Modulatory effect of heat stress on viability of primary cultured chicken satellite cells and expression of heat shock proteins ex vivo.

Satellite cells promote muscle repairing and muscle growth. Thereby the intention of the present study was to investigate the beneficial effects of heat stress at different time intervals on chicken satellite cells' viability. Satellite cells were isolated from 1-day-old chicks and treated at two different temperatures (37 °C and 41 °C) for various time periods (6 h, 12 h, 24 h, 48 h, and 72 h). Both temperatures significantly increased cell viability after 24 h and 48 h. After 12 h, cell viability was significantly increased at 41 °C compared to 37 °C. However, more apoptotic cells were observed at end of the experiment of 41 °C compared to 37 °C. In addition, more live cells were found at early of experimental period at 41 °C than 37 °C. Additionally, protein and mRNA expression of HSP70, HP60 and HSP47 were significantly upregulated throughout the experimental period at temperature of 41 °C compared to those at 37 °C. These results indicate that cell viability and expression of heat stress related proteins/genes are induced by high temperature of 41 °C via heat stress pathway whereas activation of heat stress related proteins/genes are lower at 37 °C. Thus, 41 °C can trigger satellite cells' viability essential for better cell survival than 37 °C at early incubation time.

Direct exposure to mild heat stress stimulates cell viability and heat shock protein expression in primary cultured broiler fibroblasts.

Fibroblasts produce collagen which is mainly essential for repairing tissue damage and maintaining the structural integrity of tissues. However, studies have given scientific evidence about harmful effect of thermal manipulation in fibroblast. Therefore, the aim of this study was to determine the mild heat stress temperature which increased broiler fibroblast viability. The experiment was divided into two groups (37 °C and 41 °C), and each group was divided into five subgroups based on different incubation times (6 h, 12 h, 24 h, 48 h, and 72 h) with three replications. In experimental group (41 °C), fibroblast viability increased significantly in 12 h but decreased in 72 h compared with control (37 °C). At 41 °C, live cell increased significantly in 24 h and then declined in 48 h as well as 72 h than control. Moreover, the S phase lengthened in shorter incubation time of experimental group compared with control. Protein and mRNA (HSP70, HSP60, and HSP47) expressions were significantly higher at 41 °C compared with 37 °C, but at the end of the experiment, HSP expression level was higher in both groups. Finally, this study recommended 41 °C as a mild heat stress temperature for increasing broiler fibroblast viability.

Effects of In Ovo Supplementation with Nanonutrition (L-Arginine Conjugated with Ag NPs) on Muscle Growth, Immune Response and Heat Shock Proteins at Different Chicken Embryonic Development Stages.

The aim of the study was to analyze the in ovo injection of inorganic and organic synthesized silver nanoparticles (Ag NPs) using Brassica oleracea L. var. capitate F. rubra (BOL) conjugation with L-Arginine (L-Arg) on the immune, muscle growth, survivability and hatchability of broiler chickens. The conjugation of L-Arg (100 µg) with 1000 µg of Ag NPs synthesized by (BOL)-extract and L-Arg (100 µg) conjugated with 100 µg of Ag NPs inorganic synthesized were injected into fertile eggs at 8 d, 14 d and 18 d of incubation. Survival and hatching rate were significantly improved in the dose of L-Arg (100 µg) with 1000 µg (BOL-Ag NPs) and L-Arg (100 µg) with 100 µg (C-Ag NPs) on 14 d injection whereas it was decreased on 8 d or 18 d injection. Moreover, the protein expression of muscle development markers such as myogenin and myoD were significantly uprelated in 14 d of incubation whereas the heat shock proteins (HSPs), such as HSP-60 and HSP-70, were significantly upregulated in 18 d incubation. In addition, the liver function marker of serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) were significantly decreased and the immunoglobulin (IgM) levels were increased in a 14 d incubation period in serum at the same concentration.

Molecular and Cellular Response of Co-cultured Cells toward Cobalt Chloride (CoCl2)-Induced Hypoxia.

Cobalt chloride (CoCl2) is a well-known hypoxia mimetic mediator that induces hypoxia-like responses. CoCl2, a mediator confirmed to alleviate hypoxia-inducible factor-1 (HIF-1), has been associated with a variety of hypoxic responses. HIF-1 is the foremost transcriptionfactor that is particularly activated during hypoxia and regulates various genes. Therefore, this study aimed to investigate the cellular and molecular responses of the co-cultured cells under the influence of the CoCl2-induced hypoxic condition. Mono- and co-cultured C2C12 and 3T3-L1 cells were exposed to CoCl2, and a significant induction in HIF-1, reactive oxygen species and lipid peroxidase and a reduction in glutathione and catalase were observed. The expressions of proapoptotic genes like Bax, p53, caspase-9, and caspase-3 were notably increased, whereas the antiapoptotic gene, i.e., Bcl2, was downregulated during hypoxia in mono- as well as co-cultured C2C12 cells. However, the co-cultured C2C12 cells show significantly lower induction in oxidative stress and expression of apoptotic genes in comparison to monocultured C2C12 cells. Whereas, the co-cultured 3T3-L1 cells show comparatively higher oxidative stress and apoptotic event in comparison to monocultured 3T3-L1 cells. The reason may be the communication between the cells and some soluble factors that help in cell survival/death from hypoxia. Moreover, it may also be due to the fact that fat and muscle cells interact and communicate via proximity and mutual ability when growing together. Therefore, the co-culture system provides a unique approach to intercellular communication between the two different cell types.

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

A phenoxazine-based fluorescence chemosensor 4PB [(4-(tert-butyl)-N-(4-((4-((5-oxo-5H-benzo[a]phenoxazin-6-yl)amino)phenyl)sulfonyl)phenyl)benzamide)] was designed and synthesized by a simple synthetic methods. The 4PB fluorescence chemosensor selectively detects Ba2+ in the existence of other alkaline metal ions. In addition, 4PB showed high selectivity and sensitivity for Ba2+ detection. The detection limit of 4PB was 0.282 µM and the binding constant was 1.0 × 106 M-1 in CH3CN/H2O (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 4PB and its capability for specific detection of Ba2+ in living cells.

Effect of In Ovo Injection of L-Arginine in Different Chicken Embryonic Development Stages on Post-Hatchability, Immune Response, and Myo-D and Myogenin Proteins.

The aim of this study was to evaluate the effect of in ovo injection with different ratios of L-arginine (L-Arg) into Ross broiler eggs at three different embryonic developmental stages (eighth day (d), 14th day, and 18th day) on the survival, hatchability, and body weight (BW) of one-day-old hatched chicks. Additionally, we have analyzed the levels of serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT), the protein expression of heat shock proteins (HSPs), and we have also determined micronuclei (MN) and nuclear abnormality (NA). In addition, the genotoxic effect was observed in peripheral blood cells such as the presence of micronuclei and nuclear abnormalities in the experimental groups. The results showed that survival and hatching rates as well as body weight were increased on the 14th day of incubation compared to the eighth and 18th day of incubation at lower concentrations of L-Arg. Moreover, the levels of SGOT and SGPT were also significantly (p < 0.05) increased on the 14th day of incubation at the same concentration (100 µg/µL/egg) of injection. In addition, immunoglobulin (IgM) levels were increased on the 14th day of incubation compared to other days. The protein expressions of HSP-47, HSP-60, and HSP-70 in the liver were significantly down-regulated, whereas the expression of myogenin and myoblast determination protein (MyoD) were significantly up-regulated on the 14th day after incubation when treated with all different doses such as 100 µg, 1000 µg, and 2500 µg/µL/egg, namely 3T1, 3T2, and 3T3, respectively. However, the treatment with low doses of L-Arg down-regulated the expression levels of those proteins on the 14th day of incubation. Histopathology of the liver by hematoxylin and eosin (H&E) staining showed that the majority of liver damage, specifically intracytoplasmic vacuoles, were observed in the 3T1, 3T2, and 3T3 groups. The minimum dose of 100 µg/mL/egg on the 14th day of incubation significantly prevented intracytoplasmic vacuole damages. These results demonstrate that in ovo administration of L-Arg at (100 µg/µL/egg) may be an effective method to increase chick BW, hatch rate, muscle growth-related proteins, and promote the immune response through increasing IgM on the 14th day of the incubation period.

Synthesis and Anticancer Evaluation of 1,4-Naphthoquinone Derivatives Containing a Phenylaminosulfanyl Moiety.

1,4-Naphthoquinones are exceptional building blocks in organic synthesis and have been used to synthesize several well-known pharmaceutically active agents. Herein we report the synthesis, structural characterization, and biological evaluation of new phenylaminosulfanyl-1,4-naphthoquinone derivatives. We evaluated the cytotoxic activity of the synthesized compounds against three human cancer cell lines: A549, HeLa, and MCF-7. Most of the synthesized compounds displayed potent cytotoxic activity. Specifically, compounds 5 e [3,5-dichloro-N-(4-((4-((1,4-dioxo-3-(phenylthio)-1,4-dihydronaphthalen-2-yl)amino)phenyl)sulfonyl)phenyl)benzamide], 5 f [N-(4-((4-((1,4-dioxo-3-(phenylthio)-1,4-dihydronaphthalen-2-yl)amino)phenyl)sulfonyl)phenyl)-3,5-dinitrobenzamide], and 5 p [N-(4-((4-((1,4-dioxo-3-(phenylthio)-1,4-dihydronaphthalen-2-yl)amino)phenyl)sulfonyl)phenyl)thiophene-2-carboxamide] showed remarkable cytotoxic activity. The synthesized compounds showed low toxicity in normal human kidney HEK293 cells. The cytotoxic mechanism of compounds 5 e, 5 f, and 5 p was explored in MCF-7 cells. The results confirmed that these three compounds induce apoptosis and arrest the cell cycle at the G1 phase. In addition, compounds 5 e, 5 f, and 5 p were found to induce apoptosis via upregulation of caspase-3 and caspase-7 proteins as well as by upregulation of the gene expression levels of caspases-3 and -7. Our findings demonstrate that compounds 5 e, 5 f, and 5 p could be potent agents against a number of cancer types.

Highly Flexible Electrospun Hybrid (Polyurethane/Dextran/Pyocyanin) Membrane for Antibacterial Activity via Generation of Oxidative Stress.

A hybrid nanofibrous mat consisting of polyurethane, dextran, and 10 wt % of biopigment (i.e., pyocyanin) was facilely fabricated using a direct-conventional electrospinning method. The field emission scanning electron microscopy showed the bead-free fibers with a twisted morphology for the pyocyanin-loaded mat. The addition of pyocyanin enables the unprecedented approach to tailor the hydrophilicity of hybrid mat, as verified from the water contact measurement. Thermomechanical stabilities of electrospun mats were investigated in terms of thermogravimetric analysis, differential scanning calorimetry, and dynamic mechanical analysis. The bacterial inhibition test revealed that the antibacterial activity of electrospun mat containing pyocyanin was 98.54 and 90.2% toward Escherichia coli and Staphylococcus aureus, respectively. By the combined efforts of rapid release of pyocyanin and oxidative stress, the PU-dextran-pyocyanin (PUDP) electrospun mat significantly declined the viable cell number that disrupts the cell morphology. Hence, the proposed PUDP electrospun mat must meet the requirements of efficient antimicrobial material in various applications such as disinfectant wiping, food packaging, and textile industries.

One-Pot Facile Synthesis of Pt Nanoparticles Using Cultural Filtrate of Microgravity Simulated Grown P. chrysogenum and Their Activity on Bacteria and Cancer Cells.

Platinum nanoparticles (Pt NPs) was synthesized via a facile and cost competitive ont-pot green mediated synthesis using cell free cultural filtrate (microgravity simulated grown Penicillium chrysogenum) as a reducing agent. The toxicity effect of synthesized Pt NPs toward myoblast C2C12 carcinoma cells was then investigated. The particle size analyzer (DLS) and transmission electron microscopy (TEM) results demonstrates that both NG-Pt NPs and MG-Pt NPS are spherical in shape with an average diameter of 15 nm and 8.5 nm, respectively. The results from UV-visible (UV-vis) spectroscopy and X-ray diffraction (XRD) analysis show a characteristic strong resonance centered at 265 nm and a single crystalline nature, respectively. The results derived from in vitro cytotoxicity showed a significant concentration-dependent decrement in cell viability when C2C12 cells were exposed to Pt NPs. Such decrement in cell viability is because of increased reactive oxygen species (ROS) generation. Cell apoptosis was proved by acridine orange-ethidium bromide (AO/EtBr) dual staining, annexin V-FITC/PI-staining and immunocytochemistry. Moreover, the protein expression of both (i) apoptosis related proteins such as cas-3 and cas-9, (ii) inflammatory response proteins such as TNF-α, TGF-ß and NF-κB were significantly upregulated in MG-Pt NPs treated cells than NG-Pt NPs treated cells. Uptake and intracellular localization of MG-Pt NPs caused by accumulation of autophagosomes in C2C12 cells and bacterial cells, indicate that synthesized MG-Pt NPs enable for the swift cell apoptosis than NG-Pt NPs. Interestingly, At the concentration of 40 and 80 µg/ml MG-Pt NPs showed more potent cytotoxicity toward cancer cells while, under identical concentration, NG-Pt NPs exhibited rather lower cytotoxicity. Overall, our results demonstrated that MG-Pt NPs could be selectively inhibit the growth of cancer cells via ROS-mediated nucleus NF-κB and caspases activation when compared to NG-Pt NPs.

Biological Differences between Hanwoo longissimus dorsi and semimembranosus Muscles in Collagen Synthesis of Fibroblasts.

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 different fibroblast cells derived from Hanwoo m. longissimus dorsi (LD) and semimembranosus (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 muscle. 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.

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).

Cell-Cell Communication Between Fibroblast and 3T3-L1 Cells Under Co-culturing in Oxidative Stress Condition Induced by H2O2.

The present study was carried out to understand the interaction between fibroblast and 3T3-L1 preadipocyte cells under H2O2-induced oxidative stress condition. H2O2 (40 µM) was added in co-culture and monoculture of fibroblast and 3T3-L1 cell. The cells in the lower well were harvested for analysis and the process was carried out for both cells. The cell growth, oxidative stress markers, and antioxidant enzymes were analyzed. Additionally, the mRNA expressions of caspase-3 and caspase-7 were selected for analysis of apoptotic pathways and TNF-α and NF-κB were analyzed for inflammatory pathways. The adipogenic marker such as adiponectin and PPAR-γ and collagen synthesis markers such as LOX and BMP-1 were analyzed in the co-culture of fibroblast and 3T3-L1 cells. Cell viability and antioxidant enzymes were significantly increased in the co-culture compared to the monoculture under stress condition. The apoptotic, inflammatory, adipogenic, and collagen-synthesized markers were significantly altered in H2O2-induced co-culture of fibroblast and 3T3-L1 cells when compared with the monoculture of H2O2-induced fibroblast and 3T3-L1 cells. In addition, the confocal microscopical investigation indicated that the co-culture of H2O2-induced 3T3-L1 and fibroblast cells increases collagen type I and type III expression. From our results, we suggested that co-culture of fat cell (3T3-L1) and fibroblast cells may influence/regulate each other and made the cells able to withstand against oxidative stress and aging. It is conceivable that the same mechanism might have been occurring from cell to cell while animals are stressed by various environmental conditions.