Familial Accumulation of Social Anxiety Symptoms and Maladaptive Emotion Regulation

BackgroundSocial anxiety is thought to be strongly related to maladaptive emotion regulation (ER). As social anxiety symptoms accumulate in families, we hypothesize that maladaptive ER is also more prevalent in families with anxious children. Thus, we analyze differences in emotion regulation of both child and mother in relation to social anxiety, as well as both their ER strategies in dealing with anxiety. Further, a positive relation between child and maternal ER strategies is assumed.MethodChildren (aged 9 to 13 years) with social, anxiety disorder (SAD; n = 25) and healthy controls (HC, n = 26) as well as their mothers completed several measures of social anxiety and trait ER strategies towards anxiety. As ER of children is still in development, age is considered as covariate.ResultsSAD children and their mothers reported more maladaptive ER strategies than HC dyads. Maternal maladaptive ER was related negatively to child adaptive ER which was further moderated by the child’s age.DiscussionMaladaptive ER strategies seem to contribute to the exacerbation of social anxiety in both mother and child. Mothers reporting maladaptive ER may have difficulties supporting their child in coping with social anxiety while simultaneously also experiencing heightened levels of anxiety. Deeper understanding of interactional processes between mothers and children during development can assist the comprehension of factors maintaining SAD. Implications for future research and possible consequences for interventions are discussed.     

Extracellular esterase activity as an indicator of endoplasmic reticulum calcium depletion

Abstract

Context: Endoplasmic reticulum (ER) calcium depletion is associated with diverse diseases, including cardiac, hepatic, and neurologic diseases.

Objective: The aim of the present study was to identify and characterize an endogenous protein that could be used to monitor ER calcium depletion comparably to a previously described exogenous reporter protein.

Materials and methods: The use of a selective esterase-fluorescein diester pair allowed for carboxylesterase activity in extracellular fluid to be measured using a fluorescent readout. Cell culture media from three different cell lines, rat plasma, and human serum all possess quantifiable amounts of esterase activity.

Results: Fluorescence produced by the interaction of carboxylesterases with a fluorescein diester substrate tracks with pharmacological and physiological inducers of ER calcium depletion. The fluorescence measured for in vitro and in vivo samples were consistent with ER calcium depletion being the trigger for increased esterase activity.

Discussion: Decreased luminal ER calcium causes ER resident esterases to be released from the cell, and, when assessed concurrently with other disease biomarkers, these esterases may provide insight into the role of ER calcium homeostasis in human diseases.

Conclusion: Our results indicate that carboxylesterases are putative markers of ER calcium dysfunction.     

Agreement of self-reported hormone receptor status with cancer registry data in young breast cancer patients

BackgroundThough breast cancer subtype is a key determinant of treatment choice and prognosis, few studies have assessed breast cancer patients’ knowledge of estrogen and progesterone receptor (ER/PR) status.MethodsWomen diagnosed with invasive breast cancer at age 18–64 years in 2007 were recruited from the Pennsylvania Cancer Registry, and mailed a questionnaire that asked respondents to identify their ER/PR status. There were 2191 respondents included in the analysis. Agreement between self-report and cancer registry ER/PR status was assessed using kappa statistic. Logistic regression was used to assess the association of demographic, socioeconomic, and tumor factors with inaccurate self-report of ER/PR status.ResultsFifty-nine percent of respondents reported ER/PR positive status, 15% reported ER/PR negative status, 17% responded ‘don’t know’, and 9% did not respond. Overall, there was 69% agreement between self-report and cancer registry data, and fair agreement as measured by kappa (0.36). After excluding women who did not know or did not report their ER/PR status, there was 93% agreement, and substantial agreement as measured by kappa (0.76). Women who were older, non-white, less educated, lower income, and had ER/PR negative disease were significantly more likely to inaccurately report their ER/PR status.ConclusionsThough a significant proportion of women do not know their hormone receptor status, women who reported their ER/PR status were accurate. Our results suggest room for improvement in patient knowledge of tumor subtypes, but also that self-reported ER/PR status may be a useful surrogate when medical record or cancer registry data is unavailable.     

研究快报：锻炼缓解内质网还原应激从而促进健康衰老

目的 锻炼是延缓衰老的有效策略,本工作的目的在于探索锻炼是如何在细胞器水平影响内质网的氧化还原状态,以及内质网氧化还原状态是否影响个体衰老。方法 利用定位于内质网响应过氧化氢的Hyperion探针检测线虫衰老过程中及经过游泳运动后体壁肌肉内质网的氧化还原状态。通过在线虫内质网中特异过表达哺乳动物过氧化氢酶的同源基因ctl-1构建内质网特异的还原应激模型,研究了内质网还原应激对个体衰老的影响。线虫的健康状态以线虫寿命、身体摆动次数及对压力的响应能力为判断指标进行表征。结果 用Hyperion_ER探针检测发现,衰老线虫的内质网中过氧化氢水平相比与年轻线虫显著降低,表明内质网在衰老过程中发生了还原应激。线虫经过短时90 min游泳运动及长时期4 d （3次+3次+2次+2次,90 min/次）的游泳运动都可以增加内质网的氧化力。相比于对照,内质网还原应激的线虫寿命缩短,身体摆动次数降低,应对压力的响应能力下降,表明内质网还原应激加速线虫衰老。进一步研究发现,长时期的锻炼可以提高内质网的氧化力,缓解衰老相关的内质网还原应激,经过锻炼的第8天的线虫运动活力显著高于未锻炼的第...     

Mechanistic insights in recov-ER-phagy: micro-ER-phagy to recover from stress

ABSTRACT

Physiological and pathological stresses may cause swelling of the endoplasmic reticulum (ER), a biosynthetic organelle in eukaryotic cells. Upon conclusion of the stress, ER size and content return to physiological levels. The translocon component SEC62 decorates the portions of excess ER that must be cleared from cells. Our recent paper highlights the role of endosomal sorting complex required for transport (ESCRT)-III-driven micro-ER-phagy in ER remodeling during cell recovery from ER stress.     

Repeat estradiol exposure differentially regulates protein expression patterns for estrogen receptor and E-cadherin in older mouse ovarian surface epithelium: Implications for replacement and adjuvant hormone therapies?

BackgroundEstrogen replacement therapy increases risk for ovarian epithelial cancer, a cancer of mainly older women, yet the response of older ovarian surface epithelium (OSE) to repeat estrogen exposure overtime has not been studied. We have previously reported significant reductions in estrogen receptor (ER) protein expression, particularly the ERβ1 isoform, in older mouse OSE following a single depot estradiol injection. The current study examined OSE from older mice following a single, and repeat estradiol injection, given 14 days apart over 28 days.MethodsCohorts of mice were sacrificed 48 hours following each estradiol injection, and at three other equidistant time points. Serum and ovarian tissue estradiol concentration was correlated to immunohistochemical and morphometric parameters used to identify evidence of OSE hyperplasia and hypertrophy. Using immunohistochemistry, E-cadherin expression was investigated in OSE 48 hours following both estradiol injections, while ERα and ERβ1 expression was examined in OSE following repeat estradiol exposure only.ResultsFirst exposure to exogenous estradiol resulted in OSE hypertrophy and hyperplasia, and high levels of E-cadherin expression. In contrast, repeat estradiol exposure resulted in no OSE hyperplasia or hypertrophy, low levels of E-cadherin expression, high ERα and reduced ERβ1 protein expression in OSE, and low stromal ERα expression. Blood and ovarian tissue estradiol levels following repeat estradiol injection were half those recorded after a first dose equivalent injection, but remained significantly elevated above controls.ConclusionRepeat estradiol exposure leads to accumulation of estradiol in ovarian tissue, differentially regulating protein expression patterns for E-cadherin in OSE and ER in OSE and stroma.     

Clinicopathologic Characteristics of Oestrogen Receptor-Positive/Progesterone Receptor-Negative/Her2-Negative Breast Cancer According to a Novel Definition of Negative Progesterone Receptor Status: A Large Population-Based Study from China

PurposeA lack of progesterone receptor (PgR) expression in oestrogen receptor-positive (ER+) tumours is associated with worse survival. PgR status is usually defined as positive or negative using 1% positive nuclei as a cut-off point. In this study, we aimed to assess the clinicopathologic characteristics of ER+/PgR-/HER2- tumours by comparing them with ER+/PgR+/HER2- tumours using a PgR cut-off point of 20% as a divisive criterion.MethodsWe analysed 1,522 patients with primary breast cancer who had undergone surgery at the Cancer Center of Fudan University between 2012 and 2014. Age, grade, tumour size, lymph node status and lymphovascular invasion were assessed. Multinomial logistic regression, linear regression and chi-square test models were applied to assess associations between ER, PR and clinical features.ResultsER+/PgR-/HER2- tumours showed poorer clinicopathologic characteristics relative to ER+/PgR+/HER2- tumours using a PgR threshold of 20% instead of 1%. The clinicopathologic characteristics did not differ between tumours with purely negative PgR expression and tumours with a PgR percentage ranging from 1% to 19%. The prognostic significance of PR expression appeared more pronounced in patients under a high Ki-67 status than those under a low Ki-67 status.ConclusionsBased on these findings, we propose the use of a novel threshold of 20% to define PgR status. Nevertheless, the impact of this new criterion on patient management and clinical treatment requires additional study.     

Transverse False Tendons in the Left Ventricular Cavity Are Associated with Early Repolarization

BackgroundLeft ventricular false tendons (LVFTs) are related to precordial murmurs, ventricular arrhythmias and some repolarization abnormalities. Early repolarization (ER) is a specific type of repolarization abnormality.ObjectiveThe aim of the present study was to investigate the relationship between LVFTs and ER.MethodsThis study retrospectively included 99 consecutive healthy subjects and 33 patients with ER. Early repolarization was defined as an elevation of the QRS-ST junction of >0.1 mV from baseline in at least 2 inferior or lateral leads, manifested as QRS slurring or notching. Each participant was examined using echocardiography with second harmonic imaging, and the attachments of the LVFTs were recorded.ResultsA total of 93 LVFTs were present in 82 (83%) of the 99 healthy subjects. Of these 93 LVFTs, the majority (79/93, or 84.9%) were longitudinal-type LVFTs, which originated from the basal interventricular septum (IVS) and progressed toward the apical segment of the left ventricular free wall. There were significant differences in the positioning of the LVFTs between the ER patients and control (P < 0.0001). LVFTs between mid-IVS to the middle of the LV free wall were found more common in patients with ER compared with control subjects (47.5% vs. 6.5%, P < 0.0001). In the ER group, LVFTs between the basal IVS to the apical segment of LV free wall were only identified in 21% of the LVFTs, compared to a value of 84.9% for the control group (P < 0.0001). The distribution of LVFT trends in the ER group was also significantly different from that in the control group (P < 0.05).ConclusionsLVFTs are commonly visualized using echocardiography. An LVFT from the basal IVS to the apical segment of the left ventricular free wall may be a normal anatomical structure in the left ventricular cavity. On the contrary, transverse false tendons in the left ventricular cavity may be associated with ER.     

Icariin ameliorates estrogen-deficiency induced bone loss by enhancing IGF-I signaling via its crosstalk with non-genomic ERα signaling

BackgroundRapid, non-genomic estrogen receptor (ER) signaling plays an integral role in mediating the tissue selective properties of ER modulators. Icariin, a bone bioactive flavonoid, has been reported to selectively activate non-genomic ERα signaling in in vitro and in vivo studies.PurposeThe mechanisms underlying the estrogen-like bone protective effects of icariin are not fully understood, especially those that are related to insulin-like growth factor I (IGF-1) signaling. The bone protective effects of icariin were investigated in female mature ovariectomized (OVX) rats and the signaling of IGF-IR- ERα cross-talk was determined in osteoblastic cells.Study design and methodsIcariin at 3 different dosages (50, 500 and 3000 ppm) were orally administrated to rats for 3 months through daily intake of phytoestrogen-free animal diets containing icariin. Bone marrow stromal cells (BMSCs) and osteoclast precursors from femurs were harvested for experiments and RNA-sequencing. The interactions between IGF-IR and non-genomic ERα signaling were examined in pre-osteoblastic MC3T3-E1 cells and mature osteoblasts differentiated from BMSCs.ResultsOur results show that chronic administration of icariin to OVX rats significantly protected them against bone loss at the long bone and lumbar spine without inducing any uterotrophic effects. Ex vivo studies using BMSCs and osteoclast precursors confirmed the stimulatory effects of icariin on osteoblastogenesis and its inhibitory effects on osteoclastogenesis, respectively. RNA-sequencing analysis of mRNA from BMSCs revealed that icariin at 500 ppm significantly altered IGF-1 signaling as well as PI3K-Akt pathways. Our results demonstrated for the first time the rapid induction of interactions between IGF-IR and ERα as well as IGF-IR signaling and the downstream Akt phosphorylation by icariin in MC3T3-E1 cells. The activation of ERα and Akt phosphorylation by icariin in MC3T3-E1 cells and the osteogenic effects of icariin on ALP activity in mature osteoblasts were shown to be IGF-IR-dependent.ConclusionOur findings reveal that icariin activates both ERα and Akt via enhancing rapid induction of IGF-1 signaling in osteoblastic cells for osteogenesis and might be regarded as a novel pathway-selective phytoestrogen for management of postmenopausal osteoporosis.     

Novel triterpenoid AECHL-1 induces apoptosis in breast cancer cells by perturbing the mitochondria–endoplasmic reticulum interactions and targeting diverse apoptotic pathways

BackgroundThe modus operandi for an anti-cancer drug must allow for an efficient discrimination system between tumorigenic and non-tumorigenic cells. Targeting ER stress and mitochondrial function in cancer cells appears to be a suitable option, as these processes are dysregulated in tumor cells. AECHL-1, a novel triterpenoid, exhibits potent anticancer activity against an array of cancer cell lines however, its mechanism of action remains elusive.MethodsMolecular targets of AECHL-1 were investigated using breast adenocarcinoma cells MCF-7, MDA-MB-231 and mammary epithelial cell line MCF 10A in vitro and xenograft tumors in SCID mice in vivo. Western blotting, flow cytometry, and immunohistochemical studies were employed to delineate the molecular pathways.ResultsAECHL-1 caused a transient elevation of ER stress proteins along with a prolonged phosphorylation of eIF2α in breast cancer cells. This was accompanied by a simultaneous release of calcium from ER stores and subsequent mitochondrial accumulation. These effects could be reversed by using ER stress inhibitors. AECHL-1 brings about mitochondria mediated, caspase independent cell death via AIF in MCF-7 cells; MDA-MB-231 succumbed to caspase dependent extrinsic pathway. Xenograft studies closely echoed our in vitro results. AECHL-1 did not alter cellular and molecular parameters in MCF 10A.ConclusionThese findings reveal that, AECHL-1 targets the Achilles Heel of cancer cell, namely dysfunctional ER and mitochondria while being non toxic to normal parenchyma and can thus be further explored as a potential chemotherapeutic intervention.General significanceAggravation of ER stress by AECHL-1 uncovers a novel pathway for selective elimination of cancer cells.     

Versatility of the Endoplasmic Reticulum Protein Folding Factory

ABSTRACT

The endoplasmic reticulum (ER) is dedicated to import, folding and assembly of all proteins that travel along or reside in the secretory pathway of eukaryotic cells. Folding in the ER is special. For instance, newly synthesized proteins are N-glycosylated and by default form disulfide bonds in the ER, but not elsewhere in the cell. In this review, we discuss which features distinguish the ER as an efficient folding factory, how the ER monitors its output and how it disposes of folding failures.     

Induction of anti-tumor immunity by dendritic cells pulsed with an endoplasmic reticulum retrieval signal modifies heparanase epitope in mice

Background aimsIt has been reported that the heparanase epitope can elicit a cytotoxic T lymphocyte (CTL)-mediated anti-tumor response; however, the potential of the heparanase epitope modified by an endoplasmic reticulum (ER) retrieval signal, a C-terminal Lys–Asp–Glu–Leu sequence, is still unknown.MethodsThe heparanase epitope was modified by ER retrieval signal, and dendritic cells (DC) were pulsed with the modified peptide. The location and presentation of the modified peptide were detected, and the potential of the anti-tumor response was assessed.ResultsThe modified peptide could target the ER of DC to form stable major histocompatibility complex (MHC)–peptide complexes. In addition, vaccination with DC pulsed with the modified peptide elicited a robust, specific CTL response, significantly inhibited tumor growth and prolonged the lifespan of the mice.ConclusionsThe heparanase epitope modified by ER retrieval signal can be considered an ideal tumor vaccine, and may represent a new strategy for cancer immunotherapy in the clinic.     

In vivo and in vitro application of black soybean peptides in the amelioration of endoplasmic reticulum stress and improvement of insulin resistance

AimsHepatic endoplasmic reticulum (ER) stress plays a key role in the development of obesity-induced insulin resistance. This study evaluated the effects of peptides from black soybean (BSP) on ER stress and insulin signaling in vitro and in vivo.Main methodsUsing C2C12 myotubes or HepG2 cells, we evaluated the effects of BSP on the expression of proteins involved in insulin signaling and in the ER stress response in insulin-sensitive or insulin-resistant cells. BSP was given orally to db/db mice for 5 weeks to investigate its antidiabetic effects in vivo and the underlying mechanisms.Key findingsBSP increased GLUT4 translocation and glucose transport in myotubes and stimulated Akt-mediated glycogen synthase kinase-3β (GSK-3β) and Foxo1 phosphorylation in HepG2 cells. BSP significantly restored the suppression of insulin-mediated Akt phosphorylation in insulin-resistant cells. BSP significantly inhibited the activation of ER stress-responsive proteins by thapsigargin. BSP also significantly reduced blood glucose and improved glucose tolerance in db/db mice. The serum lipid profile (triglyceride and high-density lipoprotein concentrations) improved concomitantly with the BSP-induced downregulation of hepatic fatty acid synthase expression in db/db mice. Consistent with the results observed in HepG2 cells, BSP downregulated the elevated hepatic ER stress response in diabetic mice concomitantly with an increased expression of phospho-Foxo1.SignificanceA peptide mixture, BSP, showed beneficial effects through multiple mechanisms involving the suppression of hepatic ER stress and restoration of insulin resistance, suggesting that it has potential as an antidiabetic agent.     

A new role for a COPII cargo adaptor in autophagy

ABSTRACT

Endoplasmic reticulum (ER) homeostasis is maintained by the removal of misfolded ER proteins via different quality control pathways. Aggregation-prone proteins, including certain disease-linked proteins, are resistant to conventional ER degradation pathways and require other disposal mechanisms. Reticulophagy is a disposal pathway that uses resident autophagy receptors. How these receptors, which are dispersed throughout the ER network, target a specific ER domain for degradation is unknown. We recently showed in budding yeast, that ER stress upregulates the reticulophagy receptor, triggering its association with the COPII cargo adaptor complex, Sfb3/Lst1-Sec23 (SEC24C-SEC23 in mammals), to discrete sites on the ER. These domains are packaged into phagophores for degradation to prevent the accumulation of protein aggregates in the ER. This unconventional role for Sfb3/Lst1 is conserved in mammals and is independent of its role as a cargo adaptor on the secretory pathway. Our findings may have important therapeutic implications in protein-aggregation linked neurodegenerative disorders.     

Receptor-based virtual screening evaluation for the identification of estrogen receptor β ligands

Abstract

In this paper, a receptor-based virtual screening study for the identification of estrogen receptor β (ERβ) ligands was developed. Starting from a commercial database of 400?000 molecules, only six compounds resulted to be potential active ligands of ERβ. Interestingly, all the six molecules possess scaffolds that had already been reported in known ERβ ligands. Therefore, the results obtained herein confirm the reliability of our virtual screening procedure, thus encouraging the application of this protocol to larger commercial databases in order to identify new ERβ ligands.     

Loss of tumor susceptibility gene 101 (TSG101) perturbs endoplasmic reticulum structure and function

Tumor susceptibility gene 101 (TSG101), an ESCRT-I protein, is implicated in multiple cellular processes and its functional depletion can lead to blocked lysosomal degradation, cell cycle arrest, demyelination and neurodegeneration. Here, we show that loss of TSG101 results in endoplasmic reticulum (ER) stress and this causes ER membrane remodelling (EMR). This correlates with an expansion of ER, increased vacuolation, altered relative distribution of the rough and smooth ER and disruption of three-way junctions. Blocked lysosomal degradation due to TSG101 depletion leads to ER stress and Ca²⁺ leakage from ER stores, causing destabilization of actin cytoskeleton. Inhibiting Ca²⁺ release from the ER by blocking ryanodine receptors (RYRs) with Dantrolene partially rescues the ER stress phenotypes. Hence, in this study we have identified the involvement of TSG101 in modulating ER stress mediated remodelling by engaging the actin cytoskeleton. This is significant because functional depletion of TSG101 effectuates ER-stress, perturbs the structure, mobility and function of the ER, all aspects closely associated with neurodegenerative diseases.Summary statementWe show that tumor susceptibility gene (TSG) 101 regulates endoplasmic reticulum (ER) stress and its membrane remodelling. Loss of TSG101 perturbs structure, mobility and function of the ER as a consequence of actin destabilization.