Hsp70 Content Research Articles

Comparing Heat Shock Proteins in Angus and Brahman Cattle and their Effect On Tenderness

ObjectivesHeat shock proteins (HSP) are biomarkers of stress and perform chaperoning functions to fold, unfold, and refold proteins after heat stress. Brahman are more heat tolerant than Angus, while Angus beef has been associated with greater palatability than Brahman. The objectives were to determine if HSP content in the longissimus lumborum differs between Angus and Brahman and examine how HSP content relates to the eating quality of beef.Materials and MethodsAngus and Brahman steers (n = 12 per breed) were finished during summer in Florida and harvested at approximately 17 mo of age. Samples of longissimus lumborum were collected at 1 h after exsanguination and were immediately immersed in liquid nitrogen. Samples were then pulverized, diluted in extraction buffer, and homogenized. The protein samples were assayed to assess protein concentration and subsequently diluted to equal concentrations for loading into acrylamide gels. Proteins were separated by gel electrophoresis, and western blotting was used to evaluate content of aβ-crystallin, HSP27, HSP60, HSP70, and HSP90. Target bands were detected and quantified using LI-COR Odyssey and target signal was normalized to total protein stain. Tenderness was evaluated in 14d-aged steaks using Warner-Bratzler shear force (WBSF) and a trained sensory panel. Data were analyzed using one-way ANOVA and Pearson correlations were conducted for content of HSPs and objective and subjective tenderness.ResultsHSP27, HSP60, and HSP70 did not differ between breeds (P > 0.05); however, HSP90 and aβ-crystallin were greater (P = 0.005) in the longissimus lumborum of Angus compared to Brahman. Even though WBSF did not differ (P = 0.29) between breeds, breed affected (P < 0.0001) sensory tenderness. Content of aβ-crystallin was associated with sensory tenderness (r2 = 0.52, P = 0.0098).ConclusionLongissimus dorsi from the Angus were contained more aβ-crystallin and HSP90 than Brahman. Elevated concentrations of both aβ-crystallin and HSP90 could be breed related or may have been influenced by the season they were harvested. While WBSF was not affected by breed, panelists rated Angus steaks as more tender after aging for 14d. Content of aβ-crystallin is associated with tenderness; however further work is necessary to determine if this small HSP affects proteolysis.

Insect Hsp90 Chaperone Assists Bacillus thuringiensis Cry Toxicity by Enhancing Protoxin Binding to the Receptor and by Protecting Protoxin from Gut Protease Degradation.

Bacillus thuringiensis Cry proteins are pore-forming insecticidal toxins with specificity against different crop pests and insect vectors of human diseases. Previous work suggested that the insect host Hsp90 chaperone could be involved in Cry toxin action. Here, we show that the interaction of Cry toxins with insect Hsp90 constitutes a positive loop to enhance the performance of these toxins. Plutella xylostella Hsp90 (PxHsp90) greatly enhanced Cry1Ab or Cry1Ac toxicity when fed together to P. xylostella larvae and also in the less susceptible Spodoptera frugiperda larvae. PxHsp90 bound Cry1Ab and Cry1Ac protoxins in an ATP- and chaperone activity-dependent interaction. The chaperone Hsp90 participates in the correct folding of proteins and may suppress mutations of some client proteins, and we show here that PxHsp90 recovered the toxicity of the Cry1AbG439D protoxin affected in receptor binding, in contrast to the Cry1AbR99E or Cry1AbE129K mutant, affected in oligomerization or membrane insertion, respectively, which showed a slight toxicity improvement. Specifically, PxHsp90 enhanced the binding of Cry1AbG439D protoxin to the cadherin receptor. Furthermore, PxHsp90 protected Cry1A protoxins from degradation by insect midgut proteases. Our data show that PxHsp90 assists Cry1A proteins by enhancing their binding to the receptor and by protecting Cry protoxin from gut protease degradation. Finally, we show that the insect cochaperone protein PxHsp70 also increases the toxicity of Cry1Ac in P. xylostella larvae, in contrast to a bacterial GroEL chaperone, which had a marginal effect, indicating that the use of insect chaperones along with Cry toxins could have important biotechnological applications for the improvement of Cry insecticidal activity, resulting in effective control of insect pests.IMPORTANCEBacillus thuringiensis took advantage of important insect cellular proteins, such as chaperones, involved in maintaining protein homeostasis, to enhance its insecticidal activity. This constitutes a positive loop where the concentrations of Hsp90 and Hsp70 in the gut lumen are likely to increase as midgut cells burst due to Cry1A pore formation action. Hsp90 protects Cry1A protoxin from degradation and enhances receptor binding, resulting in increased toxicity. The effect of insect chaperones on Cry toxicity could have important biotechnological applications to enhance the toxicity of Cry proteins to insect pests, especially those that show low susceptibility to these toxins.

Heat shock response to exercise in pancreatic islets of obese mice

Chronic obesity imposes an organismal state of low-grade inflammation because the physiological resolution of inflammation is progressively repressed giving rise to cellular senescence and its accompanying Senescence-Associated Secretory Phenotype (SASP), which avoids apoptosis but perpetuates the relay of inflammatory signals from adipose tissue toward the rest of the body. Conversely, resolution of inflammation depends on the integrity of heat shock response (HSR) pathway that leads to the expression of cytoprotective and anti-inflammatory protein chaperones of the 70 kDa family (HSP70). However, chronic exposure to the aforementioned injuring factors leads to SASP, which, in turn, suppresses the HSR. A main metabolic tissue severely jeopardized by obesity-related dysfunctions is the endocrine pancreas, particularly β-cells of the islets of Langerhans. Because exercise is a powerful inducer of HSR and predicted to alleviate negative health outcomes of obesity, we sought whether obesity influence HSP70 expression in pancreatic islets and other metabolic tissues (adipose tissue and skeletal muscle) of adult B6.129SF2/J mice fed on a high-fat diet (HFD) for 13 weeks since the weaning and whether acute exercise as well as moderate-intensity exercise training (8 weeks) could interfere with this scenario. We showed that acute exercise of moderate intensity protects pancreatic islets against cytokine-induced cell death. In addition, acute exercise challenge time-dependently increased islet HSP70 that peaked at 12 h post-exercise in both trained and untrained mice fed on a control diet, suggesting an adequate HSR to exercise training. Unexpectedly, however, neither exercise training nor acute exercise challenges were able to increase islet HSP70 contents in trained mice submitted to HFD, but only in untrained HFD animals. In parallel, HFD disrupted glycemic status which is accompanied by loss of muscular mass resembling sarcopenic obesity that could not be rescued by exercise training. These results suggest that exercise influences HSR in pancreatic islets but obesity undermines islet, muscle and adipose tissue HSR, which is associated with metabolic abnormalities observed in such tissues.

Nilotinib ameliorates folic acid-induced acute kidney injury through modulation of TWEAK and HSP-70 pathways

PurposeAcute Kidney Injury (AKI) is a global health concern associated with high morbidity and mortality. Experimental folic acid-induced AKI is comparable to human AKI and is repeatedly reported to study the pathogenic pathway of human AKI and study the therapeutic efficacy of different agents against AKI. In the present study, nilotinib has been investigated for its possible ameliorative potential against folic acid-induced AKI. MethodsAKI was induced by IP injection of single dose (250 mg/kg) folic acid in mice. Nilotinib (15 mg/kg and 25 mg/kg) was administered orally by different treatment regimens (pre, post, and pre + post). ResultsFolic acid injection induced marked AKI in mice. Nilotinib (25 mg/kg) successfully alleviated folic acid-induced AKI. Nilotinib significantly decreased folic acid-induced elevation in serum Cr, BUN, LDH and urine MTP, kidney MDA content and significantly increased folic acid-induced reduction in serum albumin, CrC, urine urea, kidney SOD activity and GSH content. Kidney TWEAK, IL-18, IL-1β, TNF-α, NF-κB, Caspase 3 and Bax contents significantly decreased upon nilotinib administration while, kidney contents of Bcl2 and HSP-70 significantly increased. ConclusionAnti-inflammatory, anti-apoptotic and anti-oxidant potentials are the main mechanisms by which nilotinib can ameliorate folic acid-induced AKI. Modulation of TWEAK and HSP 70 pathways are thought to be the main contributor to the observed ameliorative potential.

First detection of heat shock protein 60 and 70 in the serum of early pregnant bitches.

Heat shock proteins (HSPs) belong to a group of cellular stress proteins. Heat shock protein 10 immunoregulates and promotes growth during early gestation in humans, while HSP70 is considered to regulate autophagy and apoptosis during pregnancy and parturition. Both HSPs are detectable in the serum and placentas of early pregnant women and considered to contribute to the establishment of pregnancy. Within this pilot study we aimed (1) to assess whether HSPs 10, 60 and 70 are measurable in the serum of healthy early pregnant and non-pregnant bitches, and (2) to explore whether measurable differences between groups indicate pregnancy. Blood was collected from 31 bitches on days 7, 14 and 21 after mating. At 21 days post mating, all bitches were examined for pregnancy by ultrasonography; 23 were pregnant, and the eight non-pregnant bitches served as controls. Pregnant bitches had normal parturitions and gave birth to healthy puppies. The serum concentrations of HSPs 10, 60 and 70 were measured by electrophoresis and western blot. Serum HSP10 was not detectable. Average serum HSP70 concentration was significantly (d7, P = 0.030; d14, P = 0.023; d21, P = 0.030) lower in pregnant animals at all days investigated, while serum HSP60 was significantly lower at day 21 of gestation (P = 0.024) when compared to the controls. HSP 60 and HSP70 concentrations correlated positively (d7, r = +0.386, P = 0.021; d14, r = 0.450, P = 0.008; d21, r = +0.472, P = 0.006). We conclude that in pregnant bitches, serum concentrations of HSP60 and HSP70 are significantly decreased between days 7 and 21 of gestation, in comparison to non-pregnant bitches in early dioestrus, raising the question about intrauterine functions during the peri-implantation period.

HSP70 improves the viability of cryopreserved Paeonia lactiflora pollen by regulating oxidative stress and apoptosis-like programmed cell death events

Heat shock proteins (HSPs) have many positive stabilizing effects on the proteins of stressed cells. This study investigated the effect of HSP70 on cryopreserved pollen. HSP70 was differentially expressed in Paeonia lactiflora and Magnolia denudate pollen before and after exposure to liquid nitrogen (LN). In this study, cryopreserved pollen samples of P. lactiflora ‘Fen Yu Nu’, ‘Zi Feng Chao Yang’ and ‘Hong Pan Tuo Jing’ were incubated with HSP70 at six concentrations, and the pollen germination, oxidative stress and apoptosis-like programmed cell death indicators were measured after LN storage. HSP70 significantly improved pollen viability after LN storage, and the concentration of HSP70 that was required varied from cultivar to cultivar, ranging from 0.5 to 10 μg/mL. Compared to the activity of the control treatment without HSP70, the activity of superoxide dismutase (SOD) increased, the contents of reactive oxygen species (ROS) and malondialdehyde (MDA) decreased, and the Ca2+ level, apoptosis rate and caspase-3-like activity decreased depending on the appropriate concentration of HSP70 added. Pollen germination was negatively correlated with MDA content, Ca2+ level, and caspase-3-like activity and positively correlated with SOD activity. In conclusion, HSP70 improved the viability of pollen after cryopreservation by improving the activity of antioxidant enzymes, reducing the contents of ROS and MDA, and affecting the Ca2+ signal to inhibit apoptosis-like programmed cell death events. This report is the first time to describe the application of HSP70 for improving cryopreserved pollen viability by regulating oxidative and apoptosis-like programmed cell death events, and provide a novel insight into the mechanisms of HSP70 action in cryopreservation. After the pollen was rewarmed and then incubated with appropriate concentrations of exogenous HSP70, it significantly increasing the post-freezing germination of the pollen. Which by affecting oxidative stress and apoptosis-like events, the ROS level, MDA content, intracellular Ca2+ concentration, apoptosis rate and caspase-3-like enzyme activity were reduced, and SOD activity was increased, thereby significantly increasing the post-freezing germination of the pollen.

7Hsp70 serum levels in pet dogs\u2014a potential diagnostic biomarker for spontaneous round cell tumors

The concentration of circulating heat shock protein 70 (Hsp70) was measured in liquid biopsies of canine tumor patients as a potential biomarker. Compared with rodent tumor models, spontaneously occurring tumors in pet dogs reflect the clinical situation of human patients better, as dogs cohabitate with their owners in the same environment, reach a much older age than rodents, can provide blood samples much more frequently, and receive up-to-date medical care and, similar to humans, their tumors show a high genetic heterogeneity. Due to the species-specific sequence homology of human and canine Hsp70, two human enzyme-linked immunosorbent assay (ELISA) systems (R&D and lipHsp70) were used to measure canine Hsp70 concentrations in serum and plasma. In general, higher Hsp70 concentrations were found in serum compared with plasma samples of dogs, and the lipHsp70 ELISA detected higher peak concentrations of Hsp70 in a broader range than the R&D ELISA. Compared with a tumor-free control group, serum Hsp70 concentrations were higher in tumor-bearing dogs, irrespective of breed, age, body weight, and gender. A sub-classification of the different tumors according to their cytological characteristics revealed significantly elevated Hsp70 serum concentrations in dogs with round cell tumors (p < 0.01), a heterogeneous group of malignancies with hematopoietic origin such as mast cells, plasma cells, lymphocytes, histiocytes, and melanomas. Future studies with larger patient cohorts and well-defined tumor sizes are necessary to elucidate the role of serum Hsp70 as a biomarker for tumor detection and monitoring of outcome in pet animals.

Histamine Is an Inducer of the Heat Shock Response in SOD1-G93A Models of ALS.

(1) Background: Amyotrophic lateral sclerosis (ALS) is a multifactorial non-cell autonomous disease where activation of microglia and astrocytes largely contributes to motor neurons death. Heat shock proteins have been demonstrated to promote neuronal survival and exert a strong anti-inflammatory action in glia. Having previously shown that the pharmacological increase of the histamine content in the central nervous system (CNS) of SOD1-G93A mice decreases neuroinflammation, reduces motor neuron death, and increases mice life span, here we examined whether this effect could be mediated by an enhancement of the heat shock response. (2) Methods: Heat shock protein expression was analyzed in vitro and in vivo. Histamine was provided to primary microglia and NSC-34 motor neurons expressing the SOD1-G93A mutation. The brain permeable histamine precursor histidine was chronically administered to symptomatic SOD1-G93A mice. Spine density was measured by Golgi-staining in motor cortex of histidine-treated SOD1-G93A mice. (3) Results: We demonstrate that histamine activates the heat shock response in cultured SOD1-G93A microglia and motor neurons. In SOD1-G93A mice, histidine augments the protein content of GRP78 and Hsp70 in spinal cord and cortex, where the treatment also rescues type I motor neuron dendritic spine loss. (4) Conclusion: Besides the established histaminergic neuroprotective and anti-inflammatory effects, the induction of the heat shock response in the SOD1-G93A model by histamine confirms the importance of this pathway in the search for successful therapeutic solutions to treat ALS.

Protective role of zinc in Spodoptera exigua larvae under 135-generational cadmium exposure

The aim of this study was to investigate whether zinc supplementation modulates cadmium toxicity in the beet armyworm Spodoptera exigua selected for 135 generations towards cadmium tolerance. To achieve this, larvae originating from three laboratory populations of S. exigua (control strain - C; cadmium-intoxicated for 135 generations strain - Cd, and control strain intoxicated with Cd for 1 generation - CCd) were additionally exposed to zinc in three concentrations (Zn1, 400 μg Zn·g−1 dry mass of food; Zn2; 200 μg Zn·g−1 dry mass of food; Zn3, 100 μg Zn·g−1 dry mass of food). As the markers of toxicity, a life history traits (the duration of L4 and L5 stages), cellular (DNA damage indices) and biochemical parameters (ADP/ATP ratio and ATP and HSP70 concentrations) were chosen. The duration of larval stages of Zn supplemented larvae was prolonged, while cellular and biochemical indicators, in general, appeared to be lower in comparison to the insects from respective reference groups in each laboratory populations. Moreover, the range of the differences depended on zinc concentration in food. We can suspect that zinc supplementation contributed to the protection of S. exigua individuals against negative effects of cadmium intoxication, probably at the cost of growth rate. Significant differences in the response pattern between insects from different laboratory populations indicate that the influence of additional stress factors is dependent on the overall condition of animals and their previous adaptation to other stressors.

Impact of gut microbiota structure in heat-stressed broilers

Gut microbiota** play important roles in the health and disease status of both humans and animals. Little is known about whether heat stress changes the composition of the gut microbiota in chicken. The aim of this study was to investigate the effects of heat stress on changes in caecal microbiota, including changes in growth performance as well as HSP70 and cortisol levels. Sixty 14-day-old female broilers were equally divided into 2 treatment groups with different housing temperatures for 28 D: a control group (C) at 24 to 26°C and a heat stress (HS) group at 34 to 38°C. The caecal contents of the broiler chicken were then extracted on days 1, 3, 7, 14, and 28. Genomic DNA was extracted and amplified based on the V3∼V4 hypervariable region of 16S rRNA high-throughput sequence analyses. The results showed that the average daily gain and average daily feed intake were significantly decreased and that the feed conversion ratio was increased by heat stress. The concentrations of HSP70 and cortisol in the serum were significantly increased. The composition of gut microbiota was influenced by heat stress** through beta diversity analysis and taxon-based analysis. In particular, at the phylum level the composition of Firmicutes, Tenericutes, and Proteobacteria in HS group was increased than that of C group, and Bacteroidetes and Cyanobacteria in HS group were reduced than that of C group. In addition, the composition of Anaeroplasma and Lactobacillus phyla in HS group were increased than that of C group, whereas the Bacteroides, Oscillospira, Faecalibacterium, and Dorea genera in HS group were decreased than that of C group. In conclusion, the gut microbiota in broilers were changed by heat stress. And the changes of the gut microbiota could provide the basis for further research on the heat stress.

HSP72 expression is specific to skeletal muscle contraction type.

Exercise is capable of inducing the cellular stress response and increasing skeletal muscle heat shock protein (HSP) content. HSPs function as molecular chaperones and play roles in facilitating protein folding thereby contributing to muscle proteostasis. To determine the relationship between muscle contraction types, muscle damage, and HSP content, one tibialis anterior (TA) muscle from male Sprague-Dawley rats (n = 5/group) was electrically stimulated while actively lengthening (LC), shortening (SC), or remaining to stagnate (IC) for 15 repetitions (3 sets of five). Two additional LC groups underwent 5 and 10 repetitions. Maximal tetanic tension (MTT) was recorded prior to (pre) and at 5min after (post) the last contraction. Twenty-four hours after stimulation, TA muscles were removed, processed, and assessed for damage and for HSP25 and HSP72 content. Post-MTT was significantly decreased following 15 LCs, (24%; p < 0.05) but not following 15 SCs or 15 ICs. Post-MTT was also decreased by 8% (p < 0.05), and 18% (p < 0.05) for muscles subjected to 5 and 10 LCs, respectively. HSP72 content increased after all LCs conditions but not following ICs or SCs. HSP25 content remained unchanged following all contractions. Similarly, muscle damage was observed only after LCs and not after other contraction types. In conclusion, muscle HSP72 content can be increased with as few as 5 maximal lengthening contractions and appears to be related to muscle damage. This may have important implications for muscle rehabilitation and exercise training programs.

Microevolution or wide tolerance? Level of stress proteins in the beet armyworm Spodoptera eqigua hübner (Lepidoptera: Noctuidae) exposed to cadmium for over 150 generations

The aim of this study was to investigate whether the cadmium tolerance developed in the beet armyworm Spodoptera exigua selected for over 150 generations may be related to synthesis of the stress proteins metallothioneins (Mts) and 70 kDa heat shock proteins (HSP70). To achieve this, six S. exigua strains (control, k), 150-generation Cd exposure strain (cd), and four 18-generation Cd exposure strains differing in Cd concentration (cd44, cd22, cd11, cd5) were reared. Stress protein level was measured in the midgut of the 5th larval stage after 1–6, 12 and 18 generations. Cd contents was measured in the pupae. Unlike Cd concentration, which depended on metal contents in food but was not generation-dependent, the pattern of Mts and HSP70 concentrations changed in experimental strains from generation to generation. Stress protein levels in the insects exposed to the highest Cd concentration (the same as in the 150-generation Cd exposure strain), initially higher than in the control strain, after the 12th generation did not differ from the level measured in the control strains. It seems therefore that stress proteins play a protective role in insects of lower tolerance to cadmium. The tolerance developed during multigenerational exposure probably relies on mechanisms other than Mt and HSP70 synthesis.

Exosomal Signaling by Skeletal Muscle: Role in Neuromuscular Ageing

Muscle mass and function are reduced with age in mammals and this is associated with disrupted neuromuscular interactions. Skeletal muscle mounts a robust stress response to contractions by the increased expression of Heat Shock Proteins (HSPs), which facilitate muscle remodelling, promote protein folding and clearance of damaged proteins. In contrast, other tissues, including neuronal cells are unable to mount a stress response. It has been proposed that HSPs may be transferred from one cell type to another in exosomes to maintain proteostasis in the recipient cells. Further, HSP generation by muscle following contractions is attenuated in old mice and humans. We hypothesise that this inability of muscles of old mice to produce HSPs in response to contractions results in altered exosomal HSP transfer, and therefore a failure to maintain proteostasis in neighbouring cells including motor neurons, with subsequent neuronal and muscle degeneration (1).Single muscle fibres were isolated from flexor digitorum bravis (FDB) muscles of adult (6–12 month) mice, and initiated to contract using a non‐damaging electrical stimulation protocol. Media was collected at time points up to 24hrs, and exosomes were purified from the media using ultracentrifugation approaches. NanoSight analysis was used to characterize and quantify the exosomes, and HSP content analysed by western blotting.Data demonstrated that there was a significant (2–3 fold) increase in exosome release by muscle fibres of adult mice following contraction compared with non‐stimulated controls, and that the exosomal HSP content was also increased. For example, at 4hrs post contraction, there was no increase in HSP25 content of exosomes but a 2 fold increase in HSP60 was detected at this time point compared with non‐stimulated controls. Preliminary data also showed that there was a 3.5 fold increase in HSP25 exosomal content at 24hrs post stimulation.Additional data demonstrated the potential uptake of exosomes labelled with lipophilic carbocyanine fluorescent dye by immortalized neuronal cells. Additional studies will also examine the effect of age on exosome production by muscle fibres, including HSP content and neuronal uptake, by comparison with muscle from CuZn superoxide dismutase (CuZnSOD) null mice, a model of accelerated muscle ageing.Support or Funding InformationWith thanks to National Institute of Ageing (AG051442), the Physiological Society (summer studentship awarded to RR) and the University of Liverpool for their generous support.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Effect of Repeated Whole-Body Cryostimulation on the HSP-70 and Lipid Metabolisms in Healthy Subjects

The aim of this study was to evaluate the effects of exposure to 30 daily whole body cryostimulation (WBC) on lipid metabolic parameters and serum HSP-70 concentration. The study involved 45 volunteers, homogeneous in terms of diet and daily physical activity. Blood samples were collected before and after the 10(th), the 20(th), and the 30(th) session and one month after the intervention. Total cholesterol, HDL, TG concentrations and Apolipoprotein A-I, ApoB and HSP-70 protein levels were determined in serum. Additionally, the LI (Lipid Index) and the LDL level were calculated. During exposure, positive changes in the lipid profile that included a decrease in the TCh, initiated after the 20th WBC session with a simultaneous decrease in TG and LDL levels, and an increase in the HDL concentration were observed. These changes were accompanied by a downward trend in the ApoB concentration and a decrease in the ApoB:ApoA-I ratio after 30 sessions. The nature of these changes persisted for a month after the exposure. The obtained results indicate metabolic benefits that result from prolonged exposure to cryogenic temperatures, confirming the postulate of using WBC in the regulation of lipid metabolism and the prevention of cardiovascular diseases.

Immunological Response and Match Performance of Professional Tennis Players of Different Age Groups During a Competitive Season.

Kozłowska, M, Żurek, P, Rodziewicz, E, Góral, K, Żmijewski, P, Lipińska, P, Laskowski, R, Walentukiewicz, AK, Antosiewicz, J, and Ziemann, E. Immunological response and match performance of professional tennis players of different age groups during a competitive season. J Strength Cond Res 35(8): 2255-2262, 2021-We aimed to investigate the effect of physical workloads on immunological response, match performance, and iron metabolism in professional tennis players of different age groups throughout the tournament season and to determine the interdependence of vitamin D status and inflammation. Thirty-eight young, male tennis players with a top national ranking (1-25) participated in this study and were assigned to the following age groups: cadets (CG), juniors (JG), and seniors (SG). Blood samples were collected at the beginning, midpoint, and end of the tournament season to assess the proinflammatory cytokine (tumor necrosis factor-alpha [TNF-α]), anti-inflammatory myokines (interleukin [IL]-6 and IL-10), heat shock proteins (HSP70, HSP27), iron metabolism markers, and vitamin D concentrations. The total number of matches (won and lost) at the national and international events was recorded. The IL-6 and IL-10 concentrations significantly increased across all groups in the middle and end of the tournament season (effect large and very likely). The TNF-α concentration was elevated at the end of the season in CG and SG. The increase in TNF-α concentration corresponded with an increase in hepcidin concentration in these groups. The significant increase in HSP27 concentration was only noticed in SG with normal vitamin D concentrations. In JG and SG, a mild seasonal increase in vitamin D concentration was noted, but still it was insufficient. The immunological response was not affected by the number of tennis matches; however, the anti-inflammatory effect was regulated by higher concentrations of vitamin D. Unexpectedly, most tennis players had vitamin D deficiency. Iron status remained unchanged.

Effects of saccharomyces cerevisiae supplementation on milk production, insulin sensitivity and immune response in transition dairy cows during hot season

The objectives were to determine effects of feeding a live yeast supplement on the productive performance, blood metabolic profile, the immune function, and insulin sensitivity traits of transition dairy cows under the hot months of summer. From 21 d before expected date of calving until d 60 postpartum, two groups of multiparous Holstein cows (6 cows per treatment) were fed a diet without or with live yeast supplement (4 g yeast/d/head). Blood was collected on d −14, 0, 14, 28, and 60 d relative to parturition and analyzed for nonesterified fatty acids, β-hydroxybutyrate, glutamic pyruvic transaminase, glutamic oxaloacetic transaminase, creatinine, urea-N, triiodothyronine, and thyroxine. Plasma concentrations of histamine and heat shock protein 70 (Hsp70) were also determined on d –21, 14, 28, and 49 relative to parturition. An intravenous glucose challenge was made on d 14 postpartum. The cell-mediated immune and humoral responses were established through phytohemagglutinin challenge and ovalbumin immunization, respectively. Prepartum DM intake was greater in yeast-fed cows; however, this difference disappeared after parturition. Cows receiving yeast supplement produced more milk (+1.40 kg/d) and had greater concentrations of milk fat and total solid than those receiving no yeast. Loss of body condition score from calving to d 21 postpartum tended to be lower in yeast-fed cows than control cows. Yeast supplementation had no effect on the response variables to the glucose tolerance test. Plasma concentration of Hsp70 was also lower on d 14 and 28 after parturition in yeast-fed cows. Yeast supplementation enhanced cellular immune function; however, it had no effect on immunoglobulin G secretion against ovalbumin immunization. Overall, live yeast supplementation benefited milk production and milk composition, lowered plasma level of Hsp70, and enhanced the lymphocyte proliferative response in transition dairy cows, which may suggest an immunomodulatory effect of yeast supplement.

Role of heat shock protein and cytokine expression as markers of clinical outcomes with glutamine-supplemented parenteral nutrition in surgical ICU patients

Nutrients, such as glutamine (GLN), have been shown to effect levels of a family of protective proteins termed heat shock proteins (HSPs) in experimental and clinical critical illness. HSPs are believed to serve as extracellular inflammatory messengers and intracellular cytoprotective molecules. Extracellular HSP70 (eHSP70) has been termed a chaperokine due to ability to modulate the immune response. Altered levels of eHSP70 are associated with various disease states. Larger clinical trial data on GLN effect on eHSP expression and eHSP70's association with inflammatory mediators and clinical outcomes in critical illness are limited. Explore effect of longitudinal change in serum eHSP70, eHSP27 and inflammatory cytokine levels on clinical outcomes such as pneumonia and mortality in adult surgical intensive care unit (SICU) patients. Further, evaluate effect of parenteral nutrition (PN) supplemented with GLN (GLN-PN) versus GLN-free, standard PN (STD-PN) on serum eHSP70 and eHSP27 concentrations. Secondary observational analysis of a multicenter clinical trial in 150 adults after cardiac, vascular, or gastrointestinal surgery requiring PN support and SICU care conducted at five academic medical centers. Patients received isocaloric, isonitrogenous PN, with or without GLN dipeptide. Serum eHSP70 and eHSP27, interleukin-6 (IL-6), and 8 (IL-8) concentrations were analyzed in patient serum at baseline (prior to study PN) and over 28 days of follow up. eHSP70 declined over time in survivors during 28 days follow-up, but non-survivors had significantly higher eHSP70 concentrations compared to survivors. In patients developing pneumonia, eHSP70, eHSP27, IL-8, and IL-6 were significantly elevated. Adjusted relative risk for hospital mortality was reduced 75% (RR=0.25, p=0.001) for SICU patients with a faster decline in eHSP70. The area under the receiver operating characteristic curve was 0.85 (95% CI: 0.76 to 0.94) for the final model suggesting excellent discrimination between SICU survivors and non-survivors. GLN-PN did not alter eHSP70 or eHSP27 serum concentrations over time compared to STD-PN. Our results suggest that serum HSP70 concentration may be an important marker for severity of illness and likelihood of recovery in the SICU. GLN-supplemented-PN did not increase eHSP70.

Dietary selenium supplementation alleviates immune toxicity in the hearts of chickens with lead-added drinking water

ABSTRACTLead (Pb) is an environmental pollutant and can damage organisms. Selenium (Se) can alleviate Pb poisoning. The present study aimed to investigate the alleviative effect of Se on Pb-induced immune toxicity in chicken hearts. One-hundred-and-eighty Hy-line male chickens were randomly divided into four groups at 7 days of age. The control group was offered a standard commercial diet (SD) and drinking water (DW); the Se group was offered SD supplemented with sodium selenite (SeSD) and DW; the Pb + Se group was offered SeSD and DW supplemented with lead acetate (PbDW); and the Pb group was offered SD and PbDW. Relative mRNA expression of inducible nitric oxide synthase (iNOS), interleukins (IL-2, IL-4, IL-6, IL-12β, IL-17 and IFN-γ), and heat shock proteins (HSP27, HSP40, HSP60, HSP70, and HSP90) were determined by means of quantitative real-time PCR. Relative protein expression of iNOS, HSP60, HSP70, and HSP90 was assessed, as well as nitric oxide (NO) content and iNOS activity in heart tissue. The results indicated a down-regulation of interleukin (IL)-2 and IFN-γ and an up-regulation of NO, iNOS, interleukins (IL-4, IL-6, IL-12β, IL-17), and heat shock proteins (HSP27, HSP40, HSP60, HSP70, and HSP90) in Pb-damaged hearts. Se alleviated all of the above Pb-induced changes. There were time-dependent effects on NO content, iNOS activity, and mRNA levels of iNOS, IL-2, IL-4, IL-6, IL-17, HSP27, HSP40, HSP60, HSP70, and HSP90 after Pb treatment in the chicken hearts. Se alleviated Pb-induced immune toxicity in the chicken hearts.

Increased Levels of Interleukin-17A Exosomes in Psoriasis.

Exosomes are involved in modulating the immune system and mediating communication between cells. The aim of this study was to investigate the involvement of exosomes in psoriasis. Exosomes from patients with psoriasis were analysed by nanoparticle tracking analysis and protein expression was analysed by western blotting. The concentration of HSP70 was determined by an enzyme-linked immunosorbent assay, and concentrations of interleukin (IL)-1β, IL-2, IL-6, IL-10, IL-17A and tumour necrosis factor alpha (TNF-α) were determined by flow cytometry. Based on the severity of psoriasis, evaluated by body surface area (≤ 10% vs. > 10%), 2 groups of patients were compared (49 with mild psoriasis and 71 with moderate-to-severe psoriasis). The number (2.52×1011 ± 2.29×1010 vs. 1.79×1011 ± 1.93×1010, p = 0.19) and size (94.44 ± 22.00 nm vs. 96.87 ± 28.30 nm, p = 0.72) of exosomes and the concentration of HSP70 in the exosomes were not significantly different in the 2 groups of patients. IL-17A exosome levels were significantly higher in patients with moderate-to-severe psoriasis compared with those with mild psoriasis (p = 0.02). There were no significant differences in levels of TNF-α, IL-1, IL-2, IL-6 and IL-10. This study shows, for the first time, the presence of circulating exosomes in patients with psoriasis. These data confirm the involvement of circulating exosomes in psoriasis, in particular in moderate-to-severe psoriasis, through IL-17A-producing exosomes.

Sponges Lubomirskiidae as bioindicators of the state of Lake Baikal.

Due to their simple body structure, filter-feeding life style and widespread abundance sponges are ideal bioindicators of the state of the environment. For marine sponges stress response studies are conducted in the field of studying of microbial composition, gene expression and transcriptome techniques. For freshwater sponges expression of the stress protein HSP70 was studied. In Baikal sponges, it was shown that the content of HSP70 is an indicator of the stress response to temperature increase. Transcriptomic studies of the endemic baikalian sponges are in progress.