Biological Differences between Hanwoo longissi-mus dorsi and semimembranosus Muscles in Col-lagen Synthesis of Fibroblasts

Sivakumar Allur Subramaniyan,Inho Hwang 한국축산식품학회 2017 한국축산식품학회지 Vol.37 No.3

Variations in physical toughness between muscles and animals are a function of growth rate and extend of collagen type I and III. The current study was designed to investigate the ability of growth rate, collagen concentration, collagen synthesizing and degrading genes on two dif-ferent fibroblast cells derived from Hanwoo m. longissimus dorsi (LD) and semimembrano-sus (SM) muscles. Fibroblast cell survival time was determined for understanding about the characteristics of proliferation rate between the two fibroblasts. We examined the collagen concentration and protein expression of collagen type I and III between the two fibroblasts. The mRNA expression of collagen synthesis and collagen degrading genes to elucidate the molecular mechanisms on toughness and tenderness through collagen production between the two fibroblast cells. From our results the growth rate, collagen content and protein expression of collagen type I and III were significantly higher in SM than LD muscle fibroblast. The mRNA expressions of collagen synthesized genes were increased whereas the collagen degrading genes were decreased in SM than LD muscle. Results from confocal microscopical investigation showed increased fluorescence of collagen type I and III appearing stronger in SM than LD muscle fibroblast. These results implied that the locomotion muscle had higher fibroblast growth rate, leads to produce more collagen, and cause tougher than positional mus-cle. This in vitro study mirrored that background toughness of various muscles in live animal is likely associated with fibroblast growth pattern, collagen synthesis and its gene expression.

Meat Quality and Physicochemical Trait Assessments of Berkshire and Commercial 3-way Crossbred Pigs

Sivakumar Allur Subramaniyan,강다래,Shah Ahmed Belal,조은소리,정종현,정영철,최양일,심관섭 한국축산식품학회 2016 한국축산식품학회지 Vol.36 No.5

In this study, we compared qualities and physiochemical traits of meat from Berkshire (black color) pigs with those of meat from 3-way Landrace (white color) × Yorkshire (white color) × Duroc (red color) crossbred pigs (LYD). Meat quality characteristics, including pH, color, drip loss, cooking loss, and free amino acid, fatty acid, vitamin, and mineral contents of longissimus dorsi muscles, were compared. Meat from Berkshire pigs had deeper meat color (redness), higher pH, and lower drip loss and cooking loss than meat from LYD pigs. Moreover, meat from Berkshire pigs had higher levels of phosphoserine, aspartic acid, threonine, serine, asparagine, α-aminoadipic acid, valine, methionine, isoleucine, leucine, tyrosine, histidine, tryptophan, and carnosine and lower levels of glutamic acid, glycine, alanine, and ammonia than did meat from LYD pigs. The fatty acids oleic acid, docosahexaenoic acid (DHA), and monounsaturated fatty acids (MUFA) were present in significantly higher concentrations in Berkshire muscles than they were in LYD muscles. Additionally, Berkshire muscles were significantly enriched with nucleotide components (inosine), minerals (Mg and K), and antioxidant vitamins such as ascorbic acid (C) in comparison with LYD muscles. In conclusion, our results show that in comparison with LYD meat, Berkshire meat has better meat quality traits and is a superior nutritional source of all essential amino acids, monounsaturated fatty acids, vitamin C, and minerals (Mg and K).

In ovo toxicological assessment of biologically synthesized and chemically synthesized silver nanoparticles in chicken embryo–A comparative study

Sivakumar Allur Subramaniyan,Da Rae Kang,Shah Ahmed Belal,Ho Sung Choe,Kwan Seob Shim 한국가금학회 2018 한국가금학회 정기총회 및 학술발표회 Vol.2018 No.10

Antiapoptotic effect of a novel synthetic peptide from bovine muscle and MPG peptide on H2O2-induced C2C12 cells.

Sivakumar, Allur Subramaniyan,Ochirbat, Chinzorio,Cho, Soo-Hyun,Yang, Jieun,Hwang, Inho Springer 2014 In vitro cellular & developmental biology Animal Vol.50 No.7

<P>The objective of this study was to evaluate antioxidant and antiapoptotic effects of a novel peptide (T.peptide) isolated from bovine and commercially available MPG peptide. The amino acid sequences of the T.peptide were (Glu-Val-Pro-Glu-Val-His-Glu-Glu-Val). The antioxidant activities of these peptides were determined by measuring the 1,1-diphenyl-2-picrylhydrazyl (DPPH?) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS+?) radical scavenging assays. The in vitro cytotoxicity of T.peptide and MPG peptide was determined against H2O2-induced C2C12 cells. H2O2-induced apoptosis in C2C12 cells were determined by mRNA expression of caspase-3. Moreover, the mRNA expression of tumor necrosis factor alpha (TNF-α) and nuclear factor kappa B (NF-κB) were assayed by reverse transcription polymerase chain reaction. The findings of the study indicate that the mRNA expression of TNF-α and NF-κB are significantly (p?<?0.05) increased in H2O2-induced C2C12 cells, whereas a significant decrease (p?<?0.05) in these mRNA expressions are observed when pretreated with T.peptide or MPG peptide. Pretreatment with MPG or T.peptides is also found to significantly (p?<?0.05) decrease the mRNA expression of caspase-3 in H2O2-induced C2C12 cells. The results of the study demonstrate that both T.peptide and MPG peptide could reduce the DPPH? and ABTS+? radical and inhibit cytotoxicity against H2O2-induced injury, resulting in prevention of free radical generation and subsequent apoptotic cell death, which indicates the potential of bovine meat as a source of antioxidant peptides.</P>

Meat Quality and Physicochemical Trait Assessments of Berkshire and Commercial 3-way Crossbred Pigs

Subramaniyan, Sivakumar Allur,Kang, Da Rae,Belal, Shah Ahmed,Cho, Eun-So-Ri,Jung, Jong-Hyun,Jung, Young-Chul,Choi, Yang-Il,Shim, Kwan-Seob Korean Society for Food Science of Animal Resource 2016 한국축산식품학회지 Vol.36 No.5

In this study, we compared qualities and physiochemical traits of meat from Berkshire (black color) pigs with those of meat from 3-way Landrace (white color) × Yorkshire (white color) × Duroc (red color) crossbred pigs (LYD). Meat quality characteristics, including pH, color, drip loss, cooking loss, and free amino acid, fatty acid, vitamin, and mineral contents of longissimus dorsi muscles, were compared. Meat from Berkshire pigs had deeper meat color (redness), higher pH, and lower drip loss and cooking loss than meat from LYD pigs. Moreover, meat from Berkshire pigs had higher levels of phosphoserine, aspartic acid, threonine, serine, asparagine, α-aminoadipic acid, valine, methionine, isoleucine, leucine, tyrosine, histidine, tryptophan, and carnosine and lower levels of glutamic acid, glycine, alanine, and ammonia than did meat from LYD pigs. The fatty acids oleic acid, docosahexaenoic acid (DHA), and monounsaturated fatty acids (MUFA) were present in significantly higher concentrations in Berkshire muscles than they were in LYD muscles. Additionally, Berkshire muscles were significantly enriched with nucleotide components (inosine), minerals (Mg and K), and antioxidant vitamins such as ascorbic acid (C) in comparison with LYD muscles. In conclusion, our results show that in comparison with LYD meat, Berkshire meat has better meat quality traits and is a superior nutritional source of all essential amino acids, monounsaturated fatty acids, vitamin C, and minerals (Mg and K).

A Comparative Study of Biologically and Chemically Fabricated Synthesized AgNPs’ Supplementation with Respect to Heat-Shock Proteins, Survival, and Hatching Rates of Chicken Embryos: An In Ovo Study

Allur Subramaniyan, Sivakumar,Kang, Da Rae,Belal, Shah Ahmed,Choe, Ho Sung,Shim, Kwan Seob Plenum Press 2018 Journal of cluster science Vol.29 No.1

<P>The present study examined the characterization and concentration of Brassica oleracea L. var. capitata f. rubra (BOL, commonly known as red cabbage) extracted AgNPs (BOL-AgNPs) and chemically (CHE) synthesized AgNPs (CHE-AgNPs) in chicken embryos through in ovo injections. We investigated the effects of both AgNPs on the survival, hatchability, body weight, and expressions of heat-shock proteins (HSPs). The BOL-AgNPs and CHE-AgNPs at (T1) amounted 100, at (T2) 1000, at (T3) 2500, and at (T4) 5000 A mu g/100 A mu l/egg, respectively through in ovo injections. At the end of the experimental period, the body and liver weights of the chicks were altered between the groups and among the concentrations. The serum levels of SGOT and SGPT were significantly increased at BOL-T3 and -T4. Survival and hatching rates were more reduced in the BOL-T3- and -T4-treated groups than at BOL-T1 and -T2. Nevertheless, they were more diminished in the CHE-AgNPs than in the BOL-AgNPs. The CHE-AgNPs revealed the presence of micronuclei and nuclear abnormalities, in contrast to BOL-T4. The mRNA and protein expressions of HSP-60 and 70 in the liver were higher in CHE-AgNPs than in BOL-AgNPs. Hence, plant-synthesized AgNPs are superior in ovo nano-nutrition carriers of nutrients without toxicity than CHE-AgNPs.</P>

Effects of Sunphenon and Polyphenon 60 on proteolytic pathways, inflammatory cytokines and myogenic markers in H2O2-treated C2C12 cells.

Sivakumar, Allur Subramaniyan,Hwang, Inho Indian Academy of Sciences 2015 Journal of biosciences Vol.40 No.1

<P>The effect of Sunphenon and Polyphenon 60 in oxidative stress response, myogenic regulatory factors, inflammatory cytokines, apoptotic and proteolytic pathways on H2O2-induced myotube atrophy was addressed. Cellular responses of H2O2-induced C2C12cells were examined, including mRNA expression of myogenic regulatory factors, such as MyoD and myogenin, inflammatory pathways, such as TNF-alpha and NF-kB, as well as proteolytic enzymes, such as mu-calpain and m-calpain. The pre-treatment of Sunphenon (50 mu g/mL)/Polyphenon 60 (50 mu g/naL) on H2O2-treated C2C12 cells significantly down-regulated the mRNA expression of myogenin and MyoD when compared to those treated with H2O2-induced alone. Additionally, the mRNA expression of mu-calpain and m-calpain were significantly (p<0.05) increased in H2O2-treated C2C12 cells, whereas pre-treatment with Sunphenon/Polyphenon significantly down-regulated the above genes, namely mu-calpain and m-calpain. Furthermore, the mRNA expression of TNF-alpha and NF-kB were significantly increased in H2O2-treated C2C12 cells, while pre-treatment with Sunphenon (50 mu g/mL)/Polyphenon 60 (50 mu g/mL) significantly (p<0.05) down-regulated it when compared to the untreated control group. Subsequent analysis of DNA degeneration and caspase activation revealed that Sunphenon (50 mu g/mL)/Polyphenon 60 (50 mu g/mL) inhibited activation of caspase-3 and showed an inhibitory effect on DNA degradation. From this result, we know that, in stress conditions, mu-calpain may be involved in the muscle atrophy through the suppression of myogenin and MyoD. Moreover, Sunphenon may regulate the skeletal muscle genes/promote skeletal muscle recovery by the up-regulation of myogenin and MyoD and suppression of mu-calpain and inflammatory pathways and may regulate the apoptosis pathways. Our findings suggest that dietary supplementation of Sunphenon might reduce inflammatory events in muscle-associated diseases, such as myotube atrophy.</P>

Simple Fluorescence Turn-On Chemosensor for Selective Detection of Ba²⁺ Ion and Its Live Cell Imaging

Ravichandiran, Palanisamy,Subramaniyan, Sivakumar Allur,Bella, Antony Paulraj,Johnson, Princy Merlin,Kim, Ae Rhan,Shim, Kwan Seob,Yoo, Dong Jin American Chemical Society 2019 ANALYTICAL CHEMISTRY - Vol.91 No.15

<P>A phenoxazine-based fluorescence chemosensor <B>4PB</B> [(4-(<I>tert</I>-butyl)-<I>N</I>-(4-((4-((5-oxo-5<I>H</I>-benzo[<I>a</I>]phenoxazin-6-yl)amino)phenyl)sulfonyl)phenyl)benzamide)] was designed and synthesized by a simple synthetic methods. The <B>4PB</B> fluorescence chemosensor selectively detects Ba<SUP>2+</SUP> in the existence of other alkaline metal ions. In addition, <B>4PB</B> showed high selectivity and sensitivity for Ba<SUP>2+</SUP> detection. The detection limit of <B>4PB</B> was 0.282 μM and the binding constant was 1.0 × 10<SUP>6</SUP> M<SUP>-1</SUP> in CH<SUB>3</SUB>CN/H<SUB>2</SUB>O (97.5:2.5 v/v, HEPES = 1.25 mM, pH 7.3) medium. This chemosensor functioned through the intramolecular charge transfer (ICT) mechanism, which was further confirmed by DFT studies. Live cell imaging in MCF-7 cells confirmed the cell permeability of <B>4PB</B> and its capability for specific detection of Ba<SUP>2+</SUP> in living cells.</P> [FIG OMISSION]</BR>

Crosstalk Between Co-Cultured 3T3-L1 and C2C12 Cells After the Exposure of Nano-Titanium Dioxide

Tripathi, Vinay Kumar,Sivakumar, Allur Subramaniyan,Dhasmana, Anupam,Hwang, Inho American Scientific Publishers 2018 Journal of nanoscience and nanotechnology Vol.18 No.6

<P>Nanotechnology is a promptly growing field in this century, and it have been extensively used in several solicitations. Reactive oxygen species (ROS) generation is one of the important mechanism of action of nanoparticles. The excess ROS generation can induce oxidative stress, so the cells are unable to sustain the normal biological redox-regulated tasks. The high oxidative stress and ROS formation condition, damage the biological macromolecules, cell signaling pathways and finally leads to cell death or cancer initiation. The objective of the present study is to reveal the effects of TiO2 nanoparticle on co-culture system. The cell viability, oxidative stress and apoptosis were evaluated in monolayer and co-culture 3T3-L1 cells after the exposure of TiO2. Our results indicated that TiO2 significantly induces the reactive oxygen species (ROS), lipid peroxidation and decrease in the level of glutathione. Additionally, real-time PCR data analysis shown an increased in the expression of p53, Bax, caspase-9, caspase-3 and decreased the level of Bcl-2, by this means specifying that apoptosis induced by TiO2 NPs occurs via the caspase-dependent pathway. This study analytically shows that oxidative stress is the fundamental mechanism by which TiO2 causes apoptosis in a co-culture system even at very low concentrations. In the future, the use of such nanoparticles should be cautiously scrutinized.</P>

Modulatory effect of linoleic and oleic acid on cell proliferation and lipid metabolism gene expressions in primary bovine satellite cells

Shah Ahmed Belal,Allur Subramaniyan Sivakumar,강다래,조상범,최호성,심관섭 한국통합생물학회 2018 Animal cells and systems Vol.22 No.5

This study was performed to elucidate the effects of linoleic acid (LA), oleic acid (OA) and their combination (LA + OA) on cell proliferation, apoptosis, necrosis, and the lipid metabolism related gene expression in bovine satellite cells (BSCs), isolated from bovine muscles. Cell viability was significantly increased with the OA and LA treatment. Furthermore, LA + OA enhanced cell proliferation in a dose-dependent manner (10 to 100 μM), whereas it lowered at 250 μM. In addition, a cell-cycle analysis showed that 100 μM of LA and OA markedly decreased the G0/G1 phase proportion (62.58% and 61.33%, respectively), compared to controls (68.02%), whereas the S-phase cells’ proportion was increased. The ratio of G2/M phase cells was not significantly different among the groups. Moreover, analyses with AO/EtBr staining showed that no apoptosis occurred. Necrosis were determined by flow cytometry using Annexin V-FITC/PI staining which revealed no early apoptosis in the cells pretreated with LA or OA, but occurred in the LA + OA group. We also analyzed the mRNA expression of lipid metabolizing genes such as peroxisome proliferator receptor alfa (PPARα), peroxisome proliferator receptor gamma (PPARγ), acyl-CoA oxidase (ACOX), lipoprotein lipase (LPL), carnitine palmitoyl transferase (CPT-1), and fatty-acid binding protein4 (FABP4), which were upregulated in LA or OA treated cells compared to the control group. In essence, LA and OA alone promote the cell proliferation without any apoptosis and necrosis, which might upregulate the lipid metabolism related gene expressions, and increase fatty-acid oxidation in the BSCs’ lipid metabolism.