• Title/Summary/Keyword: M-sequence

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Detection of rpoB Gene Mutation in Rifampin-Resistant M. Tuberculosis by Oligonucleotide Chip (Oligonucleotide chip을 이용한 Rifampin 내성 결핵균의 rpoB 유전자 돌연변이 검출)

  • Park, Soon-Kew;Lee, Min-Ki;Chung, Byung-Seon;Kim, Cheol-Min;Chang, Chul-Hun L.;Park, Hee-Kyung;Jang, Hyun-Jung;Park, Seung-Kyu;Song, Sun-Dae
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.5
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    • pp.546-557
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    • 2000
  • Background : Oligonucleotide chip technology has proven to be a very useful tool in the rapid diagnosis of infectious disease. Rifampin resistance is considered as a useful marker of multidrug-resistance in tuberculosis. Mutations in the rpoB gene coding $\beta$ subunit of RNA polymerase represent the main mechanism of rifampin resistance. The purpose of this study was to develop a diagnosis kit using oligonucleotide chip for the rapid and accurate detection of rifampin-resistance in Mycobacterium tuberculosis. Method : The sequence specific probes for mutations in the rpoB gene were designed and spotted onto the glass slide, oligonucleotide chip. 38 clinical isolates of Mycobacterium were tested. A part of rpoB was amplified, labelled, and hybridized on the oligonucleotide chip with probes. Results were analyzed with a laser scanner. Direct sequencing was done to verify the results. Result : The low-density oligonucleotide chip design어 to determine the specific mutations in the rpoB gene of M. tuberculosis accurately detected rifampin resistance associated with mutations in 28 clinical isolates. Mutations at codons 531, 526, and 513 were confirmed by direct sequencing analysis. Conclusion : Mutant detection using oligonucleotide chip technology is a reliable and useful diagnostic tool for the detection of multidrug-resistance in M. tuberculosis.

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Isolation and Characterization of mas1+ of Schizosaccharomyces pombe, a Homologue of Human CIP29/Hcc-1 Involved in the Regulation of Cell Division (세포분열에 관여하는 인간의 CIP29/Hcc1 유전자와 상동성을 가지는 분열형 효모의 새로운 유전자 mas1+의 특성분석)

  • Cha, Jae-Young;Shin, Sang-Min;Ha, Se-Eun;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1666-1677
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    • 2011
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

A Case Study(II) on Development and Application of 'Literature-Art-Science' Integrated Education Programs ('문학-미술-과학' 융합교육 프로그램의 개발 및 적용 사례 연구(II))

  • Choi, Byung Kil
    • Korea Science and Art Forum
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    • v.32
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    • pp.319-334
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    • 2018
  • This research is a case study to make sure the enhancement of students' imagination and creativity through developing and applying the Literature-Art-Science Integrated Education Program. Its research object was totally 25 persons of 29 students of the 1st to the 4 th Grades from Gunsan Sulsan Elementary School. Its research period lasted for 4 months from September to December, 2017, and I, as the research place, used the art room at Gunsan Sulsan Elementary School. The programs were totally 10 sessions with a unit of 1 session per each grade for 2 hours from 1:00 to 3:00 in the afternoon from Monday through Friday. I fixed ten themes of this program-eight plane modeling, and two solid modeling, and finished the work of storytelling during summer vacation. And I arranged their levels as low:middle:high(3:5:2) ones. The former was 'A Film of Monster Gorilla'(L), 'Learning the Spirit of Gyeongju Choi's Family'(M), 'A Tale of My Friend Made of Natural Materials'(L), 'The Reading of My Dream'(M), 'Gathering the Objects in My Mobile'(M), 'A Mock Trial of Marrying Off'(M), 'Painting My Favorite Children's Poem'(H), and 'Painting My Favorite Children's Song'(H), and the latter was 'Seeking for a Bluebird in My Mind'(L), and 'Making My Cherished Object' (M). Then I used the unique art expression technique per each theme, which were in sequence marbling, Korean paper art, combine painting, collage, imaginary painting, imaginary painting, play dough art, imaginary painting techniques. And I delivered to the students the scientific knowledge in terms of growing or manufacturing processes of materials used for making artworks. Prior to and after the processing this program, I surveyed about the students' ability of integrated thinking and emotional experience by 'Figure B Type' and 'Figure A Type' of The Torrance Tests of Creative Thinking, and took statistics with the resultant data. And I executed a paired t-test in order to verify the significance of mean difference in the result of investigation with those data. From the analyzed result according to the elements of creativity and the mean quotients of creativity, there showed a significant difference (t=3.47, p<.01) in 'fluency', and also a significant difference(t=3.59, p<.01) in 'creativity.' Judging from the statistic values of two fields such as the student's ability of integrated thinking and emotional experience, I estimate that over the majority of the students showed the enhancement in self-confident creative expression as well as higher interest and concern through this program. The result that I arranged and analyzed the making process of artworks, the photos of the resultant, etc. as such is as follows : Firstly, from this program being proceeded as art-centered STEAM class, the student's systematic problem-solving ability was improved in his ability of integrated thinking to transform the literary contents into artistic one. Secondly, the student obtained the emotional experience such as interest in the class, self-confidence, intellectual satisfaction, self-fulfillment, etc. through art-centered STEAM class using ten art expression techniques. Thirdly, the student's mind willing to cooperate, communicate with his friends, and care for them was ripened in the process of problem-solving. Fourth, the student's self-confidence was further instilled when presenting famous artists and their artworks in the introduction and finale of ten art expression techniques. Likewise, the statistic values on the fields of student's ability of integrated thinking and emotional experience illustrate that over the majority of the students showed improvement in the ability of creative expression with confidence as well as higher interest and concern upon this program.

Keyhole Imaging Combined Phase Contrast MR Angiography Technique (Keyhole Imaging기법을 적용한 위상대조도 자기공명 혈관조영기법)

  • Lee, D.H.;Hong, C.P.;Han, B.S.;Lee, M.W.
    • Journal of Biomedical Engineering Research
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    • v.33 no.2
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    • pp.72-77
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    • 2012
  • Phase Contrast MR Angiography(PC MRA) is excellent MRA technique for measuring the velocity of vessels in the human body. PC MRA need to at least four images for angiogram reconstruction and it caused longer scan time. Therefore, we used keyhole imaging combined PC MRA to reduce the scan time. However, keyhole imaging can lead the erroneous effects as loss of phase information or frequency discontinuous. In this study, we applied the keyhole imaging combined 2D PC MRA for improving the temporal resolution and also measured the velocity to evaluate the accuracy of phase information. We used 0.32T MRI scanner(Magfinder II, Scimedix, Korea). Using the 2D PC MRA pulse sequence, the vascular images for a human brain targeted on the Superior Sagittal Sinus(SSS) were obtained. We applied tukey window function for keyhole images to minimize the ringing artifact and erroneous factors that are induced frequency discontinuous and phase information loss. We also applied zero-padded algorithm to peripheral missing k-space lines to compare keyhole imaging results and the artifact power(AP) value was measured on the complex difference images to validate the image quality. Consider as based on our results, heavy image distortions and artifacts were shown until using at least 50% keyhole factor. Using above the 50% keyhole factors are shown well reconstructed and matched for magnitude images and velocity information measurements. In conclusion, we confirmed the image quality and velocity information of keyhole technique combined 2D PC MRA. Especially, measured velocity information through the keyhole imaging combination was similar to the velocity information of full sampled k-space image despite of frequency discontinuous and phase information loss in the keyhole imaging reconstruction process. Consequently, the keyhole imaging combined 2D PC MRA will give some clinical usefulness and advantages as improving the temporal resolution and measuring the velocity information via selecting the appropriate keyhole factor at low tesla MRI system.

A New Sterol Regulatory Element Binding Protein, SrbB Is Critical for Hypoxia Adaptation and Virulence in the Human Fungal Pathogen Aspergillus fumigatus

  • Chung, Dawoon;Barker, Bridget M.;Carey, Charles C.;Merriman, Brittney;Werner, Ernst R.;Lechner, Beatrix E.;Dhingra, Sourabh;Cheng, Chao;Xu, Wenjie;Blosser, Sara J.;Morohashi, Kengo;Mazurie, Aurelien;Mitchell, Thomas K.;Haas, Hubertus;Mitchell, Aaron P.;Cramer, Robert A.
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.15-15
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    • 2015
  • Aspergillus fumigatus is a major cause of invasive aspergillosis (IA), a significant health issue worldwide with high mortality rates up to 95%. Our lab is interested in how A. fumigatus adapts to low oxygen conditions 'hypoxia', which is one of the important host microenvironments. A. fumigatus SrbA is a basic helix-loop-helix (bHLH) transcriptional regulator and belongs to sterol regulatory element binding protein (SREBP) family members. Loss of SrbA completely blocks growth in hypoxia and results in avirulence in murine models of IA suggesting an essential role of SrbA in hypoxia adaptation and virulence in A. fumigatus. We conducted chromatin immunoprecipitation sequencing (ChIP-seq) with A. fumigatus wild type using a SrbA specific antibody, and 97 genes were revealed as SrbA direct targets. One of the 'SrbA regulons' (AFUB_099590) was a putative bHLH transcriptional regulator whose sequence contained a characteristic tyrosine substitution in the basic portion of the bHLH domain of SREBPs. Therefore, we designated AFUB_099590 SrbB. Further characterization of SrbB demonstrated that SrbB is important for radial growth, biomass production, and biosynthesis of heme intermediates in hypoxia and virulence in A. fumigatus. A series of quantitative real time PCR showed that transcription of several SrbA regulons is coordinately regulated by two SREBPs, SrbA and SrbB in hypoxia. This suggests that SrbA and SrbB have both dependent and independent functions in regulation of genes responsible for hypoxia adaptation in A. fumigatus. Together, our data provide new insights into complicated roles of SREBPs in adaptation of host environments and virulence in pathogenic fungi.

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Molecular Genetic Identification of Yeast Strains Isolated from Egyptian Soils for Solubilization of Inorganic Phosphates and Growth Promotion of Corn Plants

  • Hesham, Abd El-Latif;Mohamed, Hashem M.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.55-61
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    • 2011
  • Forty yeast strains isolated from soils taken from different locations in Egypt were tested for their P-solubilizing activities on the basis of analyzing the clear zone around colonies growing on a tricalcium phosphate medium after incubation for 5 days at $25^{\circ}C$, denoted as the solubilization index (SI). Nine isolates that exhibited P-solubilization potential with an SI ranging from 1.19 to 2.76 were genetically characterized as five yeasts belonging to the genus Saccharomyces cerevisiae and four non-Saccharomyces, based on a PCR analysis of the ITS1-26S region amplied by SC1/SC2 species-specific primers. The highest P-solubilization efficiency was demonstrated by isolate PSY- 4, which was identified as Saccharomyces cerevisiae by a sequence analysis of the variable D1/D2 domain of the 26S rDNA. The effects of single and mixed inoculations with yeast PSY-4 and Bacillus polymyxa on the P-uptake and growth of corn were tested in a greenhouse experiment using different levels of a phosphorus chemical fertilizer (50, 100, and 200 kg/ha super phosphate 15.5% $P_2O_5$). The results showed that inoculating the corn with yeast PSY-4 or B. polymyxa caused significant increases in the shoot and root dry weights and P-uptake in the shoots and roots. The P-fertilization level also had a significant influence on the shoot and root dry weights and P-uptake in the shoots and roots when increasing the P-level from 50 up to 200 kg/ha. Dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 200 kg/ha gave higher values for the shoot and root dry weights and P-uptake in the shoots and roots, yet these increases were nonsignificant when compared with dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha. The best increases were obtained from dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha, which induced the following percentage increases in the shoot and root dry weights, and P-uptake in the shoots and roots; 16.22%, 46.92%, 10.09%, and 31.07%, respectively, when compared with the uninoculated control (fertilized with 100 kg/ha).

Characterization of Phosphoinositide-3-kinase, Class 3 (PIK3C3) Gene and Association Tests with Quantitative Traits in Pigs

  • Kim, J.H.;Choi, B.H.;Lim, H.T.;Park, E.W.;Lee, S.H.;Seo, B.Y.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1701-1707
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    • 2005
  • This study deals with the characterization of porcine PIK3C3 and association tests with quantitative traits. PIK3C3 belongs to the class 3 PI3Ks that participate in the regulation of hepatic glucose output, glycogen synthase, and antilipolysis in typical insulin target cells such as those in the such as liver, muscle system, and fat. On the analysis of full-length mRNA sequence, the length of the PIK3C3 CDS was recorded as 2,664 bps. As well, nucleotide and amino acid identities between human and pig subjects were 92% and 99%, respectively. Five SNPs were detected over 5 exons. We performed genotyping by using a SNP C2604T on exon24 for 145 F$_2$ animals (from a cross between Korean native boars and Landrace sows) by PCR-RFLP analysis with Hpy8I used to investigate the relationship between growth and fat depot traits. In the total association analysis, which doesn' consider transmission disequilibrium, the SNP showed a significant effect (p<0.05) on body weight and carcass fat at 30 weeks of age as well as a highly significant effect (p<0.01) on back fat. In an additional sib-pair analysis, C allele still showed positive and significant effects (p<0.05) on back fat thickness and carcass fat. Moreover, the effects of C allele on the means of within-family components for carcass fat and back fat were estimated as 2.76 kg and 5.07 mm, respectively. As a result, the SNP of porcine PIK3C3 discovered in this study could be utilized as a possible genetic marker for the selection of pigs that possess low levels of back fat and carcass fat at the slaughter weight.

Metagenomic analysis of bacterial community structure and diversity of lignocellulolytic bacteria in Vietnamese native goat rumen

  • Do, Thi Huyen;Dao, Trong Khoa;Nguyen, Khanh Hoang Viet;Le, Ngoc Giang;Nguyen, Thi Mai Phuong;Le, Tung Lam;Phung, Thu Nguyet;Straalen, Nico M. van;Roelofs, Dick;Truong, Nam Hai
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.5
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    • pp.738-747
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    • 2018
  • Objective: In a previous study, analysis of Illumina sequenced metagenomic DNA data of bacteria in Vietnamese goats' rumen showed a high diversity of putative lignocellulolytic genes. In this study, taxonomy speculation of microbial community and lignocellulolytic bacteria population in the rumen was conducted to elucidate a role of bacterial structure for effective degradation of plant materials. Methods: The metagenomic data had been subjected into Basic Local Alignment Search Tool (BLASTX) algorithm and the National Center for Biotechnology Information non-redundant sequence database. Here the BLASTX hits were further processed by the Metagenome Analyzer program to statistically analyze the abundance of taxa. Results: Microbial community in the rumen is defined by dominance of Bacteroidetes compared to Firmicutes. The ratio of Firmicutes versus Bacteroidetes was 0.36:1. An abundance of Synergistetes was uniquely identified in the goat microbiome may be formed by host genotype. With regard to bacterial lignocellulose degraders, the ratio of lignocellulolytic genes affiliated with Firmicutes compared to the genes linked to Bacteroidetes was 0.11:1, in which the genes encoding putative hemicellulases, carbohydrate esterases, polysaccharide lyases originated from Bacteroidetes were 14 to 20 times higher than from Firmicutes. Firmicutes seem to possess more cellulose hydrolysis capacity showing a Firmicutes/Bacteroidetes ratio of 0.35:1. Analysis of lignocellulolytic potential degraders shows that four species belonged to Bacteroidetes phylum, while two species belonged to Firmicutes phylum harbouring at least 12 different catalytic domains for all lignocellulose pretreatment, cellulose, as well as hemicellulose saccharification. Conclusion: Based on these findings, we speculate that increasing the members of Bacteroidetes to keep a low ratio of Firmicutes versus Bacteroidetes in goat rumen has resulted most likely in an increased lignocellulose digestion.

Synthesis and Characterization of Tetradentate N2O2 Schiff Base Ligand and its Rare Earth Metal Complexes (사배위 N2O2 Schiff 염기 리간드와 그 희토류 금속착물의 합성 및 특성)

  • Shelke, Vinod A.;Jadhav, Sarika M.;Shankarwar, Sunil G.;Munde, chut S.;Chondhekar, Trimbak K.
    • Journal of the Korean Chemical Society
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    • v.55 no.3
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    • pp.436-443
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    • 2011
  • The solid complexes of La(III), Ce(III), Pr(III), Nd(III), Sm(III) and Gd(III) with 4-hydroxy-3-(1-{2-(2,4-dihydroxy-benzylidene)-amino phenylimino}-ethyl)-6-methyl-pyran-2-one ($H_2$L) derived from o-phenylenediamine, 3-acetyl-6-methyl-(2H)pyran,2,4(3H)-dione (dehydroacetic acid or DHA) and 2, 4-dihydroxy benzaldehyde have been synthesized and characterized by elemental analysis, conductometry, magnetic susceptibility, UV-visible, FT-IR, $^1H$-NMR, X-ray diffraction, thermal analysis study, and screened for antimicrobial activity. The FT-IR spectral data suggest that the ligand behaves as a dibasic tetradentate ligand with ONNO donor atoms sequence towards central metal ion. From the microanalytical data, the stoichiometry of the complexes has been found to be 1:1 (metal: ligand). The physico-chemical data suggests distorted octahedral geometry for La(III), Ce(III), Pr(III), Nd(III), Sm(III) and Gd(III) complexes. The X-ray diffraction data suggests monoclinic crystal system for La(III) and Ce(III) and orthorombic crystal system for Pr(III) and Nd(III) complexes. Thermal behavior (TG/DTA) of the complexes was studied and kinetic parameters were determined by Horowitz-Metzger and Coats-Redfern methods. The ligand and its metal complexes were screened for antibacterial activity against Staphylococcus aureus, Escherichia coli, Bacillus Sp. and fungicidal activity against Aspergillus Niger, Trichoderma and Fusarium oxysporum.

Synthesis and Characterization of Some Transition Metal Complexes of Unsymmetrical Tetradentate Schiff Base Ligand (비대칭 Tetradentate Schiff 염기 리간드의 전이금속 착물에 대한 합성 및 특성)

  • Munde, A. S.;Jagdale, A. N.;Jadhav, S. M.;Chondhekar, T. K.
    • Journal of the Korean Chemical Society
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    • v.53 no.4
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    • pp.407-414
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    • 2009
  • The solid complexes of Cu(II), Ni(II), Co(II), Mn(II) and Fe(III) with 4-hydroxy-3-(1-{2-(2-hydroxybenzylidene)- amino phenylimino}-ethyl)-6-methy-pyran-2-one (H2L) derived from o-phenylenediamine, 3-acetyl- 6-methyl-(2H) pyran, 2,4 (3H)-dione (dehydroacetic acid or DHA) and salicylic aldehyde have been synthesized and characterized by elemental analysis, conductometry, magnetic susceptibility, UV-visible, IR, $^1H$-NMR spectra, X-ray diffraction, thermal analysis, and screened for antimicrobial activity. The IR spectral data suggest that the ligand behaves as a dibasic tetradentate ligand with ONNO donor atoms sequence towards central metal ion. From the microanalytical data, the stoichiometry of the complexes has been found to be 1:1 (metal: ligand). The physico-chemical data suggests square planar geometry for Cu(II) and Ni(II) complexes and octahedral geometry for Co(II), Mn(II) and Fe(III) complexes. The x-ray differaction data suggests orthorhombic crystal system for Cu(II) complex, monoclinic crystal system for Ni(II), Co(II) and Fe(III) and tetragonal crystal system for Mn(II) complex. Thermal behaviour (TG/DTA) of the complexes was studied and kinetic parameters were determined by Coats-Redfern method. The ligand and their metal complexes were screened for antibacterial activity against Staphylococcus aureus and Escherichia coli and fungicidal activity against Aspergillus Niger and Trichoderma.