor. And, our research team could know that the main stretcher needed average force of 32.55 (${\pm}1.01$) kg, CPR board of 27.23 (${\pm}1.08$) kg, long spine board of 27.13 (${\pm}1.18$) kg, Scoop Stretcher of 27.38 (${\pm}1.05$) kg and Floor of 27.24 (${\pm}0.93$) kg. CPR board must be necessary in the case of CPR on main stretcher in a moving ambulance. But if the condition of patient's back surface is the removable stretcher and the long spine plate, the patient doesn't have to be spent time to use a CPR board. Furthermore, this research suggests to consider that how to take advantage of the education to students for the equipment to check in real time the energy(kg) requirement of chest compressions.

Biocidal Activity of Metal Nanoparticles Synthesized by Fusarium solani against Multidrug-Resistant Bacteria and Mycotoxigenic Fungi

  • Sayed, Manal T. El;El-Sayed, Ashraf S.A.
    • Journal of Microbiology and Biotechnology
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    • v.30 no.2
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    • pp.226-236
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    • 2020
  • Antibiotic resistance by pathogenic bacteria and fungi is one of the most serious global public health problems in the 21st century, directly affecting human health and lifestyle. Pseudomonas aeruginosa and Staphylococcus aureus with strong resistance to the common antibiotics have been isolated from Intensive Care Unit patients at Zagazig Hospital. Thus, in this study we assessed the biocidal activity of nanoparticles of silver, copper and zinc synthesized by Fusarium solani KJ 623702 against these multidrug resistant-bacteria. The synthesized Metal Nano-particles (MNPs) were characterized by UV-Vis spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and Zeta potential. The Fourier transform infrared spectroscopy (FTIR) result showed the presence of different functional groups such as carboxyl, amino and thiol, ester and peptide bonds in addition to glycosidic bonds that might stabilize the dispersity of MNPs from aggregation. The antimicrobial potential of MNPs by F. solani against the multidrug-resistant (MDR) P. aeruginosa and S. aureus in addition to the mycotoxigenic Aspergillus awamori, A. fumigatus and F. oxysporum was investigated, based on the visual growth by diameter of inhibition zone. Among the synthesized MNPs, the spherical AgNPs (13.70 nm) displayed significant effect against P. aeruginosa (Zone of Inhibition 22.4 mm and Minimum Inhibitory Concentration 21.33 ㎍/ml), while ZINC oxide Nano-Particles were the most effective against F. oxysporum (ZOI, 18.5 mm and MIC 24.7 ㎍/ml). Transmission Electron Microscope micrographs of AgNP-treated P. aeruginosa showed cracks and pits in the cell wall, with internalization of NPs. Production of pyocyanin pigment was significantly inhibited by AgNPs in a concentration-dependent manner, and at 5-20 ㎍ of AgNPs/ml, the pigment production was reduced by about 15-100%, respectively.

Characterization of Acetobacter pomorum KJY8 Isolated from Korean Traditional Vinegar

  • Baek, Chang Ho;Park, Eun-Hee;Baek, Seong Yeol;Jeong, Seok-Tae;Kim, Myoung-Dong;Kwon, Joong-Ho;Jeong, Yong-Jin;Yeo, Soo-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1679-1684
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    • 2014
  • Acetobacter sp. strains were isolated from traditional vinegar collected in Daegu city and Gyeongbuk province. The strain KJY8 showing a high acetic acid productivity was isolated and characterized by phenotypic, chemotaxonomic, and phylogenetic inference based on 16S rRNA sequence analysis. The chemotaxonomic and phylogenetic analyses revealed the isolate to be a strain of Acetobacter pomorum. The isolate showed a G+C content of 60.8 mol%. It contained $\small{LL}$-diaminopimelic acid ($\small{LL}$-$A_2pm$) as the cell wall amino acid and ubiquinone $Q_9$ (H6) as the major quinone. The predominant cellular fatty acids were $C_{18:1}w9c$, w12t, and w7c. Strain KJY8 grew rapidly on glucose-yeast extract (GYC) agar and formed pale white colonies with smooth to rough surfaces. The optimum cultivation conditions for acetic acid production by the KJY8 strain were $20^{\circ}C$ and pH 3.0, with an initial ethanol concentration of 9% (w/v) to produce an acetic acid concentration of 8% (w/v).