Expression and N-glycan analysis of human 90K glycoprotein in Drosophila S2 cells

Kim, K.R.,Kim, Y.K.,Cheong, H.,Kim, J.Y.H.,Cha, H.J. IPC Science and Technology Press ; Elsevier Scienc 2013 Enzyme and microbial technology Vol.53 No.3

Human 90K (h90K; Mac-2-binding protein) glycoprotein is a potential pharmaceutical due to its inhibitory activity against cancer metastasis and expansion. Here, h90K glycoprotein was produced in insect Drosophila S2 cell system, and its N-glycan pattern was analyzed. A plasmid encoding h90K gene, fused with a hexahistidine tag under the control of Drosophila metallotionein promoter, was stably transfected into S2 cells. After copper sulfate induction, transfected S2 cells secreted recombinant h90K at a good expression level of 28mg/L in a 150-mL spinner flask culture. The purified recombinant h90K showed an apparent molecular weight of ~78kDa which was much smaller than that (~97kDa) of the natural h90K. Because de-N-glycosylated h90K appeared at ~60kDa protein band, it was suggested that the recombinant h90K from S2 cells has small N-glycans with about half the molecular weight (~18kDa) of N-glycans of the natural h90K. Through detail analyses using high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, the S2-derived recombinant h90K was confirmed that it has simple paucimannosidic structures containing two or three mannose residues with core fucose as the major (~79%) N-glycans.

A comparative study on the performance of different advanced oxidation processes (UV/O3/H2O2) treating linear alkyl benzene (LAB) production plant's wastewater

H. Zangeneh,A.A.L. Zinatizadeh,M. Feizy 한국공업화학회 2014 Journal of Industrial and Engineering Chemistry Vol.20 No.4

A detailed investigation on photooxidation of linear alkyl benzene (LAB) industrial wastewater is presented in this study. The process analysis was performed by varying four significant independent variables including two numerical factors (initial pH (3–11) and initial H2O2 concentration (0–20 mM)) and two categorical factors (UV irradiation and ozonation). The experiments were conducted based on a central composite design (CCD) and analyzed using response surface methodology (RSM). To assess the process performance, two parameters viz. TCOD removal efficiency and BOD5/COD were measured throughout the experiments. A maximum reduction in TCOD was 58, 53, 51, and 49%, respectively for UV/H2O2/O3, H2O2/O3, UV/O3 and UV/H2O2 processes at the optimum conditions (initial pH of 7, initial H2O2 concentration of 100 mM, and reaction time of 180 min). A considerable increase in BOD5/COD ratio was obtained in the combined processes (0.46, 0.51, 0.53, and 0.55 for UV/H2O2, UV/O3, H2O2/O3 and UV/H2O2/O3, respectively) compared to the single oxidant process (0.35). The results showed that mineralization of the LAB industrial wastewater in neutral pH is more favored than in acidic and basic pH. Gas chromatography–mass spectrometry (GC–MS) was applied to show the fate of organic compounds. In conclusion, the photooxidation process (UV/H2O2/O3, H2O2/O3, UV/O3 and UV/H2O2) could be an appropriate pretreatment method prior to a biological treatment process.

Oxygen-dependent enhancement of hydrogen production by engineering bacterial hemoglobin in Escherichia coli

Jo, B.H.,Kim, J.Y.H.,Seo, J.H.,Cha, H.J. Pergamon Press ; Elsevier Science Ltd 2014 International journal of hydrogen energy Vol.39 No.20

H<SUB>2</SUB> production under aerobic conditions has been proposed as an alternative method to overcome the fundamentally low yield of H<SUB>2</SUB> production by fermentative bacteria by maximizing the number of electrons that are available for H<SUB>2</SUB>. Here, we engineered Vitreoscilla hemoglobin (VHb) in Escherichia coli to study the effects of this versatile oxygen (O<SUB>2</SUB>)-binding protein on oxic H<SUB>2</SUB> production in a closed batch system that was supplemented with glucose. The H<SUB>2</SUB> yields that were obtained with the VHb-expressing E. coli were greatly enhanced in comparison to the negative control cells in culture that started with high O<SUB>2</SUB> tensions. The formate hydrogen lyase (FHL) activity of oxically cultured, VHb-expressing cells was also much higher than that of the negative control cells. Through inhibitor studies and time-course experiments, VHb was shown to contribute to the improved H<SUB>2</SUB> yield primarily by increasing the efficiency of cellular metabolism during the aerobic phase before the onset of H<SUB>2</SUB> production and not by working as an O<SUB>2</SUB>-scavenger during H<SUB>2</SUB> production. This new approach allowed more substrate to remain to be further utilized for the production of more H<SUB>2</SUB> from limited resources. We expect that VHb can be successfully engineered in potential aerobic H<SUB>2</SUB>-producing microbial systems to enhance the overall H<SUB>2</SUB> production yield. In addition, the remarkably high FHL activity of oxically grown, VHb-expressing cells may make this engineered strain an attractive whole-cell biocatalyst for converting formate to H<SUB>2</SUB>.

Selection for Duration of Fertility and Mule Duck White Plumage Colour in a Synthetic Strain of Ducks (Anas platyrhynchos)

H.C. Liu,J.F. Huang,S.R. Lee,H.L. Liu,C.H. Hsieh,C.W. Huang,M.C. Huang,C. Tai,J.P. Poivey,R. Rouvier,Y.S. Cheng 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.5

A synthetic strain of ducks (Anas platyrhynchos) was developed by introducing genes for long duration of fertility to be used as mother of mule ducklings and a seven-generation selection experiment was conducted to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. Reciprocal crossbreeding between Brown Tsaiya LRI-2 (with long duration of fertility) and Pekin L-201 (with white plumage mule ducklings) ducks produced the G0. Then G1 were intercrossed to produce G2 and so on for the following generations. Each female duck was inseminated 3 times, at 26, 29, and 32 weeks of age. The eggs were collected for 14 days from day 2 after AI. Individual data regarding the number of incubated eggs (Ie), the number of fertile eggs at candling at day 7 of incubation (F), the total number of dead embryos (M), the maximum duration of fertility (Dm) and the number of hatched mule ducklings (H) with plumage colour were recorded. The selection criterion was the breeding values of the best linear unbiased prediction animal model for F. The results show high percentage of exhibited heterosis in G2 for traits to improve (19.1% for F and 12.9% for H); F with a value of 5.92 (vs 3.74 in the Pekin L-201) was improved in the G2. Heritabilities were found to be low for Ie (h2 = 0.07±0.03) and M (h2 = 0.07±0.01), moderately low for Dm (h2 = 0.13±0.02), of medium values for H (h2 = 0.20±0.03) and F (h2 = 0.23±0.03). High and favourable genetic correlations existed between F and Dm (rg = 0.93), between F and H (rg = 0.97) and between Dm and H (rg = 0.90). The selection experiment showed a positive trend for phenotypic values of F (6.38 fertile eggs in G10 of synthetic strain vs 5.59 eggs in G4, and 3.74 eggs in Pekin L-201), with correlated response for increasing H (5.73 ducklings in G10 vs 4.86 in G4, and 3.09 ducklings in Pekin L-201) and maximum duration of the fertile period without increasing the embryo mortality rate. The average predicted genetic response for F was 40% of genetic standard deviation per generation of selection. The mule ducklings’ feather colour also was improved. It was concluded that this study provided results for a better understanding of the genetics of the duration of fertility traits in the common female duck bred for mule and that the selection of a synthetic strain was effective method of improvement.

Experimental infection of mandarin duck with highly pathogenic avian influenza A (H5N8 and H5N1) viruses

Kang, H.M.,Lee, E.K.,Song, B.M.,Heo, G.B.,Jung, J.,Jang, I.,Bae, Y.C.,Jung, S.C.,Lee, Y.J. Elsevier Scientific Pub. Co 2017 Veterinary microbiology Vol.198 No.-

<P>A highly pathogenic avian influenza (HPAI) H5N8 virus was first detected in poultry and wild birds in South Korea in January 2014. Here, we determined the pathogenicity and transmissibility of three different clades of 1-15 viruses in mandarin ducks to examine the potential for wild bird infection. H5N8 (Glade 2.3.4.4) replicated more efficiently in the upper and lower respiratory tract of mandarin ducks than two previously identified H5N1 virus clades (clades 2.2 and 2.3.2.1). However, none of the mandarin ducks infected with H5N8 and H5N1 viruses showed severe clinical signs or mortality, and gross lesions were only observed in a few tissues. Viral replication and shedding were greater in H5N8-infected ducks than in H5N1-infected ducks. Recovery of all viruses from control duck in contact with infected ducks indicated that the highly pathogenic H5 viruses spread horizontally through contact. Taken together, these results suggest that H5N8 viruses spread efficiently in mandarin ducks. Further studies of pathogenicity in wild birds are required to examine possible long-distance dissemination via migration routes. (C) 2016 Elsevier B.V. All rights reserved.</P>

A multi-virus detectable microfluidic electrochemical immunosensor for simultaneous detection of H1N1, H5N1, and H7N9 virus using ZnO nanorods for sensitivity enhancement

Han, J.H.,Lee, D.,Chew, C.H.C.,Kim, T.,Pak, J.J. Elsevier Sequoia 2016 Sensors and actuators. B Chemical Vol.228 No.-

This paper describes a multi-detectable and nano-flow immunosensor based on ZnO nanorods (NRs) grown on the inner surface of PDMS sensor region for sensing H1N1, H5N1, and H7N9 influenza viruses simultaneously using electrochemical method. Nanostructured ZnO NRs with a high isoelectric point (IEP ~9.5) tend to interact electrostatically with proteins with lower IEP such as H1N1, H5N1, and H7N9 antibodies. ZnO NRs were hydrothermally grown on the upper inner surface of the nano-flow PDMS sensor region. The forementioned three influenza viruses were successfully detected from three separate sensing regions by measuring the oxidation current of 3,3',5,5'-tetramethylbenzidine (TMB) by horseradish peroxidase (HRP) conjugated on capture antibody of those influenza viruses when proper potential was applied. The proposed immunosensors were evaluated using 1pg/ml, 10pg/ml, 100pg/ml, 1ng/ml, and 10ng/ml of H1N1, H5N1, and H7N9 antigens by amperometry. These immunosensors showed high selectivity toward H1N1, H5N1, and H7N9, which was successfully confirmed by distinguishing the target virus individually from a mixture of three virus antigens. A low limit of detection was demonstrated by detecting as low as 1pg/ml of each virus and it is believed that this was possible by enhancing the sensitivity with the ZnO NRs grown on the PDMS surface in the sensing region. The steady-state oxidation current output linearly increased with respect to the logarithm of the H1N1, H5N1, and H7N9 virus concentrations in the range of 1-10ng/ml.

돼지 H-FABP 유전자의 다형성 및 경제 형질과의 연관성 구명

최봉환,김태헌,이지웅,조용민,이혜영,조병욱,정일정 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.5

The purpose of this study was to detect association between genetic variation and economic trait in the porcine heart type fatty acid-binding protein gene as a candidate gene for the traits related with growth and meat quality in pigs. The H-FABP is a 15-kDa protein expressed in several tissues with high demand for fat metabolism such as cardiac and skeletal muscle and lactating mammary gland. H-FABP is small intracellular protein involved in fatty acid transport from the plasma membrane to the site of β-oxidation and/or triacylglycerol or phospholipid synthesis. In this study, H-FABP PCR-RFLP was performed in F_(2) population composed of 214 individuals form an intercross between Korean Native Boars and Landrace sows. PCR products form tow primer sets within H-FABP gene were amplified in 850bp and 700bp. Digestion of PCR products with the restriction digestion enzymes HaeⅢ and Hinf Ⅰ, revealed fragment length polymorphisms(RFL. Ps). The genotype frequencies from H-FABP/HaeⅢ was .29 for genotype DD, .53 for genotype Dd, and .15 for genotype dd, respectively. The genotype frequencies of HH, Hh, and hh from H-FABP(hinf Ⅰ was .38, .41, and .20, respectively, in the population.Relationships between their genotypes and economic traits were estimated. In H-FABP/HaeⅢ locus, there were specific genotypes(Dd and dd) associated with economic traits such as body weight. In H-FABP/Hinf Ⅰ Iocus, Genotypes of HH and Hh associated with growth traits such as body weights at 5, 12, and 30 week of age (p<.05 or p<.001) and back fat thickness, body fat including abdominal and trimmed fat (p<.001) and intramuscular fat(p<.05). The 'H'allele was positivecly associated with gaining of body weight and fatness deposition. In conclusion, a significant association of the H-FABP gene from its genetic variation was found on body weight, intramuscular fat and backfat thickness.

On the zeolitic imidazolate framework-8 (ZIF-8) membrane for hydrogen separation from simulated biomass-derived syngas

Yin, H.,Lee, T.,Choi, J.,Yip, A.C.K. Elsevier 2016 Microporous and mesoporous materials Vol.233 No.-

<P>Hydrogen separation from biomass-derived syngas is a critical step in the utilization of gasification technology. Compared with the traditional methods, membrane technology provides an effective and low-cost solution for adjusting the gas composition and collecting H-2 in syngas environments. In this study, a zeolitic imidazolate framework-8 (ZIF-8)-based membrane, which is a potential candidate for H-2 separation from biomass-derived syngas, was successfully fabricated through the seeded (secondary) growth method and the subsequent post-treatments. The prepared ZIF-8 membrane exhibited a modest H-2 separation performance for H-2/CO2 and H-2/CO, with separation factors of 4.95 and 6.08, respectively, and a H-2 permeance of 7.81 x 10(-8) mol m(-2) s(-1) Pa-1 at 200 degrees C in the simulated biomass-derived syngas environments (H-2/CO2/CO) with the presence of steam. In particular, the H-2/CO2 and H-2/CO separation factors were increased by 36% and 97%, respectively, with respect to those obtained through bare supports. Despite the promising H2 perm-selectivity, the H-2/CO2 and H-2/CO separation factors of ZIF-8 membranes at 200 degrees C under water-containing syngas environments were maintained up to 10 h but the longer exposure led to the gradual degradation and eventual reduction toward those of bare supports after 15 h seemingly due to the water-involved membrane degradation. This study provides the availability and limitation of ZIF-8 membranes for H-2 separations in stimulated biomass-derived syngas environments. (C) 2015 Elsevier Inc. All rights reserved.</P>

The H <small>i</small> environment of counter‐rotating gas hosts: gas accretion from cold gas blobs

Chung, Aeree,Bureau, Martin,van Gorkom, J. H.,Koribalski, Bä,rbel Blackwell Publishing Ltd 2012 Monthly notices of the Royal Astronomical Society Vol.422 No.2

<P><B>ABSTRACT</B></P><P>We probe the H <SMALL>i</SMALL> properties and the gas environments of three early‐type barred galaxies harbouring counter‐rotating ionized gas: NGC 128, NGC 3203 and NGC 7332. Each system has one or more optically identified galaxy at a similar or as yet unknown redshift within a 50‐kpc projected radius. Using H <SMALL>i</SMALL> synthesis imaging data, we investigate the hypothesis that the counter‐rotating gas in these galaxies has been accreted from their neighbours. In NGC 128 and NGC 3203, we find 9.6 × 10<SUP>7</SUP> and 2.3 × 10<SUP>8</SUP> M<SUB>⊙</SUB> of H <SMALL>i</SMALL>, respectively, covering almost the entire stellar bodies of dwarf companions that appear physically connected. Both the H <SMALL>i</SMALL> morphology and kinematics are suggestive of tidal interactions. In NGC 7332, we do not find any directly associated H <SMALL>i</SMALL>. Instead, NGC 7339, a neighbour of a comparable size at about 10 kpc, is found with 8.9 × 10<SUP>8</SUP> M<SUB>⊙</SUB> of H <SMALL>i</SMALL> gas. More recently in a single dish observation, however, another group discovered a large H <SMALL>i</SMALL> structure which seems to be an extension of NGC 7339’s H <SMALL>i</SMALL> disc and also covers NGC 7332. All these observations thus suggest that H <SMALL>i</SMALL> gas is being accreted in these three galaxies from their companions, which is likely responsible for the kinematically decoupled gas component present in their central region. In particular, the dynamical friction time‐scales of the nearest neighbours with H <SMALL>i</SMALL> gas of NGC 128 and NGC 3203 are comparable to their orbital time‐scales around the counter‐rotators, several ∼10<SUP>8</SUP> yr, implying that those neighbours will likely soon merge with the primary galaxies, fuelling them with gas. NGC 7332 also appears to be in the merging process with its neighbour through the common H <SMALL>i</SMALL> envelope. Besides, we find some other potential gas donors around NGC 128 and NGC 7332: two H <SMALL>i</SMALL>‐rich galaxies with <IMG src='/wiley-blackwell_img/equation/MNR_20679_mu1.gif' alt ='inline image'/> and 2.5 × 10<SUP>9</SUP> M<SUB>⊙</SUB> at a distance of ≈67 kpc from NGC 128 and two dwarf systems with <I>M</I><SUB>HI</SUB>= 3.9 × 10<SUP>7</SUP> and 7.4 × 10<SUP>7</SUP> M<SUB>⊙</SUB> at ≲100 kpc from NGC 7332. Among the seven H <SMALL>i</SMALL> features identified in this study, three of them are associated with dwarf galaxies, two of which have only been recently identified in a blind survey, while the third one is still not catalogued at optical wavelengths. Considering the incompleteness of existing studies of the faint dwarf galaxy population both in the optical and in H <SMALL>i</SMALL>, accretion from cold gas blobs, presumably gas‐rich dwarfs, is expected to occur even more frequently than what is inferred from such cases that have been observed to date.</P>

Neural stem cells injured by oxidative stress can be rejuvenated by GV1001, a novel peptide, through scavenging free radicals and enhancing survival signals

Park, H.H.,Yu, H.J.,Kim, S.,Kim, G.,Choi, N.Y.,Lee, E.H.,Lee, Y.J.,Yoon, M.Y.,Lee, K.Y.,Koh, S.H. Elsevier BV 2016 NeuroToxicology Vol.55 No.-

<P>Oxidative stress is a well-known pathogenic mechanism of a diverse array of neurological diseases, and thus, numerous studies have attempted to identify antioxidants that prevent neuronal cell death. GV1001 is a 16-amino-acid peptide derived from human telomerase reverse transcriptase (hTERT). Considering that hTERT has a strong antioxidant effect, whether GV1001 also has an antioxidant effect is a question of interest. In the present study, we aimed to investigate the effects of GV1001 against oxidative stress in neural stem cells (NSCs). Primary culture NSCs were treated with different concentrations of GV1001 and/or hydrogen peroxide (H2O2) for various time durations. The H2O2 decreased the viability of the NSCs in a concentration-dependent manner, with 200 mu M H2O2 significantly decreasing both proliferation and migration. However, treatment with GV1001 rescued the viability, proliferation and migration of H2O2 injured NSCs. Consistently, free radical levels were increased in rat NSCs treated with H2O2, while co-treatment with GV1001 significantly reduced these levels, especially the intracellular levels. In addition, GV1001 restored the expression of survival-related proteins and reduced the expression of death associated ones in NSCs treated with H2O2. In conclusion, GV1001 has antioxidant and neuroprotective effects in NSCs following treatment with H2O2, which appear to be mediated by scavenging free radicals, increasing survival signals and decreasing death signals. (C) 2016 Elsevier B.V. All rights reserved.</P>