Global climatic change and increasing climatic instability threaten crop productivity. Due to climatic change, drought stress is occurring more frequently in crop fields. In this study, we investigated the effect of treatment with hydrogen peroxide (H2O2) before leaf development on the growth and yield of sorghum for minimizing the damage of crops to drought. To assess the effect of H2O2 on the growth of sorghum plant, 10 mM H2O2 was used to treat sorghum leaves at the 3-leaf stage during growth in field conditions. Plant height, stem diameter, leaf length, and leaf width were increased by 7.6%, 9.6%, 8.3% and 11.5%, respectively. SPAD value, chlorophyll fluorescence (Fv/Fm), photosynthetic rate, stomatal conductance, and transpiration rate were increased by 3.0%, 4.9%, 26.0%, 23.4% and 12.7%, respectively. The amount of H2O2 in the leaf tissue of sorghum plant treated with 10 mM H2O2 was 0.7% of the applied amount after 1 hour. The level increased to approximately 1.0% after 6 hours. The highest antioxidant activity measured by the Oxygen Radical Absorbance Capacity assay was 847.3 µmol·g-1 at 6 hour after treatment. However, in the well-watered condition, the concentration of H2O2 in the plant treated by the foliar application of H2O2 was 227.8 µmol·g-1 higher than that of the untreated control. H2O2 treatment improved all the yield components and yield-related factors. Panicle length, plant dry weight, panicle weight, seed weight per plant, seed weight per unit area, and thousand seed weight were increased by 8.8%, 18.0%, 24.4%, 24.7%, 29.9% and 7.1%, respectively. Proteomic analysis showed that H2O2 treatment in sorghum increased the tolerance to drought stress and maintained growth and yield by ameliorating oxidative stress.
The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.
Water temperature governs various biological events in many aquatic animals including fish. Temperature changes the rates of gametogenesis and development, in some cases, is even capable of reversing fish sex. Treatments of fish with unusually high temperature are known to induce the expression of HSP70 gene. Development of an effective inhibitor for HSP70 gene expression is, thus, crucial to study the role of HSP70 in the temperature sensitive biological events. We have investigated the inhibitory effect of quercetin, 3,3',4',5,7-pentahydroxyflavon, a natural flavonid, on the expressions of HSP70 gene induced by high temperature ($36^{\circ}C$) in the Nile tilapia, Oreochromis niloticus, larvae and juveniles (10~13 cm in total length). The expression of HSP 70 gene was significantly decreased in tilapia larvae immersed in 50 ${\mu}M$ or 100 ${\mu}M$ quercetin solution for 6 hours before the exposure to high temperature (P<0.05). In particular, the level of HSP70 expression in fish treated with 100 ${\mu}M$ was as low as that of fish without high temperature treatment. Juveniles of tilapia were individually injected with 0.1 $m{\ell}$ of either 0.5 mM, 5 mM or 20 mM of quercetin solution before the exposure to high temperature. As the results, the expression of HSP70 gene in the gonad and brain of juvenile fish was significantly inhibited by the injection of 0.5 mM quercetin solution (P<0.05), but not by higher concentrations. We report, for the first time in the fish, that quercetin effectively inhibits the expression of HSP70 gene induced by high temperature and 100 ${\mu}M$ for the immersion of larvae and 0.5 mM for the injection to juvenile can be used for the effective concentrations for the study of temperature sensitive biological events in tilapia.
Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bel-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MIT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG ($2{\mu}M$) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.
This study presents the production characteristics and physiological characteristics of five Korean native chicken (KNC) breeds consisting of Hwanggalsaek Jaeraejong (HJ), Korean Rhode Island Red (KR), Korean White Leghorn (KL), Korean Brown Cornish (KC), and Korean Ogye (KO). We investigated their production performances, vitalities, and stress responses. We measured the survival rate, body weight, age at first egg-laying, hen-day egg production, egg weight, amount of telomeric DNA, heterophil-lymphocyte ratio (H/L ratio), and heat shock protein (HSP)-70, HSP-90α and HSP-90β gene expression levels for 493 KNCs. The survival rate was highest in KR, and lowest in KO. Body weights were steadily high in the order of KC, KR, HJ, KO and KL. Average hen-day egg production was highest in KL, and lowest in KC. While the amount of telomeric DNA was highest in KR, and lowest in KC. Furthermore, both the H/L ratio and the HSP-90β gene expression level were highest in KC, and lowest in KR. These results indicated that the KR breed was highly resistant to stress, whereas KC was more susceptible to stress. Taken together, it is considered that with improvements the KC breed would be more suited to be used as a Korean broiler breed while KL would be more appropriately used as a Korean layer breed. In addition, it is considered that the KR breed is appropriate to be used as a maternal chicken breeder based on good production capacity and excellent robustness, while the HJ breed is desirable to be improved as a high-quality Korean meat breed based on its excellent meat quality.
Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.
Background : Anti-apoptotic proteins may be involved in tumor development, progression and the response to treatment, Bcl-2 is by far the most studied anti-apoptotic protein. A novel inhibitor of apoptosis, designated survivin, and the heat shock proteins (HSPs) have recently been found in many human cancers. Immunohistochemical methods were used to determine the expression level of survivin, HSP70 and bcl-2 in non-small cell lung cancer (NSCLC) to evaluate their clinical significance. Materials and Methods : Tissue array slides were obtained from 99 surgically resected NSCLCs. Immunohistochemical staining was performed by an immuno-peroxidase technique using an avidin-biotinylated horseradish peroxidase complex. Anti-survivin rabbit polyclonal antibodies, anti-HSP70 mouse monoclonal antibodies and anti-bcl-2 mouse monoclonal antibodies were used as the primary antibodies. Results : Positive staining of survivin was detected in 33.3% of the cases. Survivin positivity is associated with to females and recurrence. A nonstatistically significant trend toward increased survivin expression was observed in non-smokers, and its expression inversely correlated with the number of cigarettes smoked in smokers. HSP70 was detected in 84.8% but this did not correlated with the clinicopathologic characteristics. Bcl-2 was detected in 18.2% and its expression correlated to tumor recurrence. No significant difference in the median survival time was noted in a comparison of all cases with survivin expression and those without. There was no association between HSP70 or bcl-2 expression and survival. Conclusion : Survivin expression was significantly associated with females and tumor recurrence. In addition its expression was inversely associated with the number of cigarettes smoked. However, HSP70 and bcl-2 expression were not associated with the clinical parameters or survival. This suggests that measuring the survivin levels may be useful in identifying patients at high risk for disease recurrence. Therefore, survivin might be a new diagnostic/therapeutic target in cancer.
Kim Tae-Hyung;Kim Kyung-Ju;Choe Mi-Kyung;Yeo In-Kyu
Journal of Aquaculture
/
v.19
no.2
/
pp.77-83
/
2006
This study was conducted to investigate changes of hemolymph count, antioxidant enzyme activities (catalase: CAT and superoxide dismutase: SOD) and Heat Shock Protein 70 (HSP70) mRNA in hemolymph, hepatopancreas and gill of abalone (Haliotis sieboldii) exposed to various water temperatures. Abalones were exposed to 10, 15, 20, 25 or $30^{\circ}C$ for 0, 6, 12, 24 or 48 hours. Survival rate of abalone was 100% at 10, 15, 20 and $25^{\circ}C$, but 0% at $30^{\circ}C$. Hemolymph counts increased at lower water temperatures (10 and $15^{\circ}C$) and decreased at $30^{\circ}C$. SOD activity decreased immediately after exposure to lower or higher water temperatures compared to the control ($20^{\circ}C$) with an exception at $30^{\circ}C$ where the activity increased. At lower temperatures, SOD activity rose high after 24 hours, but decreased again at 48 hours. At $25^{\circ}C$, it decreased compared to the control. CAT activity decreased immediately after exposure to 10 or $25^{\circ}C$ compared to the control, and then was recovered to the initial level after increment. At $15^{\circ}C$, CAT activity was high after 6 hours, and then was recovered to the initial level after increment. At $30^{\circ}C$, the activity decreased throughout the experiment. The HSP70 mRNA expression in gill increased at lower temperatures compared to the control ($20^{\circ}C$) and $25^{\circ}C$. In this study, rapid change of wale, temperature caused stress response in abalone which had been raised at $20^{\circ}C$. At molecular level, HSP70 was expressed rapidly, but antioxidant enzymes like SOD and CAT were expressed later than HSP70. At 15 and $25^{\circ}C$ of water temperatures, the HSP70, SOD and CAT expression were stable with time. However, at $30^{\circ}C$, all abalone died possibly because they could not develop resistance to high temperature.
Park, Ji Ae;Cho, Eun Jung;Choi, Eun Sik;Hong, Yeong Ho;Choi, Yeon Ho;Sohn, Sea Hwan
Korean Journal of Poultry Science
/
v.43
no.3
/
pp.177-189
/
2016
This study was conducted to verify the relationships between the expression values of stress-related markers and their production performances in 25 strains of Korean domestic chicken breeds. For stress response markers, the amount of telomeric DNA; expression levels of heat shock protein (HSP)-70, $HSP-90{\alpha}$, and $HSP-90{\beta}$; and comet scores were analyzed. Production performances were measured by the survival rate, body weights, days at first egg laying, egg weight and hen housed egg production. The results showed that the production traits and values of stress-related markers showed significant differences between strains. In general, the stress response of pure bred chickens with heavy weights was relatively high, while that of hybrid chickens with light weights was relatively low. The correlation coefficients between telomere contents and body weights showed that there were weak negative relationships. However, the correlations of telomere content with the survival rate and egg production were weakly positive after 20 weeks old. The expression levels of HSP genes and DNA damage rate (comet scores) were positively correlated to body weight, but were negatively correlated to the survival rate and egg production. The results implied that increasing body weight was associated with increasing HSPs expression and the DNA damage rate was associated with decreasing telomere content. In addition, increasing HSPs expression and the DNA damage rate decreased the survival rate and egg production, but the relationships with the telomere content was the reverse. Correlations among the stress-related markers showed that there were significant correlation coefficients between all of the marker values. HSPs expression was negatively correlated to the telomere content, while it was positively correlated to the DNA damage rate. There was a highly negative correlation between the telomere content and DNA damage rate. In conclusion, increasing the HSP values and DNA damage rate can promote telomere reduction, which led to a decrease in disease resistance and robustness of the chicken. Thus, increasing the stress response was verified to adversely affect the laying performance and viability of chickens.
Mutations in the APP gene lead to enhanced cleavage by ${\beta}-$ and ${\gamma}-secretase$, and increased $A{\beta}$ formation, which are closely associated with Alzheimer's disease (AD)-like neuropathological changes. Recent studies have shown that exercise training can ameliorate pathogenic phenotypes ($A{\beta}-42$, BDNF, GLUT-1 and HSP70) in experimental models of Alzheimer's disease. Here, we have used NSE/APPsw transgenic mice to investigate directly whether exercise training ameliorates pathogenic phenotypes within Alzheimer's brains. Sixteen weeks of exercise training resulted in a reduction of $A{\beta}-42$ peptides and also facilitated improvement of cognitive function. Furthermore, GLUT -1 and BDNF proteins produced by exercise training may protect brain neurons by inducing the concomitant expression of genes that encode proteins (HSP-70) which suppress stress induced neuron cell damages from APPsw transgenic mice. Thus, the improved cognitive function by exercise training may be mechanistically linked to a reduction of $A{\beta}-42$ peptides, possibly via activation of BDNF, GLUT-1, and HSP-70 proteins. On the basis of the evidences presented in this study, exercise training may represent a practical therapeutic management strategy for human subjects suffering from Alzheimer's disease.
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