Hsp70 Gene Expression Research Articles

Role of the heat shock protein family in chlorpromazine-induced cardiotoxicity.

Chlorpromazine (CPZ), a first-generation antipsychotic, is widely used in treating schizophrenia and other psychiatric disorders. However, CPZ is also associated with an increased likelihood of sudden cardiac death, and the underlying mechanisms remain unclear. In our study, we aimed to determine the CPZ-induced changes in some members of the heat shock protein family in rat hearts and further explore the possible mechanisms of CPZ-induced cardiotoxicity. Twenty-four Sprague Dawley rats were randomly divided into three groups (n = 8 per group): control, low dose (33.216 mg/kg) and high dose (94.211 mg/kg). CPZ administration induced hypothermia in rats. Pathological changes, including ischaemia and hypoxia, were observed in rat hearts. Furthermore, the serum levels of cardiac Troponin T (c-TN-T) and brain natriuretic peptide (BNP) were elevated in the CPZ-exposed groups. Meanwhile, the protein and gene expression of HSP70, HSP60, HSP27 and HSP10 significantly differed between the CPZ-exposed and control groups. We conclude that acute CPZ exposure could lead to myocardial injury in rats, in which HSPs might play a crucial role. Further investigations are required to elucidate the underlying mechanisms.

Effect of Quercetin Nanoparticles on Hepatic and Intestinal Enzymes and Stress-Related Genes in Nile Tilapia Fish Exposed to Silver Nanoparticles.

Recently, nanotechnology has become an important research field involved in the improvement of animals' productivity, including aquaculture. In this field, silver nanoparticles (AgNPs) have gained interest as antibacterial, antiviral, and antifungal agents. On the other hand, their extensive use in other fields increased natural water pollution causing hazardous effects on aquatic organisms. Quercetin is a natural polyphenolic compound of many plants and vegetables, and it acts as a potent antioxidant and therapeutic agent in biological systems. The current study investigated the potential mitigative effect of quercetin nanoparticles (QNPs) against AgNPs-induced toxicity in Nile tilapia via investigating liver function markers, hepatic antioxidant status, apoptosis, and bioaccumulation of silver residues in hepatic tissue in addition to the whole-body chemical composition, hormonal assay, intestinal enzymes activity, and gut microbiota. Fish were grouped into: control fish, fish exposed to 1.98 mg L-1 AgNPs, fish that received 400 mg L-1 QNPs, and fish that received QNPs and AgNPs at the same concentrations. All groups were exposed for 60 days. The moisture and ash contents of the AgNP group were significantly higher than those of the other groups. In contrast, the crude lipid and protein decreased in the whole body. AgNPs significantly increased serum levels of ALT, AST, total cholesterol, and triglycerides and decreased glycogen and growth hormone (*** p < 0.001). The liver and intestinal enzymes' activities were significantly inhibited (*** p < 0.001), while the oxidative damage liver enzymes, intestinal bacterial and Aeromonas counts, and Ag residues in the liver were significantly increased (*** p < 0.001, and * p < 0.05). AgNPs also significantly upregulated the expression of hepatic Hsp70, caspase3, and p53 genes (* p < 0.05). These findings indicate the oxidative and hepatotoxic effects of AgNPs. QNPs enhanced and restored physiological parameters and health status under normal conditions and after exposure to AgNPs.

Functional and molecular immune responses in Octopus vulgaris skin mucus and haemolymph under stressful conditions

The common octopus (Octopus vulgaris) is a cephalopod species with a high economic market value, hence one of the species in spotlight to be reared in aquaculture. The increasing importance of animal welfare has ethical considerations and also implications for improving quality standards. Unfortunately, defence mechanisms in cephalopods are largely unexplored and a comprehensive insight on cephalopod physiological and immune responses in relation to stress becomes particularly prominent. Therefore, the aim of the present study was to gather deeper insights on functional and molecular stress biomarkers in the common octopus. For this purpose, adult individuals were exposed to a stressful condition by limiting movements in the aquarium for 17 days. Innate immune parameters were assessed in skin mucus and plasma samples whereas the expression level of genes coding for antioxidant enzymes, related to immune functions, or involved in folding proteins was analysed in haemocytes at several sampling times. Our results indicated that most of the innate immune parameters measured (haemolytic, lysozyme, esterase, peroxidase, and protease activities) in skin mucus showed substantial variations at middle term of experiment (i.e., 11 days). After 11 days of movement limitations, the haemolytic and protease activities decreased in the octopus plasma. At the transcriptional level, lyz, hsp90 and ire1 gene expression values were particularly down-regulated while C1-like, prdx and sod transcripts showed up-regulation over an early stage (i.e., 5 days) in haemocytes from octopuses under stressful conditions. Moreover, C1-like, prdx and sod mRNA expression decreased at 11 and 17 days whereas lyz and hsp90 gene expression level augmented at 11 days in exposed octopus compared to unexposed specimens, with the subsequent increase at plasma and skin mucus level at long term. Overall, results from the present study showed potential functional and molecular biomarkers which could be of assistance in studies comprising stressful conditions in the common octopus. Moreover, biomarkers from skin mucus could also be considered as a low-invasive approach to assess and improve cephalopods welfare.

HSP70 Expression Signature in Renal Cell Carcinoma: A Clinical and Bioinformatic Analysis Approach.

Heat shock proteins (HSPs) are cytoprotective against stressful conditions, as in the case of cancer cell metabolism. Scientists proposed that HSP70 might be implicated in increased cancer cell survival. This study aimed to investigate the HSP70 (HSPA4) gene expression signature in patients with renal cell carcinoma (RCC) in correlation to cancer subtype, stage, grade, and recurrence, combining both clinicopathological and in silico analysis approaches. One hundred and thirty archived formalin-fixed paraffin-embedded samples, including 65 RCC tissue specimens and their paired non-cancerous tissues, were included in the study. Total RNA was extracted from each sample and analyzed using TaqMan quantitative Real-Time Polymerase Chain Reaction. Correlation and validation to the available clinicopathological data and results were executed. Upregulated HSP70 (HSPA4) gene expression was evident in RCC compared to non-cancer tissues in the studied cohort and was validated by in silico analysis. Furthermore, HSP70 expression levels showed significant positive correlations with cancer size, grade, and capsule infiltration, as well as recurrence in RCC patients. The expression levels negatively correlated with the overall survival (r = -0.87, p < 0.001). Kaplan-Meier curves showed lower survival rates in high HSP70 expressor group compared to the low expressors. In conclusion, the HSP70 expression levels are associated with poor RCC prognosis in terms of advanced grade, capsule infiltration, recurrence, and short survival.

Performance, blood biochemistry, carcass fatty acids, antioxidant status, and HSP70 gene expressions in Japanese quails reared under high stocking density: the effects of grape seed powder and meal.

Japanese quails reared under high stocking density (SD) were evaluated for the effects of grape seed powder (GSP) and meal (GSM) supplementation on performance, blood biochemistry, thigh and breast muscle fatty acids, antioxidant status, and HSP70 gene expression. We randomly assigned 288 (15-day-old) quail chicks to six treatment groups in a factorial design (2 × 3) with four replicates, involving two density levels [160 cm2/bird (LD) and 80 cm2/bird (HD)] and three feed forms (FFs) [no supplementation, grape seed powder (3% GSP), grape seed meal (3% GSM)]. SD had a significant effect on live weight, but not on weekly feed intake, daily weight gain, and feed conversion ratio. Serum creatinine and aspartate aminotransferase levels were significantly affected by FF and SD × FF (p < 0.05). A high SD reduced the n-3/n-6 ratio of breast muscle and a significant interaction was found between FF (p < 0.001). The SD × FF interaction reduced the Σn-6 ratio in HDM's thigh muscle, whereas in LDM, the ratio increased (p < 0.01). At high SD, neither GSP nor GSM reduced biological markers of oxidative stress (p > 0.05). Compared to GSP, GSM had higher efficacy at reducing HSP70 levels related to high SD levels. Despite this, at high SD, a diet containing 3% of GSP and GSM was not effective in overcoming oxidative stress. Therefore, more studies using different doses of GSM and GSP in quail diets would be beneficial.

Effect of Air Exposure and Re-Submersion on the Histological Structure, Antioxidant Response, and Gene Expression of Procambarus Clarkii.

Air exposure is an important environmental stressor during the transportation and cultivation of Procambarus clarkii. We evaluated the effect of re-submersion for 24 h after dry transportation for 24 h on the histological structure, antioxidant activity, and gene expression of crayfish. The antioxidant parameters of catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and lactate dehydrogenase (LDH), and the relative expression of CAT, SOD, HSP70, and ferritin genes were subsequently measured in the hepatopancreas and gills at both stages. Histopathology found that air exposure led to vacuolation of the hepatopancreas and disorderly arrangement of respiratory epithelial cells (REC) in the gills. The activities of catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA), and lactic dehydrogenase (LDH) in the hepatopancreas and gills increased with short-term air exposure. The relative expression of genes (CAT, SOD, HSP70, and Ferritin) were induced after short-term air exposure. During re-submersion, MDA content and CAT and SOD activities in the hepatopancreas and gills were restored after 24 h, however, LDH activity and hepatopancreatic tissue damage were not repaired. Our results indicate that air exposure can cause oxidative damage to P. clarkii, and CAT and SOD can be used to determine the response of crayfish exposed to air, in addition to some damage that can be eliminated after re-submersion to a limited degree. This study provides foundational data that re-submersion can improve crayfish performance under hypoxic stress to a certain extent and will lead to the development of more effective transportation strategies and decrease economic losses in the future.

Influences of Bacillus subtilis and fructooligosaccharide on growth performances, immune responses, and disease resistance of Nile tilapia, Oreochromis niloticus.

The present study investigated the effects of Bacillus subtilis and fructooligosaccharide (FOS) on growth performances, immunity improvement, and disease resistance of Nile tilapia (Oreochromis niloticus). The fish (24.5 ± 1.6 g) were fed a basal diet (G1), diets supplemented with 1 g/kg (G2), 3 g/kg (G3) and 5 g/kg (G4) of FOS as well as diets supplemented with 1 × 109 CFU/g (G5), 3 × 109 CFU/g (G6) and 5 × 109 CFU/g (G7) of B. subtilis for 56 days. After the feeding trial, the complement C3, IL-1β, TNF-α, IFN-γ, hsp70 gene expression in the liver was then analyzed by a quantitative Real-time PCR. Then, fish were infected with Streptococcus agalactiae, and the survival rate was recorded. The results showed that FOS and B. subtilis had no significant effect (P > 0.05) on growth performances and survival rate. Lysozyme activity was significantly greater in the G4, G5, G6, and G7 groups. Also, all fish fed FOS and B. subtilis showed significantly (P < 0.05) higher respiratory burst activity than other groups. The expressions of complement C3, IL-1β, TNF-α, IFN-γ, and hsp-70 in the liver were significantly higher for fish fed 5 g/kg of FOS as well as for fish that received any concentration level of B. subtilis (P < 0.05) used in the study. After the S. agalactiae challenge test, the survival rate of fish-fed diets supplemented with FOS and B. subtilis was slightly higher than for the control group. The results indicated that FOS and B. subtilis could stimulate immune responses and immune-related genes in tilapia. However, further investigation of other prebiotics or herbs in combination with B. subtilis is encouraged at molecular levels and screening for beneficial metabolites that may increasingly improve digestive enzymes, growth performances, and health benefits in tilapia. In addition, on-farm experiments are needed.

Acute effects of a single moderate-intensity exercise bout performed in fast or fed states on cell metabolism and signaling: Comparison between lean and obese rats

AimsAlthough the benefits of exercise can be potentiated by fasting in healthy subjects, few studies evaluated the effects of this intervention on the metabolism of obese subjects. This study investigated the immediate effects of a single moderate-intensity exercise bout performed in fast or fed states on the metabolism of gastrocnemius and soleus of lean and obese rats. Main methodsMale rats received a high-fat diet (HFD) for twelve weeks to induce obesity or were fed standard diet (SD). After this period, the animals were subdivided in groups: fed and rest (FER), fed and exercise (30 min treadmill, FEE), 8 h fasted and rest (FAR) and fasted and exercise (FAE). Muscle samples were used to investigate the oxidative capacity and gene expression of AMPK, PGC1α, SIRT1, HSF1 and HSP70. Key findingsIn relation to lean animals, obese animals' gastrocnemius glycogen decreased 60 %, triglycerides increased 31 %; glucose and alanine oxidation decreased 26 % and 38 %, respectively; in soleus, triglycerides reduced 46 % and glucose oxidation decreased 37 %. Exercise and fasting induced different effects in glycolytic and oxidative muscles of obese rats. In soleus, fasting exercise spared glycogen and increased palmitate oxidation, while in gastrocnemius, glucose oxidation increased. In obese animals' gastrocnemius, AMPK expression decreased 29 % and SIRT1 increased 28 % in relation to lean. The AMPK response was more sensitive to exercise and fasting in lean than obese rats. SignificanceExercise and fasting induced different effects on the metabolism of glycolytic and oxidative muscles of obese rats that can promote health benefits in these animals.

High Thermosensitivity of MDA-MB-231 Cells as a Prerequisite for Thermoradiosensitization of Triple-negative Breast Cancer in Clinical Practice

The work is dedicated to the analysis of thermal sensitivity and efficiency of thermosensitization to γ-radiation of three cell lines of breast cancer of different molecular subtypes. It has been established that the cells of the studied lines differ in the sensitivity to hyperthermia and, as a result, in the effectiveness of thermoradiosensitization, which raises the question of finding criteria for assessing the rationale of using hyperthermia in each specific case. It has been shown that the level of heat-responsive expression of HSP70 gene and also the level of cell genomic instability can serve as such criteria. It was found that the efficiency of thermoradiosensitization is significantly higher in MDA-MB-231 cells compared to the cells of other studied lines. The data obtained suggest that the use of hyperthermia as a radiotherapy-sensitizing co-treatment may be particularly promising toward triple negative breast cancer.

Adjusted Thr: Lys Ratio Improved the Performance and Efficiency of Japanese Quail

Two hundred and forty Japanese quail aged 125 days were randomly allocated to five treatment groups based on laying (%) and quail’s weight (90.71 ± 1.8% egg/day × 100 and 178.05 ± 9.38 g, respectively), each of which included six replicates of eight quails. The diets were formulated based on corn, soybean meal, and industrial amino acids. An optimal diet achieves 100% of amino acids required by the quail requirements, except for threonine. Five treatments were made: 20% less amino acid; 10% less amino acid; optimal diet; 10% more amino acid; and 20% more amino acids than those in the optimal diet. The increase in amino acid levels in a fixed Lys: amino acid ratio led to histological alterations in the liver and uterine epithelium, reduction in blood lipid peroxidation, lower hepatic HSP70 gene expression, and the performance of laying Japanese quail. The optimal diet based on the NRC with an adjusted Thr: Lys 78 ratio (Lys 1.0%) improved the performance and efficiency of Japanese quail from 125 to 230 days of age.

Effect of Leishmania RNA virus 2 on virulence factors and cytokines gene expression in a human macrophage infected with Leishmania major: A preliminary study

Cutaneous leishmaniasis (CL) is one of the most important infectious parasitic diseases in the world caused by the Leishmania parasite. In recent decades, the presence of a virus from the Totiviridae family has been proven in some Leishmania species. Although the existence of LRV2 in the Old world Leishmania species has been confirmed, almost no studies have been done to determine the potential impact of LRV2 on the immunopathogenicity of the Leishmania parasite.In this preliminary study, we measured the expression of target genes, including Glycoprotein 63 (gp63), Heat Shock Protein 70 (hsp70), Cysteine Protease b (cpb), Interleukin 1 beta (IL-1β), IL8 and IL-12 in LRV2 positive Leishmania major strain (LRV2+L. major) and LRV2 negative L. major strain (LRV2-L. major). We exposed THP-1, a human leukemia monocytic cell line, to promastigotes of both strains. After the initial infection, RNA was extracted at different time points, and the relative gene expression was determined using a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Findings showed that the presence of LRV2 in L. major was able to increase the expression of gp63, hsp70, and cpb genes; also, we observed lower levels of expression in cytokine genes of IL-1β, IL-8, IL-12 in the presence of LRV2+, which are critical factors in the host's immune response against leishmaniasis. These changes could suggest that the presence of LRV2 in L. major parasite may change the outcome of the disease and increase the probability of Leishmania survival; nevertheless, further studies are needed to confirm our results.

Effect of emodin on growth performance, antioxidant status, intestinal morphology and heat shock protein gene expressions of freshwater prawn (Macrobrachium rosenbergii)

The purpose of this study was to assess the effect of emodin on biochemical variables, antioxidant activity, heat shock protein gene expression and prawn intestinal morphology (Macrobrachium rosenbergii). Three feeds containing 0 mg kg–1 (T1, control group), 25 mg kg–1 (T2) and 50 mg kg–1 (T3) of emodin were formulated and fed for eight weeks. Alanine aminotransferase, total cholesterol and triglyceride contents were significantly lowered although glucose tended to increase following the emodin remedy. Superoxide dismutase variables in haemolymph were dramatically upgraded, whilst malondialdehyde and superoxide anion situations were significantly lowered when compared to control. Emodin supplementation had no influence on glutathione peroxidase contents. Emodin remedy dropped the expression of HSP60 and HSP90 genes in comparison to the control. In T2, the intestinal epithelial cells were complete, well-organised and tightly affixed to the basement membrane, in discrepancy to the slightly disunited basement membrane in the control group. The emodin-treated prawn individuals had a thicker and full intestinal morphology than the control groups. Supplementation with emodin at dosages of 25 mg kg–1 improved growth performance, blood biochemical, antioxidant capacity and immune gene expression.

Electrospun silk fibroin and gelatin blended nanofibers functionalized with noble metal nanoparticles for enhanced biomedical applications

The combination of highly anticancer active platinum group metals and biocompatible natural fibers exhibits significant anticancer activity while also allowing cell regeneration and repair to the damaged region. In this research, platinum and palladium nanoparticles anchored silk fibroin and gelatin (SF/GL/Pd-Pt) hybrid composite nanofiber was fabricated through the electrospinning method. The amino groups in SF and GL act as a reducingd stabilizing agent for the resulting Pd and Pt nanoparticles. Morphological and functional studies confirmed that crystalline Pd and Pt nanoparticles with average sizes of 5.11 and 3.03 nm are anchored on a 154.93 ± 34.19 nm SF/GL nanofiber matrix. The atomic force microscopic (AFM) images show that the SF/GL/Pd-Pt composite nanofiber has a thickness of 4.74 nm and a roughness area (Ra) of 258.5 nm. X-ray photoelectron spectroscopic (XPS) results confirmed the presence of C, N, O, Pt, and Pd in the composite fiber with an atomic percentage of 67.56%, 11.05%, 17.72%, 2.01% and 1.66%. respectively. This SF/GL/Pd-Pt composite nanofiber is more effective than its counterparts in terms of anticancer activity (IC50 value of 28.82 µg/mL−1) against A549 human lung carcinoma cells. Furthermore, the β-actin and Hsp90 gene expression in A549 cells confirmed its superior anticancer activity.

The effects of overwintering temperature on the survival of female adult mud crab, Scylla paramamosain, under recirculating aquaculture systems as examined by histological analysis of the hepatopancreas and expression of apoptosis-related genes

The effects of the overwintering temperature (8, 12, 16, and 20 °C) on survival, antioxidant capacity, and heat shock protein (HSP) gene expression in Scylla paramamosain were examined during an experimental period of eight weeks. The effects of the temperature on survival were parabolic, with the highest and lowest survival rates occurring at 16 °C and 8 °C, respectively. Histological examination of the hepatopancreas revealed that the hepatopancreas tissue structure remained intact and that cell boundaries were clearly observed at 16 °C and 20 °C. However, hepatopancreas cells in the 12 °C and 8 °C groups appeared to be vacuolated and indistinct, respectively. Furthermore, high levels of reactive oxygen species (ROS) and malondialdehyde (MDA) and low levels of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in the hepatopancreas were observed in the 8 °C group. Hsp60 and hsp70 gene expression were dramatically upregulated, but hsp10, hsp40, hsp90, and glucose regulatory protein 78 (grp78) gene expression were downregulated at 8 °C. The terminal deoxynucleotidyl transferase deoxy-UTP nick end labeling (TUNEL) method showed that there was more DNA damage at 8 °C and 12 °C than at 16 °C. Moreover, apoptosis was significantly induced with decreasing temperature, and the 8 °C group had the highest apoptosis index. Overall, these results suggest that the overwintering temperature greatly impacts the survival of mud crabs and that these effects may be mediated through antioxidant, HSP response, and apoptosis pathways. Significantly, the optimal overwintering temperature for S. paramamosain was extrapolated to be 14.2–16.1 °C. This suggests that temperature control should be considered in the process of mud crab overwintering.

Differential expression and regulation of HSP70 gene during growth phase in ruminants in response to heat stress

Heat shock proteins regulate the physiological mechanism of heat stress adaptation at cellular level. The present investigation was carried out to analyse the HSP70 gene regulation in various growth stage in ruminants in peripheral blood mononuclear cells (PBMCs). The relationship between HSP gene expression and thermotolerance in age-specific manner in ruminants has not been analysed. Therefore m-RNA HSP70 expression level was examined in different age groups of Jamunpari goat during hot climatic conditions. The experiment was carried out in 32 animals of Jamunapari goat belonging to the age groups of 3-months, 9-months, 12-months, and adults (2–3 year). Total RNA was isolated from peripheral blood mononuclear cells. The physiological response such as rectal temperature (RT), respiration rate (RR) and heart rate (HR) was used as indicator to heat stress. Temperature Humidity Index (THI) was used as an indicator of severity of environmental stress. The THI range varied from 82.00–92.08 during experimental period. The m-RNA HSP70 expression level at 9-month age of animals was up-regulated and significantly higher than other age groups. It was observed that the level of HSP70 transcripts in PBMCs was highest at 9-month age group, and age-related decline in HSP70 expression was observed in adult age. Based on the physiological response, the contrasting heat-stress phenotypes were recognised as heat stress susceptible (HSS) and heat stress tolerant (HST) individuals and the expression of m-RNA HSP70 was analysed at different ages in response to chronic heat stress. The differential mRNA expression of HSS individuals at 3 and 9-month of age showed the highest fold expression than HST. Age and phenotype had significant effect (p < 0.01) on the crossing point (CP) value. The m-RNA HSP70 gene expression in different age groups was correlated with heat stress tolerance and this could be used as biomarker for breeders to analyse the HSP response in -vivo in ruminants.

Are Marine Heatwaves Responsible for Mortalities of Farmed Mytilus galloprovincialis? A Pathophysiological Analysis of Marteilia Infected Mussels from Thermaikos Gulf, Greece.

Simple SummaryClimate change effects strongly and negatively influence the health and productivity of marine bivalves and particularly bivalve aquaculture. It is therefore of critical importance to study the physiological response of bivalves against marine heatwaves under field conditions. Thus, in the present study, we evaluated the pathophysiological pathways of the Mediterranean mussel originating from mortality events attributed to extreme seawater temperatures in the context of Marteilia refrigens infection. Our study is focused on the Thermaikos Gulf, the most important mussel cultivation area in Greece, comparing the traditional bouchot-like mussel farming system with the modern long-line system. Heatwaves increased all examined molecular, biochemical and pathophysiological markers in M. galloprovincialis mussels, which were reinforced in Marteilia infected individuals. Therefore, these results enlighten us on the biological impacts of heatwaves, providing valuable information regarding the underlying mechanisms. Thus, insights for the future management of the marine aquatic sector could be provided and subsequently address measures for the decrease or elimination of this phenomenon and restoration of marine production.Marine heatwaves (excessive seawater temperature increases) pose high risk to bivalves’ health and farming. The seawater temperature increase is responsible for various pathogen population expansions causing intense stress to marine organisms. Since the majority of knowledge so far derives from laboratory experiments, it is crucial to investigate stress responses in field conditions in order to understand the mechanisms leading to bivalves’ mortality events after exposure to temperature extremes. Thus, we evaluated the pathophysiological response of the Mediterranean mussel Mytilus galloprovincialis originating from mortality events enhanced by intense heatwaves in Thermaikos Gulf, north Greece, along with Marteilia refrigens infection. Mussels that have been exposed to high environmental stressors such as high temperature were examined for various molecular and biochemical markers, such as hsp70, bax, bcl-2, irak4 and traf6 gene expression, as well as the enzymatic activity of the hsp70, hsp90, bax, bcl-2, cleaved caspases, TNFa and ll-6 proteins. Furthermore, histopathology and molecular positivity to Marteilia sp. were addressed and correlated with the gene expression results. Our findings elucidate the molecular and biochemical pathways leading to mortality in farmed mussels in the context of Marteilia infection, which according to the results is multiplied by heatwaves causing a significant increase in pathophysiological markers.

Molecular insights into the heat shock proteins of the human parasitic blood fluke Schistosoma mansoni

BackgroundHeat shock proteins (HSPs) are evolutionarily conserved proteins, produced by cells in response to hostile environmental conditions, that are vital to organism homeostasis. Here, we undertook the first detailed molecular bioinformatic analysis of these important proteins and mapped their tissue expression in the human parasitic blood fluke, Schistosoma mansoni, one of the causative agents of the neglected tropical disease human schistosomiasis.MethodsUsing bioinformatic tools we classified and phylogenetically analysed HSP family members in schistosomes, and performed transcriptomic, phosphoproteomic, and interactomic analysis of the S. mansoni HSPs. In addition, S. mansoni HSP protein expression was mapped in intact parasites using immunofluorescence.ResultsFifty-five HSPs were identified in S. mansoni across five HSP families; high conservation of HSP sequences were apparent across S. mansoni, Schistosoma haematobium and Schistosoma japonicum, with S. haematobium HSPs showing greater similarity to S. mansoni than those of S. japonicum. For S. mansoni, differential HSP gene expression was evident across the various parasite life stages, supporting varying roles for the HSPs in the different stages, and suggesting that they might confer some degree of protection during life stage transitions. Protein expression patterns of HSPs were visualised in intact S. mansoni cercariae, 3 h and 24 h somules, and adult male and female worms, revealing HSPs in the tegument, cephalic ganglia, tubercles, testes, ovaries as well as other important organs. Analysis of putative HSP protein-protein associations highlighted proteins that are involved in transcription, modification, stability, and ubiquitination; functional enrichment analysis revealed functions for HSP networks in S. mansoni including protein export for HSP 40/70, and FOXO/mTOR signalling for HSP90 networks. Finally, a total of 76 phosphorylation sites were discovered within 17 of the 55 HSPs, with 30 phosphorylation sites being conserved with those of human HSPs, highlighting their likely core functional significance.ConclusionsThis analysis highlights the fascinating biology of S. mansoni HSPs and their likely importance to schistosome function, offering a valuable and novel framework for future physiological investigations into the roles of HSPs in schistosomes, particularly in the context of survival in the host and with the aim of developing novel anti-schistosome therapeutics.

Physiological and gene expression responses of diploid and triploid Pacific oyster ( Crassostrea gigas ) to heat acclimation

High temperature is considered to be one of the important causes of mass mortality of diploid and triploid oysters in summer. In order to compare the tolerance adaptability of diploid and triploid oysters to heat stress, the activities of superoxide dismutase activity (SOD), catalase activity (CAT) and contents of malondialdehyde (MDA) and the expression of Hsp70 and metallothionein (MT) genes in diploid and triploid oysters under acute and chronic heat stress were studied. The results showed that the survival rate of diploid oysters was significantly higher than that of triploid oysters under acute and chronic heat stress (p < 0.05). Under acute heat stress, the SOD levels in gills of both diploid and triploid oysters increased significantly from hour 3 (p < 0.05), but there was no significant difference in SOD levels between diploid and triploid oysters at all time points. The SOD level in hepatopancreas of triploid oysters was significantly higher than that of diploid oysters at all time points except 48 h (p < 0.05). The CAT level of diploid and triploid oysters decreased sharply at 48 h and that of diploid oysters was significantly higher than that of triploid oysters (p < 0.05). Under acute heat stress, the increase in MDA content in triploid oysters was significantly higher than that in diploid oysters (p < 0.05). Under acute heat stress, the expression levels of Hsp70 in the gills and hepatopancreas were significantly increased in diploid and triploid oysters (p < 0.05), but increased varies between gills and hepatopancreas. The expression levels of MT in the gills and hepatopancreas were significantly decreased in diploid and triploid oysters (p < 0.05). The results obtained in this study provide physiological and immunological evidence to explain differences in high-temperature tolerance between diploid and triploid oysters and help us better understand the mass mortality of oysters that occurs during high temperatures in summer.

Both AMP-activated and cAMP-dependent protein kinase regulate the expression of heat shock protein 70-2 gene in Neopyropia yezoensis

The upregulation of heat shock protein 70 (hsp70) gene under high temperature stress is a common phenomenon. Although heat shock protein-mediated stress responses play an important role in intertidal Neopyropia yezoensis, the detailed regulatory mechanism of the hsp70 gene expression is still unclear. Here, a full-length sequence of the hsp70-2 gene was cloned and its’ expression regulation was analyzed. There was an activating transcription factors element (ATFE) of cAMP-dependent protein kinase (PKA) was found at the gene promoter region and a highly conserved deduced amino acid sequence with calmodulin-binding activity was detected. Reagents implicated in the induction of the hsp70 gene were then selected to treat the algal samples at 24°C, and the photosynthetic parameters, transcription and translation of this gene were determined. Results showed that quercetin inhibited the transcription of the hsp70-2 gene, significantly decreased the synthesis of the HSP70-2 protein, and lowered the photosynthetic activity of N. yezoensis under high temperature stress conditions. Although the addition of trifluoperazine (TFP), an inhibitor of calmodulin (CAM), downregulated the photosynthetic parameters, the transcription of the hsp70-2 gene was not influenced at high temperature treatment, implying that CAM was not involved in the transcription of the hsp70-2 gene but involved in the heat stress reponding pathways. 5′-aminoimidasole-4-carboxamide-1-β-d-ribofuranoside (AICAR) promoted the synthesis of hsp70-2 transcripts significantly and HSP70-2 protein slightly, which indicated that AMPK regulated the induction of the hsp70-2 gene in N. yezoensis. Forskolin also promoted the upregulation of the hsp70-2 gene. Thus, both AMPK and cAMP-dependent protein kinase (PKA) could phosphorylate HSF and activate the expression of the hsp70-2 gene in N. yezoensis. However, there was no strict correlation between transcripts of the hsp70-2 gene and HSP70-2 protein contents. It was proposed that the posttranscriptional mechanisms were involved in regulating the synthesis of the HSP70-2 protein. Which allowed it to adapt to the extremely variable intertidal environments with an instant responding manner, since the accumulated transcripts could be translated rapidly when needed.

Dietary supplementation of Nile tilapia (Oreochromis niloticus) with β-glucan and/or Bacillus coagulans: Synergistic impacts on performance, immune responses, redox status and expression of some related genes.

A 14-week feeding study was conducted to assess the effects of feed supplementation with prebiotics β-glucan (BG group) and/or probiotics Bacillus coagulans (BC group) on O. niloticus growth performance, body analysis, intestinal structure, immunological response, and antioxidant status. The fish were equally divided into six groups, as follows: the fish group fed an un-supplemented diet served as a control group; the other fish groups were fed supplemented diets with 0.1 g β-glucan kg−1; 1 g Bacillus coagulans kg−1; 2 g B. coagulans kg−1; 0.1 g β-glucan combined with 1 g B. coagulans kg−1; 0.1 g β-glucan combined with 2 g B. coagulans kg−1. The findings revealed that supplementing B. coagulans and the β-glucan mixture improved growth performance and feed efficiency parameters (RGR and SGR) more than the other groups. The fish flesh analysis revealed increased crude protein and dry matter content and lower lipid and ash levels in the BG and BC supplemented groups than in the Control group. On the other hand, β-glucan and B. coagulans supplementation significantly boosted antioxidant activity and immunological responses in serum as determined by CAT, MDA, lysozyme, and phagocytic activity. Dietary β-glucan and B. coagulans supplementation remarkedly enhanced anterior intestine villus histomorphometry characteristics. Furthermore, B. coagulans, alone or in combination with β-glucan, could reduce HSP70 and IL-1β gene expression while increasing IL-8 and GH gene expression. According to the findings, B. coagulans and/or BG increased growth performance by increasing gut health and morphology. Furthermore, β-glucan and B. coagulans supplementation enhanced Tilapia's body composition, immunological responses, and antioxidant status.