Concurrent Elevation Research Articles

0159 LABORATORY-BASED SLEEP RESTRICTION ENHANCES THE CORTISOL AWAKENING RESPONSE IN HEALTHY ADULTS

Sleep restriction is related to risk for stress-related symptoms and disease processes. The cortisol awakening response (CAR) is the increase in cortisol observed immediately post-awakening in the morning. CAR is an indicator of an individual’s readiness to maintain alertness and psychological function in response to demands of the upcoming day. CAR is affected by insufficient sleep, though the direct effect of extended sleep restriction on CAR is not known. We examined profiles of CAR of healthy good-sleeping young adults in response to extended moderate sleep restriction and subsequent sleep recovery. Healthy participants [N=48; M(SD) age: 25.4(3.9); 39.4% Female] underwent sleep satiation (5 nights/10h time in bed [TIB]), extended sleep restriction (5 nights/5 h TIB; 0200-0700), and sleep recovery (3 nights/8h TIB). Salivary cortisol was collected 5-, 30-, and 45-minutes post-awakening following sleep satiation (baseline), and every morning following sleep restriction and recovery evenings. CAR was quantified using area under the curve relative to ground (AUC-G). Linear mixed effects models were used to examine baseline-to-sleep restriction and baseline-to-recovery AUC-G trends. AUC-G increased linearly from baseline across sleep restriction (b=.06, SE=.02, t=2.8, p=.007), and remained elevated relative to baseline following the sleep recovery (b=.03, SE=.01, t=2.4, p=.02). Model fit was not improved including age and sex as covariates. Increased CAR across the restricted sleep period supports the notion that the CAR may represent an adaptive response to restricted sleep associated with arousal and anticipation to meet the demands of the upcoming day. We also observed a lack of recovery to baseline following 3 nights of recovery sleep. While acute increases in CAR may be related to the ability to effectively cope with the demands of a day following sleep loss, more prolonged or chronic sleep restriction and concurrent elevations in CAR may be related to health-compromising hypothalmic-pituitary-adrenocortical (HPA) function. Department of Defense Military Operational Medicine Research Program (MOMRP).

Perinatal programming of the orexinergic (hypocretinergic) system in hypothalamus and anterior pituitary by testosterone

Orexins A/B derived from hypothalamic prepro-orexin (PPO) are agonists for orexin receptors 1 (OX1) and 2 (OX2). Previously, we showed clear sex differences in the hypothalamic-pituitary-gonadal orexinergic system in adult rodents. Here, we studied the effect of sexual brain differentiation on the orexinergic system in neuroendocrine structures regulating reproduction. We evaluated: a: proestrous and neonatally androgenized female rats; b: adult males, untreated or gonadectomized in adulthood and injected with oil or estradiol and progesterone (E2/P4); c: control and demasculinized males (perinatally treated with flutamide and later castration) injected either with oil or E2/P4 in adulthood. Rats were sacrificed at 12:00 and 18:00h; blood samples and brains were collected. Hormones were measured using radioimmunoassay. PPO, OX1 and OX2 mRNAs were quantified by qPCR in medial basal hypothalamus, anterior hypothalamus, adenohypophysis, and cortex. Western blots for OX1 were done in the same structures. In normal females, gonadotropins surged at 18:00h coinciding with significant elevations of PPO, OX1 and OX2 mRNAs and OX1 protein in hypothalamus and pituitary; no increases were observed at noon. Afternoon changes were absent in masculinized females. Demasculinized males when treated with E2/P4 showed high PPO, OX1 and OX2 mRNAs and OX1 protein expression in hypothalamus and pituitary at 12:00 and 18:00h compared vehicle-treated controls. The same steroid treatment was ineffective in males with normal brain masculinization. Here we show that neonatal testosterone shapes the sexual differences in the hypothalamic-pituitary orexinergic system in synchronicity to establishing the brain sex differences of the reproductive axis. The female brain controls gonadotropin surges and concurrent elevations of all studied components of the orexinergic system, suggesting its participation as a possible link between food intake, behavior and hormonal control of reproduction.

Prolactin regulation of oxytocin neurone activity in pregnancy and lactation.

During lactation, prolactin promotes milk synthesis and oxytocin stimulates milk ejection. In virgin rats, prolactin inhibits the activity of oxytocin-secreting neurones. We found that prolactin inhibition of oxytocin neurone activity is lost in lactation, and that some oxytocin neurones were excited by prolactin in lactating rats. The change in prolactin regulation of oxytocin neurone activity was not associated with a change in activation of intracellular signalling pathways known to couple to prolactin receptors. The change in prolactin regulation of oxytocin neurone activity in lactation might allow coordinated activation of both populations of neurones when required for successful lactation. Secretion of prolactin for milk synthesis and oxytocin for milk secretion is required for successful lactation. In virgin rats, prolactin inhibits oxytocin neurones but this effect would be counterproductive during lactation when secretion of both hormones is required for synthesis and delivery of milk to the newborn. Hence, we determined the effects of intracerebroventricular (i.c.v.) prolactin on oxytocin neurones in urethane-anaesthetised virgin, pregnant and lactating rats. Prolactin (2μg) consistently inhibited oxytocin neurones in virgin and pregnant rats (by 1.9±0.4 and 1.8±0.5spikess-1 , respectively), but not in lactating rats; indeed, prolactin excited six of 27 oxytocin neurones by >1spikes-1 in lactating rats but excited none in virgin or pregnant rats (χ22 =7.2, P=0.03). Vasopressin neurones were unaffected by prolactin (2μg) in virgin rats but were inhibited by 1.1±0.2spikess-1 in lactating rats. Immunohistochemistry showed that i.c.v. prolactin increased oxytocin expression in virgin and lactating rats and increased signal transducer and activator of transcription 5 phosphorylation to a similar extent in oxytocin neurones of virgin and lactating rats. Western blotting showed that i.c.v. prolactin did not affect phosphorylation of extracellular regulated kinase 1 or 2, or of Akt in the supraoptic or paraventricular nuclei of virgin or lactating rats. Hence, prolactin inhibition of oxytocin neurones is lost in lactation, which might allow concurrent elevation of prolactin secretion from the pituitary gland and activation of oxytocin neurones for synthesis and delivery of milk to the newborn.

Embryonic mosaic deletion of APP results in displaced Reelin-expressing cells in the cerebral cortex

It is widely accepted that amyloid precursor protein (APP) plays a central role in the pathogenesis of Alzheimer's disease. In addition, APP has been proposed to have functions in numerous biological processes including neuronal proliferation, differentiation, migration, axon guidance, and neurite outgrowth, as well as in synapse formation and function. However, germline knockout of APP yields relatively subtle phenotypes, and brain development appears grossly normal. This is thought to be due in part to functional compensation by APP family members and other type I transmembrane proteins. Here, we have generated a conditional mouse knockout for APP that is controlled temporally using CreER and tamoxifen administration. We show that total cortical expression of APP is reduced following tamoxifen administration during embryonic time points critical for cortical lamination, and that this results in displacement of Reelin-positive cells below the cortical plate with a concurrent elevation in Reelin protein levels. These results support a role for APP in cortical lamination and demonstrate the utility of a conditional knockout approach in which APP can be deleted with temporal control in vivo. This new tool should be useful for many different applications in the study of APP function across the mammalian life span.

Cancer cell-secreted IGF2 instigates fibroblasts and bone marrow-derived vascular progenitor cells to promote cancer progression

Local interactions between cancer cells and stroma can produce systemic effects on distant organs to govern cancer progression. Here we show that IGF2 secreted by inhibitor of differentiation (Id1)-overexpressing oesophageal cancer cells instigates VEGFR1-positive bone marrow cells in the tumour macroenvironment to form pre-metastatic niches at distant sites by increasing VEGF secretion from cancer-associated fibroblasts. Cancer cells are then attracted to the metastatic site via the CXCL5/CXCR2 axis. Bone marrow cells transplanted from nude mice bearing Id1-overexpressing oesophageal tumours enhance tumour growth and metastasis in recipient mice, whereas systemic administration of VEGFR1 antibody abrogates these effects. Mechanistically, IGF2 regulates VEGF in fibroblasts via miR-29c in a p53-dependent manner. Analysis of patient serum samples showed that concurrent elevation of IGF2 and VEGF levels may serve as a prognostic biomarker for oesophageal cancer. These findings suggest that the Id1/IGF2/VEGF/VEGFR1 cascade plays a critical role in tumour-driven pathophysiological processes underlying cancer progression.

SKP2 loss destabilizes EZH2 by promoting TRAF6-mediated ubiquitination to suppress prostate cancer.

EZH2 is crucial for the progression of prostate cancer (PCa) and castration-resistant prostate cancer (CRPC) through upregulation and activation of progenitor genes, as well as androgen receptor (AR)-target genes. However, the mechanisms by which EZH2 is regulated in PCa and CRPC remain elusive. Here we report that EZH2 is post-transcriptionally regulated by SKP2 in vitro in cultured cells and in vivo in mouse models. We observed aberrant upregulation of Skp2, Ezh2 and histone H3 lysine 27 trimethylation (H3K27me3) in both Pten null mouse embryonic fibroblasts (MEFs) and Pten null mouse prostate tissues. Loss of Skp2 resulted in a striking decrease of Ezh2 levels in Pten/Trp53 double-null MEFs and in prostate tumors of Pten/Trp53 double-null mutant mice. SKP2 knockdown decreased EZH2 levels in human PCa cells through upregulation of TRAF6-mediated and lysine(K) 63-linked ubiquitination of EZH2 for degradation. Ectopic expression of TRAF6 promoted the K63-linked ubiquitination of EZH2 to decrease EZH2 and H3K27me3 levels in PCa cells. In contrast, TRAF6 knockdown resulted in a reduced EZH2 ubiquitination with an increase of EZH2 and H3K27me3 levels in PCa cells. Furthermore, the catalytically dead mutant TRAF6 C70A abolished the TRAF6-mediated polyubiquitination of recombinant human EZH2 in vitro. Most importantly, a concurrent elevation of Skp2 and Ezh2 was found in CRPC tumors of Pten/Trp53 mutant mice, and expression levels of SKP2 and EZH2 were positively correlated in human PCa specimens. Taken together, our findings revealed a novel mechanism on EZH2 ubiquitination and an important signaling network of SKP2-TRAF6-EZH2/H3K27me3, and targeting SKP2-EZH2 pathway may be a promising therapeutic strategy for CRPC treatment.

Latent classes of emotional and behavioural problems in epidemiological and referred samples and their relations to DSM-IV diagnoses

Researchers’ interest have recently moved toward the identification of recurrent psychopathological profiles characterized by concurrent elevations on different behavioural and emotional traits. This new strategy turned to be useful in terms of diagnosis and outcome prediction. We used a person-centred statistical approach to examine whether different groups could be identified in a referred sample and in a general-population sample of children and adolescents, and we investigated their relation to DSM-IV diagnoses. A latent class analysis (LCA) was performed on the Child Behaviour Checklist (CBCL) syndrome scales of the referred sample (N = 1225), of the general-population sample (N = 3418), and of the total sample. Models estimating 1-class through 5-class solutions were compared and agreement in the classification of subjects was evaluated. Chi square analyses, a logistic regression, and a multinomial logistic regression analysis were used to investigate the relations between classes and diagnoses. In the two samples and in the total sample, the best-fitting models were 4-class solutions. The identified classes were Internalizing Problems (15.68%), Severe Dysregulated (7.82%), Attention/Hyperactivity (10.19%), and Low Problems (66.32%). Subsequent analyses indicated a significant relationship between diagnoses and classes as well as a main association between the severe dysregulated class and comorbidity. Our data suggested the presence of four different psychopathological profiles related to different outcomes in terms of psychopathological diagnoses. In particular, our results underline the presence of a profile characterized by severe emotional and behavioural dysregulation that is mostly associated with the presence of multiple diagnosis.

The role of morphine on rat neural stem cells viability, neuro-angiogenesis and neuro-steroidgenesis properties

A lack of comprehensive data exists on the effect of morphine on neural stem cell neuro-steroidogenesis and neuro-angiogenesis properties. We, herein, investigated the effects of morphine (100μM), naloxone (100μM) and their combination on rat neural stem cells viability, clonogenicity and Ki-67 expression over a period of 72h. Any alterations in the total fatty acids profile under treatment protocols were elucidated by direct transesterification method. We also monitored the expression of p53, aromatase and 5-alpha reductase by real-time PCR assay. To examine angiogenic capacity, in vitro tubulogenesis and the level of VE-cadherin transcript were investigated during neural to endothelial differentiation under the experimental procedure.Cells supplemented with morphine displayed reduced survival (p<0.01) and clonogenicity (p<0.001). Flow cytometric analysis showed a decrease in Ki-67 during 72h. Naloxone potentially blunted morphine-induced all effects. The normal levels of fatty acids, including saturated and unsaturated were altered by naloxone and morphine supplements. Following 48h, the up-regulation of p53, aromatase and 5-alpha reductase genes occurred in morphine-primed cells. Using three-dimensional culture models of angiogenesis and real time PCR assay, we showed morphine impaired the tubulogenesis properties of neural stem cells (p<0.001) by the inhibition of trans-differentiation into vascular cells and led to decrease of in VE-cadherin expression. Collectively, morphine strongly impaired the healthy status of neural stem cells by inducing p53 and concurrent elevation of aromatase and 5-alpha reductase activities especially during early 48h. Also, neural stem cells-being exposed to morphine lost their potency to elicit angiogenesis.

Intermittent Pattern of Detection of Hepatitis B Viral DNA After Achieving Undetectable Viral Levels

Introduction: Intermittent detection of hepatitis B viral DNA after suppression to undetectable viral level raises concerns about inadequate viral suppression, viral breakthrough, resistance, or noncompliance. This retrospective review was conducted in order to characterize the frequency and clinical significance of this laboratory finding, which will be referred to as intermittency pattern (IP). Methods: Patients with hepatitis B seen in an outpatient hepatology practice from April 2015 to 2016 who had been treated or were undergoing treatment with oral nucleos(t)ides were included for review. Patients who did not attain viral suppression < 20 IU/ml by PCR amplification for quantitative HBV DNA were excluded. Results: A total of 61 patients with HBV were identified. 10 did not require treatment, another 10 did not achieve viral suppression to < 20 IU/ml. The final study included 41 patients, ranging in age from 33 to 96 years (median 53), 51% female, and HBeAg positive in 27% at time of initiation of therapy. Thirtyeight (93%) were treated with a regimen that included either tenofovir (N=29) or entecavir (N=8) or both (N=1), the remainder were treated with lamivudine or adefovir (N=3). 83% (N=34) reached undetectable viral load, while the remainder had detectable HBV DNA but < 20 IU/ ml at nadir. Of these 34 patients, 50% (N=17) demonstrated IP, with median of 2 occasions of detectable DNA after reaching undetectable level. In all of these cases, viral load was < 20 IU/ml but detectable on PCR. None had concurrent elevation in ALT or symptoms at times of viral detectability. In those with IP, median age was 55 years, 58% were female, HBeAg was present in 23%, 6% had cirrhosis, and median time to reach viral suppression < 20 IU/ml was 2 years. In those without IP (N=17) median age was 52 years, 41% were female, HBeAg was present in 29%, and 17% had cirrhosis. Conclusion: 50% of patients with long term viral suppression demonstrated IP by PCR. There was a slight female predominance and fewer patients with cirrhosis in patients with IP compared to those without. Possible explanations for the IP phenomenon include low level of viral replication fluctuating around the level of detectability in patients whose virus is adequately suppressed by treatment, incomplete medication compliance, and false positivity of the assay. Given that IP was not associated with concurrent elevation of ALT, symptoms, or quantifiable viral level, IP appears to have little clinical significance.

Calcium-Sensing Receptor and Transient Receptor Ankyrin-1 Mediate Emesis Induction by Deoxynivalenol (Vomitoxin).

The common foodborne mycotoxin deoxynivalenol (DON, vomitoxin) can negatively impact animal and human health by causing food refusal and vomiting. Gut enteroendocrine cells (EECs) secrete hormones that mediate DON's anorectic and emetic effects. In prior work utilizing a cloned EEC model, our laboratory discovered that DON-induced activation of calcium-sensing receptor (CaSR), a G-coupled protein receptor (GPCR), and transient receptor ankyrin-1 (TRPA1), a transient receptor potential (TRP) channel, drives Ca2+-mediated hormone secretion. Consistent with these in vitro findings, CaSR and TRPA1 mediate DON-induced satiety hormone release and food refusal in the mouse, an animal model incapable of vomiting. However, the roles of this GPCR and TRP in DON's emetic effects remain to be determined. To address this, we tested the hypothesis that DON triggers emesis in mink by activating CaSR and TRPA1. Oral gavage with selective agonists for CaSR (R-568) or TRPA1 (allyl isothiocyanate; AITC) rapidly elicited emesis in the mink in dose-dependent fashion. Oral pretreatment of the animals with the CaSR antagonist NPS-2143 or the TRP antagonist ruthenium red (RR), respectively, inhibited these responses. Importantly, DON-induced emesis in mink was similarly inhibited by oral pretreatment with NPS-2143 or RR. In addition, these antagonists suppressed concurrent DON-induced elevations in plasma peptide YY3-36 and 5-hydroxytryptamine-hormones previously demonstrated to mediate the toxin's emetic effects in mink. Furthermore, antagonist co-treatment additively suppressed DON-induced emesis and peptide YY 3-36 release. To summarize, the observations here strongly suggest that activation of CaSR and TRPA1 might have critical roles in DON-induced emesis.

Concurrent elevation of CO2, O3 and temperature severely affects oil quality and quantity in rapeseed.

Plant oil is an essential dietary and bio-energy resource. Despite this, the effects of climate change on plant oil quality remain to be elucidated. The present study is the first to show changes in oil quality and quantity of four rapeseed cultivars in climate scenarios with elevated [CO2], [O3] and temperature (T) combined and as single factors. The combination of environmental factors resembled IPCC's 'business as usual' emission scenario predicted for late this century. Generally, the climate scenarios reduced the average amounts of the six fatty acids (FAs) analysed, though in some treatments single FAs remained unchanged or even increased. Most reduced was the FA essential for human nutrition, C18:3-ω3, which decreased by 39% and 45% in the combined scenarios with elevated [CO2]+T+[O3] and [CO2]+T, respectively. Average oil content decreased 3-17%. When [CO2] and T were elevated concurrently, the seed biomass was reduced by half, doubling the losses in FAs and oil content. This corresponded to a 58% reduction in the oil yield per hectare, and C18:3-ω3 decreased by 77%. Furthermore, the polyunsaturated FAs were significantly decreased. The results indicate undesirable consequences for production and health benefits of rapeseed oil with future climate change. The results also showed strong interactive effects of CO2, T and O3 on oil quality, demonstrating why prediction of climate effects requires experiments with combined factors and should not be based on extrapolation from single factor experiments.

Blood biomarkers of endocrine, immune, inflammatory, and metabolic systems in obstructive sleep apnea

Objective/backgroundObstructive sleep apnea (OSA) is a common disorder, affecting over 100 million adults. Untreated OSA leads to serious health consequences and perturbations in endocrine, immune, inflammatory, and metabolic systems. Study objectives are to evaluate the association between OSA and biomarkers, and to test the hypothesis that a combination of markers may be useful in screening for OSA. Patients/methodsA multicenter trial was conducted enrolling symptomatic male patients with suspected OSA. All subjects underwent in-laboratory overnight polysomnography. A non-symptomatic control group was also obtained. Eleven biomarkers were tested: HbA1c, CRP, EPO, IL-6, uric acid, cortisol, hGH, prolactin, testosterone, DHEA (Beckman Coulter UniCel DxC 600i Synchron® Access® Clinical Systems), IGF-1. Results73 male subjects were enrolled; 26 had moderate/severe OSA. ROC curve analysis showed HbA1c, CRP, EPO, IL-6, and Uric Acid (AUCs: 0.76, 0.73, 0.65, 0.65, 0.61) were superior to the Epworth Sleepiness Scale (AUC: 0.52). Concurrent elevation of HbA1c and CRP provide even greater predictive power. A combination of elevated HbA1c, CRP, and EPO provided 0.08 increase in AUC (0.84 [0.75 - 0.94]) over individual markers (p<0.05), with high sensitivity (85%), and specificity (79%) for moderate/severe OSA. ConclusionsOSA induces characteristic endocrine, immune, inflammatory, and metabolic disturbances that can be detected with blood biomarkers. These biomarkers are superior to standard screening questionnaires. Various clusters of these biomarkers have an even greater association with OSA and thus may represent physiologic signatures of the disorder that may have value in initial screening for OSA as well as for follow-up of therapy response.

Concurrent Elevations of VEGF, Osteopontin and MCP-1 Serum Levels Are Independent Predictors of Survival in Patients with Diffuse Large B-Cell Lymphoma

Background: Diffuse large B-cell lymphomas (DLBCL) are heterogeneous diseases, and the identification of additional DLBCL risk factors is especially important. Methods: In this pilot study, we determined pretreatment serum levels of vascular endothelial growth factor (VEGF), osteopontin (OPN) and macrophage chemotactic protein-1 (MCP-1) in 67 newly diagnosed DLBCL patients before treatment with standard chemoimmunotherapy and in 30 healthy persons. Results: Serum levels of all three cytokines were significantly elevated in untreated patients compared to controls. VEGF and OPN concentrations were higher in patients with advanced Ann Arbor stage, B symptoms, Eastern Cooperative Oncology Group score ≥2, International Prognostic Index (IPI) ≥3 and partial/no remission. A high MCP-1 level was associated with advanced stage, increased IPI and bone marrow infiltration. In univariate analysis, elevated OPN and VEGF, and concurrent elevation of all three biomarkers, were identified as significant predictors of poor survival. Multivariate Cox analysis revealed that elevated OPN combined with elevated VEGF levels was one of the best parameter subsets predicting poorest survival. Conclusion: According to our preliminary results, serum levels of VEGF and OPN before treatment predict response to therapy and survival after chemoimmunotherapy, and may help to further stratify DLBCL patients into risk groups.

Abstract TP440: Heart Failure is Associated With Elevated von Willebrand Factor (vWF) Antigen but not Factor VIII (FVIII) in Acute Ischemic Stroke (AIS)

Introduction: von Willebrand Factor (vWF) and factor VIII (FVIII) contribute to thrombosis and are important risk factors in acute ischemic stroke (AIS). Elevated vWF has been correlated with depressed left ventricular (LV) function, but has not been examined in a cohort of patients with AIS, particularly with regard to concurrent elevation in FVIII. Hypothesis: We hypothesized that vWF is associated with HF and that combined elevation of vWF and FVIII would strengthen this association. Methods: From our prospective stroke registry, AIS patients &gt;18 years of age admitted from 09/2011 to 01/2015 were included if both FVIII and vWF were measured during hospitalization and an echocardiogram was performed. Comparisons were done between the following groups: patients with normal (-) vs. elevated (+) vWF, patients with (-) vs. (+) FVIII, and patients with (-) vs. (+) levels of both factors. Results: Of 1,091 patients in the study period, 212 patients met inclusion criteria. In all groups, patients differed significantly according to history of DM. +vWF had higher frequency of female gender than -vWF, but did not differ according to any other demographic or baseline characteristics. +vWF patients had higher frequency of HF (p=0.007) and higher frequency of depressed LV function (p=0.028) compared to those with -vWF. vWF level correlated with HF (r=0.189, p=0.006) and reduced LV function (r=0.187, p=0.006). Similar findings were found with dual elevation, but we found no relationship between FVIII and HF. After adjustment for HTN, CAD, and DM, vWF remained an independent predictor of HF (OR 1.003, 95%CI 1.000-1.007, p=0.036). While we found no relationship between LV function and FVIII level, median vWF levels increased with decreasing LV function (p=0.027). Conclusion: The results of our study suggest an association between HF and elevated vWF in the setting of AIS, irrespective of FVIII elevation. These results suggest the potential utility of vWF as an independent biomarker for heart failure in the AIS patient population.

Protein kinase CK2 regulates metal toxicity in neuronal cells.

Protein kinase CK2 is a pleiotropic tetrameric enzyme, regulating numerous biological processes from cell proliferation to stress response. This study demonstrates for the first time that CK2 is involved in the regulation of metal uptake and toxicity in neuronal cells. After the determination of inhibitory concentrations (IC50) for a range of metal salts (ZnSO4, Al(mal)3, CoCl2, CrO3, NaAsO2 and CaCl2) in Neuro-2a mouse neuroblastoma cells, the effect of CK2 on metal toxicity was investigated by three lines of experiments using CK2 inhibitors, metal ion specific fluorophores and siRNA-mediated knockdown of CK2 expression. The results showed that both CK2 inhibitors, 4,5,6,7-tetrabromobenzotriazole (TBB) and quinalizarin, markedly reduced the toxicity of Zn(ii), Al(iii), Co(ii), Cr(vi) and As(iii). Confocal microscopy imaging revealed that Zn(ii) uptake was accompanied by the increase of intracellular Ca(ii) in Neuro-2a cells treated with IC50 of ZnSO4 (240 μM), and such concurrent elevation of intracellular Zn(ii) and Ca(ii) was blocked by TBB and quinalizarin. The role of CK2 in metal uptake was further characterised using specific siRNA against each of the three subunits (CK2α, α' and β) and the data demonstrate that CK2α' is the prominent subunit regulating the metal toxicity. Finally, the role of CK2 in metal toxicity was found to be conserved in the distant species-Saccharomyces cerevisiae by employing the complete deletion mutants of CK2 (cka1Δ, cka2Δ, ckb1Δ and ckb2Δ). Taken together, these findings shed light on a new facet of CK2 functionality and provide a basis for further research on the regulation of Zn(ii) and Ca(ii) homeostasis by CK2.

Eugenia caryophyllus Extract Exerts Hypocholesterolemic and Antioxidant Effects in High-Cholesterol-Fed Rats

Background: Eugenia caryophyllus (clove) is an important aromatic household spice. The plant is believed to possess medicinal properties and is commonly used in herbal preparations by traditional healers in the treatment of different ailments and diseases. Objectives: We investigated the lipid-lowering and antioxidant effects of aqueous extract of Eugenia caryophyllus on high-cholesterol-fed rats. Materials and Methods: Cholesterol (40 mg/0.3 mL) was administered to induce hypercholesterolemia in rats by oral gavage, and Eugenia caryophyllus (100 or 200 mg/kg) and Questran (0.26 g/kg) were administered five times a week for eight consecutive weeks. Serum lipid-profiles, lipid peroxidation (LPO), and antioxidant parameters were examined in liver and heart post mitochondrial fraction (PMF). Aspartate and alanine aminotransferase (ALT and AST) activities and liver tissue histology were used to evaluate tissue toxicity. Results: Hypercholesterolemia produced a noticeable decrease in serum HDL-c, whereas a concurrent elevation in serum LDL-c, total cholesterol, and triglycerides as well as serum ALT and AST was observed. Furthermore, hypercholesterolemia remarkably decreased antioxidant status, but LPO content was increased. These indices were significantly attenuated in hypercholesterolemic rats treated with E. caryophyllus extract (100 or 200 mg/kg of body weight). Specifically, reduced glutathione (GSH) concentration was increased in a dose dependent manner in liver and heart PMF. Conclusions: These results demonstrate that the hypolipidemic and antioxidative effects of aqueous extract of E. caryophyllus might be due to its ability to ameliorate lipid profiles, enhance antioxidant activities, and delay the lipid peroxidation process. This confirms the previously identified protective roles of E. caryophyllus in human health.

Abstract A1-58: Revisiting cancer energetics: Establishing a link between altered fatty-acid metabolism and metastasis

Abstract Identifying genetic alterations that drive metastatic transformation of cancer cells with enhanced invasiveness and stem cell-like properties is crucial for developing novel therapeutics. Reprogramming of cellular metabolism is an important hallmark of cancer that sustains uncontrolled growth and proliferation. While alterations resulting in elevated glucose uptake and switch to aerobic glycolysis (Warburg effect) have received much attention in recent years, the relevance of fatty-acid (FA) metabolism in cancer progression has remained unclear. Highly aggressive, metastatic tumors resemble stem cells at the genetic and physiological level, and recent studies have revealed a critical role of FA metabolism in providing energy and maintaining the undifferentiated state of stem cells. Moreover, obesity and concurrent elevation in FA levels has been linked to increased mortality rates in cancer patients. These observations raise the possibility of a multi-faceted link between increased availability of FA, altered FA metabolism and cancer progression. Here we investigated the changes in gene expression , mutations and copy number variations in primary and metastatic tumors using a literature-curated set of genes involved in regulating cellular FA uptake, lipogenesis, FA oxidation and the Warburg-effect. Using the pan-cancer TCGA dataset, we showed that &amp;gt;70% of FA oxidation and lipogenesis genes exhibit a significantly higher mutation frequency in metastatic tumors compared to primary tumors. Moreover, &amp;gt;40% of the genes involved in FA uptake showed a significant copy-number gain in metastatic tumors. Next, we performed an association analysis between the gene expression levels of FA metabolism and Warburg effect genes with markers of metastatic progression in the TCGA RNAseq datasets for breast, colorectal, kidney, liver, lung, ovarian and prostate cancers. To score the metastatic potential of individual tumors, we assigned a previously established epithelial-mesenchymal transition gene signature score (EMT score) and then identified a set of co-expressed FA-metabolism genes in each cancer type that shared a concordant association with EMT score. Next, we applied a regression modeling approach utilizing the FA metabolism and Warburg-effect genes to predict the aggressiveness of tumors in each cancer type. We further demonstrated the ability of the gene signature derived from the regression model to accurately predict the aggressiveness of tumors in independent gene expression datasets. Our study established a metabolic gene signature that can predict metastatic potential and may serve as putative therapeutic targets for aggressive tumors. Importantly, this work highlights the crucial association of genetic alterations in FA metabolism with metastasis. Citation Format: Aritro Nath, Christina Chan. Revisiting cancer energetics: Establishing a link between altered fatty-acid metabolism and metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Translation of the Cancer Genome; Feb 7-9, 2015; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(22 Suppl 1):Abstract nr A1-58.

Topical treatment with coenzyme Q10-containing formulas improves skin's Q10 level and provides antioxidative effects.

Ubiquinone (coenzyme Q10, Q10) represents an endogenously synthesized lipid‐soluble antioxidant which is crucial for cellular energy production but is diminished with age and under the influence of external stress factors in human skin. Here, it is shown that topical Q10 treatment is beneficial with regard to effective Q10 replenishment, augmentation of cellular energy metabolism, and antioxidant effects. Application of Q10‐containing formulas significantly increased the levels of this quinone on the skin surface. In the deeper layers of the epidermis the ubiquinone level was significantly augmented indicating effective supplementation. Concurrent elevation of ubiquinol levels suggested metabolic transformation of ubiquinone resulting from increased energy metabolism. Incubation of cultured human keratinocytes with Q10 concentrations equivalent to treated skin showed a significant augmentation of energy metabolism. Moreover, the results demonstrated that stressed skin benefits from the topical Q10 treatment by reduction of free radicals and an increase in antioxidant capacity. © 2015 BioFactors, 41(6):383–390, 2015

Viral transduction of renin rapidly establishes persistent hypertension in diverse murine strains.

Mice provide a unique platform to dissect disease pathogenesis, with the availability of recombinant inbred strains and diverse genetically modified strains. Leveraging these reagents to elucidate the mechanisms of hypertensive tissue injury has been hindered by difficulty establishing persistent hypertension in these inbred lines. ANG II infusion provides relatively short-term activation of the renin-angiotensinogen system (RAS) with concomitant elevated arterial pressure. Longer-duration studies using renin transgenic mice are powerful models of chronic hypertension, yet are limited by the genetic background on which the transgene exists and the exposure throughout development. The present studies characterized hypertension produced by transduction with a renin-coding adeno-associated virus (ReninAAV). ReninAAV mice experienced elevated circulating renin with concurrent elevations in arterial pressure. Following a single injection of ReninAAV, arterial pressure increased on average +56 mmHg, an increase that persisted for at least 12 wk in three distinct and widely used strains of adult mice: 129/S6, C56BL/6, and DBA/2J. This was accomplished without surgical implantation of pumps or complex breeding and backcrossing. In addition, ReninAAV mice developed pathophysiological changes associated with chronic hypertension, including increased heart weight and albuminuria. Thus ReninAAV provides a unique tool to study the onset of and effects of persistent hypertension in diverse murine models. This model should facilitate our understanding of the pathogenesis of hypertensive injury.

Effect of concurrent elevation of serum creatinine and C-reactive protein values on the long-term outcome in patients with ST-elevation acute myocardial infarction

Effect of concurrent elevation of serum creatinine and C-reactive protein values on the long-term outcome in patients with ST-elevation acute myocardial infarction