Levels Of Hsps Research Articles

The regulation of ovarian degeneration in Pampus argenteus by heat shock protein genes under low‐temperature stress

AbstractTemperature is a crucial environmental factor that significantly impacts the growth, development, metabolism, and physiological functions of fish. To study the effects of low‐temperature on the gonadal development of silver pomfret (Pampus argenteus), a cold period by gradually lowering the water temperature from 18°C to 9°C was simulated. The results showed that hsp70, hsp90a, and hsp90b were widely expressed in the tissues of P. argenteus, with hsp70 primarily expressed in the pituitary, hsp90a and hsp90b mainly expressed in the lateral. The hsps were involved in the development process of P. argenteus from 1 to 27 days post‐hatching (dph). The expression levels of hsp70 and hsp90b were highest at 17 dph, while the levels of hsp90a were at 25 dph. Under the condition of 9°C, regressed oocyte were observed in the ovaries, the oocyte diameter significantly decreased, and the ovaries degenerated 100% after low‐temperature stress. The expression levels of hsps in the ovaries were significantly higher than in the control group, while the expression levels in the testes were significantly lower than in the control group. Taken together, hsps may regulate the ovarian degeneration under low‐temperature stress. Male fish rapidly completed meiosis and maintained the testes in the prophase of meiosis to resist the low‐temperature stress.

SlWRKY55 coordinately acts with SlVQ11 to enhance tomato thermotolerance by activating SlHsfA2.

High temperature (HT) severely restricts plant growth, development, and productivity. Plants have evolved a set of mechanisms to cope with HT, including the regulation of heat stress transcription factors (Hsfs) and heat shock proteins (Hsps). However, it is not clear how the transcriptional and translational levels of Hsfs and Hsps are controlled in tomato. Here, we reported that the HT-induced transcription factor SlWRKY55 recruited SlVQ11 to coordinately regulate defense against HT. SlWRKY55 directly bound to the promoter of SlHsfA2 and promoted its expression, which was increased by SlVQ11. Moreover, both SlWRKY55 and SlVQ11 physically interacted with SlHsfA2 to enhance the transcriptional activity of SlHsfA2. Thus, our results revealed a molecular mechanism that the SlWRKY55/SlVQ11-SlHsfA2 cascade enhanced thermotolerance and provided potential target genes for improving the adaptability of crops to HT.

Modulating responses of indicator genes in cellular homeostasis, immune defense and apoptotic process in the Macrophthalmus japonicus exposed to di(2-ethylhexyl) phthalate as a plastic additive

Di(2-ethylhexyl) phthalate (DEHP), have been increasingly used as plasticizers to manufacture soft and flexible materials and ubiquitously found in water and sediments in the aquatic ecosystem. The aim of the present study was to evaluate the effect of DEHP exposure on cellular homeostasis (HSF1 and seven HSPs), immune responses (ILF), and apoptotic responses (p53, BAX, Bcl-2). DEHP exposure upregulated the expression of HSF1 and ILF. Moreover, it altered the expression levels of HSPs (upregulation of HSP70, HSP90, HSP40, HSP83, and HSP67B2 and downregulation of HSP60 and HSP21) in conjunction with HSF1 and ILF in the gills and hepatopancreas of M. japonicus exposed to DEHP. At the protein level, DEHP exposure changed apoptotic signals in both tissues of M. japonicus. These findings indicate that chronic exposures to several DEHP concentrations could disturb cellular balance, damage the inflammatory and immune systems, and induce apoptotic cell death, thereby affecting the survival of M. japonicus.

The development of abamectin resistance in Liriomyza trifolii and its contribution to thermotolerance.

Liriomyza trifolii is an economically significant, invasive pest of horticultural and vegetable crops. The larvae form tunnels in foliage and hasten senescence and death. Outbreaks of L. trifolii often erupt in hot weather and are driven by thermotolerance; furthermore, the poor effectiveness of pesticides has made outbreaks more severe. But it is still unclear whether the development of insecticide tolerance will contribute to thermotolerance in L. trifolii. To explore potential synergistic relationships between insecticide exposure and thermotolerance in L. trifolii, we first generated an abamectin-resistant (AB-R) strain. Knockdown behavior, eclosion and survival rates, and expression levels of genes encoding heat shock proteins (Hsps) in L. trifolii were then examined in AB-R and abamectin-susceptible (AB-S) strains. Our results demonstrated that long-term selection pressure for abamectin resistance made L. trifolii more prone to develop cross-resistance to other insecticides containing similar ingredients. Furthermore, the AB-R strain exhibited enhanced thermotolerance and possessed an elevated critical thermal maximum temperature, and upregulated expression levels of Hsps during heat stress. Collectively, our results indicate that thermal adaptation in L. trifolii was accompanied by emerging abamectin resistance. This study provides a theoretical basis for investigating the synergistic or cross-adaptive mechanisms that insects use to cope with adversity and demonstrates the complexity of insect adaptation to environmental and chemical stress. © 2023 Society of Chemical Industry.

Elucidation of Site-Specific Ubiquitination on Chaperones in Response to Mutant Huntingtin.

Huntington's disease (HD) is one of the prominent neurodegenerative diseases, characterized by the progressive decline of neuronal function, due to the accumulation and aggregation of misfolded proteins. Pathological progression of HD is hallmarked by the aberrant aggregation of the huntingtin protein (HTT) and subsequent neurotoxicity. Molecular chaperones (heat shock proteins, HSPs) play a pivotal role in maintaining proteostasis by facilitating protein refolding, degradation, or sequestration to limit the accumulation of misfolded proteins during neurotoxicity. However, the role of post-translational modifications such as ubiquitination among HSPs during HD is less known. In this study, we aimed to elucidate HSPs ubiquitin code in the context of HD pathogenesis. In a comprehensive proteomic analysis, we identified site-specific ubiquitination events in HSPs associated with HTT in HD-affected brain regions. To assess the impact of ubiquitination on HSPs during HD, we quantified the abundance of ubiquitinated lysine sites in both the rat cortex/striatum and in the mouse primary cortical neurons. Strikingly, we observed highly tissue-specific alterations in the relative ubiquitination levels of HSPs under HD conditions, emphasizing the importance of spatial perturbed post-translational modifications (PTMs) in shaping disease pathology. These ubiquitination events, combined with other PTMs on HSPs, are likely to influence the phase transitions of HTT. In conclusion, our study uncovered differential site-specific ubiquitination of molecular chaperones and offers a comprehensive view of the intricate relationship between protein aggregation, and PTMs in the context of Huntington's disease.

Melatonin-regulated heat shock proteins and mitochondrial ATP synthase induce drought tolerance through sustaining ROS homeostasis in H2S-dependent manner

Drought is thought to be one of the major global hazards to crop production. Understanding the role of melatonin (Mel) during plant adaptive responses to drought stress (DS) was the aim of the current investigation. Involvement of hydrogen sulfide (H2S) was also explored in Mel-regulated mechanisms of plants’ tolerance to DS. A perusal of the data shows that exposure of tomato plants to DS elevated the activity of mitochondrial enzymes viz. pyruvate dehydrogenase, malate dehydrogenase, and citrate synthase. Whereas the activity of ATP synthase and ATPase was downregulated under stress conditions. Under DS, an increase in the expression level of heat shock proteins (HSPs) and activation level of antioxidant defense system was observed as well. On the other hand, an increase in the activity of NADPH oxidase and glycolate oxidase was observed along with the commencement of oxidative stress and accompanying damage. Application of 30 μM Mel to drought-stressed plants enhanced H2S accumulation and further elevated the activity of mitochondrial enzymes, activation level of the defense system, and expression of HSP17.6 and HSP70. Positive effect of Mel on these attributes was reflected by reduced level of ROS and related damage. However, application of H2S biosynthesis inhibitor DL-propargylglycine reversed the effect of Mel on the said attributes and again the damaging effects of drought were observed even in presence of Mel. This observation led us to conclude that Mel-regulated defense mechanisms operate through endogenous H2S under DS conditions.

Characterization of genes encoding heat shock proteins reveals a differential response to temperature in two geographic populations of Liriomyza trifolii (Diptera: Agromyzidae)

Liriomyza trifolii is a significant, invasive pest that damages horticultural crops and vegetables. The distribution of L. trifolii is influenced by temperature, and prior research has demonstrated that variations in thermal adaptability differ among geographic populations of the insect. Heat shock proteins (Hsps) are involved in adaptation to temperatures; however, the underlying molecular mechanism for thermal adaption in different L. trifolii populations remains unclear. This study examines the temperature adaptability of two L. trifolii populations from Hainan (HN) and Jiangsu (JS) provinces. The results indicate that the HN population has a higher survival rate and a higher critical thermal maximum (CTmax) than the JS population under high temperature stress. Transcriptome data at 42 °C revealed that the JS population has more differentially expressed genes (DEGs) than the HN population, while the HN population has more upregulated DEGs. The two populations were similar in functional annotation of DEGs, and a large number of Hsps were upregulated. However, the HN population had larger numbers and higher expression levels of Hsps during heat stress as compared to the JS population. Additionally, the expression patterns of differentially expressed Hsps varied between the HN and JS populations in response to different elevated temperatures. Notably, the transcription levels of Hsp70s were higher in the HN population as compared to the JS population, while the expression level of genes encoding small heat shock proteins was higher in the JS population. These findings have significant scientific value in understanding the underlying mechanism of temperature adaption in L. trifolii and provide a fresh perspective on the distribution of this invasive pest.

Thermal stress impaired expression of insulin-like growth factor binding proteins in pigs

This study aimed to investigate the correlation of IGFs, IGFBPs, and IGFBPRs of grower pigs with change in temperature humidity index (THI). THI was calculated to know the months of the year when pigs feel discomfort and samples of feed offered were analyzed to know the nutritional availability/deficit. The calculated average THI was above thermal comfort zone for seven months in a year. It was observed that crude protein (CP) and metabolizable energy (ME) availability to pigs were deficient. Gilts of two months age and similar body weight were randomly divided into two groups (n=6); viz. Con-fed animals (feed offered by the farmer) and For-fed animals (formulated feed supplement + Con-fed). The composition of formulated feed supplement was dried leaf of Moringa oleifera, soybean meal, rice mill waste, and mineral mixture. The plasma cortisol level increased in the experimental animals with accelerated THI as evidenced by the strong positive correlation of THI with HSP70 and HSP90. The level of leptin and ghrelin did not have significant relation with the expression level of HSPs in the present study, however, in the control animals, a low level of leptin was observed. The correlation between the growth factors and HSPs was not significant. During the season, where the THI was above the thermal comfort zone, the IGFBPR and IGFBP concentrations were non-significantly increased along with the increased THI.

Heat shock proteins as diagnostic and prognostic markers in malignant tumors of the central nervous system

This review overview current epidemiology data of malignant tumors of the central nervous system, and determines the significance of molecular diagnostics and expression of heat shock proteins in tumorigenesis. Particular attention is paid to the description of molecular chaperones as diagnostic and prognostic markers, as well as the prospects for chaperones using in personalized therapy the central nervous system tumors. The search for literature was carried out on the database platforms Medline (via PubMed) and Scopus, Cochrane Library, The Lancet Public Health Journal published between 1988 and 2022. The material was searched for keywords and terms, including “heat shock proteins”, “tumors of the central nervous system”, “brain tumors”, “molecular diagnostics”, “expression of molecular chaperones”, Hsp110, Hsp90, Hsp70, Hsp60, Hsp40, small HSPs. Molecular chaperones, due to their important role in physiological processes in the cell, are highly expressed in brain tumors, and the expression level of HSPs strongly correlates with the degree of malignancy, invasive potential, and resistance to radiochemotherapy. For some HSPs (i.e. HSP10, HSPB1, DNAJC10, HSPA7, HSP90) a direct correlation has been found between the level of protein expression (based on IHC analysis) and a poor overall survival prognosis for patients with glial tumors. This indicates the prognostic value of these markers, which in the future may be included in the diagnostic panel when examining a tumor sample.

Profiling of key heat shock proteins and their relationship with male sexual behavior and seminal characteristics in Kankrej (Bos indicus) breeding bulls during different seasons.

The goal of this study is to use indirect ELISA to determine the concentration of major heat shock proteins (Hsps) in Kankrej (Bos indicus) breeding bulls and their relationship with certain male phenotypic traits including sexual behavior, sperm quality, and bull fertility in different seasons. The seasonal fluctuation in the concentration of three major Hsps (60, 70, and 90) was determined using an indirect enzyme-linked immunosorbent assay (ELISA). According to the findings, Hsps levels are significantly higher during the summer season and are associated with both fresh and post-thawed semen quality traits in Kankrej breeding bulls. The better sexual behavior of bulls and seminal parameters of fresh or thawed semen was observed in the winter season together with the lower concentrations of HSPs. These could suggest negative association between HSPs with bull sexual behavior and seminal parameters. As a result, the concentration of Hsps in breeding bulls may be a useful indicator for determining fertility traits.

Neuroepigenetic Changes in DNA Methylation Affecting Diabetes-Induced Cognitive Impairment.

Chronic diabetic conditions have been associated with certain cerebral complications, that include neurobehavioral dysfunctional patterns and morphological alterations of neurons, especially the hippocampus. Neuroanatomical studies done by the authors have shown decreased total dendritic length, intersections, dendritic length per branch order and nodes in the CA1 hippocampal region of the diabetic brain as compared to its normal control group, indicating reduced dendritic arborization of the hippocampal CA1 neurons. Epigenetic alterations in the brain are well known to affect age-associated disorders, however its association with the evolving diabetes-induced damage in the brain is still not fully understood. DNA hypermethylation within the neurons, tend to silent the gene expression of several regulatory proteins. The findings in the study have shown an increase in global DNA methylation in palmitic acid-induced lipotoxic Neuro-2a cells as well as within the diabetic mice brain. Inhibiting DNA methylation, restored the levels of HSF1 and certain HSPs, suggesting plausible effect of DNMTs in maintaining the proteostasis and synaptic fidelity. Neuroinflammation, as exhibited by the astrocyte activation (GFAP), were further significantly decreased in the 5-azadeoxycytidine group (DNMT inhibitor). This was further evidenced by decrease in proinflammatory cytokines TNF⍺, IL-6, and mediators iNOS and Phospho-NFkB. Our results suggest that changes in DNA methylation advocate epigenetic dysregulation and its involvement in disrupting the synaptic exactitude in the hippocampus of diabetic mice model, providing an insight into the pathophysiology of diabetes-induced neuroepigenetic changes.

Influence of dietary vitamin E and selenium supplementation on broilers subjected to heat stress, Part II: oxidative stress, immune response, gut integrity, and intestinal microbiota

This study evaluated the effects of vitamin E (Vit E) and selenium (Se) supplementation on mRNA abundance of antioxidant, immune response, and tight junction genes, as well as taxonomic and functional profiles of ileal microbiota of broilers exposed to daily 4-h elevated temperature during d 28 to 35. A total of 640-day-old Cobb male broiler chicks were randomly allocated to 32 floor pens in a 2 × 2 factorial arrangement that included ambient temperature (thermoneutral [TN] or heat stress [HS]) and dietary treatments (basal diet or Vit E + Se). Vit E and organic Se were added to the basal diet at the rate of 250 mg/kg and 1 mg/kg, respectively. Liver and jejunum tissue samples were taken on d 27 (1 bird/pen), d 28 and d 35 (2 birds/pen) from birds for qPCR analysis. Data were subjected to a 2-way ANOVA using the GLM procedure of JMP. Ileal contents were taken on d 27 and d 35 for microbial profiling. Microbiota data were analyzed in QIIME 2 and significance between treatments identified linear discriminant analysis effect size (LEfSe, P < 0.05). Dietary Vit E/Se significantly downregulated the mRNA levels of HSPs in liver and jejunal tissues of the HS-challenged birds both on d 28 and d 35. Moreover, mRNA abundance of TLR2, TNFα, IFNγ, IL-1β, IL-10, and iNOS in the liver were significantly downregulated in birds fed the Vit E/Se diet on d 35. However, dietary treatment had no significant impact on oxidative stress, immunity, and gut integrity related genes analyzed in jejunal tissues on d 28 and d 35, except downregulation of IFNγ on d 35 (P = 0.052). LEfSe analysis revealed that Lachnospiraceae FE2018 and Ruminococcaceae NK4A214 groups was enriched in the Vit E/Se birds on d 35. Moreover, PICRUSt analysis predicted significant functional differences among the treatment groups. In conclusion, dietary supplementation of Vit E/Se mitigated the negative effects of HS potentially via improving antioxidant status, regulating cytokine responses and modifying ileal microbiota and its function.

Thermal Responses in Japanese Pearl Oysters, Pinctada fucata martensii

In order to examine the thermal responses in pearl oysters, both the mRNA expression levels of heatshock protein (Hsp) in their tissues and changes in the mineral levels of their hemolymph were analyzed using heattreated and heat-untreated pearl oysters. Pearl oysters that live in Tsukumo Bay on the Noto Peninsula were used as experimental materials. Pearl oysters kept at 25 C in artificial seawater were exposed to high temperatures (30 C or 35 C) for 1 h. After exposure, the pearl oysters were transferred into the seawater at 25 C and kept for 4 h. Thereafter, the hemolymph was collected by inserting a syringe into the adductor muscle of the heat-treated pearl oyster, and then hemocytes were separated from hemolymph by centrifugation. In addition to hemocytes, the mantles and gills were taken from heat-treated pearl oysters to analyze Hsp mRNA expression. The collected hemolymph was used to analyze its mineral concentration. The data of pearl oysters treated at a high temperature (30 C or 35 C) were compared with those of pearl oysters kept at the initial temperature (25 C) as a heat-untreated control. First, the mRNA expressions of several Hsps (Hsp20, Hsp40, Hsp70, and Hsp90) were analyzed using the hemocytes, mantles, and gills to examine the thermal responses of pearl oysters. With heat treatment, the mRNA expressions of all Hsps (Hsp20, Hsp40, Hsp70, and Hsp90) used in the present study were significantly increased. Especially, Hsp20 mRNA expression in hemocytes, mantles, and gills was remarkably increased by heat treatment. The responsiveness of Hsps mRNA expression was greatly higher at the rate of increase at 35 C than at the rate of increase at 30 C. Among the tissues used in the present study, the mRNA expression levels of Hsp20, Hsp40, Hsp70, and Hsp90 were higher in gills than in hemocytes and mantles. Second, we analyzed the mineral concentrations of hemolymph in the heat-treated pearl oysters. The calcium, magnesium, sodium, chlorine, and potassium ion levels in the hemolymph of pearl oysters kept at 35 C increased significantly as compared with those in the initial hemolymph of pearl oysters kept at 25 C. This suggests that heat treatment influences excretory organs such as gills and kidneys. The fact that the expression level of Hsps was highest in gills, which function as respiratory and International Journal of Zoological Investigations Contents available at Journals Home Page: www.ijzi.net Editor-in-Chief: Prof. Ajai Kumar Srivastav Published by: Saran Publications, Gorakhpur, India 372 excretory organs, supported the influences of the excretory system. This study is the first report to demonstrate changes in both the monovalent and divalent ions of hemolymph in response to heat treatment in Japanese pearl oysters. We plan to investigate the detailed mechanism of the above mineral regulation together with HSP expressions in the near future.

Breast Cancer Revisited: Stress Induced Conspiracy, Sensitivity and Specificity

Introduction: Cancer and related disorders remain as one of the most devastating ailments worldwide. It signifies the importance to devise particular techniques, which could help in attenuating the effects of breast cancer and related outcomes. Primary aim of the current study is to revisit the role of inflammatory and stress markers in the development and progression of breast cancer and to determine their sensitivity and specificity. Material and Methods: Local sample of premenopausal Pakistani women (age 40 ± 3.6) diagnosed with breast cancer (n = 498) at Jinnah Hospital Lahore, and equal number of controls (age 36 ± 5.2) were selected for the current case control study after informed consent. Serum samples were separated and estimated for MMPs, ILs, HSPs, TNF-α, INOS and Vitamin D on ELISA, whereas, MDA, SOD, NO, CAT, GSH, Vitamin A and D were estimated on Spectrophotometry. Results: Levels of MDA, ILs, MMPs, TNF-α, NO, INOS and HSPs were significantly higher in the breast cancer patients, but the levels of SOD, CAT, GSH, Vitamins A, C and D were significantly lower, determining the relatively positive relationship of oxidants and negative relationship of antioxidants with the increased incidence of pathology among the patients. Furthermore, HSP-27, MMP-9 and IL-1 levels revealed remarkable sensitivity and specificity on ROC analysis. Conclusion: It concludes that HSP-27, MMP-9 and IL-1 may serve as diagnostic and prognostic markers. Furthermore, it can be extrapolated how the increased levels of oxidative and nitrosative stress species along with inflammatory cytokines predispose to the incidence of cell invasion, cell migration and epithelial-mesenchymal transition. Therefore, it infers that increase in growth factors i.e., VEGF, TGF-β and Bcl2 under the influence of said variables play a crucial role in the metastasis of breast cancer. Funding Information: None to declare. Declaration of Interests: Authors declare no conflict of interest. Ethics Approval Statement: All of the patients signed the informed consent and approval was obtained from the Ethical Review Committee, Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore-Pakistan.

Differential expression of heat shock protein genes in Rhipicephalus sanguineus (Acari: Ixodidae) exposed to temperature and humidity variations.

The brown dog tick (Rhipicephalus sanguineus), a well-recognized parasite that is an important vector of many pathogens, widely distributed in the world, can be found in dogs and many other host animals, including humans. However, the response of this species to climate changes is not fully understood yet. Therefore, this study aimed to analyze the expression of the genes encoding for hsp20 and hsp70 of nymphs and eggs of R. sanguineus in response to different relative humidity and temperature. While no significant differences were observed in hsp70 levels in nymphs, striking variations were found for hsp20. On the contrary, significant differences were found on the expression levels of both HSPs when exposed at different treatments. Expression of hsp20 was more affected by relative humidity than by temperature, which could imply that hsp20 may have a more critical protective role during vulnerable developmental stages of the tick than hsp70.

Chronic heat stress regulates the relation between heat shock protein and immunity in broiler small intestine

Chronic heat stress is considered to decrease the immune functions which makes negative effect on broiler growth performance. Here, we investigated the relationship between chronic heat stress, growth performance, and immunity in the small intestine of broilers. The study included two groups (control and heat stressed group) with eight replications per group. Ten broilers of 20-day aged were allocated in each replication. On day 35, the treatment group was subdivided into two groups based on their body weights (heavy and low body weight). Although, there was only the control and treatment group on day 28. The growth performance decreased and expression of heat shock protein 70 (HSP70), HSP60, and HSP47 increased on days 28 and 35 in the chronic heat stress group as compared with those in the control group. The expression levels of HSPs were significantly higher in the low body weight group than in the control group. The genes HSP70 and HSP60 were significantly associated with pro- and anti-inflammatory cytokines in the small intestine of the broilers of the treatment group. Thus, HSP70 and HSP60 activated the adaptive immunity in the small intestines of the broilers from the treatment group to allow adaptation to chronic heat stress environment.

Identification of single nucleotide polymorphism in heat shock protein HSP70 and HSP90 after four selection generations in two lines of chickens

Climatic changes have affected the entire world resulting in high temperatures, consequently leading to a negative impact on poultry production. Therefore, we conducted this study to detect the single nucleotide polymorphisms (SNPs), which is responsible for producing heat shock protein HSP70 and HSP90 in two selected lines of chicken, Dandarawi and Leghorn that were formally selected for four generations, for achieving a better growth rate under heat pressure. At age 16 weeks, body weight and body measurements were evaluated for Dandarawi and Leghorn males. mRNA levels of HSP70 and HSP90 genes were measured, and the polymorphism of these two genes was examined via DNA sequencing. Results showed that after four generations under heat stress conditions, the Dandarawi and Leghorn lines of chicken were distinguishable in terms of body weight and body measurements. Additionally, HSP70 and HSP90 a highly significant increase in gene expression for the Dandarawi line compared to the Leghorn line. These results indicate that the Dandarawi breed has genes that enable it to adapt under hot climate conditions. Hence, when selected to increase production under high temperatures, the gene expression is twice that of the Leghorn strain under the same conditions of selection. The Dandarawi chicken had better heat tolerance than the Leghorn chicken. The difference was observed on chromosome 5 in the region 52784621: 52784671 in exon 1 in HSP70 that was caused by the amino acid cysteine instead of arginine because of the substitution of T nucleotide with C, indicating the alteration in the gene expression level of HSPs. Therefore, we recommend using chromosome 5 in the region 52784621: 52784671 in exon 1 in HSP70 as a candidate gene in selection programs for evaluating heat stress tolerance.

Biomarkers of Planarian Dugesia japonica in Response to Herbicide Glyphosate Exposure.

As the worldwide top-selling herbicide, glyphosate is ubiquitously distributed in the natural environment, and its influence on the ecological safety and human health has being increasingly concerned. In this study, mRNA expressions of GPX and three heat shock protein genes in freshwater planarian Dugesia japonica in response to glyphosate were determined, and two oxidative stress parameters were measured. The results suggested that GPX activity can be used as a more sensitive biomarker in contrast with GPX gene expression, and mRNA expressions of Hsp70, Hsp90 genes are more sensitive than Hsp40 for planarians in response to glyphosate stress. Besides, the deduced T-AOC as well as varied GPX activity and mRNA expression levels of Hsps also indicated that glyphosate exposure would inhibit antioxidation and induce oxidative stress in D. japonica, while specific antioxidant systems and stress proteins tried to protect cells by their own regulation. The results of this study will be helpful to elucidate the stress response mechanisms of freshwater planarians to herbicide glyphosate.

Abstract P3-01-18: Kinase D interacting substrate 220 (Kidins220) and disease progression of breast cancer, the role of heat shock protein90 (HSP90)

Abstract Background: Kidins220 (Kinase D Interacting Substrate 220) is a transmembrane scaffold protein and was initially found as the substrate of Protein Kinase D. It has several binding partners and plays an important role in neuronal activities and attribute to the occurrence of neuronal disease. Moreover, emerging evidence has revealed the involvement of Kidins220 in regulating cancer development including melanoma, neuroblastoma, and prostate cancer. Our current study investigated the role of Kidins220 in breast cancer and the biological implications on breast cancer cells. Method: A panel of human breast cancer cell lines were used in the biological studies. Kidin220 Knockdown cell model was established using lentivirus anti-Kidins220 shRNA and the knockdown of Kidins220 was verified in gene and protein levels. Cell matrix attachment and cell migration assay were determined using an automated cell analyser (ECIS). The growth of breast cancer cells was evaluated using colorimetric method. Human breast tumours (n=106) together with adjacent background tissues were collected immediately after surgery. Transcription level of Kidins220 and HSPs in the fresh tissues was determined using quantitative transcript analyses. Results: In the clinical cohort of breast cancer, an increase in expression of Kidins220 was found in tumour tissues in comparison with adjacent normal tissues. Patients with poor prognosis (NPI&amp;gt;5.4) had the lowest expression of Kidins220 compared with patients with good and moderate prognosis. Likewise, high grade tumours had markedly reduced level of Kidins220. Furthermore, patients remain disease free had the highest Kidins220 level when compared with those who had breast cancer-related incidence. Kaplan-Meier plot showed high level of Kidins220 was linked to longer survival. Our database also revealed that the levels of expression of Kidins200 were significantly correlated with that of HSP90A (Heat Shock Protein-90A) (r=0.37, p&amp;lt;0.001), but not HSP90B. Moreover, breast cancer cells (MDA MB-231 and MCF-7) after losing Kidins220 by way of knockdown showed an increase in cellular migration and cell matrix adhesion, with little changes in cell growth compared with control cells. In these cell based models, HSP90 inhibitor was found to affect Kidins220 mediated changes in matrix adhesion. It was also noteworthy that cells with Kidins220 knockdown had dramatically reduced expression of HSP90, particularly HSP90A. Conclusion: Kidins220 may act as a tumour suppressor in regulating the disease progression of breast cancer and has a correlation with the survival of patients with breast cancer. Kidins220 regulates the aggressive property including migration and cell matrix attachment of breast cancer with the involvement of HSP90A. Citation Format: Zhe Zhao, Tracey A. Martin, Johannes Benedikt, Shuo Cai, Lin Ye, Eleri Davies, Robert E Mansel, Wen G. Jiang. Kinase D interacting substrate 220 (Kidins220) and disease progression of breast cancer, the role of heat shock protein90 (HSP90) [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P3-01-18.

Selenium Deficiency Induces Inflammation via the iNOS/NF-κB Pathway in the Brain of Pigs.

Selenium (Se) is an essential trace element to maintain homeostasis in humans and animals. The aim of the present study was to clarify the mechanism of Se deficiency-induced inflammation in the pig's brain. Twenty-four healthy pigs were randomly divided into two groups (n = 12/group): control group (group C) was fed diet with 0.3mg/kg inorganic Se, and Se-deficient group (group L) was fed diet with 0.007mg/kg inorganic Se. At the 90th day of the experiment, the histology in the pig's brain was observed by the microscope, the NO levels and iNOS activity were assayed, and the mRNA and protein expression levels of inflammatory cytokines (iNOS, COX-2, NF-κB, and PTGEs) and HSPs (HSP27, HSP40, HSP60, HSP70, and HSP90) were detected by real-time quantitative PCR and Western blot. Compared with group C, both of NO levels and iNOS activity were increased in group L, and the mRNA and protein expression levels of inflammatory cytokines (iNOS, COX-2, NF-κB, and PTGEs) and HSPs (HSP27, HSP40, HSP60, HSP70, and HSP90) were also upregulated; histological observation displayed inflammatory response in the brain of pig. In summary, diet with Se deficiency can activate the iNOS/NF-κB pathway to upregulate the expression of inflammatory cytokines, thereby leading to inflammatory lesions in the pig's brain, and HSPs are involved in the compensatory regulation of inflammation. This study provides a reference for the prevention of pig brain inflammation from the perspective of nutrition.