Stress-induced Increase Research Articles

Adipocyte Adrb3 Receptor Mediates Mechanical Hypersensitivity in Chronic Overlapping Pain Conditions

Chronic overlapping pain conditions (COPCs), such as temporomandibular disorder and fibromyalgia, are recognized as a significant healthcare problem. While the underlying mechanisms are not fully understood, the origins of COPCs are linked to genetic and environmental factors resulting in enhanced catecholaminergic tone. Patients with COPCs have genetic variants that lead to reduced activity of catechol-O-methyltransferase (COMT), a ubiquitously expressed enzyme that metabolizes NE, and corresponding increases in basal and stress-induced levels of norepinephrine (NE) in circulation and. We established a novel clinically relevant mouse model of COPCs that integrates COMT genetic background, stress, and minor surgery. Using this novel model, here we show an essential role of the adipocyte adrenergic receptor beta 3 (Adrb3) in driving mechanical hypersensitivity that affects multiple body sites. Results from in vitro studies of differentiated mature primary adipocytes, demonstrates that stimulation of Adrb3 with NE induces the release of pro-inflammatory cytokine interleukin-6 (IL-6) evokes calcium influx in a dose-dependent manner. To demonstrate the specific role of adipocyte Adrb3 in our mouse model of COPCs, we selectively deleted Adrb3 in adipocytes by crossing Adrb3 flox mice with AdipoQ-Cre mice to generate Adrb3 conditional knockout in adipocyte (Adrb3 fl/fl;AdipoQ-Cre mice). We then crossed these mice with COMT-/- mice to get Adrb3 fl/fl;AdipoQ-Cre; COMT+/- mice. Our results demonstrate that compared to WT:COMT+/- mice, the AdipoQAdrb3+/-:COMT+/- mice exhibit reduced stress-induced increases in NE and reduced mechanical hypersensitivity. In summary, our findings suggest that adipocyte Adrb3 promotes mechanical hypersensitivity through NE-dependent increases in cytokines/adipokines. Further, this work provides the first direct evidence that adipocytes (fat cells) contribute to multi-site body pain. Unraveling the adipocyte Adrb3 mechanisms involved in COPCs may facilitate the discovery of peripherally-targeted therapeutics that effectively and safely alleviate pain in patients with COPCs. Grant support from NIH/NINDS R01 NS109541 to AN. Chronic overlapping pain conditions (COPCs), such as temporomandibular disorder and fibromyalgia, are recognized as a significant healthcare problem. While the underlying mechanisms are not fully understood, the origins of COPCs are linked to genetic and environmental factors resulting in enhanced catecholaminergic tone. Patients with COPCs have genetic variants that lead to reduced activity of catechol-O-methyltransferase (COMT), a ubiquitously expressed enzyme that metabolizes NE, and corresponding increases in basal and stress-induced levels of norepinephrine (NE) in circulation and. We established a novel clinically relevant mouse model of COPCs that integrates COMT genetic background, stress, and minor surgery. Using this novel model, here we show an essential role of the adipocyte adrenergic receptor beta 3 (Adrb3) in driving mechanical hypersensitivity that affects multiple body sites. Results from in vitro studies of differentiated mature primary adipocytes, demonstrates that stimulation of Adrb3 with NE induces the release of pro-inflammatory cytokine interleukin-6 (IL-6) evokes calcium influx in a dose-dependent manner. To demonstrate the specific role of adipocyte Adrb3 in our mouse model of COPCs, we selectively deleted Adrb3 in adipocytes by crossing Adrb3 flox mice with AdipoQ-Cre mice to generate Adrb3 conditional knockout in adipocyte (Adrb3 fl/fl;AdipoQ-Cre mice). We then crossed these mice with COMT-/- mice to get Adrb3 fl/fl;AdipoQ-Cre; COMT+/- mice. Our results demonstrate that compared to WT:COMT+/- mice, the AdipoQAdrb3+/-:COMT+/- mice exhibit reduced stress-induced increases in NE and reduced mechanical hypersensitivity. In summary, our findings suggest that adipocyte Adrb3 promotes mechanical hypersensitivity through NE-dependent increases in cytokines/adipokines. Further, this work provides the first direct evidence that adipocytes (fat cells) contribute to multi-site body pain. Unraveling the adipocyte Adrb3 mechanisms involved in COPCs may facilitate the discovery of peripherally-targeted therapeutics that effectively and safely alleviate pain in patients with COPCs. Grant support from NIH/NINDS R01 NS109541 to AN.

A Potential Link Between Negative Affective Responsivity to Daily Stress and Blood Pressure Reactivity to Acute Stress in College‐Aged Adults

Daily stress, defined as the routine challenges of day‐to‐day living and the unexpected small hassles that disrupt everyday life, is predictive of future cardiovascular disease (CVD)‐related health outcomes; however, the underlying mechanisms remain unclear. Given that heightened blood pressure (BP) reactivity to stress is also purported to increase future CVD risk, we examined a potential link between daily stress and acute stress‐induced increases in BP. We hypothesized that increased frequency of daily stressor exposures and greater negative emotional responsiveness to daily stress (i.e., negative affective responsivity) would be positively related to increased BP reactivity to acute stressors in the laboratory setting. Thirty adults (23±5 yrs; 21 female) completed the Daily Inventory of Stressful Events (DISE) interview every day for 8 consecutive days to quantify objective (e.g., frequency) and subjective appraisal characteristics (e.g., emotions) of naturally occurring daily stressors (e.g., stressful event at home/work). On DISE Day 8, mean arterial pressure (MAP; finger photoplethysmography) was measured during two distinct laboratory‐based acute stress stimuli: the cold pressor test (CPT; generalized sympathoexcitatory stimulus) and the Stroop Color Word Test (SCWT; cognitive stress stimulus). Daily stress exposure was calculated as the total number of stressors across the 8‐day protocol, and negative affective responsivity was calculated as the magnitude of the increase in negative affect on stressor exposure days (at least 1 stressor reported) compared to negative affect on non‐stressor exposure days (no stressors reported). BP reactivity was quantified as the peak acute stress‐induced increase in MAP to the CPT and SCWT. Participants experienced a total of 3±3 stressors (range: 0‐9) over the 8‐day interview timeframe. Negative affect was greater on stressor exposure days (0.68±0.50 a.u.) compared to non‐exposure days (0.37±0.36 a.u.; p<0.01). Both the CPT (∆21±7 mmHg) and the SCWT (∆9±6 mmHg) elicited increases in MAP (both p<0.05). The total number of stressors was not related to the peak increase in MAP during either the CPT (slope=‐0.10; r=0.04; p=0.85) or the SCWT (slope=‐0.43; r=19; p=0.31). Negative affective responsivity was positively related to the peak increase in MAP during the CPT (slope=12.41; r=0.45; p=0.03) but not the SCWT (slope=3.00; r=0.14, p=0.50). These preliminary data demonstrate that daily stressor exposure by itself is not related to BP reactivity to laboratory stressors in college‐aged adults. However, greater negative affective responsivity is associated with greater acute stress‐induced increases in BP during the CPT, but not the SCWT, perhaps suggesting that the link between daily stress processes and BP reactivity may be dependent on the stress‐responsive neurocircuitry being activated.

Cortisol reactivity impairs suppression-induced forgetting

To some extent, we can shape our recollections by intentionally remembering certain experiences while trying to forget others, for example, by intentional suppression. Acute stress impairs suppression-induced forgetting of memories. It is unclear, however, whether these deficits are a direct consequence of the acute stress-induced cortisol response. The present study was designed to examine the stress-induced impairment in suppression-induced forgetting in a subgroup of cortisol responders. We exposed healthy participants to a stress (n = 55) or no-stress control (n = 32) version of the Maastricht Acute Stress Test before they performed the Think / No-Think task. Here, participants aimed to repeatedly retrieve or suppress memories of previously learned cue-target video clips to mimic the complexity of episodic memories. Results on the subsequent memory test revealed that, while the no-stress controls and cortisol non-responders demonstrated suppression-induced forgetting, this effect was absent in cortisol responders. Moreover, the magnitude of suppression-induced forgetting was negatively correlated to stress-induced cortisol increases. The current study extends findings on stress-induced impairments in suppression-induced forgetting by specifically focusing on cortisol reactivity. Furthermore, our findings show the importance of individual differences in cortisol responses as a driving mechanism behind stress-induced alterations in our capacity to actively control our memory.

Fluid Shear Stress Ameliorates Prehypertension-Associated Decline in Endothelium-Reparative Potential of Early Endothelial Progenitor Cells.

This study investigated the effects of prehypertension and shear stress on the reendothelialization potential of human early EPCs and explored its potential mechanisms. Early EPCs from the prehypertensive patients showed reduced migration and adhesion in vitro and demonstrated a significantly impaired in vivo reendothelialization capacity. Shear stress pretreatment markedly promoted the in vivo reendothelialization capacity of EPCs. Although basal CXCR4 expression in early EPCs from prehypertensive donors was similar to that from healthy control, SDF-1-induced phosphorylation of CXCR4 was lower in prehypertensive EPCs. Shear stress up-regulated CXCR4 expression and increased CXCR4 phosphorylation, and restored the SDF-1/CXCR4-dependent JAK-2 phosphorylation in prehypertensive EPCs. CXCR4 knockdown or JAK-2 inhibitor treatment prevents against shear stress-induced increase in the migration, adhesion and reendothelialization capacity of the prehypertensive EPCs. Collectively, CXCR4 receptor profoundly modulates the reendothelialization potential of early EPCs. The abnormal CXCR4-mediated JAK-2 signaling may contribute to impaired functions of EPCs from patients with prehypertension.

Oxidative Stress-Induced TRPV2 Expression Increase Is Involved in Diabetic Cataracts and Apoptosis of Lens Epithelial Cells in a High-Glucose Environment.

Cataracts are a serious complication of diabetes. In long-term hyperglycemia, intracellular Ca2+ concentration ([Ca2+]i) and reactive oxygen species (ROS) are increased. The apoptosis of lens epithelial cells plays a key role in the development of cataract. We investigated a potential role for transient receptor potential vanilloid 2 (TRPV2) in the development of diabetic cataracts. Immunohistochemical and Western blotting analyses showed that TRPV2 expression levels were significantly increased in the lens epithelial cells of patients with diabetic cataracts as compared with senile cataract, as well as in both a human lens epithelial cell line (HLEpiC) and primary rat lens epithelial cells (RLEpiCs) cultured under high-glucose conditions. The [Ca2+]i increase evoked by a TRPV2 channel agonist was significantly enhanced in both HLEpiCs and RLEpiCs cultured in high-glucose media. This enhancement was blocked by the TRPV2 nonspecific inhibitor ruthenium red and by TRPV2-specific small interfering (si)RNA transfection. Culturing HLEpiCs or RLEpiCs for seven days in high glucose significantly increased apoptosis, which was inhibited by TRPV2-specific siRNA transfection. In addition, ROS inhibitor significantly suppressed the ROS-induced increase of TRPV2-mediated Ca2+ signal and apoptosis under high-glucose conditions. These findings suggest a mechanism underlying high-glucose–induced apoptosis of lens epithelial cells, and offer a potential target for developing new therapeutic options for diabetes-related cataracts.

In This Issue

HomeCirculation ResearchVol. 130, No. 5In This Issue Free AccessIn BriefPDF/EPUBAboutView PDFView EPUBSections ToolsAdd to favoritesDownload citationsTrack citationsPermissions ShareShare onFacebookTwitterLinked InMendeleyRedditDiggEmail Jump toFree AccessIn BriefPDF/EPUBIn This Issue Ruth Williams Ruth WilliamsRuth Williams Search for more papers by this author Originally published3 Mar 2022https://doi.org/10.1161/RES.0000000000000536Circulation Research. 2022;130:691is related toEro1α-Dependent ERp44 Dissociation From RyR2 Contributes to Cardiac ArrhythmiaGLP1R Attenuates Sympathetic Response to High Glucose via Carotid Body InhibitionInhibition of IL11 Signaling Reduces Aortic Pathology in Murine Marfan SyndromeGLP1R Attenuates Sympathetic Response to High Glucose via Carotid Body Inhibition (p 694)Gut hormone receptors in the carotid body may be treatment targets for metabolic syndrome, say Pauza et al.Download figureDownload PowerPointHypertension and diabetes are risk factors for cardiovascular disease (CVD) and yet, for many patients with these two conditions, lowering blood pressure and blood sugar is insufficient for eliminating the risk. Because both hypertension and diabetes are linked to increased sympathetic nerve activation, and because the carotid body (CB)—a clusters of sensory cells in the carotid artery—regulates sympathetic nerve activity, Pauza and colleagues investigated the CB’s possible involvement. They performed a transcriptome analysis of CB tissue from rats with and without spontaneous hypertension and found, among many differently expressed genes, that the transcript encoding glucagon-like peptide-1 receptor (GLP1R) was considerably less abundant in the hypertensive animals. This was of particular interest because the gut hormone GLP1 promotes insulin secretion and tends to be suppressed in type 2 diabetes. Moreover, GLP1R agonists are already used as diabetes treatments. The team now show that treating rat CBs with such an agonist suppresses sympathetic nerve activation and arterial blood pressure, suggesting such drugs may provide benefits in more ways than one, and that the CB could be a novel target for lowering CVD risk in metabolic syndrome.Ero1α-Dependent ERp44 Dissociation from RyR2 Contributes to Cardiac Arrhythmia (p 711)Blocking oxidoreductase Ero1α could reduce arrhythmia risk in cardiovascular disease, say Hamilton et al.Download figureDownload PowerPointThe oxidative stress associated with cardiovascular disease (CVD) can lead to heart arrhythmia and sudden death. At the cellular level, the increased oxidation in the sarcoplasmic reticulum (SR) triggers aberrant release of calcium—via the ryanodine receptor (RyR2) channel—and this in turn ramps up muscle contractions. However, the precise molecular details of the pathology are lacking. Cellular stress is known to raise the level of SR oxidoreductase Ero1α, which interacts with an inhibitor of calcium release called ERp44. Now, Hamilton and colleagues complete the molecular puzzle, showing that ERp44 normally binds and inhibits RyR2 in rat cardiomyocytes. The stress-induced increase in Ero1α expression (in hypertrophic rat cardiomyocytes) causes dissociation of ERp44 from RyR2, they report, thus boosting calcium release. Importantly, the team also showed that inhibiting Ero1α reduced oxidation in the SR, prevented ERp44-RyR2 disassociation and reduced RyR2-mediated calcium release. It also curbed arrhythmia in isolated hypertrophic hearts. Because myocardium from heart failure patients also exhibited increased Ero1α expression compared with controls, the authors say, Ero1α inhibition could be a clinical strategy for preventing CVD-associated arrhythmia.Inhibition of IL11 Signaling Reduces Aortic Pathology in Murine Marfan Syndrome (p 728)Lim et al suggest cytokine IL-11 as a new target for the treatment of Marfan syndrome.Download figureDownload PowerPointPeople with the genetic connective tissue disorder Marfan syndrome (MFS) are typically tall and thin with long limbs, and are prone to skeletal, eye and cardiovascular problems including a life-threatening weakening of the aorta. While MFS patients commonly take blood-pressure lowering treatments to minimize the risk of aortic aneurysm and dissection, there is no cure for MFS nor even a targeted therapy. With the aim of finding novel MFS treatments, Lim and colleagues focused their attention on IL-11. This cytokine is strongly induced in vascular smooth muscle cells (VSMCs) upon treatment with the growth factor TGFβ—itself overactivated in MFS patients and considered a key feature of the syndrome’s molecular pathology. The team now show that IL-11 is indeed strongly upregulated in the aortas of MFS model mice and that genetically eliminating IL-11 in these animals can protect them against the aortic pathology they normally exhibit—including dilation, fibrosis, inflammation, elastin degradation and loss of VSMCs. Furthermore, treating MFS mice with an anti-IL-11 neutralizing antibody had the same beneficial effects. The results therefore suggest that inhibiting IL-11 activity could be a novel approach for protecting the aortas of MFS patients. Previous Back to top Next FiguresReferencesRelatedDetailsRelated articlesEro1α-Dependent ERp44 Dissociation From RyR2 Contributes to Cardiac ArrhythmiaShanna Hamilton, et al. Circulation Research. 2022;130:711-724GLP1R Attenuates Sympathetic Response to High Glucose via Carotid Body InhibitionAudrys G. Pauza, et al. Circulation Research. 2022;130:694-707Inhibition of IL11 Signaling Reduces Aortic Pathology in Murine Marfan SyndromeWei-Wen Lim, et al. Circulation Research. 2022;130:728-740 March 4, 2022Vol 130, Issue 5Article InformationMetrics © 2022 American Heart Association, Inc.https://doi.org/10.1161/RES.0000000000000536 Originally publishedMarch 3, 2022 PDF download Advertisement

Stress-induced increase of monoclonal antibody production in CHO cells.

Monoclonal antibodies (mAbs) are of great interest to the biopharmaceutical industry due to their widely used application as human therapeutic and diagnostic agents. As such, mAb require to exhibit human‐like glycolization patterns. Therefore, recombinant Chinese hamster ovary (CHO) cells are the favored production organisms; many relevant biopharmaceuticals are already produced by this cell type. To optimize the mAb yield in CHO DG44 cells a corelation between stress‐induced cell size expansion and increased specific productivity was investigated. CO2 and macronutrient supply of the cells during a 12‐day fed‐batch cultivation process were tested as stress factors. Shake flasks (500 mL) and a small‐scale bioreactor system (15 mL) were used for the cultivation experiments and compared in terms of their effect on cell diameter, integral viable cell concentration (IVCC), and cell‐specific productivity. The achieved stress‐induced increase in cell‐specific productivity of up to 94.94.9%–134.4% correlates to a cell diameter shift of up to 7.34 μm. The highest final product titer of 4 g/L was reached by glucose oversupply during the batch phase of the process.

Psychological stress disrupts intestinal epithelial cell function and mucosal integrity through microbe and host-directed processes

ABSTRACT Psychological stress alters the gut microbiota and predisposes individuals to increased risk for enteric infections and chronic bowel conditions. Intestinal epithelial cells (IECs) are responsible for maintaining homeostatic interactions between the gut microbiota and its host. In this study, we hypothesized that disruption to colonic IECs is a key factor underlying stress-induced disturbances to intestinal homeostasis. Conventionally raised (CONV-R) and germ-free (GF) mice were exposed to a social disruption stressor (Str) to ascertain how stress modifies colonic IECs, the mucosal layer, and the gut microbiota. RNA sequencing of IECs isolated from CONV-R mice revealed a robust pro-inflammatory (Saa1, Il18), pro-oxidative (Duox2, Nos2), and antimicrobial (Reg3b/g) transcriptional profile as a result of Str. This response occurred concomitant to mucus layer thinning and signs of microbial translocation. In contrast to their CONV-R counterparts, IECs from GF mice or mice treated with broad spectrum antibiotics exhibited no detectable transcriptional changes in response to Str. Nevertheless, IECs from Str-exposed GF mice exhibited an altered response to ex vivo bacterial challenge (increased dual Oxidase-2 [Duox2] and nitric oxide synthase-2 (Nos2)), indicating that STR primes host IEC pro-oxidative responses. In CONV-R mice stress-induced increases in colonic Duox2 and Nos2 (ROS generating enzymes) strongly paralleled changes to microbiome composition and function, evidencing Str-mediated ROS production as a primary factor mediating gut-microbiota dysbiosis. In conclusion, a mouse model of social stress disrupts colonic epithelial and mucosal integrity, a response dependent on an intact microbiota and host stress signals. Together these preclinical findings may provide new insight into mechanisms of stress-associated bowel pathologies in humans.

Lactobacillus rhamnosus CNCM I-3690 decreases subjective academic stress in healthy adults: a randomized placebo-controlled trial

ABSTRACT Psychological stress negatively affects the intestinal barrier function in animals and humans. We aimed to study the effect of Lactobacillus rhamnosus CNCM I-3690 on intestinal permeability and stress-markers during public speech. Healthy students were randomized to L. rhamnosus-containing (test) or acidified (placebo) milk consumed twice daily for 4 weeks, with 46 subjects per treatment group. Small intestinal permeability was quantified by a 2 h urinary lactulose–mannitol ratio (LMR, primary outcome), fractional excretion of lactulose (FEL) and mannitol (FEM). Salivary cortisol, State-Trait Anxiety Inventory (STAI) and Perceived Stress scores (PSS) were collected. No between-treatment differences were found for LMR (p = .71), FEL or FEM. Within-treatment analyses showed similar LMR and FEL but a stress-induced increase of FEM with the placebo (p < .05) but not test product. Despite a similar increase in salivary cortisol, the stress-induced increase in STAI was significantly lower with the test product vs. placebo (p = .01). Moreover, a stress-preventative effect of the probiotic was found for PSS and more pronounced in subjects with high stress-induced cortisol (p = .01). While increased FEM was mediated by salivary cortisol levels, the effect of the test product on subjective stress was not mediated by changes in FEM. No serious adverse events occurred. In conclusion, we demonstrated that L. rhamnosus CNCM I-3690 prevented stress-induced hyperpermeability to mannitol. Subjective but not objective stress-markers were reduced with L. rhamnosus vs. placebo, suggesting anxiolytic effects, which were independent of barrier stabilization and attractive for the reduction of stress in both health and disease. Clinicaltrials.gov, number NCT03408691.

ANT participation in mitochondrial permeability transition depends on the induction mechanism

Mitochondrial Permeability Transition (PT) refers to the phenomenon of stress-induced increase in permeability of the inner mitochondrial membrane caused by the opening of the nonspecific channel PT pore. Adenine Nucleotide Translator (ANT) is an important component of PT. However, the precise role of ANT remains controversial with some researchers proposing its primary role in pore formation while others suggesting modulatory role. We hypothesized that ANT involvement in PT might depend on the specific pathway of PT activation.

Acute stress anticipates and amplifies the luteinizing hormone pre-ovulatory surge in rats: Role of noradrenergic neurons

The stress experienced during rape seems to facilitate ovulation since the pregnancy rate in raped women is higher than that resulting from consensual intercourse. Adrenal progesterone, as well as central norepinephrine, is released in stressful situations. At adequate estrogenic levels, one of the main actions of progesterone is to anticipate the preovulatory LH surge through noradrenaline release. We aimed to investigate whether acute stresses that mimic those of rape (exposure to predator, restraint and cervix stimulation) applied on the proestrus morning in female rats could release progesterone, activate the noradrenergic neurons and facilitate the occurrence of the LH surge. Female rats were submitted to jugular vein cannulation immediately following acute stress: restraint (R), exposure to cat (P), uterine cervix stimulation (CS) applied individually or in association (SA). Non-stressed rats were used as control. Blood samples were collected from 11:00–18:00 h for LH, progesterone, corticosterone and estradiol measurements. Double labeling for c-Fos and tyrosine hydroxylase (TH) was examined in A1, A2 and A6 noradrenergic neurons after stresses. The SA group showed a greater stress-induced increase in progesterone compared to the other groups and the preovulatory LH surge was anticipated and amplified. This effect of SA seems to be related to the higher number of c-Fos/TH + neurons in the A1 and A2. The effect of anticipating the preovulatory surge of LH could in part elucidate why, in raped women, conception can occur in phases of the menstrual cycle other than the ovulatory phase facilitating the occurrence of pregnancies.

Crosstalk between Yeast Cell Plasma Membrane Ergosterol Content and Cell Wall Stiffness under Acetic Acid Stress Involving Pdr18.

Acetic acid is a major inhibitory compound in several industrial bioprocesses, in particular in lignocellulosic yeast biorefineries. Cell envelope remodeling, involving cell wall and plasma membrane composition, structure and function, is among the mechanisms behind yeast adaptation and tolerance to stress. Pdr18 is a plasma membrane ABC transporter of the pleiotropic drug resistance family and a reported determinant of acetic acid tolerance mediating ergosterol transport. This study provides evidence for the impact of Pdr18 expression in yeast cell wall during adaptation to acetic acid stress. The time-course of acetic-acid-induced transcriptional activation of cell wall biosynthetic genes (FKS1, BGL2, CHS3, GAS1) and of increased cell wall stiffness and cell wall polysaccharide content in cells with the PDR18 deleted, compared to parental cells, is reported. Despite the robust and more intense adaptive response of the pdr18Δ population, the stress-induced increase of cell wall resistance to lyticase activity was below parental strain levels, and the duration of the period required for intracellular pH recovery from acidification and growth resumption was higher in the less tolerant pdr18Δ population. The ergosterol content, critical for plasma membrane stabilization, suffered a drastic reduction in the first hour of cultivation under acetic acid stress, especially in pdr18Δ cells. Results revealed a crosstalk between plasma membrane ergosterol content and cell wall biophysical properties, suggesting a coordinated response to counteract the deleterious effects of acetic acid.

Yokukansan, a traditional Japanese medicine, suppresses stress-induced sympathetic activation and central responses to stress in rats

Stress exposure activates the sympathetic nervous system. We previously reported that restraint stress and stress-related neuropeptides increases plasma catecholamine levels and that brain prostanoids medicate these responses in rats. Yokukansan (YKS), a traditional Japanese medicine, has been used for neurosis, night crying and irritability. Additionally, it has been reported that YKS is also effective against the behavioral and psychological symptoms of dementia in patients with Alzheimer&apos;s disease, stress-related anxiety-like behavior and stress-induced increase in plasma corticosterone in rodents. These previous findings raise the possibility that YKS can exert its action in the brain and has suppressive effects on stress responses. In this study, we examined effects of YKS on stress-induced sympathetic activation and stress responses in the brain using rats. Repeated administration with YKS suppressed stress-induced increase in plasma adrenaline level. YKS also suppressed stress-induced elevation of prostaglandin E2 and thromboxane B2 levels in the paraventricular hypothalamic nucleus (PVN). In addition, YKS suppressed stress-induced increases of acetylcholine, GABA and serotonin in the PVN. Our results suggest that YKS can ameliorate stress-induced sympathetic activation via inhibition of stress responses in the brain.

Inhibitory effects of pine nodule extract on mental stress-induced increases in sympathetic nervous activity in young students

Inhibitory effects of pine nodule extract on mental stress-induced increases in sympathetic nervous activity in young students

Juvenile stress exerts sex-independent effects on anxiety, antidepressant-like behaviours and dopaminergic innervation of the prelimbic cortex in adulthood and does not alter hippocampal neurogenesis

Stress, particularly during childhood, is a major risk factor for the development of depression. Depression is twice as prevalent in women compared to men, which suggests that biological sex also contributes to depression susceptibility. However, the neurobiology underpinning sex differences in the long-term consequences of childhood stress remains unknown. Thus, the aim of this study was to determine whether stress applied during the prepubertal juvenile period (postnatal day 27–29) in rats induces sex-specific changes in anxiety-like behaviour, anhedonia, and antidepressant-like behaviour in adulthood in males and females. The impact of juvenile stress on two systems in the brain associated with these behaviours and that develop during the juvenile period, the mesocorticolimbic dopaminergic system and hippocampal neurogenesis, were also investigated. Juvenile stress altered escape-oriented behaviours in the forced swim test in both sexes, decreased latency to drink a palatable substance in a novel environment in the novelty-induced hypophagia test in both sexes, and decreased open field supported rearing behavior in females. These behavioural changes were accompanied by stress-induced increases in tyrosine hydroxylase immunoreactivity in the prefrontal cortex of both sexes, but not other regions of the mesocorticolimbic dopaminergic system. Juvenile stress did not impact anhedonia in adulthood as measured by the saccharin preference test and had no effect hippocampal neurogenesis across the longitudinal axis of the hippocampus. These results suggest that juvenile stress has long-lasting impacts on antidepressant-like and reward-seeking behaviour in adulthood and these changes may be due to alterations to catecholaminergic innervation of the medial prefrontal cortex.

Abstract 9526: Fto Demethylase Mediates Altered N6-methyladenosine (m6a)-rna Modification in Human Failing Heart and Stressed-Induced Rat Cardiomyocytes

Background: N6-methyladenosine (m6A) methylation is one of the important posttranscriptional modification of RNA, which affects RNA splicing, translation and stability. These modifications are dynamically regulated by writers, readers and erasers, and their alteration affects m6A modification and have shown to play key regulatory role in numerous biological processes. Fat mass- and obesity-associated (FTO) gene, known as a nucleic acid demethylase removes the methyl group from RNA and has been shown to be associated with progression of heart diseases. In this study, we determined m6A methylation in human failing heart, and evaluated if FTO regulates m6A-methylation of transcripts related to inflammation in cultured rat cardiomyocytes under various stress conditions. Methods and Results: m6A RNA-methylation was increased in human failing hearts as compared to heart samples from non-failing subjects. Interestingly, failing hearts were associated with significant decrease in FTO mRNA and protein expression. Furthermore, cultured rat cardiomyoblasts (H9C2) cells exposed to stressors (hypoxia or isoproterenol) for 24hrs showed significant decrease of FTO and an associated increase in m6A-RNA methylation (dot blot analysis). To further test if stress-induced increase in m6A-RNA modification is mediated through FTO, we knocked-down FTO, treated with isoproterenol for 24hrs and evaluated m6A methylation status of RNA. Interestingly, dot blot analysis showed significant increase in m6A methylation levels and proinflammatory cytokines (IL6 and TNFa) in FTO knockdown cells as compared to scramble, both at basal level and after exposure to stress. Furthermore, we evaluated the effect of FTO on methylation of RNA transcripts that encode inflammatory cytokines. MeRIP assay (Methylated RNA immunoprecipitation using m6A antibody and analysis of m6A-transcript by qRT-PCR) demonstrated that the gene expression of proinflammatory cytokines, IL6 and TNF-alpha was significantly increased in FTO knockdown cells as compared to scramble control cells . Conclusion: This data suggests stress-induced loss of FTO in the heart might mediate m6A-RNA methylation and therefore potential leading to upregulation of inflammatory response in the myocardium.

Glucocorticoid Receptor Antagonist Alters Corticosterone and Receptor-sensitive mRNAs in the Hypoxic Neonatal Rat.

Hypoxia, a common stressor with preterm birth, increases morbidity and mortality associated with prematurity. Glucocorticoids (GCs) are administered to the preterm infant to improve oxygenation; prolonged use of GCs remains controversial. We evaluated a selective glucocorticoid receptor (GR) antagonist (CORT113176) in our neonatal rat model of human prematurity to assess how fasting and hypoxia-induced increases in neonatal corticosterone affects endogenous hormones and endocrine pancreas function. Neonatal rat pups at postnatal day (PD) 2, PD8, and PD15 were pretreated with CORT113176 and, after 60 minutes of separation and fasting, exposed to hypoxia (8% O2) or control (normoxia) for 30 or 60 minutes while fasting was continued. Plasma corticosterone, ACTH, glucose, and insulin were measured and fasting Homeostatic Model Assessment of Insulin Resistance was calculated. Glucocorticoid and insulin receptor-sensitive gene mRNAs were analyzed in liver, muscle, and adipose to evaluate target tissue biomarkers. CORT113176 pretreatment augmented baseline and hypoxia-induced increases in corticosterone and attenuated hypoxia-induced increases in insulin resistance at PD2. Normoxic and hypoxic stress increased the hepatic GR-sensitive gene mRNAs, Gilz and Per1; this was eliminated by pretreatment with CORT113176. CORT113176 pretreatment decreased baseline insulin receptor-sensitive gene mRNAs Akt2, Irs1, Pik3r1, and Srebp1c at PD2. We show that CORT113176 variably augments the stress-induced increases in corticosterone concentrations (attenuation of negative feedback) and that GR is critical for hepatic responses to stress in the hypoxic neonate. We also propose that measurement of Gilz and Per1 mRNA expression may be useful to evaluate the effectiveness of GR antagonism.

Retrospective analysis of influencing factors on the efficacy of mechanical ventilation in severe and critical COVID-19 patients.

The novel coronavirus disease 2019 (COVID-19) has spread widely around the world with strong infectivity, rapid mutation and a high mortality rate. Mechanical ventilation has been included in the Diagnosis and Treatment Protocol for Novel Coronavirus Pneumonia (Trial Version 8) as an important treatment for severe and critical COVID-19 patients, but its clinical efficacy in COVID-19 patients is various. Therefore, it is necessary to study the influencing factors on the efficacy of mechanical ventilation in severe and critical COVID-19 patients. The aim of this study was to determine the influencing factors on the efficacy of mechanical ventilation in severe and critical COVID-19 patients. A total of 27 severe and critical COVID-19 patients were enrolled in this study and treated with mechanical ventilation at the Optical Valley Campus of Hubei Maternal and Child Health Care Hospital (Wuhan, Hubei Province) from February 20, 2020 to April 5, 2020. According to the final treatment outcomes, the patients were divided into the "effective group" and "death group." The clinical data of the two groups, such as the treatment process and final outcome, were retrospectively analyzed in order to determine the specific curative effects on the two groups and the reasons for the differences in such curative effects, as well as to explore the factors related to death. This study enrolled 27 severe and critical COVID-19 patients, including 17 males (63.0%) and 10 females (37.0%). Their ages were 74.41 ± 11.73-years-old, and 19 patients (70.4%) were over 70-years-old. Severe COVID-19 patients over 70-years-old who were treated with mechanical ventilation died in 14 cases (82.4%); thus, this was the peak age. A total of 17 patients died of basic disease, 16 of whom had more than two basic diseases. The basic diseases were hypertension, diabetes, and cardiovascular and cerebrovascular diseases. At the same time, 13 patients (76.5%) died from an abnormal increase in blood glucose. Among them, eight had diabetes before contracting COVID-19 and five had a stress-induced increase in blood glucose after contracting COVID-19. Diabetic ketoacidosis occurred in one case. The use of tocilizumab may be a double-edged sword that carries a certain risk in clinical usage. Among the patients who died, 16 (94.1%) went into septic shock at the end. There were significant differences in the degree of infection, cardiac and renal function, and blood glucose between the death group and effective group. Age, blood glucose, cardiac and renal function, and inflammatory reaction are important indicators of poor prognosis for mechanical ventilation in severe and critical COVID-19 patients.

FKBP51 in the Oval Bed Nucleus of the Stria Terminalis Regulates Anxiety-Like Behavior.

The cochaperone FKBP51, encoded by the Fkbp5 gene, has been identified as central risk factor for anxiety-related disorders and stress system dysregulation. In the brain, the oval bed nucleus of the stria terminalis (ovBNST) has been implicated in stress-induced anxiety. However, the role of Fkbp5 in the ovBNST and its impact on anxiety-like behavior have remained unknown. Here, we show in mice that Fkbp5 in the ovBNST is reactive to acute stress and coexpressed with the stress-regulated neuropeptides Tac2 and Crh Subsequently, results obtained from viral-mediated manipulation indicate that Fkbp5 overexpression (OE) in the ovBNST results in an anxiolytic-like tendency regarding behavior and endocrinology, whereas a Fkbp5 knock-out (KO) exposed a clear anxiogenic phenotype, indicating that native ovBNST expression and regulation is necessary for normal anxiety-related behavior. Notably, our data suggests that a stress-induced increase of Fkbp5 in the ovBNST may in fact have a protective role, leading to a transient decrease in anxiety and suppression of a future stress-induced hypothalamic-pituitary-adrenal (HPA) axis activation. Together, our findings provide a first insight into the previously unknown relationship and effects of Fkbp5 and the ovBNST on anxiety-like behavior and HPA axis functioning.

Cation transporters in cell fate determination and plant adaptive responses to a low-oxygen environment.

Soil flooding creates low-oxygen environments in root zones and thus severely affects plant growth and productivity. Plants adapt to low-oxygen environments by a suite of orchestrated metabolic and anatomical alterations. Of these, formation of aerenchyma and development of adventitious roots are considered very critical to enable plant performance in waterlogged soils. Both traits have been firmly associated with stress-induced increases in ethylene levels in root tissues that operate upstream of signalling pathways. Recently, we used a bioinformatic approach to demonstrate that several Ca2+ and K+ -permeable channels from KCO, AKT, and TPC families could also operate in low oxygen sensing in Arabidopsis. Here we argue that low-oxygen-induced changes to cellular ion homeostasis and operation of membrane transporters may be critical for cell fate determination and formation of the lysigenous aerenchyma in plant roots and shaping the root architecture and adventitious root development in grasses. We summarize the existing evidence for a causal link between tissue-specific changes in oxygen concentration, intracellular Ca2+ and K+ homeostasis, and reactive oxygen species levels, and their role in conferring those two major traits enabling plant adaptation to a low-oxygen environment. We conclude that, for efficient operation, plants may rely on several complementary signalling pathway mechanisms that operate in concert and 'fine-tune' each other. A better understanding of this interaction may create additional and previously unexplored opportunities to crop breeders to improve cereal crop yield losses to soil flooding.