Changes In Related Proteins Research Articles

Resveratrol improves glucose uptake in insulin-resistant adipocytes via Sirt1

Insulin resistance serves as “common soil” for promoting the development of metabolic diseases; however, the precise pathological factors leading to insulin resistance are not well clarified. Resveratrol (Res) is a natural polyphenolic compound with anti-inflammatory and antioxidative effects. However, effects and mechanisms of Res on glucose metabolism in adipocytes remain largely unknown. In this study, we show Res treatment significantly increases glucose uptake in insulin-resistant 3T3-L1 adipocytes in vitro. Mechanistically, Res up-regulates the protein level of Sirt1 that improves insulin signaling pathway and promotes cellular membrane Glut4 accumulation. Meanwhile, Sirt1 enhances phosphorylation level of AMPK which elevates p-AKT level. Consequently, the transcription factor FOXO1 translocalizes from nucleus to cytoplasm where protein degradation occurs. Therefore, the gene expression of resistin, a direct transcriptional target of FOXO1, is reduced and insulin sensitivity is improved. Importantly, we recapitulate the similar pattern of related protein changes in epididymal adipose tissues of insulin-resistant mice after Res intervention in vivo, reinforcing the hypothesis of Res being involved in regulation of glucose uptake via Sirt1–AMPK axis. Our findings clarify the beneficial effects of Res on glucose transportation in insulin-resistant adipocytes and involved pathway including Sirt1-AMPK, suggesting its potential therapeutic application in the treatment or prevention of insulin-resistance-related metabolic symptoms.

Protective Effect of Right Ventricular Mitochondrial Damage by Cyclosporine A in Monocrotaline-induced Pulmonary Hypertension.

Background and ObjectivesMitochondria play a key role in the pathophysiology of heart failure and mitochondrial permeability transition pore (MPTP) play a critical role in cell death and a critical target for cardioprotection. The aim of this study was to evaluate the protective effects of cyclosporine A (CsA), one of MPTP blockers, and morphological changes of mitochondria and MPTP related proteins in monocrotaline (MCT) induced pulmonary arterial hypertension (PAH).MethodsEight weeks old Sprague-Dawley rats were randomized to control, MCT (60 mg/kg) and MCT plus CsA (10 mg/kg/day) treatment groups. Four weeks later, right ventricular hypertrophy (RVH) and morphological changes of right ventricle (RV) were done. Western blot and reverse transcription polymerase chain reaction (RT-PCR) for MPTP related protein were performed.ResultsIn electron microscopy, CsA treatment prevented MCT-induced mitochondrial disruption of RV. RVH was significantly increased in MCT group compared to that of the controls but RVH was more increased with CsA treatment. Thickened medial wall thickness of pulmonary arteriole in PAH was not changed after CsA treatment. In western blot, caspase-3 was significantly increased in MCT group, and was attenuated in CsA treatment. There were no significant differences in voltage-dependent anion channel, adenine nucleotide translocator 1 and cyclophilin D expression in western blot and RT-PCR between the 3 groups.ConclusionsCsA reduces MCT induced RV mitochondrial damage. Although, MPTP blocking does not reverse pulmonary pathology, it may reduce RV dysfunction in PAH. The results suggest that it could serve as an adjunctive therapy to PAH treatment.

Effect and mechanism of SS31 peptide on autophagy after spinal cord injury in rats

Objective To explore the effect of SS31 peptide on autophagy after spinal cord injury (SCI) and possible mechanism. Methods Allen method was used to construct the spinal cord injury model in rats. Sprague-Dawley rats were randomly divided into sham surgery group (sham group), SCI group and SS31 peptide group, with 30 rats in each group. The sham group only received laminectomy. The rats in SCI group were sustained SCI and were given no intervention. The rats in SS31 group received SS31 peptide injection after SCI. Scores of Basso Beattie Bresnahan (BBB) motor functions were assessed at 6 h, 1, 3, 7 and 14 d after the injury. The changes in related proteins of Beclin-1 and LC3-II were also detected. Results Scores of BBB scale at 6 h and at days 1, 3, 7 and 14 after injury in SCI group (0, 1.7±0.4, 3.5±0.6, 6.1±0.7, 10.1±0.6) and SS31 peptide group (0, 2.5±0.7, 4.1±0.7, 9.3±0.6, 13.4±0.6) were lower than that in sham group (21 at all time points) (P 0.05). Compared with sham group, the expression of Beclin-1 in SCI group and SS31 peptide group was increased, reached a peak at day 3 (1.478±0.030, 1.841±0.051), remained high level at day 7 (1.302±0.049, 1.551±0.032) and showed high expression at day 14 (1.252±0.048, 1.471±0.062) (P 0.05). Conclusion SS31 peptide can improve motor function and enhance the autophagy of nerve cells after SCI in rats, which may be one of the mechanisms for SS31 peptide treating spinal cord injury. Key words: Spinal cord injuries; Autophagy; Beclin-1

PTEN Gene Induces Cell Invasion and Migration via Regulating AKT/GSK-3β/β-Catenin Signaling Pathway in Human Gastric Cancer.

Abnormality of PTEN gene and Wnt/β-catenin signaling have been strongly implicated in various malignant cancers. Recently, it has been noted that a functional interaction/cross-talk was found between the PTEN/PI3K/AKT and Wnt/β-catenin, which plays a key role in the development of cancers. However, few related studies on gastric cancer are available. We examined the expression of PTEN and β-catenin in gastric cancer tissues and detected whether down-regulation of PTEN promotes the migration and invasion in gastric cancer cells along with its underlying mechanism. Immunocytochemistry, a wound healing assay, a Matrigel invasion assay, an immunofluorescence staining were performed to detect expression of PTEN and β-catenin in gastric cancer and adjacent normal tissues, cell migration, cell invasion, and the effects of PTEN knockdown on β-catenin in cells, respectively. Further, MMP-2 and MMP-9 activities were analyzed by zymography assay. The changes in related proteins were further quantified by western blotting. Low expression of PTEN was found in majority of gastric cancer tissues, which showed significant associations with differentiation grade in gastric cancer patients. Further, a negative correlation was revealed between PTEN and β-catenin protein expression in gastric cancer tissues (r=- 0.546, P<0.01). Additionally, PTEN knockdown promoted the migration and invasion of cells and caused an obvious increase in p-AKT, p-GSK-3β, β-catenin, E-cadherin, MMP-7, MMP-2, and MMP-9 in gastric cancer cells. Our results indicated PTEN gene might induce cell invasion and migration via regulating AKT/GSK-3β/β-catenin signaling pathway, playing a vital role in the progression of gastric cancer.

Effects of chlorogenic acid on capacity of free radicals scavenging and proteomic changes in postharvest fruit of nectarine.

To study how chlorogenic acid affects changes of reactive oxygen species (ROS) and the proteins involved in ROS scavenging of nectarine during storage time, the fruits were treated with chlorogenic acid (CHA) then stored at 25°C for further studies. The CHA-treatment significantly reduced O2-· production rate, H2O2 content, and membrane permeability of nectarine fruit during storage. The key proteins related the nectarine fruit senescence during storage were identified by two-dimensional electrophoresis and MALDI-TOF/TOF. Level and enzymatic activity of peroxidase were reduced, while both the protein levels and the enzymatic activities of superoxide dismutase, glutathione reductase, glutathione-s-transferase and monodehydroascorbate reductase were enhanced in nectarine fruit treated with CHA. In addition, levels of several pathogen-related proteins were also enhanced by CHA-treatment. Taking together, the present study showed that CHA could influence changes in defense related proteins and reduced oxidative damage in nectarine fruit during postharvest ripening.

The dynamic changes of interferon lambdas related genes and proteins in JAK/STAT pathway in both acute and chronic HIV-1 infected patients

BackgroundHost immune responses during acute HIV-1 infection can influence the viral setpoint, which is a predictor of disease progression. Interferon (IFN)-lambdas are newly classified type III interferons, which use JAK-STAT pathway. Currently, the dynamics of IFN-lambdas related genes and proteins expression in the signaling pathway have not been well elaborated, especially in acute HIV-1-infected patients.ObjectivesTo evaluate the dynamic changes of IFN-lambdas related genes and proteins in JAK/STAT pathway in acute HIV-1-infected patients, and analyze their correlation with CD4 T cell counts and HIV-1 viral loads.Study designReal-time PCR and flow cytometry methods were used to evaluate the dynamic changes of IFN-lambdas related genes and proteins in JAK/STAT pathway in both acute and chronic HIV-1-infected patients.ResultsThe IFN-alpha receptors (R), IFN-gamma R, IFN-lambdas R and STAT1 mRNA and protein levels increased in acute HIV-1-infected patients (p < 0.01), in addition, Mx1 mRNA levels in acute HIV-1-infected patients are higher than those in HIV-negative subjects. IFN-lambdas R and IFN-alpha R mRNA levels are inversely correlated with CD4+ T-cell counts, but are positively correlated with viral loads.ConclusionsThe dynamic changes of IFNs related genes in JAK-STAT pathway in acute HIV-1 infection will deepen our understanding of the roles of IFN-lambdas in HIV pathogenesis.

Melatonin alleviates brain and peripheral tissue edema in a neonatal rat model of hypoxic-ischemic brain damage: the involvement of edema related proteins

BackgroundPrevious studies have indicated edema may be involved in the pathophysiology following hypoxic-ischemic encephalopathy (HIE), and melatonin may exhibit neuro-protection against brain insults. However, little is known regarding the mechanisms that involve the protective effects of melatonin in the brain and peripheral tissues after HIE. The present study aimed to examine the effects of melatonin on multiple organs, and the expression of edema related proteins in a neonatal rat model of hypoxic-ischemic brain damage (HIBD).MethodsOne hundred ninety-two neonatal rats were randomly divided into three subgroups that underwent a sham surgery or HIBD. After the HIBD or sham-injury, the rats received an intraperitoneal injection of melatonin or an equal volume vehicle, respectively. We investigated the effects of melatonin on brain, kidney, and colon edema via histological examination and the expression of edema related proteins, including AQP-4, ZO-1 and occludin, via qPCR and western blot.ResultsOur data indicated (1) Melatonin reduced the histological injury in the brain and peripheral organs induced by HIBD as assessed via H-E staining and transmission electron microscopy. (2) Melatonin alleviated the HIBD-induced cerebral edema characterized by increased brain water content. (3) HIBD induced significant changes of edema related proteins, such as AQP-4, ZO-1 and occludin, and these changes were partially reversed by melatonin treatment.ConclusionsThese findings provide substantial evidence that melatonin treatment has protective effects on the brain and peripheral organs after HIBD, and the edema related proteins, AQP4, ZO-1, and occludin, may indirectly contribute tothe mechanism of the edema protection by melatonin.

BMP‑7 suppresses excessive scar formation by activating the BMP‑7/Smad1/5/8 signaling pathway.

Scarring is the inevitable consequence of wound repair, which may cause significant physical and mental pain to patients when excessive. Bone morphogenetic protein-7 (BMP-7) has been proved to inhibit TGF-β-induced fibrosis in various tissues including dermal papilla cells. However, the effect of BMP-7 on hypertrophic scarring, a common proliferative disorder of dermal fibroblasts, has not been determined. To overcome this problem, the present study established a mouse model of thermal injury to investigate the inhibitory effects of BMP-7 on scar formation. The histological analysis of scar tissues was performed by H&E and Masson's trichrome staining. Western blot assay was used to determine the level changes of related proteins and TUNEL assay was performed to assess the apoptosis of scar tissues. The results demonstrated that BMP-7 promoted wound healing and inhibited scar formation when compared with untreated mice. Collagen deposition and the expression of fibrotic proteins were suppressed in the scar tissues of mice treated with BMP-7. In addition, BMP-7 induced fibroblast apoptosis in scar tissues. Furthermore, activation of the BMP-7/Smad1/5/8 signaling pathway may have been involved in the inhibitory effects of BMP-7 on scar formation. In conclusion, the results of the present study indicate that BMP-7 may inhibit excessive scar formation via activation of the BMP-7/Smad1/5/8 signaling pathway. The results present a potential alternative therapeutic strategy for the treatment of hypertrophic scarring.

Mechansim of resveratrol inhibiting interleukin-6-induced invasion of human gastric cancer SGC7901 cells

Objective To elucidate the inhibitory effect of resveratrol on interleukin (IL)-6-induced invasive ability of SGC7901 cells and the molecular mechanism. Methods The cell viability of cells treated with resveratrol was evaluated by methyl thiazolyltetrazolium (MTT) assay. The SGC7901 cells were treated with IL-6 and non-cytotoxic concentration of resveratrol. The invasion of SGC7901 cells was detected by Boyden chamber assay. Matrix metalloproteinase (MMP)-2 activity was determined by gelatine zymography assay. The changes of the related proteins were detected by Western blotting. Results Resveratrol was not toxic to SGC7901 cells at the concentration under 30 μmol/L. The invasion ability of IL-6-induced SGC7901 cells was decreased about 20% after treatment with 20 μmol/L resveratrol for 24 h, accompanied by the inhibition of MMP-2 secretion for 50%, as well as the levels of phosphorylated extracellular regulated protein kinases 1/2 (p-ERK1/2). Conclusion Resveratrol of 20 μmol/L inhibits invasion of SGC7901 cells possibly through the suppression of the activation of ERK signaling pathway, subsequently exerting inhibitory effect on MMP-2. Key words: Resveratrol; Extracellular signal regulated kinase 1/2; Gastric carcinoma

Dynamic Changes in Starch Metabolism and Endogenous Hormones during Stolon Formation in Tulipa edulis

The stolon is the main asexual reproductive organ of the medicinal plant Tulipa edulis and has special morphology. In the greenhouse experiment presented herein, the dynamic changes in carbohydrates and related enzymes, proteins, and endogenous hormones of stolons during T. edulis stolon formation were investigated. The results showed that the content of total soluble sugar, sucrose, reducing sugar, fructose, and starch were all significantly enhanced in the middle period when stolon emerged and maintained at relatively high levels until the later period of stolon formation, while protein content decreased during stolon formation. The activities of amylase (AMY), sucrose phosphate synthase (SPS), and sucrose synthase (SS) peaked in the initial period and were negatively correlated with soluble sugars. However, adenosine diphosphoglucose pyrophosphorylase (AGPase) activity increased as stolon formation progressed, and the changes in soluble starch synthase (SSS), granule-bound starch synthase (GBSS) activities presented a single peak, reaching their maximums in the middle period. AGPase, SSS, and GBSS activities were all positively related to starch content. Moreover, quantitative real-time polymerase chain reaction (qRT-PCR) verified the changes in SS and SSS activities via the expression levels of the SS, SSSI, and SSSII genes. The gibberellin (GA) and zeatin riboside (ZR) content attained their maximum in the initial period of stolon formation. Indole-3-acetic acid (IAA) and abscisic acid (ABA) remained at high levels during the initial and middle period and decreased significantly during the later period of stolon formation, inversely to the ratio of ABA:IAA. Analysis of the physiological changes in T. edulis stolon indicated that the accumulation of soluble sugars and starch via various enzymes, a high level of IAA and a low ABA to IAA ratio mainly contributed to stolon development of T. edulis. This paper explored carbohydrate levels and endogenous hormones profiles during stolon formation, which provided the theory basis for further regulating stolon growth of T. edulis.

Abstract 2696: Arenobufagin targets K-Ras downstream to enhance the efficiency of gemcitabine in human pancreatic carcinoma cells

Abstract Pancreatic carcinoma (PC) is a deadly form of cancer with poor overall survival. Currently, chemotherapeutic such as gemcitabine and 5-fluorouracil (5-FU) are the most popular medication that can slightly improve survival, but rapid drug-resistance makes it urgent to search for more effective drugs. In hunting for natural components that cure PC, we found that arenobufagin, a cardiac glycosides-like compound, showed significant effects on the gemcitabine-resistant cell line Panc-1, the gemcitabine-sensitive cell line ASPC-1 and K-Ras activating HPNE cells at nanomolar concentrations. We used MTT and clonogenic survival assays to examine survival and proliferation and western blotting to assess changes of K-Ras downstream and apoptosis related proteins. We also detected the cell cycle and migration by flow cytometry and wound healing assay. Arenobufagin inhibited cellular survival and proliferation, decreased the phosphorylation of key proteins of K-Ras downstream such as Akt, Erk and nuclear factor kappa B (NF-κB), induced cell cycle G2/M arrest and apoptosis, and downregulated the level of phosphorylated epidermal growth factor receptor (EGFR). Most importantly, our data showed that arenobufagin can enhance the sensitivity of PC cells to gemcitabine and 5-fluorouracil (5-FU). Arenobufagin could enhance the effect of gemcitabine and 5-FU on PC cells through targeting multiple key proteins. All above makes arenobufagin a novel agent for combination therapy of pancreatic carcinoma. Citation Format: Tianjiao Wang, Yuhui Yuan. Arenobufagin targets K-Ras downstream to enhance the efficiency of gemcitabine in human pancreatic carcinoma cells. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2696. doi:10.1158/1538-7445.AM2015-2696

Simvastatin alleviates cardiac fibrosis induced by infarction via up-regulation of TGF-β receptor III expression.

Statins decrease heart disease risk, but their mechanisms are not completely understood. We examined the role of the TGF-β receptor III (TGFBR3) in the inhibition of cardiac fibrosis by simvastatin. Myocardial infarction (MI) was induced by ligation of the left anterior descending coronary artery in mice given simvastatin orally for 7 days. Cardiac fibrosis was measured by Masson staining and electron microscopy. Heart function was evaluated by echocardiography. Signalling through TGFBR3, ERK1/2, JNK and p38 pathways was measured using Western blotting. Collagen content and cell viability were measured in cultures of neonatal mouse cardiac fibroblasts (NMCFs). Interactions between TGFBR3 and the scaffolding protein, GAIP-interacting protein C-terminus (GIPC) were detected using co-immunoprecipitation (co-IP). In vivo, hearts were injected with lentivirus carrying shRNA for TGFBR3. Simvastatin prevented fibrosis following MI, improved heart ultrastructure and function, up-regulated TGFBR3 and decreased ERK1/2 and JNK phosphorylation. Simvastatin up-regulated TGFBR3 in NMCFs, whereas silencing TGFBR3 reversed inhibitory effects of simvastatin on cell proliferation and collagen production. Simvastatin inhibited ERK1/2 and JNK signalling while silencing TGFBR3 opposed this effect. Co-IP demonstrated TGFBR3 binding to GIPC. Overexpressing TGFBR3 inhibited ERK1/2 and JNK signalling which was abolished by knock-down of GIPC. In vivo, suppression of cardiac TGFBR3 abolished anti-fibrotic effects, improvement of cardiac function and changes in related proteins after simvastatin. TGFBR3 mediated the decreased cardiac fibrosis, collagen deposition and fibroblast activity, induced by simvastatin, following MI. These effects involved GIPC inhibition of the ERK1/2/JNK pathway.

MP31-14 CHANGES IN APOPTOSIS RELATED PROTEINS IN THE BLADDER AFTER PARTIAL BLADDER OUTLET OBSTRUCTION RELIEF

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 2015MP31-14 CHANGES IN APOPTOSIS RELATED PROTEINS IN THE BLADDER AFTER PARTIAL BLADDER OUTLET OBSTRUCTION RELIEF Ki Hak Song, Chong koo Sul, Yong gil Na, Jae sung Lim, Ju Hyun Shin, and Jong mok Park Ki Hak SongKi Hak Song More articles by this author , Chong koo SulChong koo Sul More articles by this author , Yong gil NaYong gil Na More articles by this author , Jae sung LimJae sung Lim More articles by this author , Ju Hyun ShinJu Hyun Shin More articles by this author , and Jong mok ParkJong mok Park More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.1369AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Partial bladder outlet obstruction (PBOO) induces prolonged bladder over-distension, leading to bladder dysfunction and structural alterations via induction of apoptosis. The aim of present study was to investigate sequential course of the apoptosis in bladder tissue, and changes of apoptosis related proteins in PBOO and subsequent relief. METHODS The study was conducted using 60 female Sprague-Dawley rats (body weight 250–300 g), divided into sham operated group (n=20), PBOO only (n=20), and PBOO group plus subsequent relief (n=20). PBOO rats were induced for 2 weeks, the obstruction has relived by removal of ligation after 2 weeks. After PBOO and relief surgery, urodynamic studies measuring contraction interval and contraction pressure were done. The tissues of rats were assessed by histologic anaylsis (H&E and TUNNEL stain) and expression of apoptosis related receptors (such as caspase-3, Erk, Jnk, P38, Bax, Bcl-2 and Survivin) have detected by RT-PCR and Western blotting. RESULTS Filling cystometry studies showed a significant decrease in contraction interval and increase in contraction pressure in the PBOO group. TUNNEL positive cells were significantly increased in PBOO group compared to sham operated group and decreased in PBOO relief group compared to PBOO group. On RT-PCR and Western blotting, expression of Bax, caspase-3, P38 and Jnk was significantly increased in PBOO group. But, expression of Erk, Bcl-2 was significantly decreased in PBOO group. The expression of Erk and Bcl-2 was significantly increased in PBOO relief group compared to PBOO group. CONCLUSIONS PBOO induced apoptosis of bladder tissue is associated with imbalance of MAPK pathways and apoptosis related protein. These results may also imply that pro-survival Erk kinase cascade is activated in response ischemic bladder injury and associated with initiation of bladder protection after PBOO. However, the mechanism of Survivin as anti-apoptotic factor in ischemic bladder tissue remains unclear. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e361 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ki Hak Song More articles by this author Chong koo Sul More articles by this author Yong gil Na More articles by this author Jae sung Lim More articles by this author Ju Hyun Shin More articles by this author Jong mok Park More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

The inhibitory effects of a monoclonal antibody targeting neuropilin-1 on adhesion of glioma cells to fibronectin.

We investigated the inhibitory effects of anti-Neuropilin-1 monoclonal antibody (NRP-1 MAb) on adhesion of glioma cells to Fibronectin (FN) and the associated mechanisms. The effects of NRP-1 MAb on adhesion of U251 glioma cells and formation of stress fibers were studied using adhesion assays and confocal fluorescence microscopy. After treatment of U251 glioma cells with NRP-1 MAb, changes of related proteins adhering to FN were detected by co-immunoprecipitation and Western blot. Adhesion assays showed that NRP-1 MAb played an inhibitory role in adhesion of U251 glioma cells. Confocal fluorescence microscopy, co-immunoprecipitation and Western blot showed that NRP-1 MAb can inhibit the formation of stress fibers adhering to U251 cells. NRP-1 MAb can also affect the formation of Integrin α5β1-NRP-1 complex, and also reduce the level of phosphorylation of FAK and p130cas. In conclusion, NRP-1 MAb has the inhibitory effects on adhesion of glioma cells to FN. This effect is related to the inhibition of NRP-1 MAb on the formation of Integrin α5β1-NRP-1 complex, which leads to the inhibition of FAK/p130cas signaling pathway.

Mitochondrial adaptations evoked with exercise are associated with a reduction in age-induced testicular atrophy in Fischer-344 rats.

Mitochondrial dysfunction in various tissues has been associated with numerous conditions including aging. In testes, aging induces atrophy and a decline in male reproductive function but the involvement of mitochondria is not clear. The purpose of this study was to examine whether the mitochondrial profile differed with (1) aging, and (2) 10-weeks of treadmill exercise training, in the testes of young (6 month) and old (24 month) Fischer-344 (F344) animals. Old animals exhibited significant atrophy (30 % decline; P < 0.05) in testes compared to young animals. However, relative mitochondrial content was not reduced with age and this was consistent with the lack of change in the mitochondrial biogenesis regulator protein, peroxisome proliferator-activated receptor gamma coactivator 1-alpha and its downstream targets nuclear respiratory factor-1 and mitochondrial transcription factor A. No effect was observed in the pro- or anti-apoptotic proteins, Bax and Bcl-2, respectively, but age increased apoptosis inducing factor levels. Endurance training induced beneficial mitochondrial adaptations that were more prominent in old animals including greater increases in relative mtDNA content, biogenesis/remodeling (mitofusin 2), antioxidant capacity (mitochondrial superoxide dismutase) and lower levels of phosphorylated histone H2AX, an early marker of DNA damage (P < 0.05). Importantly, these exercise-induced changes were associated with an attenuation of testes atrophy in older sedentary animals (P < 0.05). Our results indicate that aging-induced atrophy in testes may not be associated with changes in relative mitochondrial content and key regulatory proteins and that exercise started in late-life elicits beneficial changes in mitochondria that may protect against age-induced testicular atrophy.

Excitation/inhibition balance and learning are modified by Dyrk1a gene dosage

Cognitive deficits in Down syndrome (DS) have been linked to increased synaptic inhibition, leading to an imbalance of excitation/inhibition (E/I). Various mouse models and studies from human brains have implicated an HSA21 gene, the serine/threonine kinase DYRK1A, as a candidate for inducing cognitive dysfunction. Here, consequences of alterations in Dyrk1a dosage were assessed in mouse models with varying copy numbers of Dyrk1a: mBACtgDyrk1a, Ts65Dn and Dp(16)1Yey (with 3 gene copies) and Dyrk1a(+/−) (one functional copy). Molecular (i.e. immunoblotting/immunohistochemistry) and behavioral analyses (e.g., rotarod, Morris water maze, Y-maze) were performed in mBACtgDyrk1a mice. Increased expression of DYRK1A in mBACtgDyrk1a induced molecular alterations in synaptic plasticity pathways, particularly expression changes in GABAergic and glutaminergic related proteins. Similar alterations were observed in models with partial trisomy of MMU16, Ts65Dn and Dp(16)1Yey, and were reversed in the Dyrk1a(+/−) model. Dyrk1a overexpression produced an increased number and signal intensity of GAD67 positive neurons, indicating enhanced inhibition pathways in three different models: mBACtgDyrk1a, hYACtgDyrk1a and Dp(16)1Yey. Functionally, Dyrk1a overexpression protected mice from PTZ-induced seizures related to GABAergic neuron plasticity. Our study shows that DYRK1A overexpression affects pathways involved in synaptogenesis and synaptic plasticity and influences E/I balance toward inhibition. Inhibition of DYRK1A activity offers a therapeutic target for DS, but its inhibition/activation may also be relevant for other psychiatric diseases with E/I balance alterations.

Cellular responses in Bacillus thuringiensis CS33 during bacteriophage BtCS33 infection

Bacillus thuringiensis (Bt) has been widely used for 50years as a biopesticide for controlling insect pests. However, bacteriophage infection can cause failures in 50%-80% of the batches during Bt fermentation, resulting in severe losses. In the present work, the physiological and biochemical impacts of Bt strain CS33 have been studied during bacteriophage infection. This study adopted a gel-based proteomics approach to probe the sequential changed proteins in phage-infected Bt cells. To phage, it depressed the host energy metabolism by suppressing the respiration chain, the TCA cycle, and the utilization of PHB on one hand; on the other hand, it hijacked the host translational machine for its own macromolecular synthesis. To host, superinfection exclusion might be triggered by the changes of S-layer protein and flagella related proteins, which were located on the cell surface and might play as the candidates for the phage recognition. More importantly, the growth rate, cell mass, and ICPs yield were significantly decreased. The low yield of ICPs was mainly due to the suppressed utilization of PHB granules. Further functional study on these altered proteins may lead to a better understanding of the pathogenic mechanisms and the identification of new targets for phage control. B. thuringiensis (Bt) has been widely used for 50years as a safe biopesticide for controlling agricultural and sanitary insect pests. However, bacteriophage infection can cause severe losses during B. thuringiensis fermentation. The processes and consequences of interactions between bacteriophage and Bt were still poorly understood, and the molecular mechanisms involved were more unknown. This study adopted a gel-based proteomics approach to probe the physiological and biochemical impacts of Bt strain CS33 after phage-infection. The interactions between phage BtCS33 and its host Bt strain CS33 occurred mainly on four aspects. First, phage synthesized its nucleic acids through metabolic regulation by increasing the amount of NDK. Second, it is reasonable to infer that a phage resistance or superinfection exclusion was triggered by several increased or decreased proteins (SLP, FliD, FlaB), which were located on the cell surface and might play as candidates for the phage recognition. Third, combining the decreased flavoproteins (SdhA and EtfB) and the down regulated Fe-S cluster biosynthesis pathway together, it can be suggested that the respiration chain was weakened after phage infection. Additionally, three key enzymes (AcnB, FumC and AdhA) involved in the TCA cycle were all decreased, indicating the TCA cycle was seriously inhibited after infection. Fourth, the growth rate, cell mass and ICPs yield of the host were significantly decreased. To the best of our knowledge, this work represents the first systematic study on the interactions of an insecticidal bacterium with its phage, and has contributed novel information to understand the molecular events in the important biological pesticide producer, B. thuringiensis, in response to phage challenge.

GW24-e1882 Quantitative proteomics analysis of changes in myocardial energy metabolism related proteins between left and right atrial appendages from patients with permanent atrial fibrillation

ObjectivesAF is usually initiated and maintained in the left atrium. There is convincing evidence demonstrating an important pathophysiological association between LA remodelling and atrial fibrillation. A recent microarray study of...

Proteomics screen to reveal molecular changes mediated by C722G missense mutation in CHRM2 gene

Previously, we reported a missense mutation (C722G) in the M2-muscarinic acetylcholine receptor (CHRM2) gene associated with familial dilated cardiomyopathy. However, the exact molecular mechanisms by the related protein changes of CHRM2-C722G mutation induced are still unclear. CHRM2 and CHRM2-C722G lentiviral vector was infected to CHO cells. Proteomic analysis by label-free shotgun strategy and the STRING 9.0 software were performed. A total of 102 proteins with at least 2-fold change in the CHRM2-C722G group were identified, 42 proteins were up-regulated, whereas 57 were down-regulated. These altered proteins belong to three broad functional categories: (i) metabolic (e.g. Cytosolic acyl coenzyme A thioester hydrolase, Malate dehydrogenase); (ii) cytoskeletal (e.g. Actin-related protein, Myosin light polypeptide 6 and Alpha-actinin-1) and (iii) stress response (e.g. heat shock protein 70, Ras-related protein Rab-10). Interestingly, the marked differences in the expression of selected eight proteins (change >4.0-fold), were connected with many proteins related to apoptosis and immune/inflammatory response such as: FOS, BAX, MYC, TP53 and IL6. This novel study demonstrated for the first time a full-scale screening of the proteomics research by CHRM2-C722G mutation and profiled 102 changed proteins, of which, eight might be critical in cardiac dysfunction for future mapping. It was a full-scale screening of the proteomics research by CHRM2-C722G mutation. These proteins might serve as valuable biomarkers that could predict the presence of a precursor field. These proteins might serve to further explore the pathophysiological mechanisms in familial DCM patients with C176W mutation.

Dynamic changes of miRNA and cancer related proteins (CRPs) expression during neoadjuvant chemotherapy (NAC) for locally advanced breast cancer (LABC).

e22099 Background: NAC is used in LABC to downsize the primary tumor and eradicate metastases. This study assessed the Dynamic changes of miRNA and CRP expression during NAC with aim to identify molecular biomarkers that may predict outcome. Methods: Tissue, blood and serum samples were collected at four time points from LABC patients undergoing NAC, including time of diagnosis (base line) after 1st and 4th cycle of Adriamycin/cyclophosphamide, and after 4th cycle of Docetaxel (end point). We used microRNA-microarray for tissue samples and Proximity Ligation Assay (PLA) for serum samples to quantify global miRNA expression and 92 CRPs in LABC patients. The miRNA and CRPS were correlated with clinico-pathological features including stage, grade and clinical response.Statistical analysis was performed using Significant Analysis of Microarray ( SAM), Student's t-test, and Pearson's correlation test. Significant p value was defined as p&lt;0.05. Results: Tissue miRNA expression profiling (n=14) revealed 58 miRNA species that were differentially expressed between NAC endpoint and the baseline (FDR &lt; 15%). Reduced miR-21 expression was associated with disease recurrence. Using PLA (n=33) we identified serumosteoprotegerin, MCP-1, Prolactin, Carbonic Anhydrase IX, CXCL10, CD30L and TGF-alpha levels were significantly associated with overall clinical response (t-test, p&lt;0.05). Moreover, Prolactin and Erythropoietin showed significant associations with the stage (t-test, p&lt;0.05), while MMP3, ErbB4, Midkine, Cathepsin D and Prostasin proteins were associated with the recurrence. Analysis is ongoing to determine the dynamic changes of these biomarkers during the NAC. Conclusions: Our study identified specific miRNAs and CRPs that were differentially expressed during NAC in LABC patients which may have clinical significance.