• Title/Summary/Keyword: merA

Search Result 239, Processing Time 0.023 seconds

Generation of single stranded DNA with selective affinity to bovine spermatozoa

  • Vinod, Sivadasan Pathiyil;Vignesh, Rajamani;Priyanka, Mani;Tirumurugaan, Krishnaswamy Gopalan;Sivaselvam, Salem Nagalingam;Raj, Gopal Dhinakar
    • Animal Bioscience
    • /
    • v.34 no.10
    • /
    • pp.1579-1589
    • /
    • 2021
  • Objective: This study was conducted to generate single stranded DNA oligonucleotides with selective affinity to bovine spermatozoa, assess its binding potential and explore its potential utility in trapping spermatozoa from suspensions. Methods: A combinatorial library of 94 mer long oligonucleotide was used for systematic evolution of ligands by exponential enrichment (SELEX) with bovine spermatozoa. The amplicons from sixth and seventh rounds of SELEX were sequenced, and the reads were clustered employing cluster database at high identity with tolerance (CD-HIT) and FASTAptamer. The enriched nucleotides were predicted for secondary structures by Mfold, motifs by Multiple Em for Motif Elicitation and 5' labelled with biotin/6-FAM to determine the binding potential and binding pattern. Results: We generated 14.1 and 17.7 million reads from sixth and seventh rounds of SELEX respectively to bovine spermatozoa. The CD-HIT clustered 78,098 and 21,196 reads in the top ten clusters and FASTAptamer identified 2,195 and 4,405 unique sequences in the top three clusters from the sixth and seventh rounds, respectively. The identified oligonucleotides formed secondary structures with delta G values between -1.17 to -26.18 kcal/mol indicating varied stability. Confocal imaging with the oligonucleotides from the seventh round revealed different patterns of binding to bovine spermatozoa (fluorescence of the whole head, spot of fluorescence in head and mid- piece and tail). Use of a 5'-biotin tagged oligonucleotide from the sixth round at 100 pmol with 4×106 spermatozoa could trap almost 80% from the suspension. Conclusion: The binding patterns and ability of the identified oligonucleotides confirms successful optimization of the SELEX process and generation of aptamers to bovine spermatozoa. These oligonucleotides provide a quick approach for selective capture of spermatozoa from complex samples. Future SELEX rounds with X- or Y- enriched sperm suspension will be used to generate oligonucleotides that bind to spermatozoa of a specific sex type.

Phylogenetic relationships of genera Grifola on the basis of ITS region sequences (rDNA의 ITS 부위 염기서열 분석에 의한 잎새버섯(Grifola)속 균주의 유전적인 유연관계 분석)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Kong, Won-Sik;Suh, Jang-Sun
    • Journal of Mushroom
    • /
    • v.10 no.2
    • /
    • pp.93-99
    • /
    • 2012
  • This study was carried to identify a correct species and asses genetic diversity within the same species of Grifola spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Grifola spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Grifola clustered into one group, most of which correlated with species-groups identified by RAPD method. Eight isolates included strain GM01 showed high similarity with Grifola frondosa. All isolates were collected in the Japan(GM01, GM02, GM03) was identified as Grifola frondosa and isolates of the China(GM05, GM06, GM08) was identified as Bjerkandera fumosa, Grifola frondosa and Dichomitus squalens, respectively. RAPD analysis of genetic polymorphisms of genus Grifola showed a very different band patterns on the isolat. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 4 isolates of Grifola spp. may be need to reclassify or eliminate from preserved catalogue.

Study on Electroluminescence of the Phosphorescent Iridium(III) Complex Prepared by Ultrasonic Wave (초음파 합성법을 이용한 이리듐계 인광 물질 합성과 합성된 인광 물질의 전계 발광 특성 분석)

  • Yu, Hong-Jeong;Chung, Won-Keun;Chun, Byung-Hee;Kim, Sung-Hyun
    • Korean Chemical Engineering Research
    • /
    • v.49 no.3
    • /
    • pp.325-329
    • /
    • 2011
  • $Ir(pmb)_{3}$(Iridium(III)Tri(1-phenyl-3-methylbenzimidazolin-2-ylidene-$C,C^{2'}$ ) was synthesized to develop a deep blue-emitting Ir(III) complex. We suggested the ultrasonic reactor to enhance the poor reaction yield of $Ir(pmb)_{3}$. The ultrasonic wave enhanced the reaction yield of $Ir(pmb)_{3}$ because the ultrasound helped non-soluble reactants disperse efficiently and produced free radial during the reaction. The maximum yield of $Ir(pmb)_{3}$ was 42.5%, which was 4 times higher than conventional method. Organic light emitting devices were fabricated with the synthesized mer-$Ir(pmb)_{3}$ which emitted at 405 nm. A range of host materials with large bandgaps (UGH2, mCP and CBP) were tested for developing a deep blue emitting device. In case of the device with mCP as the host material, it emitted deep blue and performed quite well relative to the other host materials tested.

Phylogenetic relationships of genera Polyporus on the basis of ITS region sequences (rDNA의 ITS 부위 염기서열 분석에 의한 겨울우산버섯(Polyporus)속 균주의 유전적인 유연관계 분석)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Kong, Won S.
    • Journal of Mushroom
    • /
    • v.10 no.1
    • /
    • pp.37-43
    • /
    • 2012
  • This study was carried to identify a correct species and asses genetic diversity within the same species of Polyporus spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Polyporus spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that three strains were different species and four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Polyporus clustered into five distinct group, most of which correlated with species-groups identified by RAPD method. Four isolates included strain PM02 showed high similarity with P. arcularius, four isolates included strain PM03 high similarity with P. alveolaris, three isolates included strain PM01 high similarity with P. tuberaster, and PM 06 and PM04 high similarity with P. brumalis and P. squamossus. Isolates were collected in the United States(PM10, PM11) was identified as P. alveolarius and P. arcularius. RAPD analysis of genetic polymorphisms of genus Polyporus showed a very different band patterns. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 6 isolates of Polyporus spp. may be need to reclassify or eliminate from preserved catalogue.

Diagnosis of fibroadenoma using radiolabeled bladder cancer specific peptide

  • Ha, Yeong Su;Kwak, Wonjung;Lee, Hwa Young;Lee, Byung-Heon;Hong, Il-Hwa;Jeong, Kyu-Shik;An, Gwang Il;Yoo, Jeongsoo
    • Journal of Radiopharmaceuticals and Molecular Probes
    • /
    • v.1 no.1
    • /
    • pp.38-45
    • /
    • 2015
  • Most tumors are believed to overexpress several receptors, and small peptides targeting these receptors were developed for diagnosis and tumor therapy during past decade. Here we report that fibroadenoma can be visualized by bladder cancer specific peptide. A 9-mer bladder cancer specific peptide, which was discovered from the phage display method, was synthesized by peptide synthesizer, and additional tyrosine was conjugated at the N-terminal for radioiodination (Y-BP). Y-BP was radiolabeled with $^{131/124}I$ using Iodogen tube. The rat treated with N-butyl-N-(4-hydroxybutyl)nitrosamine for 8 weeks was allowed to grow until large size tumor was developed under axilla. The tumor model was microPET imaged sequentially using [$^{18}F$]FDG and radioiodinated $^{124}I-Y-BP$. The tumor was excised and examined by immunostaining studies. Radioiodinated $^{124}I-Y-BP$ was purified using fast protein liquid chromatography (FPLC) in > 90% radiochemical purity. The whole tumor was well visualized by [$^{18}F$]FDG with several intense focal uptake within tumor. The tumor was also clearly seen with $^{124}I-Y-BP$ at 4 h post-injection, and to our surprise the tumor uptake of $^{124}I-Y-BP$ lasted up to three days. The tumor was diagnosed histologically as a fibroadenoma derived from mammary gland. In conclusion, the bladder cancer specific peptide showed the good potential as a new radiotracer for the detection of breast fibroadenoma.

$\acute{E}$tude du Processus de Morphogen$\grave{e}$se de l'$\hat{I}$le Rocheuse de Baek dans la Ville de Yeosu en Cor$\acute{e}$edu Sud (여수시 백도의 지형형성과정에 대한 고찰)

  • Lee, Jeong Hun
    • Journal of the Korean association of regional geographers
    • /
    • v.19 no.4
    • /
    • pp.627-640
    • /
    • 2013
  • Cette $\acute{e}$tude a pour objet d'analyser le processus de morphogen$\grave{e}$se de l'$\hat{I}$le rocheuse de Baek. Nous y voyons une cl$\acute{e}$ pour apprendre son relief marin et le processus de morphogen$\grave{e}$se des l'$\hat{I}$les m$\acute{e}$ridionales de Cor$\acute{e}$e du Sud. Le granit porphorique qui compose l'$\hat{I}$le rocheuse de Baek est une roche magmatique qui s'est form$\acute{e}$e il y a 60 million d'ann$\acute{e}$es. La cause principale de formation de l'$\hat{I}$le rocheuse de Baek, est une ligne de d$\acute{e}$lit vers le NE-SO et l'ENE-OSO, un soul$\grave{e}$vement de la plaque tectonique et une $\acute{e}$rosion par les vagues. L'$\hat{I}$le rocheuse de Baek pr$\acute{e}$sente un caract$\grave{e}$re d'$\acute{e}$ruption de magma de calc-alcalin par analyse g$\acute{e}$ochimique de son granit porphorique et fait partie du granit de l'arc volcanique. Il s'agit d'un magma qui s'est form$\acute{e}$ dans la subduction pr$\grave{e}$s du continent. Il est aussi n$\acute{e}$ssaire d'examiner un soul$\grave{e}$vement qui est plus $\acute{e}$lev$\acute{e}$ qu' un mouvement ascendant de la surface de la mer $\grave{a}$ l'$\grave{e}$re quaternaire environ de l'$\hat{I}$le rocheuse de Baek malgr$\acute{e}$ que, selon nous, nous y trouvions une faille et une terrasse marine.

  • PDF

Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
    • /
    • v.24 no.4
    • /
    • pp.115-120
    • /
    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan (베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발)

  • Kim, Su Cheol;Kim, Hye Soo;Cho, Yong Un;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
    • /
    • v.13 no.1
    • /
    • pp.79-83
    • /
    • 2015
  • In this study, SCAR marker that differentiates Pleurotus eryngii strains with higher ${\beta}$-glucan from control strain was developed. Genomic DNAs of 9 control strains of Pleurotus eryngii and 9 Pleurotus eryngii strains with higher ${\beta}$-glucan were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). One-hundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 91 bp was yielded by OP-R03 primer from the Pleurotus eryngii strains with higher ${\beta}$-glucan. A sequence characterized amplified region (SCAR) marker, designated as OP-R03-1-F and OP-R03-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-R03-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains with higher ${\beta}$-glucan from the control strains.

The Effects of Education for Women's Health on the Performance of Health Promoting Behavior and Self Esteem of College Women (여성건강교육이 여대생의 건강증진행위와 자아존중감에 미치는 영향)

  • Sin, Hye-Suk;Kim, Yeong-Hui
    • The Korean Nurse
    • /
    • v.37 no.4
    • /
    • pp.72-84
    • /
    • 1998
  • This study has been done for the purpose of testing the effects of education for women's health on the performance of health promoting behavior and self esteem of college women. The study was designed as simulated control group pretest-posttest design, the experimental group was composed 182 college women and the control group was composed 151 college women at D University in Seoul. The instruments used for this study were Health Promoting Behavior Scale developed by Walker et al(1987)were modified by Shin(1997) and Self Esteem Scale developed by Rosenberg(1965). The data were analyzed by chi-square test, t-test and ANCOVA using $SPSSIPC^+$ program. The results were as follows: 1. The result of the homogeneity test in terms of the demographic characteristics of two group showed that there were significant difference in major discipline($x^2=155.854$, p=.000), religion($x^2=6.325$, p=.011), and disease experience($x^2=3.949$, p=.046). 2. Hypotheses 1 that the college women who get women's health education will have a higher score of health promoting behavior than the college women who do not get women's health education was supported. The score of health promoting behavior between two group showed statistically difference(t=7.25, p= .000). Mer control of covariates(major discipline, religion, and disease experience), the score of health promoting behavior between two group showed statistically difference(F=31.817, p= .000). 3. Hypotheses 2 that the college women who get women's health education will have a higher score of self esteem than the college women who do not get women's health education was supported. The score of self esteem between two group showed statistically difference(t=4.91, p= .000). After control of covariates(major discipline, religion, disease experience), the score of self esteem between two group showed statistically difference(F=12.688, p= .000). The following suggestions are made based on the above results : 1) Replication of the research is needed to confirm effects of health education including the college man and various demographic differences. 2) More effective health education programs appropriate to subject need to be developed. 3) Nursing college or departments of nursing should make an effort to develop and carry out various health education program for health for all.

  • PDF

Percutaneous Absorption of Antisense Phosphorothioate Oligonucleotide in vitro

  • Lee, Young-Mi;Song, Kyung;Lee, Sung-Hee;Ko, Geon-Il;Kim, Jae-Baek;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
    • /
    • v.19 no.2
    • /
    • pp.116-121
    • /
    • 1996
  • Antisense oligonucleotides seem to provide a promising new tool for the therapy. Choi et al. (1995) reported antisense phosphorothioate oligonucleotides (PS-ODN, 25 mer) complementary to TGF-.betha. mRNA designed for scar formation inhibitor to eliminate scars, which was caused by undesired collagen deposition due to overexpression of TGF-.betha., in wounded skin. PS-ODN were evaluated in vitro for skin penetration using normal and tape-stripped damaged rat skin. The in vitro skin transports were carried out with partially modified PS-ODN (6S) and fully modified PS-ODN (25S). The cumulative amount of PS-ODN (6S) penetrated through normal rat skin was $0.234{\pm}0.041{\mu}g/cm^2$ and that of tape-stripped damaged rat skin was $1.077{\pm}0.301{\mu}g/cm^2$ over 8 hrs. PS-ODN (25S) can not be found in receptor medium through normal skin due to high molecular weight (Mol.Wt.=8,000) and polyanionic charge. However, the cumulative amount of PS-ODN (25S) penetrated across damaged rat skin in PBS was $0.340{\pm}0.296{\mu}g/cm^2$ over 8 hrs. The absense of dermis raised the cumulative amount of PS-ODN (6S) penetrated through rat skin. And the fluxes of PS-ODN (6S) and PSODN (25S) at 8hrs across damaged rat skin were $134.63{\pm}37.67{\mu}g/cm^2$ h, and $42.50{\pm}36.95ng/cm^2$ h, respectively. While PS-ODN (25S) was stable in 10% heat inactivated fetal bovine serum (FBS) during 24 hrs, PS-ODN (6S) was less stable than PS-ODN (25S), but was markedly stable than unmodified phosphodiester. It is suggested that the cumulative amount of PS-ODN (6S) penetrated through damaged rat skin is larger than that of PS-ODN (25S) since the former is easier to degrade by nuclease than the latter and then is apt to penetrate into skin. Thus, PS-ODN represents a logical candidate for further evaluation due to the potential for delivery into the wounded skin.

  • PDF