Hsp70 Levels Research Articles

The first reporting of prevalence Vibrio species and expression of HSP genes in rayed pearl oyster (Pinctada radiata) under thermal conditions.

The main goal of the present study was to evaluate the influence of thermal exposure on Vibrio population and HSP genes expression (HSP 90, HSP70, and HSP20) in rayed pearl oyster (P. radiata). To this end, the oysters were reared for 30 days at temperatures of 22°C (control), 25°C, 27°C, and 29°C. The results showed that five dominate Vibrio strains including Vibrio hepatarius, V. harveyi, V. alginolyticus, V. parahaemolyticus, and V. rotiferianus were identified. The highest population of V. parahaemolyticus, V. alginolyticus, and V. harveyi, was found in 29οC group. According to real-time PCR, mantle exhibited the highest expression levels of HSP20, HSP70, and HSP90 genes. A higher level of HSP20 expression was observed at high temperatures (25°C, 27°C, and 29°C) in the gonad and mantle compared to the control group (22°C) while decrease in HSP90 expression level was recorded in 25°C, 27°C, and 29°C groups. HSP20 expression level in adductor muscle was remarkably down-regulated in 27°C and 29°C groups. In this tissue, HSP70 was detected at highest levels in the 29οC group. In mantle, HSP90 gene expression was lowest at 22°C water temperature. Several Vibrio strains have been identified from pearl Gulf oyster that haven't been previously reported. The identification of dominant Vibrio species is essential for epidemiological management strategies to control and prevent Vibrio outbreaks in pearl oyster farms. The expression pattern of HSP genes differs in rayed pearl oyster tissues due to differences in their thermal tolerance capability and physiological and biological characteristics. The present study provides useful molecular information for the ecological adaptation of rayed pearl oysters after exposure to different temperature levels.

The impact of chasing stress on the metabolism of the Atlantic Ghost Crab Ocypode quadrata (Fabricius, 1787).

Ocypode quadrata, a Ghost crab species found along the western Atlantic coast, is considered a bioindicator of anthropogenic impact on sandy beaches. Ghost Crabbing, a touristic activity in which ghost crabs are chased just for fun, is a potentially threatening activity for this crab. In crustaceans, metabolites such as glucose and lactate, and the gene expression of crustacean hyperglycemic hormone (CHH) and heat shock proteins (HSPs) increase when the animals are exposed to several types of stress, including alterations in temperature, salinity, or exposure to xenobiotics. This work was developed to identify if being chased by humans would affect these markers of stress in this species of crab. The effects of chasing stress on hemolymph and tissue metabolites and the gene expression levels of CHH and HSP70 were investigated. The levels of lactate in the hemolymph of stressed crabs were six times higher than those of control crabs immediately after chasing and decreased progressively during recovery, indicating an active anaerobic metabolism during the stress. On the contrary, glucose levels in the hemolymph of the stressed crabs increased progressively from 30 to 60 min after chasing, indicating an inverse correlation between glucose and lactate and the conversion of lactate to glucose by gluconeogenesis. In stressed crabs, the levels of triglycerides in the hemolymph decreased 30 min after chasing, while the opposite tended to occur in the hepatopancreas, indicating that during recovery, the crabs use triglycerides as energy source to sustain aerobic metabolism. Finally, this study demonstrates that ghost crabs are stressed by minimum human contact and that "ghost crabbing" must not be encouraged as a tourist activity.

The implications of exercise in Drosophila melanogaster: insights into Akt/p38 MAPK/Nrf2 pathway associated with Hsp70 regulation in redox balance maintenance.

This study investigated the potential effects of exercise on the responses of energy metabolism, redox balance maintenance, and apoptosis regulation in Drosophila melanogaster to shed more light on the mechanisms underlying the increased performance that this emerging exercise model provides. Three groups were evaluated for seven days: the control (no exercise or locomotor limitations), movement-limited flies (MLF) (no exercise, with locomotor limitations), and EXE (with exercise, no locomotor limitations). The EXE flies demonstrated greater endurance-like tolerance in the swimming test, associated with increased citrate synthase activity, lactate dehydrogenase activity and lactate levels, and metabolic markers in exercise. Notably, the EXE protocol regulated the Akt/p38 MAPK/Nrf2 pathway, which was associated with decreased Hsp70 activation, culminating in glutathione turnover regulation. Moreover, reducing the locomotion environment in the MLF group decreased endurance-like tolerance and did not alter citrate synthase activity, lactate dehydrogenase activity, or lactate levels. The MLF treatment promoted a pro-oxidant effect, altering the Akt/p38 MAPK/Nrf2 pathway and increasing Hsp70 levels, leading to a poorly-regulated glutathione system. Lastly, we demonstrated that exercise could modulate major metabolic responses in Drosophila melanogaster aerobic and anaerobic metabolism, associated with apoptosis and cellular redox balance maintenance in an emergent exercise model.

Extra and intra cellular HSP70 levels in adults with and without metabolic disorders: a systematic review and meta-analysis.

Metabolic disorders, such as obesity, type 2 diabetesmellitus (T2DM), and metabolic syndrome (MS) are related to chronic pro-inflammatory conditions. Evidence suggests that heat shock proteins are linked to metabolic disorders. Intracellular HSP70 (iHSP70) is mandatory for normal insulin signalling, and proteostasis, and exerts a powerful anti-inflammatory role. On the other hand, the extracellular (eHSP72) is linked with a pro-inflammatory state and induces insulin resistance in humans. Then, we conducted a systematic review with meta-analysis to summarize the data of HSP70 in people with and without metabolic disorders. PubMed, Embase, Scopus, and Web of Science databases were used. Eligibility criteria included observational and baseline data of experimental studies that assessed iHSP70 and/or eHSP72 in adults with metabolic disorders and healthy people. The risk of bias was assessed by the Newcastle-Ottawa scale. Meta-analysis was performed using a random-effect model and the mean difference was estimated for eHSP72 and the standardized mean difference for iHSP70. A total of 11,255 articles were retrieved, 31 articles were assessed for eligibility and 15 were included for data extraction. There was no difference in eHSP72 between metabolic disorders and healthy controls (mean difference (MD) = 0.11; 95% confidence interval (CIs) = -0.05 to 0.27; I2 = 95%). Subgroup analysis showed higher levels of eHSP72 in T2DMpeople than healthy ones (MD = 0.32; 95% CIs = 0.17 to 0.47; I2 = 92%). For iHSP70 no difference was found (standardized mean difference (SMD) =-0.24; 95% CIs =-1.62 to 1.15; I2 = 86%). Our results suggest that eHSP72 levels may be dependent on metabolic condition and no difference in iHSP70 levels are attributed to high heterogeneity level between studies (PROSPERO REGISTRATION: CRD42022323514).

LncRNA MALAT1 Promoted Neuronal Necroptosis in Cerebral Ischemia-reperfusion Mice by Stabilizing HSP90.

The objective of this research was to investigate the role of lncRNA MALAT1 and HSP90 in the regulation of neuronal necroptosis in mice with cerebral ischemia-reperfusion (CIR). We used male C57BL/6J mice to establish a middle cerebral artery occlusion (MCAO) model and conducted in vitro experiments using the HT-22 mouse hippocampal neuron cell line. The cellular localization of NeuN and MLKL, as well as the expression levels of neuronal necroptosis factors, MALAT1, and HSP90 were analyzed. Cell viability and necroptosis were assessed, and we also investigated the relationship between MALAT1 and HSP90. The results showed that MALAT1 expression increased after MCAO and oxygen-glucose deprivation/re-oxygenation (OGD/R) treatment in both cerebral tissues and cells compared with the control group. The levels of neuronal necroptosis factors and the co-localization of NeuN and MLKL were also increased in MCAO mice compared with the Sham group. MALAT1 was found to interact with HSP90, and inhibition of HSP90 expression led to decreased phosphorylation levels of neuronal necroptosis factors. Inhibition of MALAT1 expression resulted in decreased co-localization levels of NeuN and MLKL, decreased phosphorylation levels of neuronal necroptosis factors, and reduced necroptosis rate in cerebral tissues. Furthermore, inhibiting MALAT1 expression also led to a shorter half-life of HSP90, increased ubiquitination level, and decreased phosphorylation levels of neuronal necroptosis factors in cells. In conclusion, this study demonstrated that lncRNA MALAT1 promotes neuronal necroptosis in CIR mice by stabilizing HSP90.

Ischemic preconditioning attenuates endoplasmic reticulum stress-dependent apoptosis of hepatocytes by regulating autophagy in hepatic ischemia-reperfusion injury

Hepatic ischemia–reperfusion injury (HIRI) usually occurs during subtotal hepatectomy and severely damages liver function during the perioperative period. Endoplasmic reticulum stress (ERS) dependent apoptosis has been suggested to play a crucial role in HIRI progression. The present study focused on the regulatory effect of autophagy activation induced by ischemic preconditioning (IPC) on ERS-dependent apoptosis of hepatocytes in HIRI. A HIRI mouse model and oxygen-glucose deprivation/reperfusion (OGD/R) AML-12 hepatocyte cell lines were constructed to evaluate the protective effect of IPC in vivo and in vitro. The protein levels of p-eIF2α, CHOP, and cleaved caspase-12 were used to evaluate the ERS-dependent apoptosis, whereas LC3-II and p62 were considered as the autophagy activation markers. The beneficial molecular chaperones GRP78, HSP60, and HSP70 were also tested to evaluate autophagy. HIRI significantly increased ERS-dependent apoptosis markers and the number of apoptotic cells and damaged liver function. The ERS inhibitor salubrinal significantly alleviated liver injury in HIRI and OGD/R hepatocytes. Furthermore, both remote IPC and direct IPC significantly alleviated liver injury and inflammatory cell infiltration. IPC also upregulated LC3-II, downregulated p62 expression, and increased the mRNA levels of GRP78, HSP60, and HSP70 in HIRI mice and OGD/R hepatocytes, indicating the activation of autophagy by IPC. The autophagy inhibitor 3-methyladenine significantly attenuated the protective effects of IPC on ERS-dependent apoptosis and liver function, whereas the autophagy activator rapamycin mimicked the protective effects of IPC on ERS-dependent apoptosis in vivo and in vitro, suggesting a regulatory role of autophagy in ERS-dependent apoptosis. These results demonstrated that IPC could induce moderate autophagy and upregulate a few molecular chaperones to strengthen endogenous defense mechanisms, which is beneficial for alleviating ERS-dependent apoptosis and protecting hepatocytes from HIRI.

Lysine butyrylation of HSP90 regulated by KAT8 and HDAC11 confers chemoresistance

Posttranslational modification dramatically enhances protein complexity, but the function and precise mechanism of novel lysine acylation modifications remain unknown. Chemoresistance remains a daunting challenge to successful treatment. We found that lysine butyrylation (Kbu) is specifically upregulated in chemoresistant tumor cells and tissues. By integrating butyrylome profiling and gain/loss-of-function experiments, lysine 754 in HSP90 (HSP90 K754) was identified as a substrate for Kbu. Kbu modification leads to overexpression of HSP90 in esophageal squamous cell carcinoma (ESCC) and its further increase in relapse samples. Upregulation of HSP90 contributes to 5-FU resistance and can predict poor prognosis in cancer patients. Mechanistically, HSP90 K754 is regulated by the cooperation of KAT8 and HDAC11 as the writer and eraser, respectively; SDCBP increases the Kbu level and stability of HSP90 by binding competitively to HDAC11. Furthermore, SDCBP blockade with the lead compound V020-9974 can target HSP90 K754 to overcome 5-FU resistance, constituting a potential therapeutic strategy.

Immunostimulation of Parasteatoda tepidariorum (Araneae: Theridiidae) in juvenile and adult stages. Immunity reactions to injury with foreign body and Bacillus subtilis infection.

To assess the immune potential of spiders, in the present study juvenile and adult females of Parasteatoda tepidariorum were exposed to Bacillus subtilis infection, injury by a nylon monofilament and a combination of both. The expression level of selected immune-related genes: defensin 1 (PtDEF1), lysozyme 1 (PtLYS1), lysozyme C (PtLYSC), lysozyme M1 (PtLYSM1), autophagy-related protein 101 (PtATG101), dynamin (PtDYN) and heat shock proteins (HSP70) (PtHSPB, PtHSPB2A, PtHSPB2B), production of lysozyme and HSP70 proteins, and hemocytes viability were measured. The obtained results indicated expression of the lysozyme, autophagy-related protein and HSP70 genes in both ontogenetic stages of P. tepidariorum. It has been also shown that the simultaneous action of mechanical and biological factors causes higher level of lysozyme and HSP70, cell apoptosis intensity and lower level of hemocytes viability than in the case of exposure to a single immunostimulant. Moreover, mature females showed stronger early immune responses compared to juveniles.

THE RELATIONSHIP BETWEEN ATTACHMENT AND SERUM OXYTOCIN AND HEAT SHOCK PROTEIN-70 LEVELS IN ADOLESCENTS OF PARENTS WITH SCHIZOPHRENIA AND BIPOLAR DISORDER.

The aim of this study was to evaluate serum heat shock protein 70 (HSP70) and oxytocin levels, attachment and perceived social support levels in adolescents with parental bipolar disorder (BD) and Schizophrenia (SCZ). This study included 9 adolescents with SCZ parents, 30 adolescents with BD parents and 31 healthy adolescents. Brief Symptom Inventory (BSI), Relationship Scale Questionnaire-Adolescent Form (RSQ-A) and Multidimensional Scale of Perceived Social Support (MSPSS) were administered to all participants. In addition, serum HSP-70 and oxytocin levels were evaluated. There was no significant difference between the groups in terms of attachment style, psychiatric symptoms and perceived social support. Serum HSP-70 levels were found to be lower in adolescents whose parents had BD. Serum oxytocin levels of the SCZ group were significantly lower than those of the BD group. HSP-70 level was found to be lower in adolescents with BD parents. Oxytocin level was found to be lower in adolescents with SCZ parents. These findings suggest that HSP-70 and oxytocin may be a marker of early life stress in adolescents with parental psychopathology. However, studies are needed to evaluate the relationship between attachment, oxytocin and HSP-70 in adolescents exposed to parental psychopathology in early life.

The effects of STA-9090 (Ganetespib) and venetoclax (ABT-199) combination on apoptotic pathways in human cervical cancer cells.

Combined chemotherapy is recommended strategy as a first-line treatment method in patients with cervical cancer. Ganetespib (STA-9090) is a second-generation heat shock protein 90 (Hsp90) inhibitor that blocks the ATPase function of Hsp90 and inhibits the proper folding of oncogenic client proteins. Venetoclax (ABT-199) is an orally bioavailable Bcl-2 (B-cell lymphoma 2) inhibitor that stimulates apoptotic signaling pathways in cancer cells. This study evaluated the anticancer effects of STA-9090 combined with Venetoclax in the human cervical cancer cell line (HeLa). The human cervical cancer cells were treated with STA-9090, Venetoclax, and Sta-9090 plus Venetoclax for 48h, and cell viability was measured using the XTT assay. The alteration of the Hsp90 protein expression level and the chaperone activity of HSP90 were detected by ELISA and luciferase aggregation assay, respectively. For the apoptotic process, qRT-PCR was applied to study Bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), Bcl-2-like protein 1 (Bcl-xL ), Cytochrome c (Cyt-c), Caspase3 (Cas-3), and Caspase7 (Cas-7) expression levels after drug treatments. Also, a colorimetric Cas-3 activity assay was performed to detect the induction of the apoptosis process. Our results demonstrated that 8 nM of STA-9090 combined with 4 µM of Venetoclax synergistically inhibited cervical cancer cell proliferation more than STA-9090 or Venetoclax alone after 48h of treatment. STA-9090 and Venetoclax combination decreased the protein expression level of Hsp90 and significantly inhibited chaperone activity of Hsp90. This combination stimulated apoptosis in cervical cancer cells by down-regulating of anti-apoptotic markers while inducing pro-apoptotic markers. Also, the STA-9090-Venetoclax combination increased Cas-3 activity in Hela cells. Collectively, these findings pointed out that the STA-9090-Venetoclax combination exhibited more activity than the individual drugs to stimulate toxicity and apoptosis in cervical cancer cells based on HSP90 inhibition.

Circulating levels of inflammatory cytokines and angiogenesis-related growth factors in patients with osteoarthritis after COVID-19.

The disease COVID-19, caused by SARS-CoV-2 infection, has a systemic effect and is associated with a number of pathophysiological mechanisms that mobilize a wide range of biomolecules. Cytokines and growth factors (GFs) are critical regulators of tissue damage or repair in osteoarthritis (OA) and are being recognized as key players in the pathogenesis of COVID-19. A clear understanding of the long-term consequences of SARS-CoV-2 infection, especially in patients with concomitant chronic diseases, is limited and needs to be elucidated. The study aimed to evaluate the degree of inflammation and levels of pro-angiogenic and hypoxic factors, as well as heat shock proteins HSP60 and HSP70 in plasma, of patients with OA after recovery from COVID-19. The research involved patients of an orthopedic specialty clinic aged 39 to 80 diagnosed with knee OA. All examined patients were divided into three groups: the Control group included conditionally healthy donors, group OA included patients with knee OA mainly stage II or III and the group of OA and COVID-19 included patients with OA who had COVID-19. The plasma levels of pro-inflammatory molecules IL-1β, IL-6, TNF-α, NF-κB, angiogenic factors VEGF, FGF-2, PDGF, hypoxic factor HIF-1α and molecular chaperones HSP60 and HSP70 were measured by enzyme-linked immunosorbent assay. The study showed that in both groups of patients, with OA and convalescent COVID-19, there was an increase in the plasma level of IL-1β and a decrease in TNF-α and NF-κB levels when compared to healthy controls. Systemic deregulation of the cytokine profile was accompanied by reduction in plasma levels of pro-angiogenic growth factors, most pronounced in cases of VEGF and PDGF. This analysis did not reveal any significant difference in the plasma level of HIF-1α. A decrease in the level of stress protein HSP60 in the blood of patients with OA, as well as those patients who have had SARS-CoV-2 infection, has been established. The results suggest the potential role pro-inflammatory cytokines and angiogenesis-related growth factors in pathogenesis of both joint pathologies and long-term systemic post-COVID-19 disorders.

A Coumarin-Imidazothiadiazole Derivative, SP11 Abrogates Tumor Growth by Targeting HSP90 and Its Client Proteins.

Despite several treatment options for blood cancer, mortality remains high due to relapse and the disease's aggressive nature. Elevated levels of HSP90, a molecular chaperone essential for protein folding, are associated with poor prognosis in leukemia and lymphoma. HSP90 as a target for chemotherapy has been met with limited success due to toxicity and induction of heat shock. This study tested the activity of an HSP90 inhibitor, SP11, against leukemic cells, mouse lymphoma allograft, and xenograft models. SP11 induced cytotoxicity in vitro in leukemic cell lines and induced cell death via apoptosis, with minimal effect on normal cells. SP11 induced cell death by altering the status of HSP90 client proteins both in vitro and in vivo. SP11 reduced the tumor burden in allograft and xenograft mouse models without apparent toxicity. The half-life of SP11 in the plasma was approximately 2 h. SP11 binding was observed at both the N-terminal and C-terminal domains of HSP90. C-terminal binding was more potent than N-terminal binding of HSP90 in silico and in vitro using isothermal calorimetry. SP11 bioavailability and minimal toxicity in vivo make it a potential candidate to be developed as a novel anticancer agent.

Joint cadmium and polypropylene microparticle action in cadmium tolerant model insect

Microplastic enlisted as a contaminant of emerging concerns in polluted environments interact with "traditional" contaminants such as metals, causing, among others, their increased accumulation in the body. Harmful effects depend on the exposed animals' possible preadaptation and/or cross-tolerance. The project aimed to assess the role of this phenomenon in the limited toxicity of polypropylene fibers (PPf) in 0%, 0.02%, 0.06, 0.18%, 0.54%, and 1.6% of Cd-supplemented food of larvae of Spodoptera exigua multigenerationally selected to cadmium tolerance. The activity of 20 digestive enzymes (API-ZYM test), defensins, and heat shock proteins, HSP70 levels in the exposed groups were used as biomarkers. PPfs caused the increase of Cd accumulation in the body, while intake of polypropylene microfibers did not change the biomarker levels. Moreover, multigenerational Cd pre-exposure, due to increased tolerance of Cd and, possibly, cross-tolerance, prepares the insects for an additional stressor (PPf) alone and in interaction with cadmium.

Evaluation of dentinal tubule occlusion and pulp tissue response after using 980-nm diode laser for dentin hypersensitivity treatment.

To evaluate the effectiveness of the 980-nm diode laser for dentinal tubule occlusion, measure the intrapulpal temperature, and investigate the dental pulp response. The dentinal samples were randomly divided into G1-G7 groups: control; 980-nm laser irradiation (0.5 W, 10s; 0.5 W, 10s × 2; 0.8 W, 10s; 0.8 W, 10s × 2; 1.0 W, 10s; 1.0 W, 10s × 2). The dentin discs were applied for laser irradiation and analyzed by scanning electron microscopy (SEM). The intrapulpal temperature was measured on the 1.0-mm and 2.0-mm thickness samples, and then divided into G2-G7 groups according to laser irradiation. Moreover, forty Sprague Dawley rats were randomly divided into the laser-irradiated group (euthanized at 1, 7, and 14days after irradiation) and the control group (non-irradiated). qRT-PCR, histomorphology, and immunohistochemistry analysis were employed to evaluate the response of dental pulp. SEM indicated the occluding ratio of dentinal tubules in the G5 (0.8 W, 10s × 2) and G7 (1.0 W, 10s × 2) were significantly higher than the other groups (p < 0.05). The maximum intrapulpal temperature rises in the G5 were lower than the standard line (5.5 ℃). qRT-PCR showed that the mRNA expression level of TNF-α and HSP-70 upregulated significantly at 1day (p < 0.05). Histomorphology and immunohistochemistry analysis showed that, compared with the control group, the inflammatory reaction was slightly higher at the 1 and 7days (p < 0.05) and decreased to the normal levels at 14days (p > 0.05). A 980-nm laser at a power of 0.8 W with 10s × 2 defines the best treatment for dentin hypersensitivity in terms of compromise between the efficacy of the treatment and the safety of the pulp. The 980-nm laser is an effective option for treating dentin sensitivity. However, we need to ensure the safety of the pulp during laser irradiation.

The Effect of Sericin on Heat Shock Proteins 60 and 90, and TLR2 and TLR4 Induced by Chronic Heat Stress in Male Mice

Background: Heat stress followed by heat stroke, which is caused by an increase in body temperature to more than 40 °C, is a medical emergency that affects many people in work situations and weather conditions and results in irreversible neurological and cognitive damage. Methods: In general, 75 male albino mice in the weight range of 28-30 g were used in the present study. The mice were divided into 5 groups of 15, including the control group, heat stress (HS) group, HS group+sericin 100, HS group+sericin 150, and HS group+sericin 200. Toll-like receptor 2 (TLR2) and Toll-like receptor (TLR4) expression levels were measured using the real-time polymerase chain reaction (RT-PCR), and the expression of hsp60 and hsp90 proteins was measured using a western blot. Results: The expression level of TLR2 and TLR4 receptors increased significantly with the passage of time as a result of stimulation. Western blot results showed that the expression levels of HSP60 and HSP90 increased as well. Conclusion: The results of RT-PCR revealed that HS significantly increases the expression of innate immune system receptors such as TLR2 and TLR4.

Characteristics of Vitamin D3 Receptor Genotypes in T2DM of Iraqi Obese Women

Objective: This study aims to examine the association between the (rs1544410) polymorphism of the VDR gene with the pathogenesis of T2DM in Iraqi obese women.&#x0D; Methods: A case-control study was performed on 50 patients with T2DM and 50 apparently healthy subjects who were admitted to Al-Hussein Teaching Hospital and Al-Hassan Center of Diabetes and Endocrinology unit / Kerbala health directorate – Iraq during (April 2022 – March 2023). The T2DM groups were divided into two groups, 25 obese and 25 non-obese; the control group was divided into 25 obese and 25 non-obese as apparently healthy groups. The ELISA Kit was used to measure serum 25(OH)D3, heat shock protein-70, VDBP, insulin and C-peptide. Also HbA1c% and insulin resistance (HOMA-IR) were evaluated. The vitamin D3 receptor gene (VDR) variant and the SNP (rs1544410) polymorphism was determined using allele specific polymerase chain reaction, 1.5% agarose gel electrophoresis and then visualized by gel photo-documentation system.&#x0D; Results: The result of vitamin D3 variants genotype (rs 1544410) was a clear band with a molecular size of 200 bps. The size of the amplicon was determined by compare with DNA ladder 100 - 1500 bp. The result of the comparison between observed and anticipated values for SNIP with (rs 1544410) in the tested population was statistically significant, P= &lt; 0.001 and the difference between demographic characteristics and (rs 1544410) SNP, age and BMI shows non-significant difference among all groups. The difference between biomarkers and (rs1544410) SNP was performed using one-way ANOVA test to compare the mean levels of HSP-70, VDBP, C-peptide, RBC and HbA1c% which shown a non-significant difference among the variants of VDBP Genotype (rs1544410) in obese women (patients and control) studied groups, p value &gt; 0.05. &#x0D; Conclusion: The logistic analysis of the (rs1544410) SNP of the patients concluded that HSP-70, VDBP, and C-peptide level was no significantly related to the also C-peptide, was shown to be a related risk factor to both CT and CT alleles (1.003, p &gt; 0.05) in comparison with CC alleles. Furthermore, HbA1c% level was demonstrated to be related as a risk factor for the CG allele in comparison with CC and GG alleles (1.009, p &lt; 0.05).

Proliferation, migration, and resistance to oxidative and thermal stresses of HT1080 cells with knocked out genes encoding Hsp90α and Hsp90β

Heat shock protein 90 (Hsp90) fulfils essential housekeeping functions in the cell associated with the folding, stabilization, and turnover of various proteins. In mammals, there exist two Hsp90 isoforms, stress-inducible Hsp90α and constitutively expressed Hsp90β. In an attempt to identify cellular processes dependent on Hsp90α and Hsp90β, we generated a panel of clones of human fibrosarcoma HT1080 cells with the knocked out HSP90AA1 or HSP90AB1 genes encoding, respectively, Hsp90α and Hsp90β. The knockout of the HSP90AA1 and HSP90AB1 genes practically did not affect cell proliferation and resistance to thermal shock and oxidative stress. The loss of Hsp90α in Hsp90α-null cell clones also did not impair cell migration, while the migration of the Hsp90β-null cell clones was prominently reduced as compared to parent HT1080 cells. This indicated the necessity of Hsp90β for efficient basal migration of HT1080 cells whereas Hsp90α seems to be dispensable for this process. The knockout of one Hsp90 isoform was invariably accompanied by an increase in the level of the other Hsp90 isoform by 30–50%, which partly or fully compensated for a decrease in the total level of Hsp90. Thus, we demonstrated the dispensability of Hsp90α and Hsp90β for HT1080 cells in several cellular processes under normal and stress conditions, which suggested the participation of the two Hsp90 isoforms in the same biological processes and full or at least partial functional substitution of one Hsp90 isoform by the other.

Knockdown of ACC1 promotes migration of esophageal cancer cell

Objective: To investigate the effect of acetyl-CoA carboxylase 1 (ACC1) knockdown on the migration of esophageal squamous cell carcinoma (ESCC) KYSE-450 cell and underlying mechanism. Methods: Lentiviral transfection was conducted to establish sh-NC control cell and ACC1 knocking down cell (sh-ACC1). Human siRNA HSP27 and control were transfected by Lipo2000 to get si-HSP27 and si-NC. The selective acetyltransferase P300/CBP inhibitor C646 was used to inhibit histone acetylation and DMSO was used as vehicle control. Transwell assay was performed to detect cell migration. The expression of HSP27 mRNA was examined by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and the expressions of ACC1, H3K9ac, HSP27 and epithelial-mesenchymal transition-related proteins E-cadherin and Vimentin were detected by western blot. Results: The expression level of ACC1 in sh-NC group was higher than that in sh-ACC1 group (P<0.01). The number of cell migration in sh-NC group was (159.00±24.38), lower than (361.80±26.81) in sh-ACC1 group (P<0.01). The protein expression levels of E-cadherin and Vimentin in sh-NC group were statistically significant compared with sh-AAC1 group (P<0.05). The migrated cell number in sh-NC+ si-NC group was (189.20±16.02), lower than (371.60±38.40) in sh-ACC1+ si-NC group (P<0.01). The migrated cell number in sh-NC+ si-NC group was higher than that in sh-NC+ si-HSP27 group (152.40±24.30, P<0.01), and the migrated cell number in sh-ACC1+ si-NC group was higher than that in sh-ACC1+ si-HSP27 group (P<0.01). The protein expression levels of E-cadherin and Vimentin in sh-NC+ si-NC group were significantly different from those in sh-ACC1+ si-NC and sh-NC+ si-HSP27 groups (P<0.01). The protein expression levels of E-cadherin and Vimentin in sh-ACC1+ si-NC group were significantly different from those in sh-ACC1+ si-HSP27 group (P<0.01). After 24 h treatment with C646 at 20 μmmo/L, the migrated cell number in sh-NC+ DMSO group was (190.80±11.95), lower than (395.80±17.10) in sh-ACC1+ DMSO group (P<0.01). The migrated cell number in sh-NC+ DMSO group was lower than that in sh-NC+ C646 group (256.20±23.32, P<0.01). The migrated cell number in sh-ACC1+ DMSO group was higher than that in sh-ACC1+ C646 group (87.80±11.23, P<0.01). The protein expressions of H3K9ac, HSP27, E-cadherin and Vimentin in sh-NC+ DMSO group were significantly different from those in sh-ACC1+ DMSO group and sh-NC+ C646 group (P<0.01). The protein expression levels of H3K9ac, HSP27, E-cadherin and Vimentin in sh-ACC1+ DMSO group were significantly different from those in sh-ACC1+ C646 group (P<0.01). Conclusion: Knockdown of ACC1 promotes the migration of KYSE-450 cell by up-regulating HSP27 and increasing histone acetylation.

The digestive enzyme activity, intestinal microbiota and immune-related genes expression of snakehead fish (Channa argus) juveniles affected by dietary cricket (Gryllus testaceus) meal

This study aimed to investigate the potential of using cricket (Gryllus testaceus) meal as the main protein source in fish feed for snakehead fish (Channa argus) juveniles. A 10-week feeding trial was conducted to assess the effects of partial replacement of fishmeal (FM) with cricket meal (CM) on growth performance, digestive enzyme activity, intestinal microbial composition, liver antioxidant capacity, immune response and determine the appropriate replacement FM level with CM in juvenile Channa argus diet. Five isonitrogenous and iso-lipid experimental diets were formulated by replacing 0 %, 15 %, 30 %, 45 % and 60 % of FM with CM, respectively named T0 (control group), T15, T30, T45 and T60 groups. The results demonstrated T45 and T60 showed a significantly decreased feed conversion ratio (FCR) than T0 (P < 0.05). Crude lipid content was significantly higher in T45 and T60 than in T0 (P < 0.05). With increasing dietary CM content, the activities of amylase and lipase were significantly increased in T60 (P < 0.05). At the phylum level, 16S rDNA sequences revealed that Proteobacteria abundance was dominant in all treatments, while Ralstonia abundance had higher significance in T60 than others at the genus level (P < 0.05). The total antioxidant capacity (T-AOC) in liver in T30 and T45 were significantly higher than in T0 (P < 0.05). Glutathione peroxidase (GPx) activities had a significant increase in fish fed groups except T0 (P < 0.05), while malondialdehyde (MDA) content was significantly lower within T45 and T60 than T0 (P < 0.05). The up-regulated gene expression level (IL-1β, IL-10, and HSP70) and down-regulated expression level (IL-8) were showed in all treatments with CM replacement. HSP90 and Iκ-Bα expression levels were increased and peaked in T30 (P < 0.05). According to the results of the current study, in Channa argus diets, FM replacement with CM can be up to 30 % without compromising growth performance, enzyme activities, antioxidant capacity, intestinal microbiota, and immune response.

The effects of Pandanus tectorius leaf extract on the resistance of White-leg shrimp Penaeus vannamei towards pathogenic Vibrio parahaemolyticus

Pandanus tectorius leaf extract effect on the White-leg shrimp Penaeus vannamei tolerance against Vibrio parahaemolyticus were investigated in this study. Thirty shrimp post-larvae measured at approximately 1 cm were exposed for 24 h to 0.5, 1, 2, 3, 4, 5 and 6 g/L leaf extract and subsequently observed for survival and immune-related genes expression (Hsp70, ProPO, peroxinectin, penaeidin, crustin and transglutaminase), followed by determination of their tolerance and histological tissue profiles upon Vibrio challenge. Survival of shrimps treated with 6 g/L of leaf extract improved by up to 95% to controls. Hsp70, crustin, and prophenoloxidase mRNA levels were observed to be 8.5, 10.4, and 1.5-fold higher, respectively. Histopathological analysis of the hepatopancreas and the muscle tissues revealed major tissue degeneration in Vibrio-challenged shrimps but not in shrimps primed with P. tectorius leaf extract. Of all the dose examined, the best pathogen resistance results were obtained with a 24 h incubation of shrimp in 6 g/L P. tectorius methanolic leaf extract. The tolerance towards V. parahaemolyticus might be associated with the increased regulation of Hsp70, prophenoloxidase and crustin upon exposure to the extract, all immune-related proteins essential for pathogen elimination in Penaeid shrimp. The present study primarily demonstrated that P. tectorius leaf extract is a viable alternative for enhancing P. vannamei post-larvae resistance against V. parahaemolyticus, a major bacterial pathogen in aquaculture.