Structural analysis of enniatin H, I, and MK1688 and beauvericin by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and their production by Fusarium oxysporum KFCC 11363P

Song, H. -H.,Lee, H. -S.,Lee, G. -P.,Ha, S. -D.,Lee, C. Taylor Francis 2009 Food additives & contaminants. Part A. Chemistry, Vol.26 No.4

<P> The molecular structures of enniatins H, I, and MK1688 and beauvericin were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). MS fragmentation occurred by loss of -CO after opening of the cyclic molecule to carbonyl carbon, and cleavage of the peptide and ester bonds in the molecular structure. Fusarium oxysporum KFCC 11363P was tested for its ability to produce beauvericin and enniatins H, I, and MK1688 on five cereal substrates: rice, barley, maize, wheat, and Indian millet kernels. Furthermore, optimal conditions for the production of the four mycotoxins by the Fusarium isolate were examined on maize at four temperatures (15, 20, 25, and 30°C) and at three moisture contents (10, 20, and 40%). Large amounts of beauvericin and enniatin H were present in maize cultures at 25°C (232.4 and 196.4 µg g-1, respectively). Enniatins I and MK1688 were maximally formed at 20°C (221.5 and 180.2 µg g-1, respectively). The optimal moisture contents for the production of enniatins H (196.4 µg g-1) and MK1688 (165.6 µg g-1), were 40%.</P>

Lactobacillus helveticus CU 631 에 의한 Helicobacter pylon 의 Urease 및 공포 생성 독소 억제활성

송의한,원병렬,윤영호,강경희,장명웅 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.6

표준균주 혹은 유제품으로부터 분리된 Lactobacillus spp.와 Bifidobacterium spp. 32균주를 사용하여 H. pylori 생장을 현저하게 억제하는 L. helveticus CU631을 선발하고, urease와 공포생성 독소의 활성을 억제하는 효과를 측정하여 다음의 결과를 얻었다. L. helveticus CU631의 억제대의 직경이 10.0±1.5㎜ 나타내어 가장 강력한 생장 억제 능력을 보였으며 L. plantarum과 L. fermentum은 직경 4.0㎜ 내외의 억제대를 나타내어 비교적 약한 억제 활성을 보였으며 Bifidobacterium spp.에서 억제 활성을 보이지 않았다. L. helveticus CU631의 배양액과 배양 상층액 모두, H. pylori NCTC11637의 urese 억제 활성을 나타내었다. L. helveticus CU631를 H. pylori G88016를 같이 배양했을시 공포생성 독소의 역가가 50%로 감소하였으며 L. helvesticus CU631의 배양 상층액과 H. pylori G88016의 배양 상층액을 5:5와 6:4 비율로 혼합하였을 때 억제 활성이 나타났다. The inhibitory effects of 32 strains of lactobacilli against Helicobacter. pylori were determined and Lactobacillus. helveticus CU631 has been selected as the strain which possessed the strongest inhibitory effect against H. pylori NCTC11637 in inhibition zone test showing inhibition zone with the average diameter of 10±1.5㎜, whereas Lactobacillus. plantarum and L. fermentum made inhibition zone with the average diameter of 4.0㎜, H. pylori G88016 revealed the highest vacuolating toxin activity among the 8 strains of H. pylori, which showed positive reaction of vacuolating toxin gene in PCR amplification test. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activity on the urease activity of H. pylori NCTC11637. The inhibitory activity of L. helveticus CU631 on the vacuolating toxin activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains.

Neopterin Levels and Indoleamine 2,3-Dioxygenase Activity as Biomarkers of Immune System Activation and Childhood Allergic Diseases

Songül Ünüvar,Duygu Erge,,Bilge Kılıçarslan,,Harika Gözde Gözükara Bağ,Ferhat Çatal,,Gözde Girgin,,Terken Baydar, 대한진단검사의학회 2019 Annals of Laboratory Medicine Vol.39 No.3

Background: Although Th2 immune activation is predominant in allergic diseases, neopterinlevels and indoleamine 2,3-dioxygenase (IDO)-1 activity (kynurenine:tryptophan ratio), which reflect Th1 immune activity, increase with interferon-gamma (IFN-γ) stimulation. We investigated neopterin, tryptophan, and kynurenine levels as biomarkersof the Th1 immune system activation and changes in IDO-1 activityin children with asthma, allergic rhinitis, and atopic dermatitis, as well as the relationship between these biomarkers and the total IgE level, age, and disease severity. Methods: We divided 205 children (80 girls and 125 boys, four months to 17 years old) into four groups: controls, patients with asthma, patients with allergic rhinitis, and patients with atopic dermatitis. Peripheral venous blood samples were collected. Neopterin levels were determined by an enzyme immunoassay. Tryptophan and kynurenine levels were analyzed using HPLC. IDO-1 enzyme activity was calculated using tryptophan and kynurenine levels. IgE levels were measured. The Mann–Whitney U test, Kruskal–Wallis test, and Conover post-hoc method were used for statistical analysis. Results: Neopterin, tryptophan, and kynurenine levels were higher and IgE levels and IDO-1 enzyme activity were lower in patients with asthma and allergic rhinitis than in controls (P<0.05). Patients with atopic dermatitis showed higher neopterin, tryptophan, and kynurenine levels, higher IDO-1 activity, and lower IgE levels thancontrols (P<0.05). Conclusions: The Th1/Th2 balance is disrupted in children with allergic diseases, concomitant with increased Th1-mediated immune response activation and reduced IgEproduction, which is promoted by Th2-type cytokines.

PEGylation and HAylation via catechol: α-Amine-specific reaction at N-terminus of peptides and proteins

Song, I.T.,Lee, M.,Lee, H.,Han, J.,Jang, J.H.,Lee, M.S.,Koh, G.Y.,Lee, H. Elsevier BV 2016 ACTA BIOMATERIALIA Vol.43 No.-

The development of chemoselective, site-specific chemistries for proteins/peptides is essential for biochemistry, pharmaceutical chemistry, and other fields. In this work, we found that catechol, which has been extensively utilized as an adhesive molecule for material-independent surface chemistry and as a crosslinker in hydrogel preparation, specifically reacts with N-terminal α-amines, avoiding the ε-amine group in lysine. A conjugate of methoxy-poly(ethylene glycol)-catechol called mPEG-cat chemoselectively reacts with N-terminal amine groups at neutral pH resulting in site-specific PEGylation. To demonstrate the versatility of this catechol chemoselective reaction, we used four proteins (lysozyme, basic-fibroblast growth factor (bFGF), granulocyte-colony stimulating factor (G-CSF), insulin, and erythropoietin (EPO)) as well as two peptides (hinge-3 and laminin-derived peptide (LDP)). All the tested macromolecules showed N-terminal site-specific modifications. Furthermore, we prepared another catechol grafted conjugate called hyaluronic acid-catechol (HA-cat) to demonstrate that this catechol-involved chemoselective chemistry is not specific for PEG conjugates. This new catechol chemoselective chemistry could be a new platform for the functionalization of proteins and peptides for a variety of purposes. Statement of Significance: Considering the fact that biological activities of proteins or peptides depend largely on their 3-dimensional conformation, the orientation-controllable reaction is very important for preserving the intrinsic functionality of them. In addition to PEG, many other bio-polymers such as oligonucleotides, antibodies, and oligosaccharides have been conjugated with proteins or peptides for various biomedical applications. Although several chemoselective conjugation chemistries have been reported, conjugation efficiencies are different depending on types of proteins or polymers, and thus there've been strong needs for the development of alternative strategy of chemoselective conjugation that can be applied for a variety of therapeutic proteins towards high biological activities. We are certain this new catechol chemoselective chemistry could be a new platform for the functionalization of proteins and peptides for various purposes.

Characteristics of cefazolin inoculum effect-positive methicillin-susceptible staphylococcus aureus infection in a multicentre bacteraemia cohort

Song, K. H.,Jung, S. I.,Lee, S.,Park, S.,Kiem, S. M.,Lee, S. H.,Kwak, Y. G.,Kim, Y. K.,Jang, H. C.,Kim, Y. S. Springer Science + Business Media 2017 European journal of clinical microbiology & infect Vol.36 No.2

<P>Cefazolin treatment failure has been observed in high-inoculum infections caused by methicillin-susceptible Staphylococcus aureus (MSSA) with a cefazolin inoculum effect (CIE). However, data on the characteristics and risk factors for the acquisition of CIE-positive MSSA infection are scarce. CIE positivity was measured as an MIC a 16 mu g/ml with a high inoculum (similar to 5 x 10(7) CFU/ml). The blaZ gene type was assessed through sequence analysis. The clinical characteristics and risk factors for the acquisition of CIE-positive MSSA infection were assessed. The association between the antimicrobial susceptibility profile and CIE positivity was evaluated. A total of 303 MSSA bacteraemia cases and their corresponding isolates were collected from ten hospitals: 61 (20.1 %) isolates showed a positive CIE; 254 (83.8 %) were positive for the blaZ gene. No significant association was found between CIE positivity and the site of infection. Metastatic cancer (aOR 2.86, 95 % CI, 1.10-7.48) and recent (a 1 month) close contact with a chronically ill patient (aOR 4.69, 95 % CI, 1.76-12.50) were identified as significant risk factors for CIE-positive MSSA infection through multivariate analyses. Resistances to clindamycin (OR 3.55, 95 % CI, 1.62-7.80) and erythromycin (OR 5.00, 95 % CI, 2.50-9.99) were associated with CIE positivity, presenting high specificity (92.9 %) and a negative predictive value (82.3 %). CIE-positive MSSA constituted approximately one-fifth of MSSA bacteraemia cases. Although CIE positivity was not clinically discernible, CIE positivity was associated with clindamycin or erythromycin susceptibility. Therefore, our findings suggest that cefazolin can be used in the treatment of high-inoculum MSSA infection if the isolates are susceptible to clindamycin or erythromycin.</P>

수핵란의 전활성화가 토끼 핵이식 수정란의 체외발달에 미치는 효과

전병균,송상현,정기화,곽대오,이효종,최상용,박충생 경상대학교 유전공학연구소 1997 遺傳工學硏究所報 Vol.16 No.-

To examine the efficiency of nuclear transplantation the influence of electrical preactivation of recipient cytoplasm on the in vitro developmental potentyl in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G₁phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgery procedure. The separated G₁phase blastomeres of 32-cell stage were out into the non-preactivated and/or the preactivated recipient cytoplasm by electrical stimulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused. The fused nuclear transplant embryos were co-cultured with rabbit oviduct for 120h, the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 and their blastomere were counted. The electrofision rate was similar to the non-preactivated and preactivated recipient cytoplasm(81.8 and 85.7%, respectively). However, the in vitro developmental rate to blastocyst stage with the non-preactivated recipient cytoplasm(163.7cells), as compared with the preactivated recipient cytoplasm(85.4 cells). These results considered better that non-preactivated oocytes, MⅡ phase oocytes, were used for recipient cytoplasms in the rabbit nuclear transplant procedure.

High-level extracellular production and characterization of Candida antarctica lipase B in Pichia pastoris

Eom, G.T.,Lee, S.H.,Song, B.K.,Chung, K.W.,Kim, Y.W.,Song, J.K. Society for Bioscience and Bioengineering, Japan ; 2013 Journal of bioscience and bioengineering Vol.116 No.2

The gene encoding lipase B from Candida antarctica (CalB) was expressed in Pichia pastoris after it was synthesized by the recursive PCR and cloned into the Pichia expression plasmid, pPICZαA. The CalB was successfully secreted in the recombinant P. pastoris strain X-33 with an apparent molecular weight of 34 kDa. For 140 h flask culture, the dry cell weight and the extracellular lipase activity reached at 5.4 g/l and 57.9 U/l toward p-nitrophenyl palmitate, respectively. When we performed the fed-batch fermentation using a methanol feeding strategy for 110 h, the dry cell weight and the extracellular lipase activity were increased to 135.7 g/l and 11,900 U/l; the CalB protein concentration was 1.18 g/l of culture supernatant. The characteristics of CalB recovered from the P. pastoris culture were compared with the commercial form of CalB produced in Aspergillus oryzae. The kinetic constants and specific activity, the effects of activity and stability on temperature and pH, the glycosylation extent, the degree of immobilization on macroporous resin and the yield of esterification reaction between oleic acid and n-butanol were almost identical to each other. Therefore, we successfully proved that the Pichia-based expression system for CalB in this study was industrially promising compared with one of the most efficient production systems.

신경내분비분화를 보인 소아 부신피질종양-1예 보고-

송상용,이승숙,명나혜,지제근,송상용,이승숙,명나혜,지제근 대한병리학회 1993 Journal of Pathology and Translational Medicine Vol.27 No.2

Gallotannin mediated silver colloidal nanoparticles as multifunctional nano platform: Rapid colorimetric and turn-on fluorescent sensor for Hg²⁺, catalytic and <i>In vitro</i> anticancer activities

Mahajan, Prasad G.,Dige, Nilam C.,Vanjare, Balasaheb D.,Phull, Abdul Rehman,Kim, Song Ja,Lee, Ki Hwan Elsevier 2019 Journal of luminescence Vol.206 No.-

<P><B>Abstract</B></P> <P>A simple approach has been developed to prepare gallotannin (GT) mediated silver nanoparticles (AgNPs) for detection of Hg<SUP>2+</SUP> ion in aqueous solution. The synthesized silver nanoparticles was characterized by different techniques such as UV–vis spectrophotometry, XRD, scanning electron microscopy, and EDX. Amongst the all metal ion tested, only Hg<SUP>2+</SUP> shows selective colorimetric and fluorimetric change to GT capped AgNPs. A quick naked eye color change of GT capped AgNPs from yellowish-brown to colorless shows high sensitivity of present method. With addition of Hg<SUP>2+</SUP> ion, the fluorescence enhancement was observed for GT capped AgNPs. The mechanism for the change in color and fluorescence enhancement of nanoparticles with addition of incremental amount of Hg<SUP>2+</SUP> is due to the change in polarization near metal surface region and internal charge transfer phenomenon. The linear range for the detection of Hg<SUP>2+</SUP> was found to be 0.1–1 nM with lower detection limit (LOD) 0.037 nM. The present approach has been successfully utilized to quantitative determination of Hg<SUP>2+</SUP> from environmental samples. The another advantages of prepared silver nanoparticles (GT-AgNPs) is that it shows significant photocatalytic activity for organic transformation and anticancer potential against cancer cell line MDA-MB-231 and A375 with an LD<SUB>50</SUB> of 111.5 µg/mL and 185 µg/mL, respectively.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Intracellular imaging of zinc ion in living cells by fluorescein based organic nanoparticles

Mahajan, Prasad G.,Dige, Nilam C.,Vanjare, Balasaheb D.,Han, Yohan,Kim, Song Ja,Hong, Seong-Karp,Lee, Ki Hwan Elsevier 2018 Sensors and actuators. B, Chemical Vol.267 No.-

<P><B>Abstract</B></P> <P>A fluorescein based organic nanoparticles were prepared by using synthesized <I>N</I>-(3′,6′-dihydroxy-3-oxospiro[isoindoline-1,9′-xanthene]-2-yl)-2-hydroxybenzamide(FB). The interaction of series of metal ions with the FB nanoparticles (FBNPs) was investigated by means of fluorescence measurement. The selective and sensitive fluorescence enhancement was observed to FBNPs by the addition of Zn<SUP>2+</SUP> ion in an aqueous medium. The interaction of FBNPs and Zn<SUP>2+</SUP> introduced chelation enhanced fluorescence that was unaltered even addition of other metal ion. The formation of excited state complexation of FBNPs-Zn<SUP>2+</SUP> was supported by UV–vis absorption titration, fluorescence lifetime and zeta particle size of FBNPs with and without addition of Zn<SUP>2+</SUP>. The mode of binding and interaction was confirmed by results procured through NMR and IR spectroscopy. The stoichiometry and binding constant estimated by Job’s plot and Benesi-Hildebrand plot. The limit of detection (LOD) for Zn<SUP>2+</SUP> was 0.0011 μg/mL. The fascinating feature of present nano probe (FBNPs) is that outstanding intracellular Zn<SUP>2+</SUP> sensor in MDA-MB-231 living cells with nominal toxicity. The exceptional photo physical and biocompatible nature of FBNPs signifying on site application of prepared fluorescent organic nanoparticles in environmental as well as biomedical field.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Novel fluorescein based nano probe (FBNPs) was designed and synthesized. </LI> <LI> Selective fluorescence enhancement of FBNPs by Zn<SUP>2+</SUP> ion. </LI> <LI> Fluorescence enhancement is an account for chelation of FBNPs and Zn<SUP>2+</SUP> in 1:1 stoichiometry. </LI> <LI> Lower detection limit for Zn<SUP>2+</SUP> = 0.0011 μg/mL in an aqueous medium. </LI> <LI> FBNPs exhibits excellent biocompatibility and non-toxicity for intracellular detection of Zn<SUP>2+</SUP> in MDA-MB-231 cells. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>