Involvement of S6K1 in mitochondria function and structure in HeLa cells

Park, J.,Tran, Q.,Mun, K.,Masuda, K.,Kwon, S.H.,Kim, S.H.,Kim, D.H.,Thomas, G.,Park, J. Pergamon Press ; Elsevier Science Ltd 2016 Cellular signalling Vol.28 No.12

The major biological function of mitochondria is to generate cellular energy through oxidative phosphorylation. Apart from cellular respiration, mitochondria also play a key role in signaling processes, including aging and cancer metabolism. It has been shown that S6K1-knockout mice are resistant to obesity due to enhanced beta-oxidation, with an increased number of large mitochondria. Therefore, in this report, the possible involvement of S6K1 in regulating mitochondria dynamics and function has been investigated in stable lenti-shS6K1-HeLa cells. Interestingly, S6K1-stably depleted HeLa cells showed phenotypical changes in mitochondria morphology. This observation was further confirmed by detailed image analysis of mitochondria shape. Corresponding molecular changes were also observed in these cells, such as the induction of mitochondrial fission proteins (Drp1 and Fis1). Oxygen consumption is elevated in S6K1-depeleted HeLa cells and FL5.12 cells. In addition, S6K1 depletion leads to enhancement of ATP production in cytoplasm and mitochondria. However, the relative ratio of mitochondrial ATP to cytoplasmic ATP is actually decreased in lenti-shS6K1-HeLa cells compared to control cells. Lastly, induction of mitophagy was found in lenti-shS6K1-HeLa cells with corresponding changes of mitochondria shape on electron microscope analysis. Taken together, our results indicate that S6K1 is involved in the regulation of mitochondria morphology and function in HeLa cells. This study will provide novel insights into S6K1 function in mitochondria-mediated cellular signaling.

Effect of Triticale Dried Distillers Grains with Solubles on Ruminal Bacterial Populations as Revealed by Real Time Polymerase Chain Reaction

Wu, R.B.,Munns, K.,Li, J.Q.,John, S.J.,Wierenga, K.,Sharma, R.,Mcallister, T.A. Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.11

Real time PCR was used in this study to determine the effect of triticale dried distillers grains with solubles (TDDGS) as a replacement for grain or barley silage in finishing diets on the presence of six classical ruminal bacterial species (Succinivibrio dextrinosolvens, Selenomonas ruminantium, Streptococcus bovis, Megasphaera elsdenii, Prevotella ruminicola and Fibrobacter succinogenes) within the rumen contents of feedlot cattle. This study was divided into a step-wise adaptation experiment (112 days) that examined the effects of adaptation to diets containing increasing levels of TDDGS up to 30% (n = 4), a short-term experiment comparing animals (n = 16) fed control, 20%, 25% or 30% TDDGS diets over 28 days, and a rapid transition experiment (56 days) where animals (n = 4) were rapidly switched from a diet containing 30% TDDGS to a barley-based diet with no TDDGS. It was found that feeding TDDGS as replacement for barley grain (control vs. 20% TDDGS) decreased 16S rRNA copy numbers of starch-fermenting S. ruminantium and S. bovis (p<0.001 and p = 0.04, respectively), but did not alter 16S rRNA copy numbers of the other rumen bacteria. Furthermore, feeding TDDGS as a replacement barley silage (20% vs. 25% and 30% TDDGS) increased 16S rRNA copy numbers of S. ruminantium, M. elsdenii and F. succinogenes (p<0.001; p = 0.03 and p<0.001, respectively), but decreased (p<0.001) the 16S rRNA copy number of P. ruminicola. Upon removal of 30% TDDGS and return to the control diet, 16S rRNA copy numbers of S. ruminantium, M. elsdenii and F. succinogenes decreased (p = 0.01; p = 0.03 and p = 0.01, respectively), but S. dextrinosolvens and S. bovis increased (p = 0.04 and p = 0.009, respectively). The results suggest that replacement of TDDGS for grain reduces 16S rRNA copy numbers of starch-fermenting bacteria, whereas substitution for barley silage increases 16S rRNA copy numbers of bacteria involved in fibre digestion and the metabolism of lactic acid. This outcome supports the contention that the fibre in TDDGS is highly fermentable.

The Requirement of Ruminal Degradable Protein for Non-Structural Carbohydrate-Fermenting Microbes and Its Reaction with Dilution Rate in Continuous Culture

Meng, Q.X.,Xia, Z.G.,Kerley, M.S. Asian Australasian Association of Animal Productio 2000 Animal Bioscience Vol.13 No.10

A continuous culture study was conducted to determine the impact of ruminal degradable soy protein (S-RDP) level and dilution rate (D) on growth of ruminal non-structural carbohydrate-fermenting microbes. Corn starch, urea and isolated soy protein (ISP) were used to formulate three diets with S-RDP levels of 0, 35 and 70% of total dietary CP. Two Ds were 0.03 and $0.06h^{-1}$ of the fermenter volume in a single-effluent continuous culture system. As S-RDP levels increased, digestibilities of dietary dry matter (DM), organic matter (OM) and crude protein (CP) linearly (p=0.001) decreased, whereas digestion of dietary starch linearly (p=0.001) increased. Increasing D from 0.03 to $0.06h^{-1}$ resulted in decreased digestibilities of dietary DM and OM, but had no effect on digestibilities of dietary starch (p=0.77) and CP (p=0.103). Fermenter pH, the concentration of volatile fatty acids (VFA) and daily VFA production were unaffected (p=0.159-0.517) by S-RDP levels. Molar percentages of acetate, propionate and butyrate were greatly affected by S-RDP levels (p=0.016-0.091), but unaffected by D (p=0.331-0.442). With increasing S-RDP levels and D, daily bacterial counts, daily microbial N production (DMNP) and microbial efficiency (MOEFF; grams of microbial N produced per kilogram of OM truly digested) were enhanced (p=0.001). The increased microbial efficiency with increasing S-RDP levels is probably the result of peptides or amino acids that served as a stimulus for optimal protein synthesis. The quantity of ruminal degradable protein from soy proteins required for optimum protein synthesis of non-structural carbohydrate-fermenting microbes appears to be equivalent to 9.5% of dietary fermented OM.

32 EFFECTS OF FUSION/ACTIVATION METHODS ON DEVELOPMENT OF EMBRYOS PRODUCED BY NUCLEAR TRANSFER OF PORCINE FETAL FIBROBLASTS

Cong, P. Q.,Song, E. S.,Kim, E. S.,Li, Z. H.,Yi, Y. J.,Park, C. S. CSIRO Publishing 2007 Reproduction, fertility, and development Vol.19 No.1

<P> Pigs have become increasingly important in the field of biomedical research, and interest has grown in the use of transgenic cloned pigs as potential xenograft donors. The present study were carried out to investigate the effects of intensity of DC pulse, number of DC pulses, and equilibration before fusion/activation on developmental ability of porcine embryos derived from nuclear transfer. Porcine cumulus-oocyte complexes (COCs) were cultured in modified TCM-199 (mTCM-199) medium for 44 h at 38.5�C, 5% CO2 in air. After in vitro maturation (IVM), metaphase II oocytes were selected for enucleation. Porcine fetal fibroblasts were obtained from a porcine fetus on Day 35 of gestation as donor cells. Oocytes were enucleated by removing, with a micropipette, the first polar body along with adjacent cytoplasm containing the metaphase plate; then a donor cell was injected in contact with the cytoplasm of each oocyte. In experiment 1, several different fusion/activation intensities (two DC pulses of 0.4, 0.8, 1.2, 1.6, and 2.0 kV cm-1 for 30 �s) were carried out to investigate the effect on the development of nuclear transfer embryos. In experiment 2, the reconstructed oocytes were fused and activated with 1, 2, or 3 DC pulses of 1.2 kV cm-1 for 30 �s. In experiment 3, reconstructed oocytes were equilibrated in mTCM-199 medium at 38.5�C, 5% CO2 for 0, 1, 2, 3, 4, 5, and 6 h. After equilibration, the reconstructed oocytes were fused and activated with one DC pulse of 1.2 kV cm-1 for 30 �s in fusion medium. The reconstructed embryos were transferred into PZM-3 medium containing 0.3% BSA for further culture. The rates of embryo cleavage and development of blastocyst stage were evaluated at 48 h and 6-7 days, respectively. The cell numbers of blastocysts were counted by using Hoechst 33342 epifluorescence staining. Data were analyzed by ANOVA and Duncan </P>

Effect of different levels of protein concentrates supplementation on the growth performance, plasma amino acids profile and mTOR cascade genes expression in early-weaned yak calves

Q.H. Peng,N.A. Khan,B. Xue,T.H. Yan,Z.S. Wang 아세아·태평양축산학회 2018 Animal Bioscience Vol.31 No.2

Objective: This study evaluated the effects of different levels of protein concentrate supplementation on the growth performance of yak calves, and correlated the growth rate to changes occurring in the plasma- amino acids, -insulin profile, and signaling activity of mammalian target of rapamycin (mTOR) cascade to characterize the mechanism through which the protein synthesis can be improved in early weaned yaks. Methods: For this study, 48 early (3 months old) weaned yak calves were selected, and assigned into four dietary treatments according to randomized complete block design. The four blocks were balanced for body weight and sex. The yaks were either grazed on natural pasture (control diet) in a single herd or the grazing yaks was supplemented with one of the three protein rich supplements containing low (17%; LP), medium (19%; MP), or high (21%; HP) levels of crude proteins for a period of 30 days. Results: Results showed that the average daily gain of calves increased (0.14 vs 0.23-0.26 kg; p<0.05) with protein concentrates supplementation. The concentration of plasma methionine increased (p<0.05; 8.6 vs 10.1-12.4 μmol/L), while those of serine and tyrosine did not change (p>0.05) when the grazing calves were supplemented with protein concentrates. Compared to control diet, the insulin level of calves increased (p<0.05; 1.86 vs 2.16-2.54 μIU/mL) with supplementation of protein concentrates. Addition of protein concentrates up-regulated (p<0.05) expression of mTOR-raptor, mammalian vacuolar protein sorting 34 homolog, the translational regulators eukaryotic translation initiation factor 4E binding protein 1, and S6 kinase 1 genes in both Longissimus dorsi and semitendinosus. In contrast, the expression of sequestosome 1 was down-regulated in the concentrate supplemented calves. Conclusion: Our results show that protein supplementation improves the growth performance of early weaned yak calves, and that plasma methionine and insulin concentrations were the key mediator for gene expression and protein deposition in the muscles.

2. De Novo Synthesis of Amino Acids and Protein as Affected by S-deprivation and Sulfate Resupply in Brassica napus L.

Q. Zhang,B. R. Lee,S. H. Park,R. Zaman,T. H. Kim 한국초지조사료학회 2014 한국초지조사료학회 학술대회논문집 Vol.2014 No.10

Tl₂ZrCl₆ crystal: Efficient scintillator for X- and γ-ray spectroscopies

Phan, Q.V.,Kim, H.J.,Rooh, G.,Kim, S.H. Elsevier 2018 JOURNAL OF ALLOYS AND COMPOUNDS Vol.766 No.-

<P><B>Abstract</B></P> <P>Crystal growth and luminescence properties of the intrinsic Tl<SUB>2</SUB>ZrCl<SUB>6</SUB> scintillation crystal are presented. The proposed crystal is a promising candidate for γ-ray detection owing to its high light yield, good energy resolution, and high effective atomic number (<I>Z</I> = 69). It has a cubic structure, which is desirable for a large-size-crystal growth. In this study, the crystal is grown from its melt using the Bridgman technique. The emission peak under X-ray excitation is found at 460 nm, with a primary decay time of 2.7 μs. The light yield and energy resolution are estimated to be 47,000 photons/MeV and 4.3% (full-width-at-half-maximum), respectively, obtained from the pulse-height spectra under a <SUP>137</SUP>Cs 662-keV γ-ray excitation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Tl<SUB>2</SUB>ZrCl<SUB>6</SUB> crystal has been grown by Bridgman technique for first time. </LI> <LI> The grown crystal was characterized under X- and γ-rays excitation. </LI> <LI> High-Z material with high light yield and good energy resolution were obtained. </LI> <LI> It has the potential to be X- and γ-ray detectors for various applications. </LI> </UL> </P>

Evaluation of VIIRS, GOCI, and MODIS Collection 6 AOD retrievals against ground sunphotometer observations over East Asia

Xiao, Q.,Zhang, H.,Choi, M.,Li, S.,Kondragunta, S.,Kim, J.,Holben, B.,Levy, R. C.,Liu, Y. Copernicus GmbH 2016 Atmospheric Chemistry and Physics Vol.16 No.3

<P>Abstract. Persistent high aerosol loadings together with extremely high population densities have raised serious air quality and public health concerns in many urban centers in East Asia. However, ground-based air quality monitoring is relatively limited in this area. Recently, satellite-retrieved Aerosol Optical Depth (AOD) at high resolution has become a powerful tool to characterize aerosol patterns in space and time. Using ground AOD observations from the Aerosol Robotic Network (AERONET) and the Distributed Regional Aerosol Gridded Observation Networks (DRAGON)-Asia Campaign, as well as from handheld sunphotometers, we evaluated emerging aerosol products from the Visible Infrared Imaging Radiometer Suite (VIIRS) aboard the Suomi National Polar-orbiting Partnership (S-NPP), the Geostationary Ocean Color Imager (GOCI) aboard the Communication, Ocean, and Meteorology Satellite (COMS), and Terra and Aqua Moderate Resolution Imaging Spectroradiometer (MODIS) (Collection 6) in East Asia in 2012 and 2013. In the case study in Beijing, when compared with AOD observations from handheld sunphotometers, 51 % of VIIRS Environmental Data Record (EDR) AOD, 37 % of GOCI AOD, 33 % of VIIRS Intermediate Product (IP) AOD, 26 % of Terra MODIS C6 3 km AOD, and 16 % of Aqua MODIS C6 3 km AOD fell within the reference expected error (EE) envelope (±0.05 ± 0.15 AOD). Comparing against AERONET AOD over the Japan-South Korea region, 64 % of EDR, 37 % of IP, 61 % of GOCI, 39 % of Terra MODIS, and 56 % of Aqua MODIS C6 3 km AOD fell within the EE. In general, satellite aerosol products performed better in tracking the day-to-day variability than tracking the spatial variability at high resolutions. The VIIRS EDR and GOCI products provided the most accurate AOD retrievals, while VIIRS IP and MODIS C6 3 km products had positive biases. </P>

Performance evaluation of an improved fish robot actuated by piezoceramic actuators

Nguyen, Q S,Heo, S,Park, H C,Byun, D Institute of Physics Publishing 2010 Smart materials & structures Vol.19 No.3

<P>This paper presents an improved fish robot actuated by four lightweight piezocomposite actuators. Our newly developed actuation mechanism is simple to fabricate because it works without gears. With the new actuation mechanism, the fish robot has a 30% smaller cross section than our previous model. Performance tests of the fish robot in water were carried out to measure the tail-beat angle, the thrust force, the swimming speed for various tail-beat frequencies from 1 to 5 Hz and the turning radius at the optimal frequency. The maximum swimming speed of the fish robot is 7.7  cm s<SUP> − 1</SUP> at a tail-beat frequency of 3.9 Hz. A turning experiment shows that the swimming direction of the fish robot can be controlled by changing the duty ratio of the driving voltage; the fish robot has a turning radius of 0.41 m for a left turn and 0.68 m for a right turn. </P>

Genomic characterization, expression analysis, and antimicrobial function of a glyrichin homologue from rock bream, Oplegnathus fasciatus

Kasthuri, S.R.,Wan, Q.,Umasuthan, N.,Bathige, S.D.N.K.,Lim, B.S.,Jung, H.B.,Lee, J.,Whang, I. Academic Press 2013 Fish & shellfish immunology Vol.35 No.5

Antimicrobial peptides are important innate effector molecules, playing a vital role in antimicrobial immunity in all species. Glyrichin is a transmembrane protein and an antibacterial peptide, exerting its functions against a wide range of pathogenic bacteria. In this study, cDNA and a BAC clone harboring the glyrichin gene were identified from rock bream and characterized. Genomic characterization showed that the OfGlyrichin gene exhibited a 3 exon-2 intron structure. OfGlyrichin is a 79-amino-acid protein with a transmembrane domain at <SUP>22</SUP>GFMMGFAVGMAAGAMFGTFSCLR<SUP>44</SUP>. Pairwise and multiple sequence alignments showed high identity and conservation with mammalian orthologues. Phylogenetic analysis showed a close relationship with fish species. Higher levels of OfGlyrichin transcripts were detected in the liver from healthy rock bream which were induced by immunogens like lipopolysaccharide, poly I:C, rock bream irido virus, Edwardsiella tarda and Streptococcus iniae. The synthetic peptide (pOf19) showed antibacterial activity against Escherichia coli, E. tarda, and S. iniae. Analysis of the bacterial morphological features after pOf19 peptide treatment showed breakage of the cell membrane, affirming that antibacterial function is accomplished through membrane lysis. The pOf19 peptide also showed antiviral activity against RBIV infection. The high conservation of the genomic structure and protein, together with the antimicrobial roles of OfGlyrichin, provide evidence for the evolutionary existence of this protein playing a vital role in innate immune defense in rock bream.