Alpha Stimulation Research Articles

Rhythmic low-field magnetic stimulation may improve depression by increasing brain-derived neurotrophic factor.

Low-field magnetic stimulation (LFMS) has mood-elevating effect, and the increase of brain-derived neurotrophic factor (BDNF) is associated with antidepressant treatment. We evaluated the effects and association with BDNF of rhythmic LFMS in the treatment of major depressive disorder (MDD). A total of 22 MDD patients were randomized to rhythmic alpha stimulation (RAS) or rhythmic delta stimulation (RDS), with 5 sessions per week, lasting for 6 weeks. Outcomes assessments included the 17-item Hamilton Depression Rating Scale (HAMD-17), the Hamilton Anxiety Rating Scale (HAMA), and the Clinical Global Impressions-Severity scale (CGI-S) at baseline and at weeks 1, 2, 3, 4, and 6. Serum BDNF level was measured at baseline and at weeks 2, 4, and 6. HAMD-17, HAMA, and CGI-S scores were significantly reduced with both RAS and RDS. RAS patients had numerically greater reductions in HAMD-17 scores than RDS patients (8.9 ± 7.4 vs. 6.2 ± 6.2, effect size [ES]=0.40), while RDS patients had greater improvement in HAMA scores (8.2 ± 8.0 vs. 5.3 ± 5.8, ES=0.42). RAS was associated with clinically relevant advantages in response (54.5% vs. 18.2%, number-needed-to-treat [NNT]=3) and remission (36.4% vs. 9.1%, NNT=4). BDNF increased significantly during the 6-week study period (p<0.05), with greater increases in RAS at weeks 4 and 6 (ES=0.66-0.76) and statistical superiority at week 2 (p=0.034, ES=1.23). Baseline BDNF in the 8 responders (24.8±9.0 ng/ml) was lower than in the 14 nonresponders (31.1±7.3 ng/ml, p=0.083, ES=-0.79), and BDNF increased more in responders (8.9±7.8 ng/ml) than in nonresponders (1.8±3.5 ng/ml, p=0.044). The change in BDNF at week 2 was the most strongly predicted response (p=0.016). Rhythmic LFMS was effective for MDD. BDNF may moderate/mediate the efficacy of LFMS.

Glycosaminoglycan-Mediated Downstream Signaling of CXCL8 Binding to Endothelial Cells.

The recruitment of leukocytes, mediated by endothelium bound chemokine gradients, is a vital process in inflammation. The highly negatively charged, unbranched polysaccharide family of glycosaminoglycans (GAGs), such as heparan sulfate and chondroitin sulfate mediate chemokine immobilization. Specifically the binding of CXCL8 (interleukin 8) to GAGs on endothelial cell surfaces is known to regulate neutrophil recruitment. Currently, it is not clear if binding of CXCL8 to GAGs leads to endothelial downstream signaling in addition to the typical CXCR1/CXCR2 (C-X-C motif chemokine receptor 1 and 2)-mediated signaling which activates neutrophils. Here we have investigated the changes in protein expression of human microvascular endothelial cells induced by CXCL8. Tumor necrosis factor alpha (TNFα) stimulation was used to mimic an inflammatory state which allowed us to identify syndecan-4 (SDC4) as the potential proteoglycan co-receptor of CXCL8 by gene array, real-time PCR and flow cytometry experiments. Enzymatic GAG depolymerization via heparinase III and chondroitinase ABC was used to emulate the effect of glycocalyx remodeling on CXCL8-induced endothelial downstream signaling. Proteomic analyses showed changes in the expression pattern of a number of endothelial proteins such as Zyxin and Caldesmon involved in cytoskeletal organization, cell adhesion and cell mobility. These results demonstrate for the first time a potential role of GAG-mediated endothelial downstream signaling in addition to the well-known CXCL8-CXCR1/CXCR2 signaling pathways in neutrophils.

Cortical nociceptive processes are reduced by visual alpha-band entrainment in the human brain.

Acute noxious stimuli induce a suppression of cortical alpha activity, yet little is known about whether increasing alpha activity affects the processing of noxious stimuli. We have previously shown that visual alpha stimulation reduces experimental pain. Here, we demonstrate that increasing alpha power causes a reciprocal suppression of acute nociceptive processing. We attempted to increase cortical alpha activity through visual entrainment at 8Hz, 10Hz and 12Hz to investigate the influence on the electrophysiological pain response. Moderately painful laser-heat stimuli were delivered following 10minutes of visual entrainment across the alpha range. Alpha power increased significantly relative to the 1Hz control condition following 8Hz and 10Hz visual stimulation. Significant reductions in the P2 peak amplitude of the laser-evoked potential were found following visual entrainment at 10Hz; the frequency stimulation resulting in the largest reduction in pain perception. Source analysis revealed that, following the 10Hz stimulation, sources of increased alpha power and decreased nociceptive processing overlapped in precuneus and posterior cingulate cortex, with further reductions in nociceptive processing in insula cortex. As far as we are aware, this is the first study to provide direct evidence that experimental induction of increased alpha power suppresses the cortical processing of acute pain. While it is known that visual stimulation can increase the brain's oscillatory alpha rhythms, here, we show that this increase in alpha power occurs alongside reduced cortical processing of nociception, as measured with EEG. This establishes an objective marker of alpha entrainment-based analgesia that may be useful in the development of neuromodulatory treatments for clinical pain.

Ovarian Granulosa Cell Tumor: A Review of Clinical Features and Histo-Pathological Findings

Granulosa cell tumor(s) (GCT[s]) of the ovary, representing 5 to 10% of all ovarian malignancies, account for 70% of the ovarian sex-cord stromal tumors. GCTs are classified into two distinct subtypes, including the adult-type GCT and the juvenile-type GCT, and the former occurs more frequently (95%). Currently, evidence from tumor biology found that Forkhead box L2 (FOXL2), a transcription factor gene involved in ovarian development and function, whose expression is reduced and mutated in the majority of adult-type GCTs. However, FOXL2 mutant seems to be rare in juvenile-type GCTs. In addition, researchers have found the stimulatory alpha subunit of a trimeric G protein might be correlated with the prognosis of adult-type GCTs. Similar to the FOXL2 mutant, it is also unclear the precise molecular basis for juvenile-type GCTs. The recent molecular analyses of juvenile-type GCTs identified AKT1 oncogene to play an important role for the development of juvenile-type GCTs, since over 60% of the juvenile-type GCTs show overexpression of AKT1 oncogene. This provides insights into tumor biology and therapeutic leads for juvenile-type GCTs. The aim of this article is to discuss the clinico-pathology and the molecular biology of GCTs.

Nicotinic acetylcholine receptor alpha 7 stimulation dampens splenic myelopoiesis and inhibits atherogenesis in Apoe−/− mice

Background and aimsMonocyte levels predict cardiovascular outcomes and play a causal role in atherogenesis. Monocytes can be produced in the spleen and track to the atherosclerotic lesion in significant numbers. The cholinergic system has been shown to have anti-inflammatory actions in the spleen. We aimed to explore whether therapeutic stimulation of the nicotinic acetylcholine receptor alpha 7 (nAChRα7) can suppress atherogenesis. MethodsApoe−/− mice were placed on a Western-type diet and treated with bi-daily injections of the nAChRα7 agonist GTS-21 or vehicle every 2–3 days for 8 weeks. ResultsGTS-21 caused a reduction in atherosclerosis in the aortic arch and proximal aorta. This also resulted in less plaque macrophages. Moreover, GTS-21 reduced the abundance of blood monocytes, which was caused by inhibition of inflammatory cytokines and extramedullary hematopoiesis in the spleen, along with splenic monocytes. ConclusionsStimulation of nAChRα7 with GTS-21 reduced atherosclerosis, which was associated with dampened splenic myelopoiesis.

Abstract 473: Differential Regulation of Sk Channels by CamKII and Pyk2 Under Adrenergic Stimulation

Introduction: Small conductance calcium (Ca)-activated K (SK) channels present in the plasmalemma of ventricular cardiomyocytes (VCMs) are dormant in health and become functional in cardiac disease. We recently showed that SK channels contribute to VCM repolarization in a rat model of hypertrophy induced by thoracic aortic banding (TAB), despite reduced expression levels, suggestive of increased activity of the channels. However, the mechanisms responsible for disease-associated functional upregulation of the channels are yet to be defined. Hypothesis: Functional recruitment of SK channels in cardiac hypertrophy is caused by enhancement of adrenergic signaling cascades. Methods: The effects of α1 adrenergic stimulation by phenylephrine (10 μM) + propranolol (100 μM) were studied in patch-clamped rat VCMs overexpressing rat SK2 after 48 hrs in culture. Calcium transients were recorded simultaneously using confocal microscopy and were used as a ramp [Ca] to assess possible changes in Ca sensitivity of the channel. Phosphorylation of SKs by Ser/Thr and tyrosine kinases, phosphorylation of Calmodulin at Thr-79, expression levels and phosphorylation of CaMKII and tyrosine kinase Pyk2 were assessed in cultured rat VCMs and freshly isolated VCMs from TABs and Shams using Western blot analysis. Results: WB analysis showed increased Ser/Thr and Tyr phosphorylation of SK2 in TABs. Alpha1 stimulation of rat VCMs overexpressing rat SK2 mimicked this pattern, increasing I SK at high voltages in parallel with the increase in Ca transient amplitude and reduction in Ca sensitivity of the channels. Overexpression of Pyk2 inhibitor CRNK did not restore I SK despite reduction in Ca transient amplitude because of attenuating α1-agonist mediated decrease in Ca sensitivity of SKs. Application of CaMKII inhibitor KN93 fully reversed α1-mediated I SK increase at high voltages without affecting Ca sensitivity of the channel. Conclusion: Upregulation of SKs in hypertrophic hearts is likely caused by CaMKII-dependent phosphorylation which reduces voltage-dependent block of the channels. Pyk2 inhibition may present therapeutic potential by attenuating negative effects on SK Ca sensitivity thereby increasing repolarization reserve diminished in cardiac disease.

NF-κB upregulation through epigenetic silencing of LDOC1 drives tumor biology and specific immunophenotype in Group A ependymoma.

Inflammation has been identified as a hallmark of high-risk Group A (GpA) ependymoma (EPN). Chronic interleukin (IL)-6 secretion from GpA tumors drives an immune suppressive phenotype by polarizing infiltrating monocytes. This study determines the mechanism by which IL-6 is dysregulated in GpA EPN. Twenty pediatric GpA and 21 pediatric Group B (GpB) EPN had gene set enrichment analysis for MSigDB Hallmark gene sets performed. Protein and RNA from patients and cell lines were used to validate transcriptomic findings. GpA cell lines 811 and 928 were used for in vitro experiments performed in this study. The nuclear factor-kappaB (NF-κB) pathway is a master regulator of IL-6 and a signaling pathway enriched in GpA compared with GpB EPN. Knockdown of NF-κB led to significant downregulation of IL-6 in 811 and 928. NF-κB activation was independent of tumor necrosis factor alpha (TNF-α) stimulation in both cell lines, suggesting that NF-κB hyperactivation is mediated through an alternative mechanism. Leucine zipper downregulated in cancer 1 (LDOC1) is a known transcriptional repressor of NF-κB. In many cancers, LDOC1 promoter is methylated, which inhibits gene transcription. We found decreased LDOC1 gene expression in GpA compared with GpB EPN, and in other pediatric brain tumors. EPN cells treated with 5AZA-DC, demethylated LDOC1 regulatory regions, upregulated LDOC1 expression, and concomitantly decreased IL-6 secretion. Stable knockdown of LDOC1 in EPN cell lines resulted in a significant increase in gene transcription of v-rel avian reticuloendotheliosis viral oncogene homolog A, which correlated to an increase in NF-κB target genes. These results suggest that epigenetic silencing of LDOC1 in GpA EPN regulates tumor biology and drives inflammatory immune phenotype.

Platitudes and Attitudes.

Platitudes and Attitudes.

P184 Individual transcranial alternating current stimulation (tACS) within the beta range during verbal working memory

Introduction Beta band brain oscillations (13–30 Hz) correlate with cognitive manipulations during language processing ( Weiss and Muller, 2012 ). In particular, they are associated with the active maintenance of stimuli in verbal working memory ( Weiss and Rappelsberger, 2000 ). Therefore, beta oscillations are suited for tACS during verbal memory encoding. Objectives We examined whether 18 Hz tACS stimulation modulates verbal WM performance in a healthy participant. We did this single-case study since most tACS results are based on group studies and we want to test tACS in individual patients with disturbances of verbal WM. Materials & Methods A right-handed female (27 y) had to memorize 600 visually presented concrete words (150 nouns, 150 verbs; randomly presented two times) and do subsequent free recall. tACS in the beta (18 Hz) and alpha (9 Hz) band as well as a sham stimulation was applied in a double-blind multiple treatment design (ABCBCA) on 6 different days with at least 24 h between sessions. tACS-intensity was 350 μA (below individual phosphene and discomfort threshold). 20 min stimulation was applied at left temporo-parietal and right fronto-central sites. Results A significant difference between the stimulation and sham condition (Pearson χ2 = 12.0, p = 0.03, two-tailed) was found which differed for nouns and verbs. During beta stimulation the participant showed a 48% increase of recalled nouns (8% for verbs), during alpha stimulation a 28% (26 % for verbs) and during sham a 20% (36% for verbs) increase. Overall error rate was higher for verbs (Pearson χ2 = 11.73, p = 0.008, two-tailed). The influence of beta vs. alpha stimulation could not be reliably distinguished with the current protocol. Conclusion Although, beta oscillations are important for linking past and present input into a coherent representation of memorized contents ( Weiss and Rappelsberger, 2000 , Kopell and Whittington, 2011 ), no significant difference was found between alpha and beta stimulation. The word category difference is in accordance with previous findings ( Fiori and Cipollari, 2013 ) who revealed better performance for verb versus noun processing when stimulating left frontal in contrast to temporal areas.

Spatial Attention and the Effects of Frontoparietal Alpha Band Stimulation.

A frontoparietal network has long been implicated in top-down control of attention. Recent studies have suggested that this network might communicate through coherence in the alpha band. Here we aimed to test the effect of coherent alpha (8–12 Hz) stimulation on the frontoparietal network. To this end, we recorded behavioral performance and electroencephalography (EEG) data while participants were engaged in a spatial attention task. Furthermore, participants received transcranial alternating current stimulation (tACS) over the right frontal and parietal cortex, which oscillated coherently in-phase within the alpha band. Compared to a group of participants that received sham stimulation, we found that coherent frontoparietal alpha band stimulation altered a behavioral spatial attention bias. Neurally, the groups showed hemispheric-specific differences in alpha coherence between the frontal and parietal-occipital cortex. These results provide preliminary evidence that alpha coherence in the frontoparietal network might play a role in top-down control of spatial attention.

SPRY1 regulates mammary epithelial morphogenesis by modulating EGFR-dependent stromal paracrine signaling and ECM remodeling

The role of the local microenvironment in influencing cell behavior is central to both normal development and cancer formation. Here, we show that sprouty 1 (SPRY1) modulates the microenvironment to enable proper mammary branching morphogenesis. This process occurs through negative regulation of epidermal growth factor receptor (EGFR) signaling in mammary stroma. Loss of SPRY1 resulted in up-regulation of EGFR-extracellular signal-regulated kinase (ERK) signaling in response to amphiregulin and transforming growth factor alpha stimulation. Consequently, stromal paracrine signaling and ECM remodeling is augmented, leading to increased epithelial branching in the mutant gland. By contrast, down-regulation of EGFR-ERK signaling due to gain of Sprouty function in the stroma led to stunted epithelial branching. Taken together, our results show that modulation of stromal paracrine signaling and ECM remodeling by SPRY1 regulates mammary epithelial morphogenesis during postnatal development.

Differential effects of 10-Hz and 40-Hz transcranial alternating current stimulation (tACS) on endogenous versus exogenous attention

ABSTRACT Previous electrophysiological studies implicate both alpha (8–12 Hz) and gamma (>30 Hz) neural oscillations in the mechanisms of selective attention. Here, participants preformed two separate visual attention tasks, one endogenous and one exogenous, while transcranial alternating current stimulation (tACS), at 10 Hz, 40 Hz, or sham, was applied to the right parietal lobe. Our results provide new evidence for the roles of gamma and alpha oscillations in voluntary versus involuntary shifts of attention. Gamma (40 Hz) stimulation resulted in improved disengagement from invalidly cued targets in the endogenous attention task, whereas alpha stimulation (10 Hz) had no effect on endogenous attention, but increased the exogenous cuing effect. These findings agree with previous studies suggesting that right inferior parietal regions may be especially important for the disengagement of attention, and go further to provide details about the specific type of oscillatory neural activity within that brain region that is differentially involved in endogenous versus exogenous attention. Our results also have potential implications for the plasticity and training of attention systems.

Combined assessment of polymorphisms in the LHCGR and FSHR genes predict chance of pregnancy after in vitro fertilization.

Can gonadotrophin receptor variants separately or in combination, be used for the prediction of pregnancy chances in in vitro fertilization (IVF) trials? The luteinizing hormone/human chorionic gonadotrophin receptor (LHCGR) variant N312S and the follicle-stimulating hormone receptor (FSHR) variant N680S can be utilized for the prediction of pregnancy chances in women undergoing IVF. The FSHR N680S polymorphism has been shown to affect the ovarian response in response to gonadotrophin treatment, while no information is currently available regarding variants of the LHCGR in this context. Cross-sectional study, duration from September 2010 to February 2015. Women undergoing IVF were consecutively enrolled and genetic variants compared between those who became pregnant and those who did not. The study was subsequently replicated in an independent sample. Granulosa cells from a subset of women were investigated regarding functionality of the genetic variants. Women undergoing IVF (n = 384) were enrolled in the study and genotyped. Clinical variables were retrieved from medical records. For replication, an additional group of n = 233 women was utilized. Granulosa cells from n = 135 women were isolated by flow cytometry, stimulated with Follitropin alpha or Menotropin, and the downstream targets 3',5'-cyclic adenosine monophosphate (cAMP) and inositol 1,4,5-trisphosphate (IP3) measured with enzyme-linked immunosorbent assay. Women homozygous for serine (S) in both polymorphisms displayed higher pregnancy rates than women homozygous asparagine (N) (OR = 14.4, 95% CI: [1.65, 126], P = 0.016). Higher pregnancy rates were also evident for women carrying LHCGR S312, regardless of FSHR variant (OR = 1.61, 95% CI: [1.13, 2.29], P = 0.008). These women required higher doses of FSH for follicle recruitment than women homozygous N (161 versus 148 IU, P = 0.030). When combining the study cohort with the replication cohort (n = 606), even stronger associations with pregnancy rates were noted for the combined genotypes (OR = 11.5, 95% CI: [1.86, 71.0], P = 0.009) and for women carrying LHCGR S312 (OR = 1.49, 95% CI: [1.14, 1.96], P = 0.004). A linear significant trend with pregnancy rate and increasing number of G alleles was also evident in the merged study population (OR = 1.34, 95% CI: [1.10, 1.64], P = 0.004). A lower cAMP response in granulosa cells was noted following Follitropin alpha stimulation for women homozygous N in both polymorphisms, compared with women with other genotypes (0.901 pmol cAMP/mg total protein versus 2.19 pmol cAMP/mg total protein, P = 0.035). Due to racial differences in LHCGR genotype distribution, these results may not be applicable for all populations. Despite that >250 000 cycles of gonadotrophin stimulations are performed annually worldwide prior to IVF, it has not been possible to predict neither the pregnancy outcome, nor the response to the hormone with accuracy. If LHCGR and FSHR variants are recognized as biomarkers for chance of pregnancy, more individualized and thereby more efficient treatment modalities can be developed. This work was supported by Interreg IV A, EU (grant 167158) and ALF governments grant (F2014/354). Merck-Serono (Darmstadt, Germany) supported the enrollment of the subjects. The authors declare no conflict of interest.

Pamidronate Down-regulates Tumor Necrosis Factor-alpha Induced Matrix Metalloproteinases Expression in Human Intervertebral Disc Cells.

BackgroundN-containing bisphosphonates (BPs), such as pamidronate and risedronate, can inhibit osteoclastic function and reduce osteoclast number by inducing apoptotic cell death in osteoclasts. The aim of this study is to demonstrate the effect of pamidronate, second generation nitrogen-containing BPs and to elucidate matrix metallo-proteinases (MMPs) mRNA expression under serum starvation and/or tumor necrosis factor alpha (TNF-α) stimulation on metabolism of intervertebral disc (IVD) cells in vitro.MethodsFirstly, to test the effect of pamidronate on IVD cells in vitro, various concentrations (10-12, 10-10, 10-8, and 10-6 M) of pamidronate were administered to IVD cells. Then DNA and proteoglycan synthesis were measured and messenger RNA (mRNA) expressions of type I collagen, type II collagen, and aggrecan were analyzed. Secondly, to elucidate the expression of MMPs mRNA in human IVD cells under the lower serum status, IVD cells were cultivated in full serum or 1% serum. Thirdly, to elucidate the expression of MMPs mRNA in IVD cells under the stimulation of 1% serum and TNF-α (10 ng/mL) In this study, IVD cells were cultivated in three dimensional alginate bead.ResultsUnder the lower serum culture, IVD cells in alginate beads showed upregulation of MMP 2, 3, 9, 13 mRNA. The cells in lower serum and TNF-α also demonstrated upregulation of MMP-2, 3, 9, and 13 mRNA. The cells with various doses of pamidronate and lower serum and TNF-α were reveled partial down-regulation of MMPs.ConclusionsPamidronate, N-containing second generation BPs, was safe in metabolism of IVD in vitro maintaining chondrogenic phenotype and matrix synthesis, and down-regulated TNF-α induced MMPs expression.

A Novel T55A Variant of Gs α Associated with Impaired cAMP Production, Bone Fragility, and Osteolysis.

G-protein coupled receptors (GPCRs) mediate a wide spectrum of biological activities. The GNAS complex locus encodes the stimulatory alpha subunit of the guanine nucleotide binding protein (Gsα) and regulates production of the second messenger cyclic AMP (cAMP). Loss-of-function GNAS mutations classically lead to Albright's Hereditary Osteodystrophy (AHO) and pseudohypoparathyroidism, often with significant effects on bone formation and mineral metabolism. We present the case of a child who exhibits clinical features of osteolysis, multiple childhood fractures, and neonatal SIADH. Exome sequencing revealed a novel de novo heterozygous missense mutation of GNAS (c.163A<G, p.T55A) affecting the p-loop of the catalytic Gsα GTPase domain. In order to further assess whether this unique mutation resulted in a gain or loss of function of Gsα, we introduced the mutation into a rat GNAS plasmid and performed functional studies to assess the level of cAMP activity associated with this mutation. We identified a 64% decrease in isoproterenol-induced cAMP production in vitro, compared to wild type, consistent with loss of Gsα activity. Despite a significant decrease in isoproterenol-induced cAMP production in vitro, this mutation did not produce a classical AHO phenotype in our patient; however, it may account for her presentation with childhood fractures and osteolysis.

Skeletal muscle interleukin 15 promotes CD8(+) T-cell function and autoimmune myositis.

BackgroundInterleukin 15 (IL-15) is thought to be abundant in the skeletal muscle under steady state conditions based on RNA expression; however, the IL-15 RNA level may not reflect the protein level due to post-transcriptional regulation. Although exogenous protein treatment and overexpression studies indicated IL-15 functions in the skeletal muscle, how the skeletal muscle cell uses IL-15 remains unclear. In myositis patients, IL-15 protein is up-regulated in the skeletal muscle. Given the supporting role of IL-15 in CD8+ T-cell survival and activation and the pathogenic role of cytotoxic CD8+ T cells in polymyositis and inclusion-body myositis, we hypothesize that IL-15 produced by the inflamed skeletal muscle promotes myositis via CD8+ T cells.MethodsExpression of IL-15 and IL-15 receptors at the protein level by skeletal muscle cells were examined under steady state and cytokine stimulation conditions. The functions of IL-15 in the skeletal muscle were investigated using Il15 knockout (Il15−/−) mice. The immune regulatory role of skeletal muscle IL-15 was determined by co-culturing cytokine-stimulated muscle cells and memory-like CD8+ T cells in vitro and by inducing autoimmune myositis in skeletal-muscle-specific Il15−/− mice.ResultsWe found that the IL-15 protein was not expressed by skeletal muscle cells under steady state condition but induced by tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) stimulation and expressed as IL-15/IL-15 receptor alpha (IL-15Rα) complex. Skeletal muscle cells expressed a scanty amount of IL-15 receptor beta (IL-15Rβ) under either conditions and only responded to a high concentration of IL-15 hyperagonist, but not IL-15. Consistently, deficiency of endogenous IL-15 affected neither skeletal muscle growth nor its responses to TNF-α and IFN-γ. On the other hand, the cytokine-stimulated skeletal muscle cells presented antigen and provided IL-15 to promote the effector function of memory-like CD8+ T cells. Genetic ablation of Il15 in skeletal muscle cells greatly ameliorated autoimmune myositis in mice.ConclusionsThese findings together indicate that skeletal muscle IL-15 directly regulates immune effector cells but not muscle cells and thus presents a potential therapeutic target for myositis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13395-015-0058-2) contains supplementary material, which is available to authorized users.

The effect of simvastatin treatment on endothelial cell response to shear stress and tumor necrosis factor alpha stimulation.

BackgroundStatin drugs are one of the most commonly prescribed pharmaceuticals by physicians. By blocking the rate-limiting step in the cholesterol biosynthesis pathway, statins inhibit cholesterol synthesis, which benefits patient health. However, since many other important cellular processes are regulated within this pathway, they may also be influenced by statin therapy. These pleiotropic effects of statins have not been fully investigated, but are believed to positively influence endothelial cells (ECs), which line the vasculature in a confluent monolayer. Few studies have considered the effect of blood flow on ECs and how this may augment EC response to statins.MethodsIn this study, the effect of statin treatment on ECs is investigated for cells stimulated with tumor necrosis factor alpha (TNF-α), an inflammatory cytokine that promotes an atheroprone endothelium. Additionally, ECs are exposed to a physiologically relevant wall shear stress (WSS) of 12.5 dynes/cm2 using a three-dimensional tissue culture model to provide a realistic hemodynamic environment. ECs are analyzed for morphology using light microscopy as well as cytoskeletal structure and alignment using confocal microscopy. Statistical analysis is performed on the results using both the one-way analysis of variance with Bonferroni post-tests and the two-tailed t test.ResultsWe have shown that statin treatment caused cells to adapt to a rounded, atheroprone morphology, with a significantly higher shape index. Oppositely, TNF-α stimulation caused cells to elongate to an atheroprotective morphology, with a significantly lower shape index. WSS and TNF-α were unable to reverse any statin-induced cell rounding or F-actin disruption.ConclusionFurther work is therefore needed to determine why statin drugs cause cells to have an atheroprone morphology, but an atheroprotective genotype, and why TNF-α stimulation causes an atheroprotective morphology, but an atheroprone genotype. Despite the morphological changes due to statins or stimulation, ECs still respond to WSS. Understanding how statins influence ECs will allow for more targeted treatments for hypercholestemia and potentially other diseases.

PPARα Stimulation Modulates Myocardial Ischemia-induced Activation of Renin-Angiotensin System.

We have recently demonstrated that peroxisome proliferator-activated receptor alpha (PPARα) stimulation lowers the production of angiotensin II while increasing the production of Ang-(1-7), both in cardiac and plasmatic level. This stimulation improves nitric oxide bioavailability, preserving cardiac histologic features and functioning. Based on these results, we decided to study the effect of PPARα stimulation on renin-angiotensin system components of ischemic myocardium. Male Wistar rats (weighing 300-350 g) were assigned to the following groups: (1) sham, (2) myocardial ischemia vehicle-treated (MI-V), and (3) myocardial ischemia clofibrate-treated. Expression of the angiotensin-converting enzyme increased during ischemia, whereas clofibrate-treated group remained comparable to control. Activation of the PPARα receptor stimulated the expression of angiotensin-converting enzyme-2; while the activity of this enzyme was increased in MI-V, clofibrate inhibited any change. The concentration of bradykinin and phospho-Akt(SER473) in homogenate increased in the animals treated with the drug. Mas receptor expression increased in MI-V rats. In conclusion, stimulation of PPARα by clofibrate prevents an increase in the activity of renin-angiotensin system and promotes the production of vasodilator substances.

Antidiabetic property of Aerva lanata (L.) Juss. ex Schult. is mediated by inhibition of alpha glucosidase, protein glycation and stimulation of adipogenesis.

Diabetes is the leading cause of morbidity and mortality, with a number currently diagnosed as high as 371 million. Plant-based therapy could be an ideal choice because of fewer side-effects and wider acceptability. Hence, the antihyperglycemic potential of Aerva lanata, a herb prescribed for diabetes in Ayurveda was evaluated to elucidate its possible mechanism of action. High performance liquid chromatography analysis was used for the characterization of 70% ethanolic (aqueous leaf extract [ALE]) and ethyl acetate (AEA) extracts. Further, they were evaluated for their antioxidant, inhibition of alpha glucosidase, protein glycation dipeptidyl peptidase IV (DPP IV), protein tyrosine phosphatase 1B (PTP1B) and stimulation of glucose uptake and glitazone like property (adipogenic potential) using in vitro models. The promising alpha glucosidase inhibitory potential of ALE was further evaluated in normal and streptozotocin (STZ) diabetic rats. ALE inhibited yeast (IC50 - 81.76 μg/mL) and rat intestinal alpha glucosidase (IC50 - 108.7 μg/mL), protein glycation, DPP IV enzyme (IC50 - 118.62 μg/mL) and PTP1B (IC50 - 94.66 μg/mL). ALE stimulated maximal adipogenesis at 50 μg/mL and enhanced insulin mediated glucose uptake (threefold of basal) at 100 μg/mL in L6 myotubes. ALE (500 mg/kg b.w.) showed a significant antihyperglycemic activity in sucrose loaded STZ normal (15.57%) and diabetic (18.44%) rats. HPLC analysis of ALE revealed the presence of bioactives like alpha amyrin, betulin and beta sitosterol. Alpha glucosidase inhibition, antiglycation, and adipogenic potential significantly contribute to the antidiabetic property of Aerva lanata. In addition, insulin sensitization and antioxidant potential also enhance its therapeutic potential.

Novel microdeletions affecting the GNAS locus in pseudohypoparathyroidism: characterization of the underlying mechanisms.

Pseudohypoparathyroidism type Ia (PHP1A) is a rare endocrine disorder characterized by hypocalcemia, hyperphosphatemia, multiple hormonal resistance, and features of Albright hereditary osteodystrophy. When the phenotype is present but not associated with hormonal resistance, it is called psedopseudohypoparathyroidism (PPHP). Both entities have been associated to GNAS haploinsufficiency, and are mostly caused by inherited inactivating mutations at GNAS gene that codes for the stimulatory alpha subunit of G protein, although the cause remains unidentified in approximately 30% of patients. The aims of our work were 1) to identify GNAS locus defects in 112 patients with clinical diagnosis of PHP1A/PPHP and no point mutations at GNAS, to improve molecular diagnostic and genetic counseling; 2) to outline the underlying molecular mechanism(s). Methylation-specific-multiplex ligation-dependent probe amplification, qPCR, array comparative genomic hybridization, and long-PCR were used to search for genomic rearrangements at chromosome 20q and to identify their boundaries. We used different bioinformatic approaches to assess the involvement of the genomic architecture in the origin of the deletions. We discovered seven novel genomic deletions, ranging from 106-bp to 2.6-Mb. The characterization of five of seven deletion breakpoints and the definition of the putative molecular mechanisms responsible for these rearrangements revealed that Alu sequences play a major role in determining the genetic instability of the region. We observed that deletions at GNAS locus represent a significant cause of PPHP/PHP1A and that such defects are mostly associated with Alu-mediated recombination events. Their investigation revealed to be fundamental as, in some cases, they could be misdiagnosed as imprinting defects.