Hsp70 Gene Expression Research Articles

Effect of Defoliation on the Defense Reactions of Silver Birch (Betula pendula) Infected with Phytophthora plurivora

In natural environments, plants develop adaptive mechanisms at the cellular and molecular levels to cope with many external factors, e.g., insects and soil pathogens. We studied physiological stress induced by different levels of foliage removal (defoliation 30% and 60%) and by infection of root rot pathogen Phytophthora plurivora on the common Polish tree species, Betula pendula, grown in an open greenhouse. This study showed that P. plurivora damaged the root system which negatively impacted all morphological parameters. However the loss of 30% of the leaves had a positive effect on defense responses. Chlorophyll-a fluorescence parameters indicated a decrease in photosynthetic efficiency in defoliated plants, but plants inoculated with the pathogen had a higher performance index showing increased vigor of the infected plant than birches. The study demonstrated the intense immunity response of birch to P. plurivora through the expression of Hsp90 and Hsp83 genes. The trees weakened by P. plurivora became more susceptible to infection by Erysiphe ornata var. ornata.

Distinct Gene Expression Patterns of Two Heat Shock Protein 70 Members During Development, Diapause, and Temperature Stress in the Freshwater Crustacean Daphnia magna

Dormancy is a lifecycle delay that allows organisms to escape suboptimal environmental conditions. As a genetically programmed type of dormancy, diapause is usually accompanied by metabolic depression and enhanced tolerance toward adverse environmental factors. However, the drivers and regulators that steer an organism’s development into a state of suspended animation to survive environmental stress have not been fully uncovered. Heat shock proteins 70 (HSP70s), which are often produced in response to various types of stress, have been suggested to play a role in diapause. Considering the diversity of the Hsp70 family, different family members may have different functions during diapause. In the present study, we demonstrate the expression of two hsp70 genes (A and B together with protein localization of B) throughout continuous and diapause interrupted development of Daphnia magna. Before and after diapause, the expression of Dmhsp70-A is low. Only shortly before diapause and during diapause, Dmhsp70-A is significantly upregulated and may therefore be involved in diapause preparation and maintenance. In contrast, Dmhsp70-B is expressed only in developing embryos but not in diapausing embryos. During continuous development, the protein of this Hsp70 family member is localized in the cytosol. When we expose both embryo types to heat stress, expression of both hsp70 genes increases only in developing embryos, and the protein of family member B is translocated to the nucleus. In this stress formation, this protein provides effective protection of nucleoplasmic DNA. As we also see this localization in diapausing embryos, it seems that Daphnia embryo types share a common subcellular strategy when facing dormancy or heat shock, i.e., they protect their DNA by HSP70B nuclear translocation. Our study underlines the distinctive roles that different Hsp70 family members play throughout continuous and diapause interrupted development.

Diurnal and seasonal hsp70 gene expression in a cryptic reef fish, the bluebanded goby Lythrypnus dalli (Gilbert 1890)

Physiological responses of marine organisms are influenced by environmental variations, particularly in dynamic habitats. These variations could be at a diel or seasonal scale and require acclimation responses that, at a molecular level, are facilitated by gene expression regulation. Circadian rhythms allow organisms to adjust to recurring environmental changes and to synchronize physiological processes. Increased Hsp70 gene expression allows organisms to protect themselves against heat and other stressors, conferring them a periodically changing thermotolerance. This study aimed at analyzing whether the variability of exogenous factors (temperature, dissolved oxygen, salinity) in diurnal or seasonal scales affects hsp70 expression in cryptic reef fishes in their natural environment. The study model was Lythrypnus dalli, an endemic cryptic reef fish inhabiting shallow waters throughout the Gulf of California. Physical parameters varied seasonally; temperature increased 6°C during the warm season, salinity and dissolved oxygen remained constant. Minimal changes in temperature (±1 °C) occurred diurnally. Expression levels of hsp70 increased significantly during the warm season (p < 0.05), perhaps related to seasonal thermal changes. No differences in hsp70 were observed during the diurnal cycle (p > 0.05). L. dalli has apparently developed phenotypic plasticity to face environmental variability at diurnal and seasonal scales.

Addition of Popular Exogenous Antioxidant Agent, PBN, to Culture Media May Be an Important Step to Optimization of Myogenic Stem/Progenitor Cell Preparation Protocol

The aim of the study was to modify human skeletal muscle-derived stem/progenitor cells (SkMDS/PCs) and demonstrate the optimal cell preparation protocol for application in post-infarction hearts. We used conditioned SkMDS/PC culture medium with α-phenyl-N-tert-butyl nitrone (PBN). SkMDS/PCs were cultured under hypoxic conditions and the results were compared to the standard ones. We observed a significant increase of CD-56 positive phenotypic marker the ability to form functional myotubes, increase in the proportion of young cells in cell primary suspensions, and a decrease in the percentage of apoptotic cells among PBN-conditioned cells in normoxia an hypoxia. We also observed significantly higher levels of SOD3 expression; maintained expression of SOD1, SOD2, and CAT; a higher level of BCL2 gene expression; and a rather significant decrease in Hsp70 gene expression in PBN-conditioned SkMDS/PCs compared to the WT population under hypoxic conditions. In addition, significant increase of myogenic genes expression was observed after PBN addition to culture medium, compared to WT population under hypoxia. Interestingly, PBN addition significantly increased the lengths of telomeres under hypoxia. Based on the data obtained, we can postulate that PBN conditioning of human SkMDS/PCs could be a promising step in improving myogenic cell preparation protocol for pro-regenerative treatment of post-infarction hearts.

Dietary Synbiotics Can Help Relieve the Impacts of Deltamethrin Toxicity of Nile Tilapia Reared at Low Temperatures.

Simple SummaryThe toxic impacts of pesticides and insecticides are strongly correlated with water temperature. Water temperature can increase or decrease the efficacy of toxins and their influence on aquatic organisms. An alternate approach to augmenting fish resistance to ambient deltamethrin (DMT) toxicity and low water temperature via synbiotic feeding was proposed. In this study, fish were allocated into four groups and kept at suboptimal water temperature (21 ± 2 °C): control, DMT, synbiotic, and DMT + synbiotic. The results illustrate that including synbiotics in the Nile tilapia diet stimulates the immunity and antioxidant systems in fish, enabling the fish reared at a suboptimal temperature to counteract the immunity suppression and oxidative stress caused by DMT exposure.The optimal water temperature for the normal growth of Nile tilapia is between 26 and 28 °C, and the toxicity of pesticides is strongly related to water temperature. An alternate approach to augmenting the resistance of fish to ambient water toxicity and low water temperature via synbiotic feeding was proposed. In this study, fish were allocated into four groups with 10 fish in each replicate, where they were fed a basal diet or synbiotics (550 mg/kg) and kept at a suboptimal water temperature (21 ± 2 °C). The prepared diets were fed to Nile tilapia for 30 days with or without deltamethrin (DMT) ambient exposure (15 μg/L). The groups were named control (basal diet without DMT toxicity), DMT (basal diet with DMT toxicity), synbiotic (synbiotics without DMT toxicity), and DMT + synbiotic (synbiotics with DMT toxicity). The results displayed upregulated transcription of catalase, glutathione peroxidase, and interferon (IFN-γ) genes caused by DMT exposure and synbiotic feeding when compared with the controls. Moreover, HSP70 and CASP3 genes displayed increased transcription caused by DMT exposure without synbiotic feeding. However, fish fed with synbiotics showed downregulated HSP70 and CASP3 gene expressions. The transcription of IL-1β and IL-8 genes were also decreased by DMT exposure, while fish fed synbiotics showed upregulated levels. DMT exposure resulted in irregular histopathological features in gills, intestine, spleen, and liver tissues, while fish fed synbiotics showed regular, normal, and protected histopathological images. Our results indicated that dietary synbiotics ameliorated histopathological damages in DMT-exposed tilapia through alleviation of oxidative stress and inflammation as well as enhancing the immunity.

Genome-Wide Identification and Characterization of Hsf and Hsp Gene Families and Gene Expression Analysis under Heat Stress in Eggplant (Solanum melongema L.)

Under high temperature stress, a large number of proteins in plant cells will be denatured and inactivated. Meanwhile Hsfs and Hsps will be quickly induced to remove denatured proteins, so as to avoid programmed cell death, thus enhancing the thermotolerance of plants. Here, a comprehensive identification and analysis of the Hsf and Hsp gene families in eggplant under heat stress was performed. A total of 24 Hsf-like genes and 117 Hsp-like genes were identified from the eggplant genome using the interolog from Arabidopsis. The gene structure and motif composition of Hsf and Hsp genes were relatively conserved in each subfamily in eggplant. RNA-seq data and qRT-PCR analysis showed that the expressions of most eggplant Hsf and Hsp genes were increased upon exposure to heat stress, especially in thermotolerant line. The comprehensive analysis indicated that different sets of SmHsps genes were involved downstream of particular SmHsfs genes. These results provided a basis for revealing the roles of SmHsps and SmHsp for thermotolerance in eggplant, which may potentially be useful for understanding the thermotolerance mechanism involving SmHsps and SmHsp in eggplant.

Characteristic and expression of Hsp70 and Hsp90 genes from Tyrophagus putrescentiae and their response to thermal stress

The development of insects is critically affected by temperature, which therefore plays an important role in the control of stored grain pests. Extreme temperature stress conditions lead to biological responses in mites, such as the synthesis of heat shock proteins. Tyrophagus putrescentiae (Tp) is a pest mite in stored grain that has negative effects on both economy and health. Since T. putrescentiae population dynamics are strongly influenced by temperature, in the present study we have cloned the cDNA of HSP70 and HSP90 (referred to as TpHSP70-1, TpHSP70-2 and TpHSP90) and determined their expression by fluorescence real time quantitative PCR. TpHSP70 and TpHSP90 showed high homology with similar genes in other species and the open reading frames of TpHSP70-1, TpHSP70-2 and TpHSP90 encoded proteins of 665, 661 and 718 amino acid residues, respectively. Under thermal stress, expression of TpHsp70-1 and TpHsp90 was up-regulated at higher temperatures, suggesting their role in the defense against thermal stress.

In Ovo Injection of GABA Can Help Body Weight Gain at Hatch, Increase Chick Weight to Egg Weight Ratio, and Improve Broiler Heat Resistance.

Simple SummaryHeat stress is a vital issue that causes severe losses to the poultry industry. A partly developed thermoregulatory mechanism during the embryonic phase is emphasized to manipulate embryos for achieving thermotolerance during rearing. The present study was conducted firstly to standardize the dosage for an in ovo manipulation, and the selective dose was used to evaluate its effects on early-age heat-stressed (HS) broilers. HS induces cholesterol while an antioxidant acts as a first line of defense under stress. However, 5% GABA supplementation had a higher hatchling weight and chick weight to egg weight ratio (CWEWR). We selected a 10% GABA dosage for HS studies due to its higher antioxidants and lower cholesterol values in hatchlings. In ovo, 10% GABA supplementation significantly increased total antioxidant capacity and reduced malondialdehyde levels, hepatic mRNA levels of HSP70, FAS, and L-FABP in broilers when subjected to HS (38 ± 1 °C; 3 h) at ten days of age. This indicates that an in ovo GABA injection improves CWEWR and antioxidant status at hatch, and creates thermotolerance by increasing antioxidant production and downregulating the expression of HSP70 and fatty acid metabolism genes in HS chicks.The aim of this study was to explore the outcomes of an in ovo GABA injection in broilers challenged with HS. In Experiment 1, 210 Arbor Acres eggs were allocated to five treatments: no-injection, and in ovo injection of 0.6 mL of 0%, 5%, 10%, or 20% of GABA. Hatchling weight and CWEWR were significantly increased in the 5% GABA group. In ovo, injection of 10% GABA solution caused a significant decrease in plasma cholesterol and increased plasma total antioxidant capacity of hatchlings. Experiment 2 was conducted with 126 fertile Arbor Acres eggs distributed into one of two groups. At 17.5 days of incubation, one received no injection, and the other was fed 0.6 mL of 10% GABA. On day 10, one subgroup (4 replicates * 3 birds) from each treatment was submitted to HS (38 ± 1 °C for 3 h) while the other was kept at a thermoneutral temperature (29 ± 1 °C). An in ovo injection of GABA significantly increased total antioxidant capacity, but reduced malondialdehyde levels, hepatic mRNA levels of HSP70, FAS, and L-FABP with HS. In conclusion, an in ovo GABA injection improves CWEWR and antioxidant status at hatch, and enhances antioxidant status while downregulating the expression of HSP70 and fatty acid metabolism-related genes in young chicks under HS.

Neuroprotective Effects of Asparagus officinalis Stem Extract in Transgenic Mice Overexpressing Amyloid Precursor Protein.

To mimic Alzheimer's disease, transgenic mice overexpressing the amyloid precursor protein (APP) were used in this study. We hypothesize that the neuroprotective effects of ETAS®50, a standardized extract of Asparagus officinalis stem produced by Amino Up Co., Ltd. (Sapporo, Japan), are linked to the inhibition of the apoptosis cascade through an enhancement of the stress-response proteins: heat shock proteins (HSPs). APP-overexpressing mice (double-transgenic APP and PS1 mouse strains with a 129s6 background), ages 6-8 weeks old, and weighing 20-24 grams were successfully bred in our laboratory. The animals were divided into 5 groups. APP-overexpressing mice and wild-type (WT) mice were pretreated with ETAS®50 powder (50% elemental ETAS and 50% destrin) at 200 mg/kg and 1000 mg/kg body weight. Saline, the vehicle for ETAS®50, was administered in APP-overexpressing mice and WT mice. ETAS®50 and saline were administered by gavage daily for 1 month. Cognitive assessments, using the Morris Water Maze, demonstrated that memory was recovered following ETAS®50 treatment as compared to nontreated APP mice. At euthanization, the brain was removed and HSPs, amyloid β, tau proteins, and caspase-3 were evaluated through immunofluorescence staining with the appropriate antibodies. Our data indicate that APP mice have cognitive impairment along with elevated amyloid β, tau proteins, and caspase-3. ETAS®50 restored cognitive function in these transgenic mice, increased both HSP70 and HSP27, and attenuated pathogenic level of amyloid β, tau proteins, and caspsase-3 leading to neuroprotection. Our results were confirmed with a significant increase in HSP70 gene expression in the hippocampus.

Evaluation of quality and gene expression of goat embryos produced in vivo and in vitro after cryopreservation

In the present study, we aimed to identify morphological and molecular changes of in vivo and in vitro-produced goat embryos submitted to cryopreservation. In vivo embryos were recovered by transcervical technique from superovulated goats, whereas in vitro produced embryos were produced from ovaries collected at a slaughterhouse. Embryos were frozen by two-steps slow freezing method, which is defined as freezing to −32 °C followed by transfer to liquid nitrogen. Morphological evaluation of embryos was carried out by assessing blastocoel re-expansion rate and the total number of blastomeres. The expression profile of candidate genes related to thermal and oxidative stress, apoptosis, epigenetic, and implantation control was measured using RT-qPCR based SYBR Green system. In silico analyses were performed to identify conserved genes in goat species and protein-protein interaction networks were created. In vivo-produced embryos showed greater blastocoel re-expansion and more blastomere cells (P < 0.05). The expression level of CTP2 and HSP90 genes from in vitro cryopreserved embryos was higher than their in vivo counterparts. Unlikely, no significant difference was observed in the transcription level of SOD gene between groups. The high similarity of CPT2 and HSP90 proteins to their orthologs among mammals indicates that they share conserved functions. In summary, cryopreservation negatively affects the morphology and viability of goat embryos produced in vitro and changes the CPT2 and HSP90 gene expression likely in response to the in vitro production process.

The effects of vitamin C and methionine hydroxy analog supplementation on performance, blood parameters, liver enzymes, thyroid hormones, antioxidant activity of blood plasma, intestine morphology, and HSP70 gene expression of broilers under heat stress

This experiment was performed to evaluate the effect of vitamin C and methionine hydroxy analog (MHA) on performance, blood parameters, liver enzymes, activity of selected hormones, antioxidant activity of blood plasma, intestine morphology, and HSP70 gene expression in broilers under heat stress. Four hundred 1-day-old male chickens of commercial Ross 308 strain were used in a completely randomized design with 4 treatments and 5 replicates of 20 chickens each. The experimental treatments were as follows: (1) base diet based on maize soybean (the control), (2) base diet with 0.2% vitamin C, (3) base diet with 0.46, 0.36, and, 0.32% methionine hydroxy analog, in the starter, grower, and finisher periods, respectively, and (4) base diet with a mixture of 0.2% vitamin C and 0.46, 0.36, and 0.32% of methionine hydroxy analog in the starter, grower, and finisher periods respectively. Although numerically higher than control, the effect of the experimental diets on performance was not significant across the experimental period (P>0.05). Glucose and LDL concentrations under all experimental treatments were significantly reduced compared to the control (P<0.05). The concentrations of liver enzymes (AST and ALT) and antioxidant activity (SOD, GPx, and MDA) of broilers were significantly different from the control under combined MHA and vitamin C supplementation (P<0.05). All experimental diets led to significantly increased T4 concentrations, compared to control (P<0.05). The length and width of intestinal villi of broilers were significantly increased in all of the experimental treatments compared to the control (P<0.05). The difference in HSP70 gene expression was significantly reduced in all of the treatments compared to the control (P<0.05). The results of this experiment showed that the use of vitamin C and methionine hydroxy analog (MHA) can be effective in reducing the negative effects of heat stress.

Molecular Characterization of Hsp47 in Grass Carp (Ctenopharyngodon idella) and Its Correlation with Type I Collagen in Response to Fish Aerobic Exercise

Heat shock protein 47 (Hsp47) is a collagen-specific molecular chaperone that is indispensable for molecular maturation of collagen. In this study, hsp47 and hsp47-like cDNAs were cloned and characterized in grass carp (Ctenopharyngodon idella). The cDNAs were 1212 and 1218 base pairs long, respectively, and included an open reading frame encoding 403 and 405 amino acids. The molecular phylogeny based on the deduced amino acid sequences indicated that the correct sequences of the hsp47 and hsp47-like cDNA were obtained and the deduced proteins clustered distinctly into teleost clades. Primary structure analysis and characterization of Hsp47 and Hsp47-like shared the basic structure and biofunctions of Hsp47 in vertebrates. The spatial pattern of gene expression revealed that hsp47 and hsp47-like were relatively ubiquitous in different tissues and highly expressed in heart and skin. The expression levels of hsp47 and hsp47-like and type I collagen mRNAs varied similarly in different tissues. Type I collagen content increased significantly with the increase of water velocity in the muscle of grass carp in response to aerobic exercise. Among the gene expression changes of hsp47, hsp47-like, col1a1 and col1a2 in muscle that occurred in response to aerobic exercise, the change of type I collagen was most strongly correlated with hsp47 expression. Additionally, col1a1 showed the highest correlation with hsp47-like and col1a2 showed the highest correlation with hsp47. These findings suggest that grass carp Hsp47 and Hsp47-like are closely related to type I collagen synthesis. This study firstly suggests fish aerobic exercise can improve type I collagen content and Hsp47 gene expression in muscle of grass carp.

Protective Effect of Emblica officinalis in Cyprinus carpio against Hepatotoxicity Induced by Malachite Green: Ultrastructural and Molecular Analysis

Malachite green (MG) dye, besides coloring is used as an effective aquaculture therapeutic. The present study assesses the mitigating ability of Emblica officinalis (EO) fruit extract against the dye induced chronic (60 days) cyto-toxicity in Cyprinus carpio. For this, four experimental groups were maintained: group I—control, group II—MG, group III—EO (positive control), group IV—MG + EO. The study was made at three tiers: detailing structural anomalies using a light microscope and transmission electron microscope (TEM), biochemical estimation of antioxidant enzymes, and lipid peroxidation and molecular analysis of expression patterns of HSP70, and CYP1A genes. MG intoxication resulted in necrosis, cytoplasmic vacuolation, glycogen depletion, abundant macrophages, loss of cell integrity and prominent nuclear alterations. Significant (p &lt; 0.05) inhibition in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) and reduced glutathione (GSH), along with an elevation in malondialdehyde (MDA) levels, occurred after 60 days of MG exposure. CYP1A and HSP70 genes presented a significant change in their expression in MG treated fish. Whereas oral supplementation with EO significantly restored the histo-architecture, normalized the altered enzymatic activity, reduced the oxidative stress level and regulated the expression of HSP70 and CYP1A genes. Thus, it can be concluded that EO acted as an effective ameliorant against malachite green induced cyto-toxicity in Cyprinus carpio.

Heat shock protein and gene regulation in goats during heat stress

Heat shock proteins (HSPs), also known as molecular chaperons are prominent stress markers. Heat shock proteins consist of highly conserved protein expressed at the time of stress, and play an important role in adaptation to the environmental stress. Although, the expression pattern of HSP70 gene is species and breed specific, variations in adaptation and thermal tolerance is due to the nature of environment and adaptive capacity of a species. The present study was conducted to evaluate the adaptive capability of different goat (Capra hircus) breeds, i.e. Jamunapari, Barbari, Jakhrana and Sirohi under peak dry summer. The targeted gene HSP70 (HSPA6) was evaluated for this purpose using specific primers. The expression of HSP70 gene and protein was estimated by RT PCR and ELISA kits respectively. The expression of HSP70 gene was found lowest in sirohi breeds implying that this breed was more adapted followed by Jakhrana, Barbari and Jamunapari during peak summer season. Whereas, the level of HSP70 protein in blood was significantly higher in Jamunapari, followed by Barbari, Jakhrana and lowest in Sirohi. These results indicated that, during adverse climatic stress the quantum of expression (HSP70 gene and protein) was more in Jamunapari. It is concluded that Sirohi breed is better adapted to heat stress than Jamunapari, Jakhrana and Barbari and HSP70 may be a potential molecular biomarker in the future for selection of climate resilient animals.

Simulated mobile communication frequencies (3.5 GHz) emitted by a signal generator affects the sleep of Drosophila melanogaster

With the rapid development of science and technology, 5G technology will be widely used, and biosafety concerns about the effects of 5G radiofrequency radiation on health have been raised. Drosophila melanogaster was selected as the model organism for our study, in which a 3.5 GHz radiofrequency radiation (RF-EMR) environment was simulated at intensities of 0.1 W/m2, 1 W/m2, and 10 W/m2. The activity of parent male and offspring (F1) male flies was measured using a Drosophila activity monitoring system under short-term and long-term 3.5 GHz RF-EMR exposure. Core genes associated with heat stress, the circadian clock and neurotransmitters were detected by QRT-PCR technology, and the contents of GABA and glutamate were detected by UPLC-MS. The results show that short-term RF-EMR exposure increased the activity level and reduced the sleep duration while long-term RF-EMR exposure reduced the activity level and increased the sleep duration of F1 male flies. Under long-term RF-EMR, the expression of heat stress response-related hsp22, hsp26 and hsp70 genes was increased, the expression of circadian clock-related per, cyc, clk, cry, and tim genes was altered, the content of GABA and glutamate was reduced, and the expression levels of synthesis, transport and receptor genes were altered. In conclusion, long-term RF-EMR exposure enhances the heat stress response of offspring flies and then affects the expression of circadian clock and neurotransmitter genes, which leads to decreased activity, prolonged sleep duration, and improved sleep quality.

Synthesis of Escherichia coli OmpA Oral Nanoparticles and Evaluation of Immune Functions against the Major Etiologic Agent of Cow Mastitis.

: BackgroundEscherichia coli is a major etiologic agent of cow mastitis, a condition that results in huge economic losses. There is a lack of an oral vaccine for cow mastitis. Previous studies have confirmed that the outer membrane protein A (OmpA) of E. coli is immunogenic and can be used for vaccine design. In the present study, OmpA was encapsulated into nanoparticles (NP-OmpA) for an oral vaccine for cow mastitis. Methods: OmpA was purified with Ni-NTA flow resin and encapsulated with chitosan (CS) to prepare NP-OmpA nanoparticles. The gastrointestinal tract was simulated in vitro (PBS, pH 1.2) to measure the protein release rate. The optimal preparation conditions for NP-OmpA were determined by analyzing the concentrations of OmpA and CS, magnetic mixing speed, mixing time, and the ratio of tripolyphosphate (TPP)/CS (w/w). NP-OmpA safety was assessed by function factors and histopathological examination of livers and kidneys. The immune activity of NP-OmpA was determined using qRT-PCR to assess immune-related gene expression, leukocyte phagocytosis of Staphylococcus aureus, ELISA to evaluate antiserum titer and immune recognition of E. coli, and the organ index. The immune protection function of NP-OmpA was assessed by the protection rate of NP-OmpA to E. coli in mice, qRT-PCR for inflammation-related gene expression, assay kits for antioxidant factors, and visceral injury in the histopathological sections. Results: NP-OmpA nanoparticles had a diameter of about 700 nm, loading efficiency (LE) of 79.27%, and loading capacity (LC) of 20.31%. The release rate of NP-OmpA (0~96 h) was less than 50% in vitro. The optimal preparation conditions for NP-OmpAs were OmpA protein concentration of 2 mg/mL, CS concentration of 5 mg/mL, TPP/CS (w/w) of 1:1, magnetic mixing speed of 150 r/min, and mixing time of 15 min. Histopathological sections and clinical analytes of uric acid (UA), creatinine (Cr), alanine aminotransferase (ALT), aspartate transaminase (AST), catalase (CAT), glutathione (GSH), and malondialdehyde (MDA) showed NP-OmpA did not damage mice livers or kidneys. NP-OmpA could enhance the immune-related gene expression of IFN-γ and HSP70 in the spleen, liver, and kidney and the leukocyte phagocytosis of S. aureus. The antiserum titer (1:3200) was obtained from mice immunized with NP-OmpA, which had an immune recognition effect to E. coli. The immune protection rate of NP-OmpA was 71.43% (p < 0.05) to E. coli. NP-OmpA could down-regulate the inflammation-related gene expression of TNF-a, IL-6, and IL-10 in the spleen, liver, and kidney, and the antioxidant factors MDA and SOD in the liver, and reduce injury in the liver and kidney of mice induced by E. coli. Conclusions: A novel NP-OmpA nanoparticle was encapsulated, and the optimal preparation conditions were determined. The NP-OmpA was safe and had good immune functions. They are expected to induce a response that resists infection with the major etiologic agent (E. coli) of cow mastitis.

Effects of dietary chromium yeast and astaxanthin on the growth performance, anti-oxidative capacity, and resistance to heat stress of abalone Haliotis discus hannai

This study was aimed at evaluating whether dietary chromium yeast (CrYst) and astaxanthin (ASTA) could enhance the growth performance, anti-oxidative capacity, and resistance to heat stress of abalone Haliotis discus hannai (initial body weight: 1.14±0.10 g; initial shell length: 19.20±0.87 mm) after a 120-day feeding trial. Based on the basal diet, the other three experimental diets were designed with supplementation of 2 mg/kg of CrYst, 80 mg/kg of ASTA, and 2 mg/kg of CrYst + 80 mg/kg of ASTA, respectively. The results showed that supplementation of 2 mg/kg of dietary CrYst significantly increased weight gain rate and the daily increment in shell length of abalone (P 0.05). Meanwhile, there were no significant differences in feed intake and feed conversion ratio among all the groups (P > 0.05). Meanwhile, adding 2 mg/kg of CrYst and/or 80 mg/kg of ASTA into the diet improved the anti-oxidative capacity in serum and hepatopancreas, increased the gene expression of hsp70 and hsp90 in the hepatopancreas, and reduced the cumulative mortality and falling rate of abalone during the heat stress. In conclusion, dietary supplementation of CrYst and/or ASTA had positive effects on growth and anti-stress of abalone.

Hazardous Effects of SiO2 Nanoparticles on Liver and Kidney Functions, Histopathology Characteristics, and Transcriptomic Responses in Nile Tilapia (Oreochromis niloticus) Juveniles.

Simple SummaryWaterborne exposure of Nile tilapia (Oreochromis niloticus) juveniles to sub-lethal concentrations of silicon dioxide nanoparticles (SiO2NPs) induced hepato-renal damage through elevation of aspartate transaminase (AST), alanine transaminase (ALT), and alkaline phosphatase (ALP) activities as well as creatinine and blood urea levels. SiO2NPs induced irreversible dose-dependent histopathological changes in the hepatopancreas, gills, and posterior kidneys, alongside modulation of the pro-inflammatory cytokines, apoptosis-related genes, and oxidative stress genes in gills and liver of exposed fish.The current investigation assessed the impacts of sub-lethal concentrations of silicon dioxide nanoparticles (SiO2NPs) on hepato-renal functions, histopathological characteristics, and gene transcription in gills and liver of Nile tilapia juveniles. Fish were exposed to 20, 40, and 100 mg/L of SiO2NPs for 3 weeks. Pairwise comparisons with the control group showed a significant dose-dependent elevation in serum ALP, ALT, and AST enzyme activities as well as blood urea and creatinine levels in SiO2NP-intoxicated groups. Exposure to 100 mg/L SiO2NPs significantly upregulated expression of HSP70, TNF-α, IL-1β, and IL-8 genes in the gills as compared to the control group. Moreover, exposure to 100 mg/L SiO2NPs significantly upregulated the expression SOD, HSP70, IL-1β, IL-8, and TNF-α genes in the hepatic tissues as compared to the control group. Exposure of fish to 20 mg SiO2NPs/L significantly increased the mRNA expression levels of IL-12 in both the gills and liver tissues. Notably, all tested SiO2NP concentrations significantly upregulated the transcription of CASP3 gene in gills and liver of Nile tilapia as compared to the control group. Interestingly, varying histopathological alterations in renal, hepatopancreatic, and branchial tissues were observed to be correlated to the tested SiO2NP concentrations. In conclusion, our results provide additional information on the toxic impacts of SiO2NPs in Nile tilapia at the hematological, tissue, and molecular levels.

The effects of astragalus polysaccharides on the growth, heat stress tolerance and related gene expression of the leech Whitmania pigra

Whitmania pigra is an aquatic annelid with broad pharmacological effects and is widely used for treatment of cardiovascular diseases in Asia. One of the main factors limiting its commercial production is its weak heat stress tolerance. To assess the effects of astragalus polysaccharide (APS) on the growth and heat stress tolerance of W. pigra and the underlying mechanisms, six APS concentration treatments (0‰, 0.01‰, 0.03‰, 0.05‰, 0.07‰ and 0.09‰) were formulated to explore their influence on growth, digestive enzymes and the expression of immune- and growth-related genes at 26°C or 35°C. The results showed that APS significantly enhanced the final weight, weight gain rate, specific growth rate, digestive enzyme and anti-reverse enzyme activity, and the relative expression level of growth hormone, insulin-like growth factor-1 and digestive enzyme-related genes. The 0.07‰ APS group showed significantly greater values than the other groups (p < 0.05). APS up-regulated the relative expression of HSP70 and immune-related genes under heat stress, and the 0.07‰ group showed the greatest modulatory effect. Overall, APS can promote growth, immunity and heat stress resistance in W. pigra, and the recommendation dietary APS level was 0.07‰.

Nanogold induces anti-inflammation against oxidative stress induced in human neural stem cells exposed to amyloid-beta peptide

Alzheimer's disease (AD) is a neurodegenerative disorder with progressive memory loss resulting in dementia. Amyloid-beta (Aβ) peptides play a critical role in the pathogenesis of the disease by promoting inflammation and oxidative stress, leading to neurodegeneration in the brains of AD patients. Numerous in vitro 3D cell culture models are useful mimics for understanding cellular changes that occur during AD under in vivo conditions. The 3D Bioprinter developed at the CELLINK INKREDIBLE was used in this study to directly investigate the influence of 3D conditions on human neural stem cells (hNSCs) exposed to Aβ. The development of anti-AD drugs is usually difficult, mainly due to a lack of therapeutic efficacy and enhanced serious side effects. Gold nanoparticles (AuNPs) demonstrate benefits in the treatment of several diseases, including AD, and may provide a novel therapeutic approach for AD patients. However, the neuroprotective mechanisms by which AuNPs exert these beneficial effects in hNSCs treated with Aβ are still not well understood. Therefore, we tested the hypothesis that AuNPs protect against Aβ-induced inflammation and oxidative stress in hNSCs under 3D conditions. Here, we showed that AuNPs improved the viability of hNSCs exposed to Aβ, which was correlated with the reduction in the expression of inflammatory cytokines, such as TNF-α and IL-1β. In addition, AuNPs rescued the levels of the transcripts of inhibitory kappa B kinase (IKK) in Aβ-treated hNSCs. The Aβ-mediated increases in mRNA, protein, and nuclear translocation levels of NF-κB (p65), a key transcription factor involved in inflammatory responses, were all significantly abrogated following co-treatment of hNSCs with AuNPs. In addition, treatment with AuNPs significantly restored iNOS and COX-2 levels in Aβ-treated hNSCs. Importantly, hNSCs co-treated with AuNPs were significantly protected from Aβ-induced oxidative stress, as detected using the DCFH-DA and DHE staining assays. Furthermore, hNSCs co-treated with AuNPs were significantly protected from the Aβ-induced reduction in the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and Nrf2 downstream antioxidant target genes (SOD-1, SOD-2, Gpx1, GSH, Catalase, and HO-1). Moreover, AuNPs reduced the aggregates and increased the proteasome activity and the expression of HSP27 and HSP70 genes in Aβ-treated hNSCs. Taken together, these findings provide the first evidence extending our understanding of the molecular mechanisms under 3D scaffold conditions by which AuNPs reverse the inflammation and oxidative stress-induced in hNSCs exposed to Aβ. These findings may facilitate the development of novel treatments for AD.