Hsp70 Gene Research Articles

Cloning and functional analysis of heat shock protein Hsp70 from Sclerotinia sclerotiorum

To clarify the roles of the heat shock protein gene Hsp70 in the sclerotial formation and pathogenicity of Sclerotinia sclerotiorum, we employed reverse transcription PCR (RT-PCR) to clone Hsp70 from S. sclerotiorum and performed sequence analysis. Quantitative real-time PCR (qRT-PCR) was employed to determine the relative expression levels of Hsp70 at different growth stages and under the stress induced by cyclic adenosine monophosphate (cAMP) and low and high temperatures. The thermal stability of Hsp70 was measured. The Agrobacterium-mediated method was employed to construct the Hsp70-silenced strain. The pathogenicity and fungicide resistance of strains were tested by inoculation in detached rapeseed leaves and cultivation in the media containing procymidone and thiophanate-methyl, respectively. The results showed that the cloned Hsp70 had a total length of 1 890 bp and close relationship with the Hsp70 gene of Ciborinia. Hsp70 showcased the highest expression level in sclerotia, which was more than 30 times higher than that in hyphae. The cAMP stress significantly induced the expression of Hsp70. The expression level of Hsp70 showed an increasing-decreasing-increasing trend at 40 ℃ and no significant change at 4 ℃. Recombinant strain with high expression of Hsp70 showed good thermal stability. The Hsp70-silenced transformant did not form sclerotia, with decreased pathogenicity and fungicide resistance. This study reveals that Hsp70 plays an important role in the sclerotial formation and stress resistance of S. sclerotium, providing reference for further in-depth research on the biological roles of Hsp70 in S. sclerotium.

Knockout of <i>Hsp70</i> genes modulates age-related transcriptomic changes in leg muscles, reduces locomotion speed and lifespan of <i>Drosophila melanogaster</i>

The study investigated the effect of knockout of six Hsp70 genes (orthologues of the mammalian genes Hspa1a, Hspa1b, Hspa2 and Hspa8) on age-related changes in gene expression in the legs of Drosophila melanogaster, which contain predominantly skeletal muscle bundles. For this, the leg transcriptomic profile was examined in males of the w1118 control line and the Hsp70– line on the 7th, 23rd and 47th days of life. In w1118 flies, an age-related decrease in the locomotion (climbing) speed (a marker of functional state and endurance) was accompanied by a pronounced change in the transcriptomic profile of the leg skeletal muscles, which is conservative in nature. In Hsp70– flies, the median lifespan was shorter and the locomotion speed was significantly lower compare to the control; at the same time, complex changes in the age-related dynamics of the skeletal muscle transcriptome were observed. Mass spectrometry-based quantitative proteomic showed that 47-day-old Hsp70– flies, compared with w1118, demonstrated multidirectional changes in the content of key enzymes of glucose metabolism and fat oxidation (glycolysis, pentose phosphate pathway, Krebs cycle, beta-oxidation and oxidative phosphorylation). Such dysregulation may be associated with a compensatory increase in the expression of other genes encoding chaperones (small Hsp, Hsp40, 60, and 70), which regulate specific sets of target proteins. Taken together, our data show that knockout of six Hsp70 genes slightly reduces the median lifespan of flies, but significantly reduces the locomotion speed, which may be associated with complex changes in the transcriptome of the leg skeletal muscles and with multidirectional changes in the content of key enzymes of energy metabolism.

Rising global temperatures and its impact on sleep behavior of male redheaded bunting (Emberiza bruniceps).

Anthropogenic global warming is one of the most pervasive threats to nature and biodiversity. The magnitude with which earths' temperature is rising is affecting every lifeform uniquely; however, the studies highlighting the impacts of global warming on avian sleep are scarce. To this end, the present study was aimed at analyzing the impact of global warming on sleep behavior of a nocturnal migrant, Emberiza bruniceps. For this purpose, the birds were divided into two groups (N = 15 each), subjected to high (35 ± 1°C) and low (19 ± 1°C) temperature schedule with concurrent exposure to 8L:16D (short day; SD) photoperiod followed by 13L:11D (long day; LD). The experiment continued till 7 cycles of zugunruhe (LD) in birds. The results reveal significant impact of temperature treatment on initiation and quality of zugunruhe. Temporal distribution of activity and rest varied according to the temperature provided. Focusing on rest and specifically on sleep of birds, high ambient temperatures resulted in greater sleep fragmentation (evident by increased awakenings during night), whereas low temperature created a sleep conducive environment (evident by abundance of back sleep). Besides postural differences, high temperature resulted in reduced sleep duration, sleep onset latency and circulating plasma melatonin levels in comparison with low temperature suggesting the negative impact of high temperature on different sleep attributes. Not only sleep, seasonal physiology of birds such as hyperphagia, gain in body mass, and fat stores showed significant reduction in high temperature condition. Besides behavioral and physiological alterations, high ambient temperature led to elevated expression of temperature sensitive (trpv4, trpm8, hspa8, and hsp70) genes. Enhanced expression of chrm3 (responsible for wakefulness) also affirms sleep fragmentation in response to high temperature. Thus, the study highlights the negative impact of high temperature on birds' sleep behavior and seasonal physiology.

Gene expression analysis of Aquaporin genes in ruminants during growth phase in response to heat stress

Aquaporins (AQPs) are trans-membrane protein involved in water transport and different cellular functions such as cell adhesion, signalling and proliferation. These membrane proteins are essential for key physiological functions such as organ development, osmoregulation, tissue regeneration and metabolism. The regulation of AQP5 gene expression in ruminants during growth phase has not been analysed in-vivo. Therefore, the gene expression pattern was analysed in Jamunapari goats during 3 months to 12 month of age and adult age group in response to heat stress. The genotyping of the AQP5 gene was carried out by High-Resolution Melting (HRM) in four different goat breeds, which indicated four distinct genotypes in the population. The nucleotide diversity for the AQP5 gene ranged from 0.315 and 0.524 across the breeds. Additionally, a close evolutionary relationship between AQP5 and the HSP70 gene was observed, indicating a shared pathway for heat stress regulation. The m-RNA expression level of AQP5 at 3, 9, 12 month and adult age group exhibited 47.24, 1140, 43.17 and 12.55-fold higher expression than control. The m-RNA expression level of the AQP5 gene was up-regulated and significantly higher (P < 0.05) at 9-month age as compared to the other age groups. Heat stress phenotypes were classified based on respiration rate and heart rate, and the m-RNA expression of AQP5 was higher in heat stress-susceptible (HSS) individuals than heat stress-tolerant (HST) individuals at 3, 9, and 12 months of age. The AQP5 plays a significant role in thermoregulation during growth phases in response to heat stress in goats, however, it is required to understand the role of aquaporins at cellular level as well as to establish the association with production performance in ruminant system in-vivo.

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.

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.

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.

Effects of Zinc Oxide Nanoparticles (ZnO NPs) on Growth, Immune Responses and Histopathological Alterations in Asian Seabass (Lates calcarifer, Bloch 1790) under Low-Salinity Conditions.

In the present study, Asian seabass (Lates calcarifer, Bloch) fingerings were used as an animal model to investigate the toxicological effects of zinc oxide nanoparticles (ZnO NPs) under 5 ppt estuarine conditions. The fish were exposed to 0, 1, 5 or 50 ppm ZnO NPs for 8 weeks. It was found that ZnO NP concentrations of 5-50 ppm negatively affected several growth rate parameters, such as the weight and total length of the fish. Additionally, 5 and 50 ppm ZnO NPs led to 32.55% and 100% mortality, respectively, after 8 weeks after exposure (WAE). Furthermore, compared with the control, exposure to 1-50 ppm ZnO NPs strongly affected hematological indices, such as total blood cells, red blood cells, leukocytes and hematocrit, and suppressed lysozyme activity, superoxide anion production and bactericidal activity. High Zn concentrations accumulated in the head kidney, gills and liver, whereas low levels were detected in the gut, skin and muscle. Expression analysis of immune-related genes via quantitative real-time RT-PCR revealed that 5 and 50 ppm ZnO NPs significantly upregulated the cc and cd4 genes at 1 WAE. In contrast, 50 ppm ZnNPs downregulated the expression levels of the cd8, cc, hsp70, hsp90, tcrα, lyz and igmh genes at 1 WAE (p < 0.05). Finally, at 8 WAE, histopathological analysis revealed that 5 and 50 ppm ZnO NPs severely induced alterations in the head kidney, gills and liver.

Esculetin Attenuates Doxorubicin-Induced Kidney Damage By Reducing Heat Shock Proteins Expression Levels

The effectiveness of Doxorobucin (DOX), a commonly used anti-cancer and immunosuppressive medication, is hindered by its potential for organ toxicity. Prolonged use of DOX is associated with severe hepatocellular toxicity. This study reveals fresh insights into the therapeutic impact of esculetin (E) on DOX-induced kidney cell damage. Esculetin demonstrates its remedial effects by modulating heat shock protein signaling pathways. In our research, we explored the impact of DOX and E on the expression of the 70 kDa HSP gene family, including Hspa1a, Hspa4, and Hspa5, which are small stress proteins in Rattus norvegicus. The study involved the assignment of five different groups (Control, DOX, E50 mg/kg, E100 mg/kg, DOX+ E50 mg/kg, and DOX+ E100 mg/kg). Subsequently, kidney tissues were collected from rats, and cDNA libraries were generated at the conclusion of the application process. The Real-Time PCR method was employed using these libraries to detect HSP70 genes. Analyses conducted on Hspa1a, Hspa4 and Hspa5 expression revealed a statistically significant increase in the DOX group compared to the control group. Additionally, the combination of DOX and esculetin demonstrates a reduction in the increase caused by DOX alone. The study suggests that esculetin could serve as a potential protective agent for shielding kidney tissue from oxidative damage and apoptosis.

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.

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.

Genome-wide analysis reveals transcriptional and translational changes during diapause of the Asian corn borer (Ostrinia furnacalis)

BackgroundDiapause, a pivotal phase in the insect life cycle, enables survival during harsh environmental conditions. Unraveling the gene expression profiles of the diapause process helps uncover the molecular mechanisms that underlying diapause, which is crucial for understanding physiological adaptations. In this study, we utilize RNA-seq and Ribo-seq data to examine differentially expressed genes (DEGs) and translational efficiency during diapause of Asian corn borer (Ostrinia furnacalis, ACB).ResultsOur results unveil genes classified as “forwarded”, “exclusive”, “intensified”, or “buffered” during diapause, shedding light on their transcription and translation regulation patterns. Furthermore, we explore the landscape of lncRNAs (long non-coding RNAs) during diapause and identify differentially expressed lncRNAs, suggesting their roles in diapause regulation. Comparative analysis of different types of diapause in insects uncovers shared and unique KEGG pathways. While shared pathways highlight energy balance, exclusive pathways in the ACB larvae indicate insect-specific adaptations related to nutrient utilization and stress response. Interestingly, our study also reveals dynamic changes in the HSP70 gene family and proteasome pathway during diapause. Manipulating HSP protein levels and proteasome pathway by HSP activator or inhibitor and proteasome inhibitor affects diapause, indicating their vital role in the process.ConclusionsIn summary, these findings enhance our knowledge of how insects navigate challenging conditions through intricate molecular mechanisms.

Improving heat tolerance in betel palm (Areca catechu) by characterisation and expression analyses of heat shock proteins under thermal stress

Context Heat shock proteins play a vital role in cellular homeostasis by protecting proteins against various environmental stresses, which facilitates the survival of plants under unfavourable conditions. Aims We aimed to provide the first comprehensive genomic and expression analysis of the HSP70 gene family in betel palm (Areca catechu) to elucidate its role in heat stress response. Methods Genomic analysis revealed 34 putative HSP70 genes distributed across 13 chromosomes. These were renamed AcatHSP70 and classified into five subfamilies (A–E) based on phylogenetic analysis. These genes are mostly localised in the chloroplast, cytoplasm, and nucleus. Gene ontology revealed that these genes are mostly involved in heat stress. The gene duplication events of HSP70 genes involved only segmental duplications. We subjected betel palm seedlings (2 years old) to heat stress under controlled conditions for 30 days at high, low, and room temperatures for expression analyses of HSP70 genes. Key results Expression analysis revealed eight putative candidate genes (AcatHSP70-3, AcatHSP70-13, AcatHSP70-22, AcatHSP70-19, AcatHSP70-21, AcatHSP70-24, AcatHSP70-25, and AcatHSP70-26) that showed significantly higher expression under high-temperature stress. AcatHSP70-5 showed higher expression under low-temperature treatment, and AcatHSP70-16 was responsive at room temperature treatment. Conclusion We conclude that the majority of AcatHSP70 genes play a crucial role under thermal stress conditions, and respond to high-temperature stress as shown by the quantitative reverse transcription polymerase chain reaction analysis. Implications This comprehensive characterisation of the HSP70 gene family provides novel insights into the thermal protection mechanisms of betel palms in changing climates.

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.

Transcriptome-wide identification of the Hsp70 gene family in Pugionium cornutum and functional analysis of PcHsp70-5 under drought stress.

The PcHsp70-5 enhances drought stress tolerance in transgenic Arabidopsis thaliana by upregulating stress tolerance genes and antioxidant enzyme activities. Heat shock proteins (HSPs) constitute a class of evolutionarily conserved proteins synthesized by organisms in response to various adverse environmental stimuli such as elevated temperatures, drought, hormonal fluctuations, high salt concentrations, and mechanical stress. However, research on HSPs has predominantly focused on model plants and crops, whereas their functions in desert plants have not been well investigated. This study analyzed the transcriptome of Pugionium cornutum and identified the complete ORFs of 25 genes of the PcHsp70 family genes. Their expression levels under drought stress were investigated using existing RNA-seq data. PcHsp70-5 genes exhibited high expression levels in both roots and leaves under drought stress. Consequently, the PcHsp70-5 genes were cloned and transformed into Arabidopsis thaliana for further analysis of their roles in drought stress response. Real-time fluorescence quantitative PCR (qRT-PCR) analysis demonstrated that both, drought stress and ABA, induced PcHsp70-5 expression. Under drought conditions, transgenic Arabidopsis plants exhibited markedly enhanced growth compared to wild-type plants, as evidenced by improved survival rates, root length, fresh weight, chlorophyll content, and reduced levels of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in leaves, indicating that PcHsp70-5 overexpression mitigated growth inhibition and oxidative damage induced by drought stress. Subsequent research revealed that PcHsp70-5 overexpression significantly augmented the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and increased the proline content in transgenic Arabidopsis under drought conditions, alongside a significant increase in the expression levels of genes related to stress tolerance. This suggests that PcHsp70-5 enhances drought stress tolerance in transgenic Arabidopsis by upregulating stress tolerance genes and antioxidant enzyme activities.

COPPER AND MICROPLASTIC EXPOSURE AFFECTS THE GILL GENE EXPRESSION OF COMMON CARP DURING SALTWATER CHALLENGE

The aim of the present study was to assess if pre-exposure to water copper and/or polyvinyl chloride microparticles affects the transcriptional responses of common carp, Cyprinus carpio, to saltwater exposure. Fish were exposed to 0.25 mg/L copper alone (Cu) or in the presence of 0.5 mg/L polyvinyl chloride microparticles (Cu-MPVC) for 14 days, followed by 72 hours of salt water exposure (0 to 13 ppt NaCl). The copper content in the gills and the expression of heat shock protein (hsp70) and cytochrome P450 family 1 (cyp1a) transcripts were examined. The results showed that gill copper levels increased significantly (P = 0.008) in the Cu and Cu-MPVC treatments after 14 days of exposure, compared to the control fish; the Cu and Cu-MPVC treatments had similar gill copper levels. After 14 days of exposure, branchial expression of the hsp70 and cyp1a genes was significantly up-regulated in the Cu and Cu-MPVC treatments. Exposure to salt water led to a significant down-regulation of the gene transcripts in all treatments after 24 h of exposure. At this point, the Cu and Cu-MPVC treatments showed transcripts similar to those of the control fish prior to saltwater exposure. The fish treated with Cu-MPVC showed significantly higher hsp70 expression 72 h after saltwater exposure than the other treatments. At this time point, the control and Cu fish had significantly lower cyp1a expression than before saltwater exposure. In conclusion, the present data suggest that copper exposure induces stress in the fish gills, and the presence of MPCV in the water hampers normal transcriptomic responses of the fish gills to saltwater exposure.

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.