Hsp70 Gene Expression Research Articles

Transcriptional Rearrangements Associated with Thermal Stress and Preadaptation in Baikal Whitefish (Coregonus baicalensis)

In this work, we describe the transcriptional profiles of preadapted and non-adapted one-month-old juvenile Baikal whitefish after heat shock exposure. Preadapted fish were exposed to a repeated thermal rise of 6 °C above the control temperature every three days throughout their embryonic development. One month after hatching, preadapted and non-adapted larvae were either kept at control temperatures (12 °C) or exposed to an acute thermal stress (TS) of 12 °C above the control temperature. In response to this acute stress, an increase in HSP gene expression (HSP-30, HSP-40, HSP-47, HSP-70, and HSP-90) and TRIM16 was detected, independent of preadaptation. The expression levels of genes responsible for the response to oxygen levels, growth factors and the immune response, HBA, HBB, Myosin VI, Myosin VII, MHC, Plumieribetin, TnI, CYP450, and LDB3 were higher in individuals that had previously undergone adaptation. Genes responsible for the regulation of metabolism, MtCK, aFGF, ARF, CRYGB, and D-DT, however, increased their activity in non-adapted individuals. This information on transcriptional profiles will contribute to further understanding of the mechanisms of adaptation of whitefish to their environment.

Exploring the Gene Expression and Plasma Protein Levels of HSP90, HSP60, and GDNF in Multiple Sclerosis Patients and Healthy Controls.

Multiple sclerosis (MS) is a chronic neurodegenerative disease characterized by immune-mediated inflammation and neurodegeneration in the central nervous system (CNS). In this study; we aimed to investigate the gene expression and plasma protein levels of three neuroprotective genes-heat shock proteins (HSP90 and HSP60) and glial cell line-derived neurotrophic factor (GDNF)-in MS patients compared to healthy controls. Forty patients with relapsing-remitting MS and 40 healthy volunteers participated in this study. Gene expression was measured using reverse transcription quantitative real-time PCR, and protein levels were assessed via ELISA. The results showed a significant increase in HSP90 (1.7-fold) and HSP60 (2-fold) gene expression in MS patients compared to controls, along with corresponding increases in protein levels (1.5-fold for both HSP90 and HSP60). In contrast, GDNF gene expression and protein levels were significantly reduced in MS patients, with a 7-fold decrease in gene expression and a 1.6-fold reduction in protein levels. Notably, a non-linear relationship between GDNF gene expression and protein concentration was observed in MS patients, suggesting complex regulatory mechanisms influencing GDNF in the disease. The upregulation of HSP90 and HSP60 in MS highlights their roles in immune regulation and stress responses, while the reduction in GDNF indicates impaired neuroprotection. These findings suggest that HSP90, HSP60, and GDNF could serve as biomarkers for disease progression and as potential therapeutic targets in MS, offering promising avenues for future research and treatment development.

Impact of Management and Nutrition on HSP70 Gene Expression in Heat-stressed Dairy Cows

Background: This research investigated the effects of management and nutrition strategies on dairy cows under heat stress. Objectives: We aimed to evaluate the effects of zinc mineral supplementation and mist spray, alone and in combination, on yield, milk production and composition, blood parameters and HSP70 gene expression in dairy cows. Methods: Sixteen Holstein lactating cows were used for our experiment in four treatment groups: Basal diet without heat stress alleviation methods (control), zinc supplementation in diet, basal diet and application of mist spray method and supplementation of zinc in basal diet with mist spray. We measured milk production and composition, blood parameters and HSP70 gene expression in all cows. Results: The results showed that the cows in the mist and zinc+mist treatments had significantly better performance and temperature-humidity index than the control group. The milk yield and its compounds were significantly affected by experimental treatments, with the best results seen in the treatment with both spray and zinc. The cows exposed to dry and lactation periods showed a significant increase in the concentration of blood biochemical factors and antioxidant indices in response to heat stress. HSP70 gene expression was significantly decreased in all treatments compared to the control. Conclusion: This experiment suggests that applying nutritional and management strategies can effectively mitigate the effects of heat stress on dairy cows. The study recommends using zinc supplementation and mist spray to alleviate heat stress effectively. Overall, this study highlights the importance of implementing management and nutrition strategies to improve the welfare and productivity of dairy cows under heat stress.

Effects of moringa (Moringa oleifera) leaf powder supplementation on growth performance, haematobiochemical parameters and gene expression profile of stinging catfish, Heteropneustes fossilis

Moringa oleifera is a popular edible protein source due to its high concentration of micronutrients, macronutrients, and antioxidants. The present study investigated the effects of M. oleifera leaf at various levels on the growth performance, haematobiochemical response, and SOD2 and HSP70 gene expression of Heteropneustes fossilis for 90 days. The fish were randomly distributed into 12 glass aquaria (66×36×36 cm) at 50 fish/aquarium in triplicate treatments Four dietary groups were used to feed the H. fossilis, each group containing graded levels of M. oleifera leaf meal (0, 10 %, 20 %, and 40 %) with a basal diet. It was observed that H. fossilis fed a diet supplemented with 10 % MOLM produced the highest final weight, specific growth rate, condition factor, and survivability. Hemoglobin and red blood cell levels were significantly higher in the fish treatment fed with a 10 % M. oleifera diet compared to the other treatments. Significantly lower levels of ALT and AST were detected in the fish that were fed a diet containing 10 % MOLM. Frequencies of ECA and ENA were significantly higher at 40 % M. oleifera inclusion. The relative mRNA expression profile of SOD2 and HSP70 gene were significantly higher at 10 % MOLM supplementation diet in comparison to the control group. The results of the PCA showed that adding 10 % MOLM supplementation had a beneficial effect on the nutritional and haematobiochemical parameters. Overall, it can be stated that 10 % MOLM treated with basal diet as a supplement on stinging catfish have positive impact on growth performance, haematobiochemical response, and immune gene expression.

Moesin contributes to heat shock gene response through direct binding to the Med15 subunit of the Mediator complex in the nucleus.

The members of the evolutionary conserved actin-binding Ezrin, Radixin and Moesin (ERM) protein family are involved in numerous key cellular processes in the cytoplasm. In the last decades, ERM proteins, like actin and other cytoskeletal components, have also been shown to be functional components of the nucleus; however, the molecular mechanism behind their nuclear activities remained unclear. Therefore, our primary aim was to identify the nuclear protein interactome of the single Drosophila ERM protein, Moesin. We demonstrate that Moesin directly interacts with the Mediator complex through direct binding to its Med15 subunit, and the presence of Moesin at the regulatory regions of the Hsp70Ab heat shock gene was found to be Med15-dependent. Both Moesin and Med15 bind to heat shock factor (Hsf), and they are required for proper Hsp gene expression under physiological conditions. Moreover, we confirmed that Moesin, Med15 and Hsf are able to bind the monomeric form of actin and together they form a complex in the nucleus. These results elucidate a mechanism by which ERMs function within the nucleus. Finally, we present the direct interaction of the human orthologues of Drosophila Moesin and Med15, which highlights the evolutionary significance of our finding.

Adaptive capability of slow-growing backyard poultry as indicated by physiological and molecular responses in a hot and humid coastal climate

Assessing the adaptability of slow-growing rural chickens for improving thermotolerance to suit the global climate change is a major research need. This work was aimed to evaluate the adaptability of CARI-Debendra chickens and to identify the polymorphism as well as expression profiling of thermotolerant genes (HSP70 and GRP78) under prevailing temperature-humidity indices and thermal stress in a coastal environment. One hundred sixty straight run chicks were reared at THI≥75 (control) and THI>80 under coastal climate till 12 weeks. Polymorphism of HSP70 and GRP78 candidate genes were explored using restriction enzymes TaqI and HaeIII to identify possible thermotolerance markers. Expression profiling of both the genes in liver, intestine and pectoralis muscle was determined through quantitative real-time PCR. Rectal and body surface temperature recorded in the neck and back showed significant differences (P < 0.01) with higher temperature in THI>80 group. Comparatively lower live weights (P < 0.05) and poor FCR were recorded in THI>80 group. The villi height in all intestinal segments was significantly lower (P < 0.01), but deeper crypt depth was observed in THI>80 than control group. A lowest thymus weight (P < 0.05) was noted with no significant differences in immune response in treatment group. Serum levels of cholesterol, activities of lactate dehydrogenase, creatinine kinase and concentration of potassium, sodium and thyroxine hormone were not different between the 2 groups. The concentration of triiodothyronine and chloride ion was lower in THI>80 group indicating adaptive changes for thermoregulation. HSP70 gene expressions in the three tissues were differentially increased (P < 0.01) by temperature-humidity indices, but the expression of GRP78 was not different between the 2 groups. The results concluded that the environmental factors interact with genetics on adaptability towards thermotolerance in slow-growing chickens.

Silicon Modifies Photosynthesis Efficiency and hsp Gene Expression in European Beech (Fagus sylvatica) Seedlings Exposed to Drought Stress.

Background: Climate change is leading to severe and long-term droughts in European forest ecosystems. can have profound effects on various physiological processes, including photosynthesis, gene expression patterns, and nutrient uptake at the developmental stage of young trees. Objectives: Our study aimed to test the hypothesis that the application of silica (SiO2) influences photosynthetic efficiency and gene expression in 1- to 2-year-old Fagus sylvatica (L.) seedlings. Additionally, we aimed to assess whether silicon application positively influences the structural properties of leaves and roots. To determine whether the plant physiological responses are genotype-specific, seedlings of four geographically different provenances were subjected to a one-year evaluation under greenhouse conditions. Methods: We used the Kruskal-Wallis test followed by Wilcoxon's test to evaluate the differences in silicon content and ANOVA followed by Tukey's test to evaluate the physiological responses of seedlings depending on treatment and provenance. Results: Our results showed a significantly higher Si content in the roots compared with the leaves, regardless of provenance and treatment. The most significant differences in photosynthetic performance were found in trees exposed to Si treatment, but the physiological responses were generally nuanced and provenance-dependent. Expression of hsp70 and hsp90 was also increased in leaf tissues of all provenances. These results provide practical insights that Si can improve the overall health and resilience of beech seedlings in nursery and forest ecosystems, with possible differences in the beneficial role of silicon application arising from the large differences in wild populations of forest tree species.

Coenzyme Q10 and vitamin E alleviate heat stress-induced mood disturbances in male mice: Modulation of inflammatory pathways and the HPA axis

Heat stress, as an environmental stressor, can lead to temperature dysregulation and neuroinflammation, causing depression and anxiety by disrupting brain physiology and functional connectivity. This study looked at how co-enzyme Q10 (Q10) and vitamin E (Vit E), alone and together, affected heat stress-caused anxiety and depression symptoms and inflammation in male mice. Five groups were utilized in the study: control, heat stress (NS), Q10, Vit E, and the combination group (Q10+Vit E). The mice were subjected for 15 min/day to a temperature of 43°C for 14 consecutive days, followed by daily treatments for two weeks with either normal saline, Q10 (500 mg/kg), Vit E (250 mg/kg), or their combination. The forced swimming test (FST) and tail suspension test (TST) were employed to evaluate despair behavior, whereas the elevated plus maze (EPM) and open field test (OFT) were used to assess anxious behaviors. Subsequently, the animals were sacrificed, and serum corticosterone levels, protein expression of inflammasome-related proteins, and hsp70 gene expression were evaluated in the prefrontal cortex (PFC). The study revealed that treatment with Vit E and Q10, alone or together, provided anxiolytic and antidepressant effects in the heat-stress-subjected animals. Also, giving Vit E and Q10 alone or together greatly lowered serum corticosterone levels. In the PFC, they also lowered the levels of hsp70 mRNA and NF-κB, caspase 1, NLRP3, and IL-1β proteins. It is speculated that treatment with Q10 and Vit E can attenuate heat stress-associated anxious and depressive responses by inhibiting the inflammatory pathways and modulating the hypothalamus-pituitary-adrenal axis.

Comparison of Gene Expression for Preimplantation Genes in Bovine Embryos Fertilized in vitro

Background: In vitro fertilization (IVF) the technique is useful for understanding early mammalian development and maximizing and speeding up genetic improvement in livestock. Genes that function as genetic markers and are involved in the pre-implantation development of embryos may be used to evaluate the quality of an embryo and the expression of these genes is correlated with the timing of embryonic genomic activation. Methods: Maturing the oocytes of bovine, then fertilizing these matured oocytes and finally culturing the fertilized oocytes to develop into embryos. Determined the gene expression of different certain genes HSP70, OCT4, GLUT1, BAX and DNMT1 in these different cell stages of embryos that were produced. Result: The mRNA expression of these genes was estimated after collecting the embryos listed above that were produced by the in vitro technique. The results showed statistically significant differences in the relative abundance between some of the genes mentioned above in each embryo stage. The timing of the zygotic genome activation process (ZGA) can result in differences in gene expression levels between the early embryonic genes in each developmental stage of in vitro-produced bovine embryos.

PSVII-5 Heat shock protein 70 gene expression and association with morphometric and heat tolerance traits in three plumage colors of Noiler chicken exposed to acute heat stress

Abstract Heat stress is one of the most difficult climatic conditions chickens faces and is a direct result of climate change. It is important to research poultry thermotolerance because it may help find genetic lines resistant to heat stress. This study investigated the influence of plumage colors on heat shock protein 70 gene expression in Noiler chickens exposed to acute heat stress. Noiler is a dual-purpose breed developed for smallholder poultry production in Nigeria. A total of 150 birds of three plumage colors (black, brown and white) of mixed sexes were used for the experiment. The birds were raised in deep litter house separated into six groups of 25 birds each and fed with commercial broiler diets from 0-8 wk and broiler finisher diets from 8 to 12 wk old. The birds were shared into treatments in 3 x 2 x 2 factorial arrangements with 3 plumage colors (black, brown, white), two temperature regimens (Control/thermoneutral, Heat stress) and sex of birds (male, female). The heat stressed birds were exposed to 41oC for 8 h from 0600 h to 1400 h for 7 d while control group maintained a thermoneutral temperature range of 30 oC to 32 oC for the same period. At 13 wk old, two birds were selected per treatment, slaughtered and the spleen removed and stored in RNA later for heat shock protein (HSP) 70 gene expression. Data collected were analyzed with Microsoft excel code for fold expression of HSP70 gene while GLM procedure in SAS was used to determine difference in expression of gene. The relative expression of HSP70 brown plumage was greater, upregulated compared with black and white. White plumage birds in control/thermoneutral group were significantly greater in HSP70 expression than other plumage colors and heat stressed birds. Association of HSP70 delta threshold cycle (DCT) values with morphometric and heat tolerance traits were investigated (Figure 1). High negative correlations emerged between HSP70DCT expression and body weight (-0.65), body temperature (-0.73) with moderate positive correlation for rectal temperature (0.55) in black birds (Table 1). HSP70 DCT positively correlated with body weight (0.70), breast/leg circumferences but negatively with wing length, suggesting reduced surface area during heat stress and a supportive role in maintaining protein integrity (Figure 2). For white Noiler birds, HSP70DCT associated negatively with body weight (-0.94) and positively with rectal temperature during heat stress (Table 2). Thermoneutral birds were positively correlated with heat tolerance traits except temperature while heat stressed birds showed low positive correlation with respiratory and pulse rates and very low positive relationship with body and rectal temperatures (Table 4). Sex-based differences emerged, with potential compromise between HSP70-mediated growth/composition in males versus muscle maintenance in females, although leg length/circumference showed shared developmental regulation (Figure 3, Table 3). These findings highlight HSP70’s versatile roles in systemic heat tolerance mechanisms influenced by plumage color, temperature regime and sex.

Terminalia bellirica and Andrographis paniculata dietary supplementation in mitigating heat stress-induced behavioral, metabolic and genetic alterations in broiler chickens

BackgroundHeat stress (HS) is one of the most significant environmental stressors on poultry production and welfare worldwide. Identification of innovative and effective solutions is necessary. This study evaluated the effects of phytogenic feed additives (PHY) containing Terminalia bellirica and Andrographis paniculata on behavioral patterns, hematological and biochemical parameters, Oxidative stress biomarkers, and HSP70, I-FABP2, IL10, TLR4, and mTOR genes expression in different organs of broiler chickens under chronic HS conditions. A total of 208 one-day-old Avian-480 broiler chicks were randomly allocated into four treatments (4 replicate/treatment, 52 birds/treatment): Thermoneutral control treatment (TN, fed basal diet); Thermoneutral treatment (TN, fed basal diet + 1 kg/ton feed PHY); Heat stress treatment (HS, fed basal diet); Heat stress treatment (HS, fed basal diet + 1 kg/ton feed PHY).ResultsThe findings of the study indicate that HS led to a decrease in feeding, foraging, walking, and comfort behavior while increasing drinking and resting behavior, also HS increased red, and white blood cells (RBCs and WBCs) counts, and the heterophile/ lymphocyte (H/L) ratio (P < 0.05); while both mean corpuscular volume (MCV), and mean corpuscular hemoglobin (MCH) were decreased (P < 0.05). In addition, HS negatively impacted lipid, protein, and glucose levels, liver and kidney function tests, and oxidative biomarkers by increasing malondialdehyde (MDA) levels and decreasing reduced glutathion (GSH) activity (P < 0.05). Heat stress (HS) caused the upregulation in HSP70, duodenal TLR4 gene expression, and the downregulation of I-FABP2, IL10, mTOR in all investigated tissues, and hepatic TLR4 (P < 0.05) compared with the TN treatment. Phytogenic feed additives (PHY) effectively mitigated heat stress’s negative impacts on broilers via an improvement of broilers’ behavior, hematological, biochemical, and oxidative stress biomarkers with a marked decrease in HSP70 expression levels while all tissues showed increased I-FABP2, IL10, TLR4, and mTOR (except liver) levels (P < 0.05).ConclusionPhytogenic feed additives (PHY) containing Terminalia bellirica and Andrographis paniculata have ameliorated the HS-induced oxidative stress and improved the immunity as well as the gut health and welfare of broiler chickens.

Effect of aflatoxin B1 and sterigmatocystin on DNA repair genes in common carp

The present study aimed to investigate the short-time (24 h) effect of aflatoxin B1 (AFB1) and sterigmatocystin (STC) on the expression of hsp70, p53, gadd45, and ogg1 genes in common carp hepatopancreas. Our results showed that aflatoxin B1 and sterigmatocystin can stimulate the expression of DNA repair genes, mainly by hour 24. This significant finding contributes to our understanding of the short-term effects of these mycotoxins on ogg1 genes in common carp hepatopancreas. One-year-old common carp juveniles were randomly distributed into five groups (Control, AFB1 0.4 mg kg−1 feed, STC1 1 mg kg−1 feed, STC2 2 mg kg−1 feed, and STC3 3 mg kg−1 feed). Hepatopancreas samples were collected three times (8, 16, and 24 h) in each group. No significant ogg1 and p53 expression changes were observed at 8 and 16 h after exposure. All measured genes were upregulated by the 24th hour in aflatoxin and STC3 groups. An increase in hsp70 gene expression was detected in all groups and all sampling. A significant decrease in gadd45aa gene expression was observed in the aflatoxin B1 group at hour 8. At hour 16, there was no significant change, while at hour 24, all treated groups were significantly different from the control. In summary, our results suggest that aflatoxin B1 and sterigmatocystin can stimulate the expression of DNA repair genes, mainly by hour 24. Further investigations are needed to get information about DNA damage parallel to the DNA repair mechanisms.

Inter-subspecies diversity of maize to drought stress with physio-biochemical, enzymatic and molecular responses.

Drought is the most significant factor limiting maize production, given that maize is a crop with a high water demand. Therefore, studies investigating the mechanisms underlying the drought tolerance of maize are of great importance. There are no studies comparing drought tolerance among economically important subspecies of maize. This study aimed to reveal the differences between the physio-biochemical, enzymatic, and molecular mechanisms of drought tolerance in dent (Zea mays indentata), popcorn (Zea mays everta), and sugar (Zea mays saccharata) maize under control (no-stress), moderate, and severe drought stress. Three distinct irrigation regimes were employed to assess the impact of varying levels of drought stress on maize plants at the V14 growth stage. These included normal irrigation (80% field capacity), moderate drought (50% field capacity), and severe drought (30% field capacity). All plants were grown under controlled conditions. The following parameters were analyzed: leaf relative water content (RWC), loss of turgidity (LOT), proline (PRO) and soluble protein (SPR) contents, membrane durability index (MDI), malondialdehyde (MDA), and hydrogen peroxide (H2O2) content, the antioxidant enzyme activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT). Additionally, the expression of heat shock proteins (HSPs) was examined at the transcriptional and translational levels. The effects of severe drought were more pronounced in sugar maize, which had a relatively high loss of RWC and turgor, membrane damage, enzyme activities, and HSP90 gene expression. Dent maize, which is capable of maintaining its RWC and turgor in both moderate and severe droughts, and employs its defense mechanism effectively by maintaining antioxidant enzyme activities at a certain level despite less MDA and H2O2 accumulation, exhibited relatively high drought tolerance. Despite the high levels of MDA and H2O2 in popcorn maize, the up-regulation of antioxidant enzyme activities and HSP70 gene and protein expression indicated that the drought coping mechanism is activated. In particular, the positive correlation of HSP70 with PRO and HSP90 with enzyme activities is a significant result for studies examining the relationships between HSPs and other stress response systems. The discrepancies between the transcriptional and translational findings provide an opportunity for more comprehensive investigations into the role of HSPs in stress conditions.

The Effect of a Single About of Resistance Exercise with Two Intensities and Different Recovery on Pro-Inflammatory Factor TNF-Α, HSP-70, And Anti-Inflammatory Factor BDNF In Female Bodybuilders

Background and purpose: resistance exercises with high loads of HLRT can cause physiological changes, including in the immune system. The purpose and credibility of qualitative research investigated the effect of a resistance activity session with two different intensities and recovery on the expression of TNF-α, HSP-70, and BDNF genes in female bodybuilders.Methods: In this Quasi-experimental study, twelve healthy female bodybuilders (age 30±5) volunteered to participate in the research. The samples performed one weekly strength training protocol session designed at two intensities of one maximum repetition in two different recovery times of one and two minutes. 36 hours after the end of the training, sampling was done after ten hours of fasting, and the samples were centrifuged and stored at minus 20℃. At the end of the four stages, the levels of TNF-α, HSP-70, and BDNF were checked in the medical laboratory. A consent form was obtained from all the participants. SPSS version 27 software was used for data analysis at a significance level of α ≤ 0.05.Findings: The results showed that HLRT resistance training compared to moderate intensity resistance training, by increasing the training intensity to 85% of a maximum repetition and recovery time of one and two minutes, significantly reduced TNF-α level (with p&lt;0.001and a significant increase in the serum levels of BDNF and HSP-70 (with p&lt;0.0001).Conclusion: It seems that increasing the intensity of HLRT resistance training has no negative effect on the immune system, and probably with the decrease of the pro-inflammatory factor TNF-α and HSP-70, the anti-inflammatory factor BDNF also increased reduces the incidence of inflammation in female bodybuilders.

Sulforaphane modulates some stress parameters in TPT-exposed Cyprinus carpio in relation to liver metabolome

This study aimed to investigate the protective effect of sulforaphane (SFN) on liver injury induced by triphenyltin (TPT) in Cyprinus carpio (C. carpio). The fish (average weight of 56.9±0.4 g) were divided into 4 groups with four replicates: the control, TPT, SFN+TPT and SFN groups. Twenty fish were selected from each tank and cultured for 8 weeks. Then, serum and liver samples were collected for physiological, biochemical and metabolomic analyses. In the present study, TPT downregulated the expression of the lysozyme gene, upregulated HSP70 and Hsp90 gene expression, and decreased the activities of serum antioxidant enzymes (SOD, CAT, and GPX). However, dietary SFN alleviated oxidative stress, and prevented changes in immune genes. Metabolomic analysis revealed that TPT exposure changed key metabolites in the main phenylalanine, fatty acid and glycerophosphatide metabolic pathways, which are related to inflammation, oxidative stress and immunity and might also lead to an imbalance of liver energy and lipid metabolism. Dietary SFN promoted amino acid metabolism and increased metabolites related to immunity, anti-inflammation, antioxidation, and protein synthesis in liver of C. carpio. In summary, dietary SFN supplementation reversed TPT-induced decreases in immunity and oxidative stress and regulated amino acid metabolism, lipid metabolism, inflammation and immunity-related metabolic pathways.

Nano titanium dioxide alleviates the toxic effects of tris (2-chloropropyl) phosphate on the digestive gland and hemolymph of thick-shell mussel Mytilus coruscus

In the marine environment, nanoparticles play a role in adsorbing and catalytically degrading organic pollutants, thereby mitigating their toxic effects on aquatic organisms. This study aimed to investigate the impact of nano titanium dioxide (nTiO2) and tris (2-chloropropyl) phosphate (TCPP) on the hemolymph and digestive function of the thick-shell mussel Mytilus coruscus. Mussels were divided into a control group, a group exposed to TCPP alone, a group exposed to a combination of TCPP and 0.5 mg/L nTiO2, and a group exposed to a combination of TCPP and 1 mg/L nTiO2. After 14 days of exposure, oxidative stress responses, including superoxide dismutase (SOD) activity and malondialdehyde (MDA) content, immune defense responses, including acid phosphatase (ACP) and alkaline phosphatase (AKP) activities, and gene expression, including HSP70 expression, were measured in the hemolymph and digestive glands of the mussels. Compared to the control group, mussels solely exposed to 100 μg/L TCPP exhibited a significant reduction in SOD activity in the hemolymph. When TCPP was co-exposed with 0.5 mg/L nTiO2, there were significant increases in MDA content and AKP activity in both the digestive gland and hemolymph compared to the control group. Upon co-exposure of TCPP with 1 mg/L nTiO2, MDA content and AKP activity in the digestive gland significantly decreased, while SOD, ACP, and AKP activity in the hemolymph significantly increased and MDA content significantly decreased, returning to the control group levels. Furthermore, in the combined exposure, HSP70 gene expression significantly decreased as the nTiO2 concentration increased from 0.5 mg/L to 1 mg/L. In summary, TCPP impacted the hemolymph and digestive function of mussels, whereas a concentration of 1 mg/L nTiO2 effectively alleviated the toxic effects of TCPP. This study is crucial for assessing the ecological risks of nanoparticles and emerging organic pollutants in marine environments, and provides new insights into the interaction between nTiO2 and TCPP, as well as the influence of nTiO2 concentration on mitigating TCPP toxicity.

Comparison of the effects of copper oxide nanoparticles (CuO-NPs) and copper (II) sulfate on oxidative stress parameters in rainbow trout hatchlings (Oncorhynchus mykiss)

Nanoparticles of copper oxide, released uncontrollably into the environment, pose a threat to living organisms. One of the mechanisms of their toxicity is the induction of oxidative stress, contributing to the disruption of antioxidant mechanisms. The aim of the present study was to assess the degree and toxic dose of copper oxide nanoparticle (CuO-NPs) associated with the induction of oxidative stress and compare them to the toxicity of copper sulfate (VI) (CuSO4) by measuring the expression of genes encoding some key enzymes involved in protection against oxidative stress: glutathione peroxidase (gpx), cytochrome P450 (cyp1a), heat shock protein (hsp70) and superoxide dismutase (sod). Hatchlings of rainbow trout (Oncorhynchus mykiss) were used as biological material. The test individuals were exposed for 2 hours to CuO-NPs and CuSO4 in the concentration range of 4–256 mg dm−3. Individuals exposed to all concentrations of CuO-NPs showed an increase in the expression of gpx, cyp1a and hsp70 genes, and a decrease in sod gene expression, while the hatchlings exposed to CuSO4 exhibited an increase in the expression of all genes. The expression of hsp70 and sod genes in the hatchlings exposed to CuSO4 was higher than in the CuO-NP treatment, while the expression levels of gpx and cyp1a were similar. Based on the obtained results, potentially higher toxicity of CuSO4 was observed in the hatchlings of Oncorhynchus mykiss. However, the elevated expression of gpx, cyp1a and hsp70 after the exposure to CuO-NPs, indicates toxicity via oxidative stress. The decrease (sod in hatchlings exposed to CuO-NPs) or increase of gene expression was not dose-dependent. The presented study confirms a need for further research regarding the toxicity of metal nanoparticles to aquatic organisms and the necessity for monitoring the release of CuO-NPs into the aquatic environment.

Enhancing Cartilage Metabolism in Rats through a Novel Thermal Stimulation Technique with Photosensitizers.

Although the moderate thermal stimulation of articular cartilage exerts chondroprotective effects, it is difficult to effectively heat deep articular cartilage with conventional methods. Photosensitizers increase the ambient temperature using near-infrared (NIR) radiation, which has high tissue permeability. We hypothesized that the intra-articular administration of photosensitizers and NIR irradiation would exert a greater heating effect on articular cartilage. We aimed to evaluate the heating effect of this method on cultured chondrocytes and rat knee cartilage. In vitro, we irradiated a photosensitizer-containing medium with NIR and measured changes in the medium temperature, cytotoxicity, and gene expression of heat shock protein (HSP) 70 and aggrecan (ACAN). In vivo, the knee joints of rats treated with photosensitizers were irradiated with NIR, and changes in intra-articular temperature and gene expression were measured, alongside histological analysis. The results showed that the medium and intra-articular temperature were raised to approximately 40 °C with no apparent disruption to articular cartilage or the immunohistochemically enhanced staining of HSP70 in chondrocytes. The gene expression of HSP70 and ACAN was increased in both cultured and articular cartilage. In summary, this method can safely heat joints and enhance cartilage metabolism by inducing HSP70 expression in articular cartilage. It presents a new hyperthermia therapy with effective cartilage protection.

Expression analysis of Hsp70 genes in the psychrophilic yeast, Glaciozyma antarctica PI12 as biomarkers for thermal stress during heat waves

Heat shock proteins 70 (Hsp70) are potential thermal stress markers as they play a pivotal role in safeguarding cells against heat shock-induced damage. The Hsp70s are present in several variants with each containing its peculiar importance due to their specific functions such as cell protection during elevated thermal stress. The present investigation evaluated the gene expression profiles of all Hsp70 genes in Glaciozyma antarctica PI12 during a heat wave condition. In this study, we exposed G. antarctica PI12 cells to a realistic heat wave to understand the impacts of the extraordinary, unprecedented heat waves that hit Antarctica at nearly 40°C above the average in 2022. The experiment was carried out through eight days where cells were exposed gradually at 0, 2, 4, 8, 12, 16, 20, 25 and 30°C. The gene expression profiles were obtained during the simulated heat wave along with non-stressed control treatments by real-time PCR. Out of the six Hsp70 genes in G. antarctica PI12, five were expressed under the conditions tested. Among the expressed genes, gahsp70-1, gahsp70-5, and gahsp70-6 showed significant upregulation. Specifically, their expression levels increased by five- to eightfold after exposure to heat shock at 4°C. Gene expression patterns at 20°C and 30°C also showed induction with the highest at 3.6 folds and 5.8 folds, respectively. These results indicate that the expression of Hsp70 genes in G. antarctica PI12 was inducible under thermal stress, indicating their importance in cells during the heat waves. These results conclude that the gene expression patterns of Hsp70 during heat waves contribute vital information on thermal adaptation in the Antarctic marine ecosystem under climate stress.

The effects of Artemisia Sieberi, Achillea Fragrantissima, and Olea Europaea leaves on the performance and physiological parameters in heat-stressed broiler chickens.

High temperatures have detrimental effects on the performance and physiology of broiler chickens. Medicinal plants have various biological activities and may enhance the heat resistance of chickens during heat waves. Therefore, this study aimed to explore the potential roles of using specific local medicinal plants to alleviate the negative impacts of heat stress (HS) in broilers. In this study, 180 day-old chicks were used to investigate the effects of HS and dietary indigenous medicinal plants on growth performance, antioxidant biomarkers, and intestinal health. The chicks were assigned to six groups (18 pens with 10 chicks per pen) with three replicates each. In the first group, the chicks were kept under thermoneutral conditions (CON) and fed a basal diet. The other five groups were exposed to recurrent heat stress and fed a basal diet (T1, HS group) or supplemented with Artemisia Sieberi (1.25 g/kg of feed; T2), Achillea Fragrantissima (15 g/kg of feed; T3), Olea europaea (10 g/kg of feed; T4), and all the previous additives (all-in-one) combined at the same dose levels mentioned above (T5). At 21 days of age, the chicks from each group were exposed to two phases of heat stress: phase 1 from days 21 to 34 (34 ± 1°C) followed by phase 2 from days 35 to 39 (37 ± 1°C). The results indicate that HS significantly increased rectal temperature and respiration rate in broiler chickens. Feed intake and body weight gain were improved in all supplemented groups, while the feed conversion ratio was decreased in response to the dietary inclusion of medicinal plants. Additionally, glutathione peroxidase and immunoglobulin G levels were increased in the T3, T4, and T5 groups compared to the other groups. HS induced significant upregulated in the mRNA levels of heat shock protein 70 and interleukin-8, while the mRNA of occludin was decreased. The T3, T4, and T5 showed significantly decreased expression of hepatic HSP70 and ileum IL-8 genes and increased ileum mRNA occludin levels relative to the CON and T1 groups. In conclusion, supplementation with these plants enhances growth performance and maintains intestinal health sustaining the productivity of broiler chickens under HS conditions.