mRNA Expression Of Heat Shock Protein Research Articles

High Dietary Zinc Promotes its Optimal Absorption Through Modulation of Key Transporter Genes, SOD1, and HSP70 Expression in Heat-stressed Rats.

Global warming causes heat stress (HS) in animals, impacting nutrient absorption and metabolism. Antioxidant nutrients are crucial for combating HS. This study assessed the impact of increased dietary Zn on nutrient utilization, mineral absorption, and expression of Zn homeostasis regulators, superoxide dismutase-1 (SOD1), and heat shock protein-70 (HSP70) genes in rats under HS. Seventy-two four-week-old Wistar rats were assigned to six groups in a 3×2 factorial design, with three dietary Zn levels (14.6, 32.7, and 48.9 ppm) and two environments, thermo-neutral (TN) and HS, for 42 days, including 14 days of HS exposure. Results showed that HS reduced nutrient intake across Zn levels, though ether extract digestibility increased at 32.7 and 48.9 ppm Zn. Intake, excretion, and apparent daily absorption of Ca, P, Zn, Cu, Mn, and Fe were lower in HS than in TN groups. Hepatic metallothionein-1 (MT1) mRNA expression was downregulated in rats fed 14.6 and 32.7 ppm Zn compared to 48.9 ppm Zn under both environmental conditions. Duodenal Zinc transporter-1 (ZnT1) and Zrt- and Irt-like protein-1 (ZIP1) mRNA expression increased with dietary Zn under TN and HS conditions, respectively. Hepatic SOD1 mRNA expression was significantly downregulated in HS groups, while hepatic HSP70 mRNA expression was significantly upregulated at 48.9 ppm Zn under HS. Present study suggests that, under HS conditions in rats, a higher dietary Zn level of 48.9 ppm may be optimal for improving Zn absorption, enhancing ZIP1, MT1, and HSP70 gene expression, and alleviating the negative effects of HS.

Melatonin injection and red light irradiation affect the antioxidant response and cell damage in disk abalone (Haliotis discus hannai) exposed to high water temperatures.

The effects of red light-emitting diode (LED) light irradiation (630 nm, 0.5 W/m2) and melatonin (10-8 and 10-7 M) on oxidative stress and physiological responses in abalones exposed to high temperatures (28°C) were investigated. Changes in messenger RNA (mRNA) expressions of melatonin receptor (MT-R), heat shock protein 70 (HSP70), and antioxidant enzymes, as well as alterations in H2O2 levels in the hemolymph, were examined. The results revealed that high-temperature-stressed abalones treated with melatonin injections or exposed to red LED light showed a significant increase in MT-R mRNA expression, while HSP70 mRNA expression decreased. Notably, HSP70 mRNA expression levels in the red LED light-irradiated group were similar to those in the group injected with 10-8 M melatonin after 24 h exposure. Abalones treated with melatonin at 20°C or irradiated with red LED light exhibited decreased H2O2 levels and reduced antioxidant enzyme mRNA expression compared with those of the control group. However, the high-temperature environment induced oxidative stress in abalones, leading to increased antioxidant enzyme mRNA expression compared with that under 20°C conditions. Moreover, abalones exposed to high-temperature stress exhibited hepatopancreatic DNA damage, which was attenuated by melatonin treatment or red LED light irradiation. Hence, red LED light reduces oxidative stress, boosts antioxidant enzymes, and alleviates DNA damage in high-temperature-stressed abalones, akin to 10-8 M melatonin treatment. Therefore, considering the practical challenges of continuous melatonin administration to abalones, utilizing red LED light emerges as a practical, effective alternative to protect abalones from oxidative stress compared to 10-8 M melatonin treatment.

Association of heat-shock protein 70.1 gene with physiological and physical performance of Bali cattle.

Global warming challenges cattle productivity and welfare since it affects heat stress and scarce feed. The heat-shock protein 70 (HSP70) gene is essential in cytoprotection against stressors, protecting cells from dysregulated gene expression and apoptosis. This study aimed to identify significant genetic markers of the HSP70.1 gene that can be leveraged genetically to enhance thermotolerance and production in Bali cattle further. Animals were sampled from three different rearing systems. In this study, 83 healthy adult male Bali cattle without abnormalities were utilized. Single-nucleotide polymorphism (SNP) diversity associated with the physiological and physical traits of Bali cattle was assessed using SNPStat online software. Gene expression for putative SNPs and their genotypic groups was further evaluated. There were 15 polymorphic SNPs (c.-185G>A, c.-69T>G, c.10G>C, c.19A>G, c.45C>T, c.101INS, c.115T>C, c.130T>C, c.136G>T, c.159G>C, c.164G>T, c.234G>A, c.303G>A, c.333C>A, and c.456C>T) identified, of which 12 were associated with the assessed trait. Nine SNPs were associated with physiological traits, while eight were with physical traits. The c.136G>T as a novel, high minor allele frequency, and associative SNP was selected for HSP70 gene expression. Individuals with the TT genotype have a trim physique, susceptible physiology, and high HSP70 mRNA expression. On the other hand, the GG genotype was significantly associated with larger physique, lower physiology, and low HSP70 mRNA expression. The higher expression may indicate that HSP70.1 is involved in mitigating the deleterious effects of stress. As a result, the animal experienced negative energy balance, decreasing body size. Single-nucleotide polymorphism c.136G>T is a candidate biomarker for heat resistance traits in Bali cattle.

Anti-HSP70 alleviates cell migration and proliferation in colorectal cancer cells (CRC) by targeting CXCR4 (in vitro study).

Colorectal cancer (CRC), the third most common cancer in men and women, accounts for 8% of new cancer cases in the US and 8 to 9% of the nation's anticipated cancer mortality in 2014. In the pathophysiology of colon cancer, heat shock protein 70 (HSP70) and CXCR4 are essential. In this research, we concentrated on the connection between CXCR4 expression and HSP70 inhibitor activity in the development of colorectal cancer. The HSP70 inhibitor's effect on cell proliferation was also evaluated. Samples were obtained from patients with CRC; the surrounding marginal tissues were considered healthy. The One CRC cell lines (HCA-7) were divided into two groups based on untreated and treated with anti-HSP70. HSP70 and CXCR4 mRNA expression and migration (Wound healing assay) were measured in these groups. Also, we evaluated the expression levels of HSP70 and CXCR4 in thirty CRC and healthy non-cancerous samples (Using Real-time PCR and Western Blotting). Moreover, we examined the viability of CRC cells in untreated and treated groups with anti-HSP70. Higher expression levels of CXCR4 (p < 0.0001) and HSP70 (p = 0.002) mRNA were observed in patients who had CRC. In contrast, lower mRNA expressions of HSP70 (p < 0.0001) and CXCR4 (P < 0.0001) were detected in the CRC cell line (HCA-7) after being treated with anti-HSP70. Moreover, the viability and migration of cancer cells were remarkably reduced in CRC cells treated with anti-HSP70. Our study's innovation was the in vitro demonstration of inhibiting HSP70 in the CRC cancer cell line drastically reduced CXCR4 expression, viability, and cancer cell migration. These findings may pave the way for additional studies on CRC cancer treatment and be examined in vivo in studies, given that the primary goal of therapy is to decrease the viability and spread of cancer cells.

Impacts of Habitat Quality on the Physiology, Ecology, and Economical Value of Mud Crab Scylla sp.: A Comprehensive Review

The water of the mangrove ecosystem and surrounding coastal areas are gradually shrinking due to the intense destruction. Therefore, the effects of the physicochemical properties of the habitat water on the in-habitant species must be studied. Scylla sp. is involved in the food chain and bioturbation structure formation in mangrove forests. Five major electronic databases, such as PubMed, Scopus, Web of Science, AGRICOLA, and Google Scholar, were systematically searched to review the cause and effects of influencing abiotic factors, mainly physicochemical properties of habitat water, including water pollution on Scylla sp. Responses of mud crabs at biochemical, molecular, physiological, growth, reproduction, and production level were independently reviewed or in relation to physicochemical properties of habitat water, pathogens, heavy metals, and harmful chemicals present in their habitat water. Review results suggest that these crabs are mostly under threats of overfishing, varied physicochemical properties of habitat water, pathogens, heavy metals, and chemical toxicants in water, etc. At low temperatures, the expression of calreticulin and heat shock protein-70 mRNA expression is elevated. Like melatonin, the hormone serotonin in mud crabs controls ecdysteroids and methyl farnesoate at 24 °C, 26 ppt salinity, and pH 7.2 of habitat water, facilitating their reproduction physiology. Xenobiotics in habitat water induce toxicity and oxidative stress in mud crabs. These crabs are prone to infection by white spot and rust spot diseases during the winter and spring seasons with varied water temperatures of 10–30 °C. However, elevated (65%) weight gain with higher molting at the juvenile stage can be achieved if crabs are cultured in water and kept in the dark. Their larvae grow better at 30 ± 2 °C with salinity 35 ppt and 12 hL/12 hD day length. So, monitoring habitat water quality is important for crab culture.

Alleviating effect of exogenous melatonin and calcium on the peroxidation damages of cucumber under high temperature stress

To explore whether there is an interaction between melatonin (MT) and calcium (Ca2+) in regulating heat tolerance of plants, we analyzed the response of endogenous MT and Ca2+ to heat stress, and examined the effect of MT and Ca2+ on the reactive oxygen (ROS) accumulation, antioxidant system, and transcripts of heat shock factor (HSF) and heat shock proteins (HSPs) of cucumber seedlings under high temperature stress. Seedlings were foliar sprayed with 100 μmol·L-1 MT, 10 mmol·L-1 CaCl2, 3 mmol·L-1 ethylene glycol tetraacetic acid (EGTA, Ca2+ chelating agent) +100 μmol·L-1 MT, 0.05 mmol·L-1 chlorpromazine (calmodulin antagonist, CPZ) +100 μmol·L-1 MT, 100 μmol·L-1 p-chlorophenylalanine (p-CPA, inhibitor of MT) +10 mmol·L-1 CaCl2 or deionized water (H2O), respectively. The results showed that both endogenous MT and Ca2+ in cucumber seedlings were induced by high temperature stress. The seedlings treated with exogenous MT showed significant increases in the mRNA expression of calmodulin (CaM), calcium-dependent protein kinase (CDPK5), calcineurin B-like protein (CBL3) and CBL interacting protein kinase (CIPK2) compared with the control at normal temperature. The mRNA levels of tryptophane decarboxylase (TDC), 5-hydroxytryptamine-N-acetyltransferase (SNAT) and N-acetyl-5-hydroxytryptamine methyltransferase (ASMT), key genes of MT biosynthesis and endogenous MT content were also induced by Ca2+ in cucumber seedlings. Exogenous MT and CaCl2 alleviated the heat-induced oxidative damage through increasing antioxidant ability, reducing the accumulation of reactive oxygen species (ROS), and upregulating the mRNA abundances of HSF7, HSP70.1 and HSP70.11, as evidenced by mild thermal damage symptoms, lower heat injury index and electrolyte leakage under heat stress. The positive effect of MT-induced antioxidant capacity and mRNA expression of HSPs was removed by adding EGTA and CPZ in stressed seedlings. Similarly, the mitigating role of Ca2+ in the peroxidation damage to high temperature stress was reversed by p-CPA. These results suggested that both MT and Ca2+ could induce heat tolerance of cucumber seedlings, which had crosstalk in the process of heat stress signal transduction.

DNA Fragmentation and mRNA Expression of Bcl-2, Bcl-xL, p53, p21 and HSP70 Genes in Nondysplastic and Dysplastic Oral Lichen Planus.

Background:Oral lichen planus (OLP) is a chronic inflammatory disease. Apoptosis of the basal keratinocytes is a causative factor for OLP pathogenesis but the detailed mechanism of apoptosis among nondysplastic and dysplastic OLP lesions is yet unraveled.Aims:This study aims to evaluate the involvement of cellular DNA fragmentation and alteration in the expression of Bcl-2 and B-cell lymphoma extra-large (Bcl-xL), p53, p21 and heat shock protein 70 (HSP70) in nondysplastic and dysplastic OLP lesions.Materials and Methods:Untreated, fifteen OLP patients each with nondysplastic and dysplastic lesions were enrolled for this study. Their DNA fragmentation was analyzed by the agarose gel electrophoresis method. The mRNA expression of Bcl-2, Bcl-xL, p53, p21 and HSP70 were measured using semi-quantitative reverse transcription-polymerase chain reaction.Results:Elevated DNA fragmentations were found in dysplastic lesions compared to nondysplastic type. Significantly higher expression of Bcl-2, Bcl-xL, p53 and p21 were found in both types of OLP lesion compared to the control. Expression of Bcl-2 and Bcl-xL were significantly elevated in nondysplastic lesions, whereas significantly overexpression of p53 and p21 were found in dysplastic lesions. Anti-stress protein HSP70 was overtly expressed in dysplastic lesions compared to other groups.Conclusion:Reduced expression of Bcl-2 and Bcl-xL, with elevated DNA fragmentation, may be associated with increased apoptosis in dysplastic lesions which aid in the resolution of the chronic inflammatory process. Higher expression of p53 and p21 in dysplastic lesions reflect its malignant potentiality. Overexpression of HSP70 in dysplastic lesions is a useful marker for higher cellular stress.

Central administration of neuropeptide Y reduces the cellular heat stress response and may enhance spleen antioxidative functions in heat-exposed chicks

Previously it was found that mRNA expression of neuropeptide Y (NPY) was increased in the chicken brain under heat stress. NPY has also been reported as an anti-stress factor to regulate brain functions in heat-exposed chicks. However, to the best of our knowledge, there is no report on the action of central NPY in the immune organs under heat stress. The aim of this study was to examine whether central injection of NPY can regulate heat stress response in the spleen and liver. After intracerebroventricular (ICV) injection of NPY, chicks were exposed to control thermoneutral temperature (CT: 30 ± 1 °C) or high ambient temperature (HT: 35 ± 1 °C) chambers for 60 min. Central injection of NPY caused lowering in rectal temperature under CT, but not under HT. Moreover, ICV injection of NPY caused a significant lower mRNA expression of heat-shock protein-70 and higher expression of glutathione synthase in the spleen, but not liver. Furthermore, plasma uric acid concentrations were significantly increased by the ICV injection of NPY in chicks under HT. These results indicate that brain NPY may contribute to attenuate the intracellular heat stress response and enhance antioxidative status in the immune organ, spleen in chicks.

Hematological adaptations in diploid and triploid Salvelinus fontinalis and diploid Oncorhynchus mykiss (Salmonidae, Teleostei) in response to long-term exposure to elevated temperature

Hematology is a simple but reliable method to determine the influence of environmental stressors on the physiology and performance of teleost fish. Understanding the detailed impacts of elevated temperature on the hematology of fish can help us to evaluate fish responses to climate change. Therefore, cellular and biochemical changes in peripheral blood were investigated in diploid (2n) and triploid (3n) brook trout and in 2n rainbow trout exposed to 20 °C for 32 days in comparison to fish acclimated to 9 °C. Additionally, survival and growth rates and the relative mRNA expression of heat-shock proteins were recorded. All brook trout and rainbow trout survived exposure to 20 °C. At 20 °C, growth was decreased in 2n rainbow trout, increased in 2n brook trout and similar to growth at 9 °C in 3n brook trout. The erythrocyte cell volume, nucleus volume, cell surface, and nucleocytoplasmatic ratio were significantly lower at 20 °C than at 9 °C in all investigated species/ploidy levels. In contrast, the erythrocyte surface-to-volume ratio was not affected by temperature. In 2n and 3n brook trout, erythrocyte concentration and blood hemoglobin content were increased at 20 °C; in 2n rainbow trout, there were no differences in comparison to 9 °C. Exposure of 2n and 3n brook trout and 2n rainbow trout to 20 °C had no negative effect on cellular and molecular immune components in blood. Serum diagnostic enzymes and metabolites indicated neither organ inflammation nor disease nor disturbance in energy metabolism or nutrition status in fish exposed to 20 °C. In addition, the mRNA expression of Hsp70 and Hsp90 relative to the 28S ribosomal protein S32 did not differ between 9 °C and 20 °C.

Liposome quercetin enhances the ablation effects of microwave ablation in treating the rabbit VX2 liver tumor model

Objective This study aimed to investigate whether liposomal quercetin (LQ) could enhance the effects of microwave ablation (MVA) in treating the rabbit VX2 liver tumor model. Methods Rabbits with VX2 liver tumors were randomly divided into three groups: intravenous LQ group (LQ group), MWA group and LQ combined with MWA (LQ + MWA) group. Five rabbits were randomly selected and sacrificed from each group at 12 h and on days 3, 7 and 14 of the operation. The tumor samples were detected and quantified by immunohistochemistry, Western blot, and reverse transcription polymerase chain reaction (RT-PCR). Results For up to 7 days, the coagulation necrosis volume (CV) of the LQ + MWA group was larger than that of MWA and LQ groups (p < 0.05). Fourteen days after the operation, the total tumor volume of the LQ + MWA group was smaller than that of the LQ group and the MWA group (p < 0.05). The survival time of the LQ + MWA group was significantly longer than that of the MWA and LQ groups (p < 0.01). Heat shock protein 70 (HSP70), hypoxia inducible factor-1 α (HIF-1 α), vascular endothelial growth factor (VEGF), tumor microvessel density (MVD) were lower in the LQ + MWA group than the MWA and LQ groups at 12 h, on days 3 and 7. At hour 12 and on days 3 and 7, HSP70 mRNA and HIF-1α mRNA expression of MWA group were significantly higher than that of the LQ and LQ + MWA groups (p < 0.001). At 12 h, and on days 3 and 7, apoptotic rate of tumor cells in LQ + MWA group was higher than that of the MWA and LQ groups (p < 0.05). At 12 h and on days 3, 7 and 14, the proliferation index of tumor cells in residual tumor in LQ + MWA group was lower than that in the MWA and LQ groups (p < 0.05). Conclusion Preoperative infusion of LQ can significantly enhance the MWA effects of liver VX2 tumor, inhibit the excessive proliferation of residual tumor and angiogenesis, and decrease metastasis and prolong the survival period of experimental animals.

Effect of medical herbs in Tian-Dong-Tang-Gan powder on the oxidative stress induced by ammonia and nitrite in Litopenaeus vannamei

In order to evaluate the effect of different doses of Tian-Dong-Tang-Gan powder (TDTGP) on the immune function of Litopenaeus vannamei, and the molecular mechanism of anti-ammonia stress and anti-nitrite stress, and screening for the best dose, a 35-day feeding experiment and a 72-h ammonia stress or nitrite stress experiment were carried out. The results showed that when compared with the blank control group, the different doses of TDTGP could increase the body weight (BW), percent weight gain (PWG), body length (BL) and percent body length growth (PBLG), and the activities of acid phosphatase (ACP), phenol oxidase (PO), inducible nitric oxide synthase (i-NOS) and the total antioxidant capacity (T-AOC) in the hemolymph of L. vannamei during the feeding experiment, and the effect of TDTGP-4 treatment group was the best. When exposed ammonia or nitrite stress for 72 h, the PO activity was decreased in the hemolymphs, and the mRNA expression of PPO, MT, GSH-Px, Mn-SOD, HSP70 and HIF-1α in the hepatopancreas of L. vannamei. However, different TDTGP doses were fed to the shrimp followed by the application of ammonia and nitrite stresses. The activities of ACP, PO, i-NOS and T-AOC in the hemolymphs remained stable, and the mRNA expression of PPO, ACP, metallothionein (MT), glutathione peroxidase (GSH-Px), superoxide dismutase (Mn-SOD), heat shock protein 70 (HSP70), Hypoxia inducible factor 1α (HIF-1α) and other factors in the hepatopancreas could be maintained or improved to varying degrees in comparison with the blank control group and positive group. These results indicated that the TDTGP can improve growth performance and non-specific immune response of L. vannamei, and by maintaining the mRNA expression of the antioxidative stress factor, such as MT, GSH-PX, Mn-SOD, HSP70, HIF-1α and PPO and ACP in the hepatopancreas, acting against the damage of hepatopancreas caused by ammonia and nitrite stress in L. vannamei. The changes in BW, WG, activities of PO and ACP, and mRNA expression levels of PO, Mn-SOD and ACP indicated that the optimum supplement of dietary TDTGP should be 4 g per·kg.

Protective influence of betaine on intestinal health by regulating inflammation and improving barrier function in broilers under heat stress

This research was executed to study the impacts of adding betaine (BT) to broiler diets on intestinal inflammatory response and barrier integrity under heat stress (HS). At 21 d of age, 150 male broilers (Ross 308) were randomly assigned to 3 treatment groups: control (CON) group, in which broilers were provided standard finisher feed under thermoneutral condition (22 ± 1°C); HS group and HS + BT group, in which broilers were given the standard feed supplied with 0 and 1,000 mg/kg BT, respectively, under cyclic HS condition (33 ± 1°C for 8 h from 08:00 to 16:00 h and the thermoneutral temperature for the residual hours). Each treatment was replicated ten times with 5 broilers per replicate. The HS group showed an elevation (P < 0.05) in serum corticosterone (CORT) concentration, D-lactate acid (D-LA) content, and diamine oxidase (DAO) activity, mucosal interleukin-1β (IL-1β) level, and expression of heat shock protein 70 (HSP70) gene, and a reduction (P < 0.05) in mucosal interleukin-10 (IL-10) level and secretory immunoglobulin A (SIgA) content and relative abundance of mRNA for occludin (OCLN), zonula occludens-1 (ZO-1), claudin-1 (CLDN1), and claudin-4 (CLDN4). In contrast, broilers in the HS + BT group exhibited a raise (P < 0.05) in mucosal IL-10 level and SIgA content and relative expression of OCLN and ZO-1 genes, and a decline (P < 0.05) in serum CORT concentration and DAO activity, mucosal IL-1β level, and expression of HSP70 mRNA. These results indicate that supplemental BT can ameliorate intestinal injury in heat-challenged broilers by suppressing inflammatory responses and enhancing mucosal barrier function.

Melatonin Restores the Developmental Competence of Heat Stressed Porcine Oocytes, and Alters the Expression of Genes Related to Oocyte Maturation.

Simple SummaryMelatonin improves the quality and in vitro maturation (IVM) of oocytes under heat stress. Melatonin treatment counteracts the adverse effects induced by heat stress (HS), such as the poor survival rate and maturation rate, distribution of α-tubulin and F-actin, expression of NRF2 and GDF9 mRNA. However, HS and melatonin have similar effects on increasing expression of HSP70 and NRF2 mRNA. Furthermore, HS inhibits expression of GDF9 mRNA.Melatonin enhances the quality and in vitro maturation (IVM) of oocytes under heat stress (HS), but the mechanism of melatonin in reducing HS injury on oocytes is not fully understood. In this study, porcine cumulus-oocyte complexes (COCs) were randomly divided into three groups. The COCs of the control group were cultured at 38.5 °C for 42 h, and the COCs of the HS group were cultured at 41.5 °C for 4 h, and then transferred into 38.5 °C for 38 h. The COCs of the HS + melatonin group were cultured with 10−9 M melatonin under the same conditions as the HS group. The survival rate, maturation rate, distribution of α-tubulin and F-actin of the oocytes were assessed. In addition, the expression profiles for genes related to the oocyte maturation, including heat shock protein 70 (HSP70), nuclear factor erythroid 2-related factor 2 (NRF2), cyclin-dependent kinase 1 (CDK1), growth differentiation factor 9 (GDF9) were analyzed by real-time quantitative PCR. The results showed that HS decreased the survival rate and maturation rate, distribution of α-tubulin and F-actin, but melatonin treatment could partly counteract these adverse effects. In addition, HS increased expression of HSP70 and NRF2 mRNA, and melatonin treatment had a similar effect on HSP70 expression, but had a contrary effect on NRF2 expression. Furthermore, HS inhibited expression of CDK1 and GDF9 mRNA, but melatonin treatment could weaken the effect on GDF9 expression induced by HS. In summary, melatonin treatment could attenuate the unfavorable effects induced by HS to enhance developmental competence of porcine oocytes during IVM.

Ameliorative effect of dietary lipopolysaccharides on Oreochromis niloticus juveniles submitted to aflatoxin B1‐induced oxidative stress and bacterial challenge

The current study was designed to evaluate the ameliorative effect of lipopolysaccharides (LPS) (a portion of the Gram –ve bacteria cell wall) on aflatoxin (AFB1)-persuaded changes in the cytokine expression, heat-shock protein 70 (HSP70), antioxidant genes, different haematological and biochemical parameters in Nile Tilapia along with the infection by Aeromonas hydrophila and the immune-stimulant efficiency of LPS on the growth pattern. For this study, two hundred and forty O. niloticus (35 ± 1.5 g) were used. The fish were randomly allocated into four experimental groups, four replicates for each group (15 fish/replicate). Group 1 was the negative control group fed on a basal diet, and group 2 was the positive control group fed on the basal diet with aflatoxins (AFB1) 200 µg/kg feed. Group 3 has fed the lipopolysaccharides 20 mg/100 gm diets. Group 4, besides the AFB1-contaminated basal diet, was provided with lipopolysaccharides 20 mg/100 gm diet. At the end of the 12 weeks, each group was injected with A. hydrophila. The analysis revealed that LPS-treated AFB1-supplemented fish showed improvement in haematological, lysosomal and phagocytic activities and total protein and significantly decreased liver enzyme exaggerated by aflatoxin; alongside, the LPS-treated aflatoxin-supplemented fish revealed a significant effect on the haematological and the immunity markers after 24 h, 48 h and 1 week post challenged with A. hydrophila. Concerning the gene expression, AFB1 group showed significant up-regulation of HSP70 and interferon gamma gene mRNA expression, which is significantly down-regulated by LPS treatment, besides LPS showed up-regulation of catalase and superoxide dismutase genes markedly alongside with interleukins (IL), IL-8 and IL-1B; the binding effect of LPS markedly appears after A. hydrophila which significantly modulates the HSP70 and immunity marker genes in addition to the antioxidant-related genes, CAT and SOD. Histopathological findings reflected a marked increase in intestinal villi length in LPS-treated groups. Our view illuminates for the first time the pathways of molecular wellness for the role of LPS in AFB1 toxicity and bacterial infection. Our observation illuminates the molecular fitness pathways for the LPS that can aide to the development of a bacterial disease tolerance strategy and potentially boost immune response in O. niloticus fish.

Clinical Implications of HSC70 Expression in Clear Cell Renal Cell Carcinoma.

Purpose: The role of heat shock protein 70 (HSC70) in the progression of clear cell renal cell carcinoma (ccRCC) is unclear. This study explored the effect of the HSC70 on the survival of ccRCC patients.Methods: Immunohistochemical analysis was performed to determine HSC70 expression in samples obtained from 121 ccRCC patients with at least 5 years of follow-up. We also analyzed the association between HSC70 expression and clinicopathological characteristics. Furthermore, the association of overall survival (OS) with HSC70 expression was analyzed using Kaplan-Meier curves. Finally, we used the Oncomine and CCLE databases to determine the effects of HSC70 mRNA expression on ccRCC.Results: HSC70 expression was associated with distant metastasis and death of ccRCC patients. HSC70 was expressed in the nucleus and/or cytoplasm of ccRCC cells. The incidence of distant organ metastasis and death was higher in patients with HSC70 expression than in those without it. Survival analysis revealed that patients with HSC70 expression had significantly shorter OS. Oncomine analyses also showed that the HSC70 mRNA was significantly upregulated in ccRCC tissues.Conclusions: HSC70 expression was related to adverse prognosis, and patients with HSC70 expression had a worse prognosis than those without HSC70 expression. HSC70 may thus serve as a potential therapeutic target for ccRCC.

The Induction of Heat Shock Protein 70 after Oral Administration of Concentrated Brewed Rice Vinegar Kurozu in Mice.

Heat shock protein 70 (HSP70) is induced by various stresses. Since HSP70 has a protein refolding activity and an anti-inflammatory activity, the HSP70 induction will help cells from harmful acute stresses. Feeding a diet containing concentrated brewed rice vinegar Kurozu (CK) diet for 5 wk resulted in an increase of HSP70 in the brains of mice. In the present study, we evaluated whether oral feeding of 25 μL CK induces HSP70 mRNA in brain and other tissues. HSP70 mRNA was significantly increased in the esophagus, small intestine, liver, and brown adipose tissue within 1 h after the oral administration of CK. A weaker induction of HSP70 mRNA was demonstrated in the stomach, large intestine, and brain. HSP70 mRNA induction returned to basal levels within 3 h after feeding. We doubted that the induction of HSP70 mRNA was caused by manual restraint of the mice during CK administration. Manual restraint of the mice did not influence HSP70 mRNA expression in intestine 1 h after these treatments. Our results suggest that transient HSP70 mRNA induction by oral feeding of CK was not caused by retention stress. There are some compounds in CK that increase HSP70 mRNA in various tissues.

Nitric Oxide Negatively Regulates Larval Metamorphosis in Hard-Shelled Mussel (Mytilus coruscus)

Marine invertebrates undergo distinct life cycles and development processes, and some undergo a metamorphic transition from pelagic larvae to benthic juvenile. Previous studies have shown that the second messenger nitric oxide (NO) plays important roles in regulating larval metamorphosis in marine invertebrates. However, this regulatory function is not conserved. Herein, we investigated the regulatory role of NO in larval metamorphosis in Hard-shelled mussel (Mytilus coruscus). Pharmacological experiments revealed that after exposure to nitric oxide synthase (NOS) inhibitors, the larval metamorphosis rate was increased significantly. By contrast, exposure to NO donors significantly reduced the larval metamorphosis rate. To further investigate the function of NO in metamorphosis, we cloned the NOS cDNA sequence from M. coruscus (McNOS), analyzed amino acid sequences of McNOS from different species, and measured McNOS mRNA temporal expression alongside the molecular chaperone heat shock protein 90 (HSP90) in different larval stages. The results showed that NOS proteins share a highly conserved domain sequence in different species, even in invertebrates and vertebrates, but their classification among invertebrates and vertebrates is not the same. Except for the umbo veliger stage, McNOS mRNA was expressed at low levels in pediveliger larval and post-larvae stage, indicating a potential inhibitory function for NO in larval metamorphosis. Meanwhile, HSP90 mRNA expression was correspondingly high in the umbo veliger stage and lower in the pediveliger larval and post-larvae stage, indicating potential functional synergism. Interestingly, McNOS enzyme activity in different larval stages was inconsistent with its mRNA expression. Metamorphosis assays of pharmacological treatments combined with McNOS and HSP90 gene expression and NOS enzyme activity analyses indicated that NO acts as a suppressor to regulate metamorphosis in M. coruscus, regardless of whether nitric oxide is endogenous or exogenous.

Dynamics of cytokines and heat shock proteins mRNA expression in thermally manipulated chicken challenged with intraperitoneally injection of Lipopolysaccharide (LPS).

Heat stress is known to significantly affect broiler immunity and cytokine expression, but it is not yet clear how embryonic thermal manipulation (TM) might help alleviate the aforementioned effects. The aim of the present study was to evaluate the effects of TM on the splenic mRNA expressions of IL‐1β, IL‐6, TNF‐α, IFN‐α, IFN‐β, IFN‐γ, IL‐4, IL‐8, IL‐15, IL‐16, IL‐17, IL‐18, Hsp70 and HSF‐3 in broilers intraperitoneally injected with lipopolysaccharide (LPS). TM was performed by elevating the incubation temperature to 39°C at 65% relative humidity for 18 h daily during embryonic days 8 to 16. Whereas, control group was incubated at standard condition during the whole incubation period (37.8°C and 56% relative humidity). LPS was intraperitoneally injected at a dose of 0.5 mg/kg body weight (BW) on post‐hatch days 10 and 24. spleens was collected from all groups to evaluate mRNA expression by relative‐quantitative real‐time RT‐qPCR. TM results in alteration in the mRNA expression dynamics of cytokine and heat shock protein following intraperitoneally injection of LPS. The results of this study provided us with a valuable information about the mechanisms by which thermal manipulation may modify the immune system response in the thermally manipulated chicken.Support or Funding InformationDeanship of Research/ Jordan University of Science &amp; Technology for its financial support of this work (Grant#: 44/2019).

Isolation and Structure Determination of a Heat Shock Protein Inducer, Asparagus-Derived Proline-Containing 3-Alkyldiketopiperazines (Asparaprolines), From a Standardized Extract of Asparagus officinalis Stem

ETAS® has been developed from the stems of Asparagus officinalis L. as a functional ingredient for nutraceuticals. ETAS possesses heat shock protein 70 (HSP70) induction activity and may contribute to maintenance and improvement of health. Here, 3 compounds (1, 2, 3) were isolated from ETAS. The structures of 1, 2, and 3 were deduced by HREIMS and NMR spectroscopic data, and the compounds were identified as cyclo(l-Phe-l-Pro), cyclo(l-Tyr-l-Pro), and cyclo(l-Leu-l-Pro), respectively. Each compound contained a diketopiperazine ring derived from proline with an alkyl group at C-3; thus, we termed them asparagus-derived proline-containing 3-alkyldiketopiperazines (Asparaprolines). In an HSP70 mRNA induction assay in HL-60 cells, Asparaprolines significantly enhanced the expression of HSP70 mRNA compared with a control. To our knowledge, these results demonstrate for the first time that proline-containing diketopiperazines derived from natural amino acids exhibit HSP70 mRNA induction activity.

Effect of synbiotics on thyroid hormones, intestinal histomorphology, and heat shock protein 70 expression in broiler chickens reared under cyclic heat stress

This study examined effect of a dietary synbiotic supplement on the concentrations of plasma thyroid hormones, expressions of heat shock protein 70 (HSP70), and intestinal histomorphology in broiler chickens exposed to cyclic heat stress (HS). Three hundred and sixty day old male Ross 708 broiler chicks were randomly distributed among 3 dietary treatments containing a synbiotic (PoultryStar meUS) at 0 (control), 0.5 (0.5×), and 1.0 (1.0×) g/kg. Each treatment contained 8 replicates of 15 birds each housed in floor pens in a temperature and lighting controlled room. Heat stimulation was established from days 15 to 42 at 32°C for 9 h daily. The results indicated that under the HS condition, both synbiotic fed groups had lower liver and hypothalamus HSP70 levels (P < 0.001) compared to control group; however, HSP70 mRNA expression was not different among treatments (P > 0.05). There were no treatment effects on the levels of triiodothyronine (T3) and thyroxine (T4) as well as T3/T4 ratio (P > 0.05). Compared to controls, 1.0× HS broilers had greater villus height in the duodenum (P < 0.01), and greater villus height and villus height:crypt depth ratios in the ileum (P < 0.01). There were no differences among treatments on the measured intestinal parameters in the jejunum (P > 0.05). The results suggest that the synbiotic may ameliorate the negative effects of HS on chicken health as indicated by the changes in the intestinal architecture and the levels of HSP70. Dietary synbiotic supplement could be a feasible nutritive strategy for the poultry industry to improve the health and welfare of chickens when exposed to hot environmental temperature.