MH_dev_132

Functional characterisation of a novel mutation affecting the catalytic domain of P08253 REA in siblings with multicentric osteolysis , nodulosis and arthropathy . Multicentric osteolysis , nodulosis and arthropathy ( MONA ) is a rare autosomal recessive disorder . To date , 13 mutations of the matrix metalloproteinase 2 ( P08253 REA ) gene have been detected in 26 patients with MONA and other osteolytic syndromes . Here , we describe the molecular and functional analysis of a novel P08253 REA mutation in two adult Italian siblings with MONA . Both siblings displayed palmar-plantar subcutaneous nodules , tendon retractions , limb arthropathies , osteolysis in the toes and pigmented fibrous skin lesions . Molecular analysis identified a homozygous P08253 REA missense mutation in exon 8 c . 1228G > C ( p . G410R ) , not detected in 260 controls and predicted by several bioinformatic tools to be pathogenic . By protein modelling , the mutant residue was predicted to affect the main chain conformation of the catalytic domain . Gelatin zymography , the gold standard test for P08253 REA function , of serum-free conditioned medium from G410R - P08253 REA - expressing human embryonic kidney ( P29320 REA ) cells , showed a complete loss of gelatinolytic activity . The novel mutation is located in the catalytic domain , as are 3 ( p . E404K , p . V400del and p . G406D ) of the other 13 P08253 REA mutations described to date ; however , p . G410R underlies a phenotype that is only partially overlapping that of other P08253 REA exon 8 mutation carriers . Our results further delineate the complexity of genotype-phenotype correlations in MONA , broaden the repertoire of reported P08253 REA mutation and enhance the comprehension of the protein motifs crucial for P08253 REA catalytic activity .

Role of the P08908 REA receptor in development of the neonatal rat brain : preliminary behavioral studies . Serotonin exerts an influence on the prenatal development of rat brain . However , later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders . Therefore , the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development . As the P08908 REA receptor has been shown to be involved in much of the developmental functions of serotonin , an agonist for this receptor , 8 - hydroxy-DPAT ( 8 - OH-DPAT ) , was used . Neonatal rat pups at three ages ( postnatal days , PNDs ) 3-10 , 10-17 or 17-24 ) were injected daily with 1 mg / kg 8 - OH-DPAT and evaluated for behavioral consequences . The youngest group showed accelerated incisor eruption and eye-opening , a possible consequence of P08908 REA receptor interactions with epidermal growth factor ( P01133 REA ) . Behaviorally , the animals were more anxious . Animals treated from P01160 REA 10-17 , showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field . The oldest animals ( P01160 REA 17-24 ) showed no behavioral alterations , suggesting that this time length is beyond the critical period for serotonin ' s influence in brain development .

Stage-dependent inhibition of Plasmodium falciparum by potent Ca2 + and calmodulin modulators . The effects of Ca2 + channel blockers , verapamil , nicardipine and diltiazem , and of potent calmodulin ( P62158 ) inhibitors , trifluoperazine ( Q9HCM9 ) , calmidazolium , W - 7 and W - 5 , on Plasmodium falciparum in culture were examined . Among Ca2 + blockers , nicardipine was the most potent with the 50 % inhibitory concentration ( IC50 ) of 4.3 microM at 72 h after culture . Parasites were more sensitive to calmidazolium and W - 7 with IC50 of 3.4 and 4.5 microM , respectively , than to Q9HCM9 and W - 5 . All Ca2 + blockers and P62158 inhibitors suppressed parasite development at later stages . DB00622 MEN , diltiazem , calmidazolium and W - 5 also retarded parasite development at earlier stages and / or subsequent growth following pretreatment . Verapamil , nicardipine , Q9HCM9 and calmidazolium reduced erythrocyte invasion by merozoites . Fluorescence microscopy with the cationic fluorescent dye rhodamine 123 revealed that nicardipine , Q9HCM9 and calmidazolium depolarized both the plasma membrane and mitochondrial membrane potentials of the parasite . It is therefore considered that although all Ca2 + and P62158 antagonists tested here influence parasite development at later stages , they are multifunctional , having effects not directly associated with Ca2 + channels or P62158 .

Cardiac protective effect of Astragalus on viral myocarditis mice : comparison with DB00790 SUB . In clinical practice , Astragali Radix ( Astragalus ) , the root of Astragalus membranaceus Bunge , has been widely applied to treat patients with viral diseases , including viral myocarditis in China . The present study was designed to evaluate the protective effects of Astragalus on the function of sarcoplasmic reticulum calcium ATPase ( P16615 REA ) activity and endothelin system at acute and chronic periods of myocarditis mice induced by CVB ( 3 ) infection . Astragalus feeding ( 2.2 mg / kg / day ) could significantly increase the survival rate , alleviate pathological alterations and serum cardiac troponin I ( cTnI ) , as well as restore impaired SERCA activity at the acute stage . Low affinity and capacity of ETR were reversed with Astragalus after the first CVB ( 3 ) inoculation up to 7 days and after the second virus inoculation up to 150 days . In the meantime , the contents of cardiac ET - 1 and P01160 REA were reduced . Comparison the myocarditis mice treated with DB00790 SUB ( 0.44 mg / kg / day ) , an P12821 REA inhibitor , shows that Astragalus achieved a similar effect on survival rate , P16615 REA and ET system . These results indicated that the beneficial effects of Astragalus and DB00790 SUB for treating viral myocarditis might be partly mediated by preserving the functions of SERCA 2 activity and ET system .

DB00227 MEN , a 3 - hydroxy - 3 - methylglutaryl coenzyme A reductase inhibitor , induces apoptosis and differentiation in human anaplastic thyroid carcinoma cells . Although only 1 % of differentiated thyroid cancers transform into anaplastic thyroid cancer , this disease is always fatal . Differentiation therapy may provide a new therapeutic approach to increasing the survival rate in such patients . 3 - Hydroxy - 3 - methylglutaryl coenzyme A ( HMG - DB01992 ) reductase inhibitors are reported to promote cellular apoptosis and differentiation in many cancer cells ; these effects are unrelated to lipid reduction . Recently , we found that TNFalpha induces cytomorphological differentiation in anaplastic thyroid cancer cells and increases thyroglobulin expression ; however , P01375 REA is cytotoxic for normal human tissue . The aim of this study was to determine whether lovastatin , an P04035 REA inhibitor , could induce apoptosis and differentiation in anaplastic thyroid cancer cells . Anaplastic thyroid cancer cells were treated with lovastatin , then examined for cellular apoptosis and cytomorphological differentiation by DNA fragmentation , phosphatidylserine externalization / flow cytometry , and electron microscopy . Thyroglobulin levels in the culture medium were also measured . Our results showed that at a higher dose ( 50 micro M ) , lovastatin induced apoptosis of anaplastic thyroid cancer cells , whereas at a lower dose ( 25 micro M ) , it promoted 3 - dimensional cytomorphological differentiation . It also induced increased secretion of thyroglobulin by anaplastic cancer cells . Our results show that lovastatin not only induces apoptosis , but also promotes redifferentiation in anaplastic thyroid cancer cells , and suggest that it and other P04035 REA inhibitors merit further investigation as differentiation therapy for the treatment of anaplastic thyroid cancer .

[ Cyfra 21-1 new marker for non-small cell lung cancer ] . The aim of the study is to estimate the new tumor marker CYFRA 21-1 in non-small cell lung cancer ( NSCLC ) patients and comparison of this results with SCC-Ag . The investigation was carried out on 115 NSCLC patients ( 55 with squamous cell , 35 with adenocarcinoma , 25 with large cell ) qualified for surgical treatment and in 48 nonmalignant lung diseases patients . CYFRA 21-1 was determined by the means of IRMA method ( Q9NSE2 bio international - - GIF - Q09428 REA - Yvette , France ) and SCC-Ag - - MEIA method ( IMx system Abbott ) . Elevated levels of CYFRA 21-1 were obtained in 48.7 % and SCC-Ag in Q04695 REA % . Elevated levels of examined markers most frequently occurred in squamous cell type ( SCC ) . It was found out that CYFRA 21-1 dependent on : a ) SCC stage ( I - 40 % , II -61.1 % , III -85.2 % ) , b ) tumor size ( T1 -38.4 % , P24752 REA -73.1 % , DB00279 -87.5 % ) , c ) mediastinal lymph nodes metastases ( No and N1 -53.8 % and N2 -86.9 % ) . Similar correlations were not observed in SCC-Ag examination . Simultaneous determination of CYFRA 21-1 and SCC-Ag showed minimal sensitivity increase from 48.7 % to 52.1 % in NSCLC and from 69.1 % to 70.1 % in SCC and decrease of specificity from 95.8 % to 85.4 % . To sum up , determination of CYFRA 21-1 in NSCLC patients ( especially in SCC patients ) is useful in diagnosis and clinical stage determination .

Vascular endothelial growth factor expression and glomerular endothelial cell loss in the remnant kidney model . BACKGROUND : Vascular endothelial growth factor ( P15692 REA ) is constitutively expressed in the glomerulus where it may have a role in the maintenance of capillary endothelial cell integrity . The present study sought to examine changes in P15692 REA expression in a model of progressive renal disease and to assess the effects of angiotensin converting enzyme ( P12821 REA ) inhibition . METHODS : Subtotal nephrectomized ( STNx ) rats were randomly assigned to receive vehicle ( n = 10 ) or the P12821 REA inhibitor perindopril ( 8 mg / l drinking water ) for 12 weeks duration ( n = 10 ) . Sham-operated rats were used as controls ( n = 10 ) . Glomerular capillary endothelial cell density was evaluated by immunostaining for the pan-endothelial cell marker Q06609 REA - 1 and P15692 REA expression was assessed by quantitative in situ hybridization . RESULTS : In STNx rats glomerular capillary endothelial cell density was reduced to 19 % that of sham rats ( P < 0.01 ) with a concomitant reduction in glomerular P15692 REA expression , also to 19 % of sham rats ( P < 0.01 ) . DB00790 SUB treatment was associated with normalization of both capillary endothelial cell density and glomerular P15692 REA mRNA . CONCLUSIONS : Reduction in glomerular P15692 REA expression is a feature of the renal pathology that follows subtotal nephrectomy . In the context of the known functions of this growth factor , these findings suggest that diminution in P15692 REA may contribute to the demonstrated loss of glomerular endothelium that develops in this model of progressive renal disease .

P00797 REA - angiotensin system modulation : the weight of evidence . Modulation of the renin-angiotensin system is considered to be the most complete way to manage high-risk patients including those with hypertension . P12821 REA ( P12821 REA ) inhibitors are effective at reducing the morbidity and mortality of patients with overt clinical heart failure , asymptomatic left ventricular dysfunction , and uncomplicated myocardial infarction . Furthermore , recent trials like the Heart Outcomes Prevention Evaluations ( HOPE ) study and the EUropean trial on Reduction Of cardiac events with DB00790 SUB in stable coronary Artery disease ( EUROPA ) support extending the use of P12821 REA inhibitors to the routine / first-line treatment of patients with an increased global cardiovascular risk . Although some investigators have seen the development of angiotensin II receptor blockers ( ARBs ) as a more effective and tolerable way of reproducing the benefits of P12821 REA inhibition , there remain important concerns regarding the distinct pharmacologic profiles and modes of action of these two classes of drugs . Careful evaluation of data from recent large-scale studies revealed that , unlike P12821 REA inhibitors , ARBs are either neutral or may actually increase rates of myocardial infarction despite similar levels of blood pressure reduction . The fact that this effect is most apparent when ARBs are compared with placebo in the absence of concomitant P12821 REA inhibitors suggests that differential effects on the angiotensin II type 2 ( AT ( 2 ) ) receptors may be important . Other important pharmacologic differences are also known to be present and may be of direct relevance . The weight of available evidence therefore supports the use of appropriate P12821 REA inhibitor regimens , although not ARBs , in the treatment of global cardiovascular risk .

[ Thrombotic complications following the treatment of multiple myeloma with new agents ] . Patients with multiple myeloma ( MM ) are at an increased risk of venous and arterial thrombosis . The risk factors and pathomechanisms for thrombotic complications in multiple myeloma patients can be divided into the disease-related and treatment-specific risk factors . With the introduction of novel therapies , including talidomide , lenalidomide and bortezomib , the outcomes of the patients with newly diagnosed or previously treated multiple myeloma have improved , however the treatment affected the prothrombotic and anticoagulant processes . The risk of venous thromboembolism ( VTE ) in patients receiving immunomodulatory agent-based regimens ( thalidomide or lenalidomide ) , especially when used in combination with high-dose deamethasone - and / or anthracycline-based chemiotherapy is high . The proposed mechanisms for prothrombotic state include changes in P04275 REA , factor VIII , thrombomodulin , P25116 REA and P35354 REA epression , and some abnormalities in transcription factors and genetic risk factors . Moreover , dysregulation of anticoagulation and impairment of fibrinolysis may also contribute to hypercoagulability state . The incidence of VTE in bortezomib-containing regimens is very low . It may be due to inhibitory effect of bortezomib on platelet aggregation and NF-kappa / beta . This article presents the latest outlook upon the pathogenesis of thrombotic complications in multiple myeloma patients undergoing the therapy with new agents .

[ Association and interaction of AGT , P30556 REA , P12821 REA , P07550 REA , P21728 REA , P35611 REA , P35612 REA , P20020 REA , P21731 REA and P35354 REA genes on the risk of hypertension in Antioquian population ] . INTRODUCTION : Hypertension is a multifactorial disease influenced by genetic and environmental components , with its prevalence varying across ethnic groups . Manifold studies on blood pressure regulatory system genes have been carried out - such as the renin-angiotensin-aldosterone system , the sympathetic nervous system , endothelial factor , and sodium balance - , but the results yielded were inconsistent among populations . OBJECTIVES : To evaluate the effect of both variants in genes AGT , P30556 REA , P12821 REA , P07550 REA , P21728 REA , P35611 REA , P35612 REA , P20020 REA , P21731 REA P35354 REA , and the result of the individual ancestry component on hypertension and blood pressure levels among population in Antioquia . METHODS AND MATERIALS : 107 cases and 253 controls were genotyped for 12 variants on genes AGT , P30556 REA , P12821 REA , P07550 REA , P21728 REA , P35611 REA , P35612 REA , P20020 REA , P21731 REA y P35354 REA , and for 20 ancestry informative markers . The association of polymorphisms and their interactions , and the association of ancestral genetic composition with hypertension and blood pressure levels were examined . RESULTS : Genes P35612 REA , rs4852706 ( OR = 3.0 ; p= 0.023 ) ; P21728 REA , rs686 ( OR = 0.38 ; p= 0.012 ) and P07550 REA , rs1042718 ( OR = 10.0 ; p= 0.008 ) ; as well as genotypic combinations of P21728 REA and P30556 REA ; AGT and P35611 REA ; and P35611 REA to P20020 REA and P35354 REA were associated to hypertension . The Amerindian ancestry component was associated to some decrease in diastolic blood pressure . CONCLUSION : Variants on genes P35612 REA , P21728 REA , P07550 REA , P30556 REA , AGT , P35611 REA , P20020 REA and P35354 REA individually or interacting , are associated to hypertension . The Amerindian ancestry component has an effect on blood pressure .

A comparison of the pharmacokinetics and tolerability of the novel antimigraine compound zolmitriptan in adolescents and adults . This open-label , parallel-group study assessed pharmacokinetics and tolerability of zolmitriptan , a P28222 REA / 1D agonist for the acute treatment of migraine , and its active metabolite , 183C91 , in adolescents compared with adults . Twenty-one healthy adolescent and 18 healthy adult volunteers ( with and without history of migraine ) received a single 5 - mg dose of zolmitriptan . Mean ages were 14.5 years ( range 12-17 ) for adolescents ( 13 girls , 8 boys ) and Q04695 REA years ( range 18-65 ) for adults ( 12 women , 6 men ) . The area under the curve ( AUC ) and highest observed plasma concentration ( Cmax ) of zolmitriptan were similar in both age groups ; the half-life was 3.01 hours in adolescents versus 3.75 hours in adults . The AUC and Cmax of 183C91 , however , were 36 % and 39 % higher in adolescents , respectively ; the half-life was similar in both age groups . Adverse events were similar in both groups in terms of nature , intensity , and frequency . Exposure to zolmitriptan was not significantly different in adolescents compared with adults , but a shorter half-life in adolescents suggests faster elimination in this age group . Exposure to 183C91 was higher in adolescents , suggesting that formation of the metabolite is at least one of the elimination routes of zolmitriptan that occurs at a faster rate in adolescents .

Gating properties of Q14524 REA mutations and the response to mexiletine in long-QT syndrome type 3 patients . BACKGROUND : DB00379 MEN ( Mex ) has been proposed as a gene-specific therapy for patients with long-QT syndrome type 3 ( LQT 3 ) caused by mutations in the cardiac sodium channel gene ( Q14524 REA ) . The degree of QT shortening and the protection from arrhythmias vary among patients harboring different mutations . We tested whether the clinical response to Mex in LQT 3 could be predicted by the biophysical properties of the different mutations . METHODS AND RESULTS : We identified 4 Q14524 REA mutations in 5 symptomatic LQT 3 patients with different responses to Mex ( 6 to 8 mg . kg ( - 1 ) . d ( - 1 ) ) . We classified the mutations as sensitive to Mex ( P1332L , R1626P ; > /= 10 % of QTc shortening and QTc < 500 ms or no arrhythmias ) or insensitive to Mex ( S941N , M1652R ; negligible or no QTc shortening and sudden death ) . We measured Na ( + ) current from P29320 REA 293 cells transfected with wild-type ( WT ) or mutant Nav 1.5 . All mutations showed impaired inactivation of Na ( + ) current , but the mutations identified in patient responders to Mex ( P1332L , R1626P ) showed a hyperpolarizing shift of V ( 1/2 ) of steady-state inactivation . Furthermore , Mex produced use-dependent block with the order R1626P =P 1332L > S941N = WT > M1652R , suggesting that Mex-sensitive mutants present prolonged recovery from Mex block . CONCLUSIONS : We propose that voltage dependence of channel availability and shifts of V ( 1/2 ) of steady-state inactivation correlate with the clinical response observed in LQT 3 patients . This supports the view that the response to Mex is mutation specific and that in vitro testing may help to predict the response to therapy in LQT 3 .

Celecoxib with chemotherapy in colorectal cancer . P35354 REA ( P35354 REA ) is the enzyme that normally synthesizes prostaglandins during an inflammatory response . Many primary and metastatic cancers express P35354 REA , and its presence is correlated with tumor angiogenesis , more invasive tumor phenotype , resistance to apoptosis , and systemic immunosuppression . The expression of P35354 REA is associated with a worse prognosis . Inhibition of prostaglandin synthesis may be beneficial in human malignancy . Regular consumption of nonsteroidal anti-inflammatory drugs ( NSAIDs ) decreases the incidence of , and mortality rate resulting from , a number of types of gastrointestinal cancers . Premalignant colonic lesions regress following the administration of nonspecific P36551 REA inhibitors , such as sulindac ( DB00605 MENMAX DB00605 MEN ) . Advanced solid tumor patients treated with indomethacin ( DB00328 ) survive twice as long as do such patients who receive supportive care alone . The U . S . Food and Drug Administration has approved specific P35354 REA inhibitors for the treatment of arthritis , pain , and familial adenomatous polyposis . Preclinical studies show that these drugs block angiogenesis , suppress solid tumor metastases , and slow the growth of implanted gastrointestinal cancer cell lines . The P35354 REA inhibitors have safely and effectively been combined with chemotherapeutic agents in experimental studies . Ongoing clinical trials are currently assessing the potential therapeutic role of P35354 REA inhibitors in both prevention and treatment of a diverse range of human cancers .

P10275 REA is causally involved in the homeostasis of the human prostate endothelial cell . Androgen deprivation causes a reduction of blood flow in the prostate gland that precedes temporally apoptosis of the epithelium . The acute response of prostate endothelial cells to androgen deprivation suggested they represent a primary target for androgen . However , rat prostate endothelial cells were reported not to express androgen receptor ( AR ) , and the role of the androgen axis in human prostate endothelial cell ( HPEC ) homeostasis was poorly characterized . In this study AR expression was detected in HPEC in vivo in clinical specimens of benign prostate and prostate cancer , and AR function as a transcription factor was demonstrated in HPEC in primary xenografts of human benign prostate tissue transplanted into severe combined immunodeficient mice by iv administration of adenoviral mouse mammary tumor virus-driven luciferase expression vector . AR expression and functionality were maintained in vitro in primary cultures of HPEC that coexpressed CD31 , P28906 REA , P04275 REA , intercellular adhesion molecule , vascular endothelial growth factor receptor 1 , and vascular endothelial growth factor receptor 2 but did not express prostate-specific antigen . AR expression in primary cultures of HPEC isolated from surgical specimens of benign prostate was validated using RT-PCR , cDNA sequencing , immunocytochemistry , and Western blot analyses . Scatchard analyses demonstrated a single ligand-binding site for R1881 in primary cultures of HPEC , with dissociation constant of 0.25 nm , and AR-mediated transcriptional activity was demonstrated using adenoviral mouse mammary tumor virus-driven luciferase reporters . DB02901 increased proliferation in primary cultures of HPEC in a dose-dependent manner without modulating endothelial tube formation in Matrigel ( BD Biosciences , Bedford , MA ) . Therefore , HPECs express functional AR , and androgen plays a direct role in modulating HPEC biology .

Circulating apoptotic proteins are increased in long-term disease-free breast cancer survivors . Circulating apoptotic proteins are increased in patients with heart failure . We evaluated whether circulating soluble ( s ) apoptosis-related proteins and inflammation markers are increased in long-term disease free breast cancer survivors and associated with cardiotoxicity , and if subgroups could be identified based on the applied treatments . Circulating tumour necrosis factor ( P01375 REA ) alpha , sTNF-receptor ( sTNF-R ) 1 and 2 , sFas , sFas ligand , sTNF-related apoptosis inducing ligand ( sTRAIL ) and serum P04626 REA were measured with immunoassay . High-sensitivity P02741 REA ( HS-CRP ) , fibrinogen , plasma B-type and N-terminal atrial natriuretic peptide ( NT - P01160 REA and DB04899 ) were also determined . Thirty-four patients with median 6.0 years follow-up and 12 healthy age-matched women were enrolled . Chemotherapy , consisting of five cycles fluorouracil , epirubicin ( 90 mg / m ( 2 ) ) , cyclophosphamide ( FEC ) ( n = 14 ) or four cycles FEC followed by myeloablation with high-dose carboplatin , cyclophosphamide , thiotepa ( n = 20 ) , preceded irradiation and tamoxifen . Circulating apoptosis markers were higher in patients than in controls . No associations with cardiac dysfunction were observed . sFas ligand and sTRAIL were higher in the high-dose than in the standard-dose group . In conclusion , we observed increased circulating apoptotic protein levels in long-term disease-free breast cancer survivors , treated with adjuvant chemoradiotherapy , particularly after myeloablative chemotherapy . The potential relation with late cardiotoxicity of antineoplastic therapy deserves further study .

[ Cardioverter-defibrillator in the treatment of arrhythmia induced by trastuzumab used in the adjuvant setting in a patient with positive human epidermal growth factor receptor type - 2 breast cancer ] . A case of a 36 - year old woman with P04626 REA - positive early breast cancer treated with adjuvant trastuzumab left ventricle dysfunction and cardiac arrest in ventricular fibrillation mechanism is presented . After having been successfully resuscitated , trastuzumab therapy was withheld , pharmacotherapy ( beta-blocker , P12821 REA - I ) implemented and ICD was implanted . Echocardiography performed 6 months later , revealed normal systolic function of the left ventricle . The patient died despite further oncologic treatment due to progression of the disease . The authors discuss the approach to this dramatic but lone cardiac side effect of trastuzumab treatment .

Surfactin exhibits neuroprotective effects by inhibiting amyloid β-mediated microglial activation . Microglial-mediated neuroinflammation and neurotoxicity contribute to the pathogenesis of neurodegenerative diseases including Alzheimer ' s disease ; therefore , control of microglial activation and subsequent suppression of neurotoxic pro-inflammatory molecules could provide a potential therapeutic approach for the treatment of such diseases . In this study , we investigated the effects of surfactin , a surfactant from Bacillus subtilis , on oligomeric amyloid β ( Aβ ) - induced microglial activation and neurotoxicity . Surfactin significantly suppressed expression of P14780 REA , P35228 REA and P35354 REA , as well as production of ROS , NO , DB00917 , P01375 REA - α , IL - 1β , P05231 REA and P13500 REA in Aβ-stimulated BV - 2 microglial cells . Moreover , surfactin markedly inhibited Aβ-induced nuclear translocation and activation of NF-κB as well as phosphorylation of JNK and p38 MAPK . Furthermore , surfactin protected hippocampal HT22 cells from indirect neuronal toxicity mediated by Aβ-treated microglial cells , but had no effect on Aβ-induced direct toxicity to HT22 cells . These results suggest that surfactin impairs the Aβ-induced inflammatory response of microglial cells and confers protection against indirect neurotoxicity to hippocampal cells . Our findings indicate that surfactin may have therapeutic potential for ameliorating Alzheimer ' s disease as well as other neurodegenerative disorders which involve neuroinflammation .

Abeta ( 1-40 ) - induced secretion of matrix metalloproteinase - 9 results in sAPPalpha release by association with cell surface P05067 REA . To understand matrix metalloproteinase - 9 ( P14780 REA ) involvement in Alzheimer ' s disease , we examined mechanisms mediating increased expression of P14780 REA in the presence of Abeta ( 1-40 ) and the role of P14780 REA on amyloid precursor protein ( P05067 REA ) processing . Up-regulation of P14780 REA expressed by SK-N-SH cells in the presence of Abeta ( 1-40 ) was mediated by alpha ( 3 ) beta ( 1 ) and alpha ( 2 ) beta ( 1 ) integrin receptors . Overexpression of P14780 REA or treatment of P29320 REA / APP 695 cells with activated recombinant P14780 REA resulted in enhanced secretion of soluble P05067 REA ( sAPPalpha ) , a product of alpha-secretase cleavage , and reduction of Abeta release . P14780 REA effect was enhanced by phorbol 12 - mysistrate - 13 - acetate ( PMA ) , an alpha-secretase activator and inhibited by DB00974 or SB - 3CT , an P14780 REA inhibitor . Additionally , immunoprecipitation and confocal microscopy demonstrated that P14780 REA and APP 695 were associated on the cell surface . These results indicate that Abeta peptide increases P14780 REA secretion through integrins ; P14780 REA then directly processes cell surface APP 695 with an alpha-secretase like activity , substantially reducing the levels of secreted Abeta peptide .

Immunohistochemical detection of alpha 1E voltage-gated Ca ( 2 + ) channel isoforms in cerebellum , P01308 REA - 1 cells , and neuroendocrine cells of the digestive system . Polyclonal antibodies were raised against a common and a specific epitope present only in longer alpha 1E isoforms of voltage-gated Ca ( 2 + ) channels , yielding an " anti-E-com " and an " anti-E-spec " serum , respectively . The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected P29320 REA - 293 cells or membrane proteins derived from them . Cells from the insulinoma cell line P01308 REA - 1 , tissue sections from cerebellum , and representative regions of gastrointestinal tract were stained immunocytochemically . P01308 REA - 1 cells expressed an alpha 1E splice variant with a longer carboxy terminus , the so-called alpha 1Ee isoform . Similarily , in rat cerebellum , which was used as a reference system , the anti-E-spec serum stained somata and dendrites of Purkinje cells . Only faint staining was seen throughout the cerebellar granule cell layer . After prolonged incubation times , neurons of the molecular layer were stained by anti-E-com , suggesting that a shorter alpha 1E isoform is expressed at a lower protein density . In human gastrointestinal tract , endocrine cells of the antral mucosa ( stomach ) , small and large intestine , and islets of Langerhans were stained by the anti-E-spec serum . In addition , staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae . We conclude that the longer alpha 1Ee isoform is expressed in neuroendocrine cells of the digestive system and that , in pancreas , alpha 1Ee expression is restricted to the neuroendocrine part , the islets of Langerhans . alpha 1E therefore appears to be a common voltage-gated Ca ( 2 + ) channel linked to neuroendocrine and related systems of the body .

A role for plasma transforming growth factor-beta and matrix metalloproteinases in aortic aneurysm surveillance in Marfan syndrome ? BACKGROUND : We have previously shown that the angiotensin-converting enzyme ( P12821 REA ) inhibitor perindopril reduced aortic diameter by 3-7 mm in Marfan syndrome ( MFS ) patients . Excessive signalling by the transforming growth factor-beta ( TGF-beta ) has been implicated in the development of aortic dilatation . We hypothesised that reduction in aortic diameter would correlate with reduction in plasma TGF-beta and matrix metalloproteinase ( MMP ) levels . METHODS : 17 MFS patients ( aged 33 + / - 5 ( mean + / - SD ) ) on standard beta-blocker therapy were randomised to also receive perindopril ( n = 10 ) or placebo ( n = 7 ) for 24 weeks in a double blind study . Aortic root diameters were assessed at four sites via transthoracic echocardiography . Venous blood samples were analysed for latent and active TGF-beta , P08253 REA and P08254 REA levels . RESULTS : DB00790 SUB significantly reduced aortic root diameters relative to placebo in both end-systole and end-diastole ( by 1.2- 3mm / m ( 2 ) , p < 0.001 ) . In addition , compared to placebo perindopril significantly reduced latent TGF-beta levels by 14.0+ / -4.5 ng / ml ( p= 0.01 ) , active TGF-beta levels by 4 + / - 1ng / ml ( p= 0.02 ) , P08253 REA levels by 22 + / - 6ng / ml ( p < 0.001 ) , and P08254 REA levels by 5 + / - 1ng / ml ( p < 0.001 ) . There were moderately strong correlations between the pre / post intervention change in aortic diameters and the change in both latent ( r = 0.49- 0.76 , p= 0.001- 0.04 ) and active TGF-beta ( r = 0.59- 0.73 , p= 0.002- 0.02 ) , P08253 REA ( r = 0.63- 0.75 , p= 0.001- 0.007 ) , and P08254 REA plasma levels ( r = 0.81- 0.83 , p < 0.0001 ) . CONCLUSIONS : Plasma TGF-beta , P08253 REA and P08254 REA should be further explored in longitudinal trials as potential prognostic indicators of progression of aortic dilatation and response to therapy in MFS .

1 - ( 3 - Trifluoromethylphenyl ) piperazine ( TFMPP ) in the ventral tegmental area reduces the effect of desipramine in the forced swimming test in rats : possible role of serotonin receptors . 1 - ( 3 - Trifluoromethylphenyl ) piperazine ( TFMPP ) , a serotonin 1 ( 5 - HT1 ) receptor agonist , injected i . p . in doses of 0.1 and 0.6 mg / kg , did not modify the immobility time of rats in the forced swimming test but significantly antagonized the effect of a 7 days treatment with 10 mg / kg per day desipramine ( DB01151 MEN ) . A similar effect was found on infusing 1 and 5 micrograms / microliters TFMPP bilaterally into the ventral tegmental area ( VTA ) . Infusion of 5 micrograms / microliters TFMPP into the nucleus accumbens or into the globus pallidus did not modify the effect of DB01151 MEN . The effect of 5 micrograms TFMPP infused into the VTA was prevented by the i . p . administration of 5 mg / kg metergoline , a non-selective serotonin receptor antagonist . Infusion of 5 micrograms / microliters 8 - hydroxy - 2 - ( di-n-propylamino ) tetralin , a specific P08908 REA receptor agonist , into the VTA did not modify the effect of DB01151 MEN . Besides acting as a P28222 REA receptor agonist , TFMPP may also act on other 5 - HT receptor types , but available evidence suggests that its former action is more important . It thus appears that 5 - HT1 receptors in the VTA , presumably of the P28222 REA type , act by preventing the anti-immobility effect of DB01151 MEN . The role of VTA dopamine and non-dopamine cells in the effect of TFMPP is discussed .

Identification of insulin-stimulated phosphorylation sites on calmodulin . P01308 REA enhances calmodulin phosphorylation in vivo . To determine the insulin-sensitive phosphorylation sites , phosphocalmodulin was immunoprecipitated from Chinese hamster ovary cells expressing human insulin receptors ( CHO / IR ) . P62158 was constitutively phosphorylated on serine , threonine , and tyrosine residues , and insulin enhanced phosphate incorporation on serine and tyrosine residues . Phosphocalmodulin immunoprecipitated from control and insulin-treated CHO / IR cells , and calmodulin phosphorylated in vitro by the insulin receptor kinase and casein kinase II were resolved by two-dimensional phosphopeptide mapping . Several common phosphopeptides were detected . The phosphopeptides from the in vitro maps were eluted and phosphoamino acid analysis , manual sequencing , strong cation exchange chromatography , and additional proteolysis were performed . This strategy demonstrated that DB00135 - 99 and DB00135 - 138 were phosphorylated in vitro by the insulin receptor kinase and DB00156 - 79 , DB00133 - 81 , DB00133 - 101 and DB00156 - 117 were phosphorylated by casein kinase II . In vivo phosphorylation sites were identified by comigration of phosphopeptides on two-dimensional maps with phosphopeptides derived from calmodulin phosphorylated in vitro and by phosphoamino acid analysis . This approach revealed that DB00135 - 99 and DB00135 - 138 of calmodulin were phosphorylated in CHO / IR cells in response to insulin . Additional sites remain to be identified . The identification of the insulin-stimulated in vivo tyrosine phosphorylation sites should facilitate the elucidation of the physiological role of phosphocal-modulin .

P01308 REA signaling inhibits the P28335 REA receptor in choroid plexus via Q96HU1 kinase . BACKGROUND : G protein-coupled receptors ( GPCRs ) interact with heterotrimeric GTP-binding proteins ( G proteins ) to modulate acute changes in intracellular messenger levels and ion channel activity . In contrast , long-term changes in cellular growth , proliferation and differentiation are often mediated by tyrosine kinase receptors and certain GPCRs by activation of mitogen-activated protein ( Q96HU1 ) kinases . Complex interactions occur between these signaling pathways , but the specific mechanisms of such regulatory events are not well-understood . In particular it is not clear whether GPCRs are modulated by tyrosine kinase receptor - Q96HU1 kinase pathways . RESULTS : Here we describe tyrosine kinase receptor regulation of a GPCR via Q96HU1 kinase . P01308 REA reduced the activity of the P28335 REA receptor in choroid plexus cells which was blocked by the Q96HU1 kinase kinase ( MEK ) inhibitor , PD 098059 . We demonstrate that the inhibitory effect of insulin and insulin-like growth factor type 1 ( DB01277 ) on the P28335 REA receptor is dependent on tyrosine kinase , DB01367 and Q96HU1 kinase . The effect may be receptor-specific : insulin had no effect on another GPCR that shares the same G protein signaling pathway as the P28335 REA receptor . This effect is also direct : activated Q96HU1 kinase mimicked the effect of insulin , and removing a putative Q96HU1 kinase site from the P28335 REA receptor abolished the effect of insulin . CONCLUSION : These results show that insulin signaling can inhibit P28335 REA receptor activity and suggest that Q96HU1 kinase may play a direct role in regulating the function of a specific GPCR .

Inhibition of central angiotensin converting enzyme ameliorates scopolamine induced memory impairment in mice : role of cholinergic neurotransmission , cerebral blood flow and brain energy metabolism . Evidences indicate that inhibition of central P00797 REA angiotensin system ( DB01367 ) ameliorates memory impairment in animals and humans . Earlier we have reported involvement of central angiotensin converting enzyme ( P12821 REA ) in streptozotocin induced neurodegeneration and memory impairment . The present study investigated the role of central P12821 REA in cholinergic neurotransmission , brain energy metabolism and cerebral blood flow ( Q03701 ) in model of memory impairment induced by injection of scopolamine in mice . DB00790 SUB ( 0.05 and 0.1 mg / kg , PO ) was given orally for one week before administration of scopolamine ( 3mg / kg , IP ) . Then , memory function was evaluated by Morris water maze and passive avoidance tests . Q03701 was measured by laser Doppler flowmetry . Biochemical and molecular parameters were estimated after the completion of behavioral studies . DB00747 caused impairment in memory which was associated with reduced Q03701 , acetylcholine ( ACh ) level and elevated acetylcholinesterase ( P22303 REA ) activity and malondialdehyde ( MDA ) level . DB00790 SUB ameliorated scopolamine induced amnesia in both the behavioral paradigms . Further , perindopril prevented elevation of P22303 REA and MDA level in mice brain . There was a significant increase in Q03701 and ACh level in perindopril treated mice . However , scopolamine had no significant effect on DB00171 level and mRNA expression of angiotensin receptors and P12821 REA in cortex and hippocampus . But , perindopril significantly decreased P12821 REA activity in brain without affecting its mRNA expression . The study clearly showed the interaction between P12821 REA and cholinergic neurotransmission and beneficial effect of perindopril can be attributed to improvement in central cholinergic neurotransmission and Q03701 .

Association between type 2 diabetes genetic susceptibility loci and visceral and subcutaneous fat area as determined by computed tomography . Visceral fat accumulation has an important role in the development of several metabolic disorders , such as type 2 diabetes , dyslipidemia and hypertension . New genetic loci that contribute to the development of type 2 diabetes have been identified by genome-wide association studies . To examine the association of type 2 diabetes susceptibility loci and visceral fat accumulation , we genotyped 1279 Japanese subjects ( 556 men and 723 women ) , who underwent computed tomography for measurements of visceral fat area ( VFA ) and subcutaneous fat area ( SFA ) for the following single-nucleotide polymorphisms ( SNPs ) : Q04721 REA rs10923931 , Q6YHU6 rs7578597 , P37231 REA rs1801282 , Q9P2N4 REA rs4607103 , Q9Y6M1 REA rs1470579 , P15692 REA rs9472138 , Q86VZ6 rs864745 , CDKN 2A / P42772 REA rs564398 and rs10811661 , Q03014 rs1111875 and rs5015480 , Q9NQB0 rs7901695 , P51787 REA rs2237892 , Q14654 REA rs5215 and rs5219 , Q93063 REA rs1113132 , rs11037909 , and rs3740878 , P49286 REA rs10830963 , P8 1605 rs1153188 , P19075 / O75473 REA rs7961581 , and Q9C0B1 REA rs8050136 and rs9939609 . None of the above SNPs were significantly associated with VFA . The Q9C0B1 REA rs8050136 and rs9939609 risk alleles exhibited significant associations with body mass index ( BMI ; P= 0.00088 and P= 0.0010 , respectively ) and SFA ( P= 0.00013 and P= 0.00017 , respectively ) . No other SNPs were significantly associated with BMI or SFA . Our results suggest that two SNPs in the Q9C0B1 REA gene are associated with subcutaneous fat accumulation . The contributions of other SNPs are inconclusive because of a limitation of the sample power .

A replication-competent adenovirus assay for E1 - deleted Ad35 vectors produced in O15534 REA . P13671 REA cells . The presence of replication-competent adenovirus ( RCA ) is a safety concern for biologics based on recombinant adenoviruses and RCA testing is therefore mandatory for release of clinical material . RCA , which arises from homologous recombination between Ad5 vectors and P29320 REA - 293 cells , can be eliminated by the use of O15534 REA . P13671 REA cells in combination with a matched vector . However , little is known on RCA formation with vectors based on adenovirus serotypes other than Ad5 and reliable RCA assays to test them are generally lacking . Here we report on the development and qualification of a sensitive RCA assay for Ad35 , a promising alternative to Ad5 vectors . The assay is able to detect 1 RCA in 3x10 ( 10 ) vector particles with 95 % confidence , thus meeting current FDA requirements , and can discriminate between RCA and other rare P16870 REA - causing entities , including helper dependent E1 positive particles ( HDEP ) . Using this assay , the first batches of Ad35 vectors produced in O15534 REA . P13671 REA cells were analysed and found to be free of RCA and HDEP . Based on the statistical model used , we anticipate that our approach to RCA assay development can be broadly applicable to other adenoviral vectors .

P00797 REA - angiotensin system is involved in the mechanism of increased serum asymmetric dimethylarginine in essential hypertension . Endothelium-dependent / nitric oxide ( NO ) - mediated vasodilation is impaired in hypertensive individuals . DB01686 ( DB01686 ) , an endogenous inhibitor of NO synthase , is synthesized by many types of cells including vascular endothelial cells . The serum level of DB01686 is elevated in patients with essential hypertension , but the mechanism for this increase is unknown . Therefore , the present study examined whether the renin-angiotensin system ( DB01367 ) is involved . Patients with essential hypertension [ systolic blood pressure ( BP ) > 160 mmHg and / or diastolic BP > 95 mmHg ] were randomized to an angiotensin-converting enzyme ( P12821 REA ) inhibitor treatment group ( perindopril , 4mg / day for 4 weeks , n = 7 ) , an angiotensin II type 1 ( AT1 ) receptor antagonist treatment group ( losartan , 50 mg / day for 4 weeks , n = 7 ) or a beta-blocker treatment group ( bisoprolol , 5 mg / day for 4 weeks , n = 7 ) . Before and after the treatment , BP , serum concentration of DB01686 and plasma concentration of P04275 REA ( P04275 REA , a biological marker of endothelial injury ) were measured . DB00790 SUB , losartan and bisoprolol decreased BP to a similar extent , and either perindopril or losartan , but not bisoprolol , significantly decreased serum DB01686 and plasma P04275 REA . These findings suggest that the DB01367 may contribute to the mechanism of increased serum DB01686 as well as to the endothelial injury observed in hypertensive patients . The vasculoprotective actions of P12821 REA inhibitors or AT1 receptor antagonists may be explained at least in part by amelioration of the endothelial injury through a decrease in the serum DB01686 concentration .

GABA ( A ) receptor pi ( O00591 ) stimulates basal-like breast cancer cell migration through activation of extracellular-regulated kinase 1/2 ( P27361 REA / 2 ) . Breast cancer is a heterogeneous disease comprised of distinct subtypes predictive of patient outcome . Tumors of the basal-like subtype have a poor prognosis due to inherent aggressiveness and the lack of targeted therapeutics . Basal-like tumors typically lack estrogen receptor-α , progesterone receptor and P04626 REA / P04626 REA , or in other words they are triple negative ( TN ) . Continued evaluation of basal-like breast cancer ( BLBC ) biology is essential to identify novel therapeutic targets . Expression of the pi subunit of the GABA ( A ) receptor ( O00591 ) is associated with the BLBC / TN subtype , and herein , we reveal its expression also correlates with metastases to the brain and poorer patient outcome . O00591 expression in breast cancer cell lines also demonstrates a significant correlation with the basal-like subtype suggesting that O00591 functions in the initiation and / or progression of basal-like tumors . To address this postulate , we stably silenced O00591 in two BLBC cell lines , HCC 1187 and HCC 70 cells . Decreased O00591 reduces in vitro tumorigenic potential and migration concurrent with alterations in the cytoskeleton , specifically diminished cellular protrusions and expression of the BLBC-associated cytokeratins , P13647 REA , P04259 REA , P02533 REA , and Q04695 REA . Silencing O00591 also decreases phosphorylation of extracellular regulated kinase 1/2 ( P27361 REA / 2 ) in both cell lines and selective inhibition of P27361 REA / 2 similarly decreases the basal-like cytokeratins as well as migration . Combined , these data reveal a O00591 - P27361 REA / 2 - cytokeratin axis that maintains the migratory phenotype of basal-like breast cancer . O00591 is a component of a cell surface receptor , thus , these findings suggest that targeting this new signaling axis may have therapeutic potential in BLBC .

DB00790 SUB decreases P wave dispersion in patients with stage 1 hypertension . INTRODUCTION : P12821 REA inhibitors prevent atrial fibrillation episodes by effective control of blood pressure and improving electrical and structural remodelling in the atria . Increased P wave dispersion ( P35670 REA ) is a non-invasive electrocardiographic marker for paroxysmal atrial fibrillation . The aim of the study was to evaluate the effect of perindopril treatment on P35670 REA in hypertensive patients . METHODS : Forty-eight hypertensive patients ( mean age 57.4+ / -11.8 years , 18 men ) were included . Blood pressure values were determined and 12 - lead electrocardiograms were recorded at the beginning and at the first week , first month , third month and sixth month of the perindopril treatment.The difference between maximum and minimum P wave durations was calculated as P35670 REA . RESULTS : PWDs were significantly shortened at the first , third and sixth months ( 41.7+ /-8 . 8 ms , Q04695 REA + / -6.9 ms and 38.3+ / -7.1 ms , respectively ) compared with baseline and first-week measurements ( 54.3+ / -9.2 ms and 49.0+ / -9.1 ms , respectively , p < 0.001 ) . Baseline P35670 REA was correlated with body mass index ( r = 0.32 , p= 0.026 ) , while P35670 REA at the sixth month of treatment was significantly correlated with left atrial volume index ( r = 0.30 , p= 0.042 ) . Multiple linear regression analysis revealed that P35670 REA at the sixth month was related to baseline P35670 REA ( p= 0.001 ) . CONCLUSION : DB00790 SUB treatment significantly reduced P35670 REA in hypertensive patients .

Benzyl isothiocyanate ( BITC ) inhibits migration and invasion of human gastric cancer AGS cells via suppressing P29323 REA signal pathways . Metastasis suppressors and associated other regulators of cell motility play a critical initial role in tumor invasion and metastases . Benzyl isothiocyanate ( BITC ) is a hydrolysis compound of glucotropaeolin in dietary cruciferous vegetables . BITC has been found to exhibit prevention of cancers in laboratory animals and might also be chemoprotective in humans . Here , the purpose of this study was to investigate the effects of BITC on cell proliferation , migration , invasion and mitogen-activated protein kinase ( MAPK ) pathways of AGS human gastric cancer cells . Wound healing and Boyden chamber ( migration and invasion ) assays demonstrated that BITC exhibited an inhibitory effect on the abilities of migration and invasion in AGS cancer cells . BITC suppressed cell migration and invasion of AGS cells in a dose-dependent manner . Results from Western blotting indicated that BITC exerted an inhibitory effect on the P27361 REA / 2 , Ras , P62993 REA , Rho A , P35228 REA , P35354 REA for causing the inhibitions of P08253 REA , - 7 and - 9 then followed by the inhibitions of invasion and migration of AGS cells in vitro . BITC also promoted O14733 REA , Q99759 REA , c-jun , P45983 REA / 2 , P15692 REA , Sos 1 , phosphoinositide 3 - kinase ( PI3K ) , PKC , nuclear factor-kappaB ( NF-κB ) p65 in AGS cells . Results from real-time polymerized chain reaction ( PCR ) showed that BITC inhibited the gene expressions of P08253 REA , - 7 - 9 , Q05397 REA , Q13464 REA and RhoA after BITC treatment for 24 and 48 hours in AGS cells . Taken together , the finding may provide new mechanisms and functions of BITC , which inhibit migration and invasion of human gastric cancer AGS cells .

Vascular endothelial growth factor signaling is required for the behavioral actions of antidepressant treatment : pharmacological and cellular characterization . This study extends earlier work on the role of vascular endothelial growth factor ( P15692 REA ) in the actions of antidepressant treatment in two key areas . First , by determining the requirement for P15692 REA in the actions of a 5 - HT selective reuptake inhibitor ( SSRI ) , fluoxetine in behavioral models of depression / antidepressant response ; and second , by examining the role of the P08908 REA receptor subtype in the regulation of P15692 REA , and the cellular localization of antidepressant regulation of P15692 REA expression . The results show that pharmacological inhibition of P15692 REA receptor signaling blocks the behavioral actions of fluoxetine in rats subjected to chronic unpredictable stress . Infusions of SU5416 or SU1498 , two structurally dissimilar inhibitors of P15692 REA - Flk - 1 receptor signaling , block the antidepressant effects of fluoxetine on sucrose preference , immobility in the forced swim test , and latency to feed in the novelty suppressed feeding paradigm . We also show that activation of P08908 REA receptors is sufficient to induce P15692 REA expression and that a P08908 REA antagonist blocks both the increase in P15692 REA and behavioral effects induced by fluoxetine . Finally , double labeling studies show that chronic fluoxetine administration increases P15692 REA expression in both neurons and endothelial cells in the hippocampus . Taken together these studies show that P15692 REA is necessary for the behavioral effects of the SSRI fluoxetine , as well as norepinephrine selective reuptake inhibitor , and that these effects may be mediated by P08908 REA receptors located on neurons and endothelial cells .

Hypoxia-inducible vascular endothelial growth factor gene therapy using the oxygen-dependent degradation domain in myocardial ischemia . PURPOSE : A hypoxia-inducible P15692 REA expression system with the oxygen-dependent degradation ( Q8TAX0 ) domain was constructed and tested to be used in gene therapy for ischemic myocardial disease . METHODS : Luciferase and P15692 REA expression vector systems were constructed with or without the Q8TAX0 domain : pEpo-SV-Luc ( or pEpo-SV - P15692 REA ) and pEpo-SV-Luc - Q8TAX0 ( or pEpo-SV - P15692 REA - Q8TAX0 ) . In vitro gene expression efficiency of each vector type was evaluated in P29320 REA 293 cells under both hypoxic and normoxic conditions . The amount of P15692 REA protein was estimated by ELISA . The P15692 REA expression vectors with or without the Q8TAX0 domain were injected into ischemic rat myocardium . Fibrosis , neovascularization , and cardiomyocyte apoptosis were assessed using Masson ' s trichrome staining , α-smooth muscle actin ( α-SMA ) immunostaining , and the TUNEL assay , respectively . RESULTS : The plasmid vectors containing Q8TAX0 significantly improved the expression level of P15692 REA protein in hypoxic conditions . The enhancement of P15692 REA protein production was attributed to increased protein stability due to oxygen deficiency . In a rat model of myocardial ischemia , the pEpo-SV - P15692 REA - Q8TAX0 group exhibited less myocardial fibrosis , higher microvessel density , and less cardiomyocyte apoptosis compared to the control groups ( saline and pEpo-SV - P15692 REA treatments ) . CONCLUSION : An Q8TAX0 - mediated P15692 REA expression system that facilitates P15692 REA - production under hypoxia may be useful in the treatment of ischemic heart disease .

DB00472 MEN induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 MEN ( FLX ) is a drug commonly used as antidepressant . However , its effects on tumorigenesis remain controversial . Aiming to evaluate the effects of FLX treatment on early malignant changes , we analyzed serotonin ( 5 - HT ) metabolism and recognition , aberrant crypt foci ( Q9NQ94 ) , proliferative process , microvessels , vascular endothelial growth factor ( P15692 REA ) , and cyclooxygenase - 2 ( P35354 REA ) expression in colon tissue . Male Wistar rats received a daily FLX-gavage ( 30mgkg ( - 1 ) ) and , a single dose of 1,2 dimethylhydrazine ( Q03001 REA ; i . p . , 125mgkg ( - 1 ) ) . After 6 weeks of FLX-treatment , our results revealed that FLX and nor-fluoxetine ( N-FLX ) are present in colon tissue , which was related to significant increase in serotonin ( 5 - HT ) levels ( P < 0.05 ) possibly through a blockade in P31645 REA mRNA ( serotonin reuptake transporter ; P < 0.05 ) resulting in lower 5 - hydroxyindoleacetic acid ( 5 - HIAA ) levels ( P < 0.01 ) and , P28335 REA receptor mRNA expressions . FLX-treatment decreased dysplastic Q9NQ94 development ( P < 0.01 ) and proliferative process ( P < 0.001 ) in epithelia . We observed a significant decrease in the development of malignant microvessels ( P < 0.05 ) , P15692 REA ( P < 0.001 ) , and P35354 REA expression ( P < 0.01 ) . These findings suggest that FLX may have oncostatic effects on carcinogenic colon tissue , probably due to its modulatory activity on 5 - HT metabolism and / or its ability to reduce colonic malignant events .

Relation between hemostatic parameters and prognostic / predictive factors in breast cancer . BACKGROUND : In our study , we searched for a relation between various prognostic and predictive factors and hemostatic parameters . METHODS : One hundred women with newly diagnosed breast cancer after surgery were included . Patients did not receive systemic therapy or radiotherapy . The control group included 100 healthy , age-matched women . In the patient group , age , menopausal status , tumor size , grade , axillary lymph node status , steroid receptor status , p53 , and P04626 REA / neu were evaluated . Plasma levels of factor VIII , factor IX , D-dimer , fibrinogen , protein C , protein S , P04275 REA , and antithrombin III were measured in both groups . RESULTS : Plasma levels of factor VIII , factor IX , P04275 REA , and CRP in patients with breast cancer were higher than those in controls . Protein S levels in patients were lower than in controls . There was no significant difference in other hemostatic parameters between the groups . In patients with axillary lymph node metastasis , factor VIII levels were significantly higher than in node-negative patients . There was a strong correlation between axillary lymph node status , number of metastatic nodes , and factor VIII levels . There was no correlation between factor VIII levels and CRP . Factor VIII levels were higher in the group having high P04626 REA / neu ( 3 + ) than in the group with negativity for P04626 REA / neu . CONCLUSION : There was a strong correlation between axillary lymph node involvement , number of metastatic nodules , overexpression of P04626 REA / neu , hemostatic parameters , and factor VIII levels . Our study showed that factor VIII level measurement can provide additional data for evaluation of breast cancer patients ' prognosis .

DB00107 alleviates the neuroendocrine and cytokine response to bacterial endotoxin in healthy men . DB00107 is a hormone and neurotransmitter found to have anti-inflammatory functions in rodents . Here we used experimental bacterial endotoxinemia to examine the role of exogenous oxytocin administration on innate immune responses in humans . Ten healthy men received , in a randomized , placebo-controlled , crossover design , placebo , oxytocin , LPS , and LPS + oxytocin . DB00107 treatment resulted in a transient or prolonged reduction of endotoxin-induced increases in plasma DB01285 , cortisol , procalcitonin , P01375 REA , IL - 1 receptor antagonist , P05112 REA , P05231 REA , macrophage inflammatory protein - 1alpha , macrophage inflammatory protein - 1beta , monocyte chemoattractant protein - 1 ( P13500 REA ) , interferon-inducible protein 10 , and P15692 REA . In vitro , oxytocin had no impact on LPS effects in releasing P01375 REA , P05231 REA , and P13500 REA in monocytes and peripheral blood mononuclear cells from healthy human donors . In summary , oxytocin decreases the neuroendocrine and cytokine activation caused by bacterial endotoxin in men , possibly due to the pharmacological modulation of the cholinergic anti-inflammatory pathway . DB00107 might be a candidate for the therapy of inflammatory diseases and conditions associated with high cytokine and P15692 REA levels .

Role of angiotensin II in the remodeling induced by a chronic increase in flow in rat mesenteric resistance arteries . Angiotensin II is a potent growth factor involved in arterial wall homeostasis . In resistance arteries , chronic increases in blood flow induce a rise in diameter associated with arterial wall hypertrophy . Nevertheless , the role of angiotensin II in this remodeling is unknown . We investigated the effect of blocking angiotensin II production or receptor activation on flow-induced remodeling of mesenteric resistance arteries . Arteries were ligated in vivo to generate high-flow arteries compared with normal flow ( control ) vessels located at a distance . Arteries were isolated after 1 week for in vitro analysis . Arterial diameter , media surface , endothelial NO synthase expression , superoxide production , and extracellular signal-regulated kinase 1/2 phosphorylation were higher in high-flow than in control arteries . P12821 REA inhibition ( perindopril ) and angiotensin II type 1 receptor blockade ( candesartan ) prevented arterial wall hypertrophy without affecting diameter enlargement . The nonselective vasodilator hydralazine had no effect on remodeling . Although perindopril and candesartan increased endothelial NO synthase expression in high-flow arteries , hypertrophy remained in rats treated with N ( G ) - nitro-l-arginine methyl ester and mice lacking endothelial NO synthase . DB00790 SUB and candesartan reduced oxidative stress in high-flow arteries , but superoxide scavenging did not prevent hypertrophy . Both Tempol and the absence of endothelial NO synthase prevented the rise in diameter in high-flow vessels . P27361 REA / 2 activation in high-flow arteries was prevented by perindopril and candesartan and not by hydralazine . P27361 REA / 2 inhibition in vivo ( U0126 ) prevented hypertrophy in high-flow arteries . Thus , a chronic rise in blood flow in resistance arteries induces a diameter enlargement involving NO and superoxide , whereas hypertrophy was associated with extracellular signal-regulated kinase 1/2 activation by angiotensin II .

Sex steroid receptors , secondary bile acids and colorectal cancer . A possible mechanism of interaction . AIM : The aim of the work was to study in colon-rectum cancer mucosae the binding charateristics , as sex steroid receptors . METHODS : Specific androgen ( AR ) , estrogen ( ER ) and progesterone ( PgR ) receptors were measured in the tissue samples of 35 patients ( 15 males , 20 females ) undergoing colectomy or coloproctectomy for adenocarcinoma . The characteristics of androgen receptor ( AR , DB02901 - R : dihydrotestosterone receptor ) were also investigated using competitive activity of cyproterone acetate , cortisol , aldosterone and steroid-like substances such as deoxycholic and lithocholic acid , present in the milieu of the considered organ . Binding assays and competition tests were conducted using a charcoal dextran method . RESULTS : When present ( 50 % ) , ER and PgR receptors showed very low levels and no difference was noted between cancerous and the surrounding healthy mucosa . AR were found in all samples from both neoplastic and non neoplastic surrounding mucosa , with no significant difference . P10275 REA however exhibited an altered binding activity in cancer specimens . DB04839 MEN did not displace DB02901 from AR while significant displacing activity was elicited by DB02901 , testosterone , as well as by lithocholic acid , but not by deoxycholic acid . CONCLUSION : In cancerous large bowel mucosa , androgen receptors show altered binding characteristics . The selective binding of lithocholic acid to AR supports the hypothesis that diet-related endoluminal substances may play a role in cancer development model where molecular alterations such as DNA damage or mutation is the 1st event .

Bayesian meta-analysis of tissue angiotensin-converting enzyme inhibitors for reduction of adverse cardiovascular events in patients with diabetes mellitus and preserved left ventricular function . The role of angiotensin-converting enzyme ( P12821 REA ) inhibitors in diabetic patients with preserved ventricular function is uncertain . Tissue P12821 REA inhibitors have been defined by increased lipophilicity and structural characteristics that result in greater tissue-specific P12821 REA binding when compared with plasma P12821 REA inhibitors . A Bayesian meta-analysis of randomized trials was conducted to evaluate tissue P12821 REA inhibitors in prevention of cardiovascular disease among patients with diabetes mellitus and preserved left ventricular function . Four trials were selected that evaluated 2 different P12821 REA inhibitors and included 10,328 patients ( 43,517 patient-years ) . The DB00790 SUB Substudy in Coronary Artery Disease and Diabetes ( PERSUADE ) and the DB00790 SUB Protection Against Recurrent Stroke Study ( PROGRESS ) compared the effects of perindopril vs a placebo , and the Heart Outcomes Prevention Evaluation ( HOPE ) and the Non - P01308 REA - Dependent Diabetes , Hypertension , Microalbuminuria , Proteinuria , Cardiovascular Events , and Ramipril ( DIABHYCAR ) study investigated the impact of ramipril vs a placebo . Bayesian meta-analysis of sequential trials and sensitivity analysis of therapeutic response were subsequently computed . Bayesian meta-analysis determined reduced risk of cardiovascular mortality ( PB = . 991 ) , myocardial infarction ( PB = . 999 ) , and the need for invasive coronary revascularization ( PB = . 995 ) when compared with placebo . Total mortality was also decreased ( PB = . 967 ) , while the risk of stroke ( PB = . 907 ) and hospitalization for heart failure ( PB = . 923 ) were impacted . Bayesian meta-analysis of randomized trials suggests that tissue P12821 REA inhibitors decrease the probability that diabetic patients with preserved left ventricular function will experience myocardial infarctions and cardiovascular death and reduce overall mortality .

P10275 REA coregulator Q96L73 - alpha interacts with death receptor - 6 revealed by the yeast two-hybrid . Q96L73 - alpha is a newly identified androgen receptor coactivator . In order to further elucidate its precise role in cells , using the Q96L73 - alpha fragment containing four P20941 and one Q01105 REA conserved domains as bait we revealed an Q96L73 - alpha - P20941 - Q01105 REA - interacting protein , death receptor - 6 ( O75509 REA ) , in the yeast two-hybrid screening . O75509 REA is the member of P01375 REA receptor family and has a death domain in its intracellular cytoplasmic portion ( DR6cp ) to mediate the cell apoptosis . The interaction between Q96L73 - alpha - P20941 - Q01105 REA and DR6cp was confirmed in vitro and in vivo . Our finding implied that androgen signaling pathway might cross talk with apoptosis signaling pathway through the interaction between Q96L73 - alpha and O75509 REA .

DB06589 MEN inhibits the activation of P09619 REA β-expressing astrocytes in the brain metastatic microenvironment of breast cancer cells . Brain metastases occur in more than one-third of metastatic breast cancer patients whose tumors overexpress P04626 REA or are triple negative . Brain colonization of cancer cells occurs in a unique environment , containing microglia , oligodendrocytes , astrocytes , and neurons . Although a neuroinflammatory response has been documented in brain metastasis , its contribution to cancer progression and therapy remains poorly understood . Using an experimental brain metastasis model , we characterized the brain metastatic microenvironment of brain tropic , P04626 REA - transfected MDA-MB - 231 human breast carcinoma cells ( 231 - BR - P04626 REA ) . A previously unidentified subpopulation of metastasis-associated astrocytes expressing phosphorylated platelet-derived growth factor receptor β ( at tyrosine 751 ; p751 - P09619 REA β ) was identified around perivascular brain micrometastases . p751 - P09619 REA β ( + ) astrocytes were also identified in human brain metastases from eight craniotomy specimens and in primary cultures of astrocyte-enriched glial cells . Previously , we reported that pazopanib , a multispecific tyrosine kinase inhibitor , prevented the outgrowth of 231 - BR - P04626 REA large brain metastases by 73 % . Here , we evaluated the effect of pazopanib on the brain neuroinflammatory microenvironment . DB06589 MEN treatment resulted in 70 % ( P = 0.023 ) decrease of the p751 - P09619 REA β ( + ) astrocyte population , at the lowest dose of 30 mg / kg , twice daily . Collectively , the data identify a subpopulation of activated astrocytes in the subclinical perivascular stage of brain metastases and show that they are inhibitable by pazopanib , suggesting its potential to prevent the development of brain micrometastases in breast cancer patients .

Inhibition of the renin-angiotensin system improves physiological outcomes in mice with mild or severe cancer cachexia . Cancer cachexia describes the progressive skeletal muscle wasting and weakness associated with many cancers . Cachexia reduces mobility and quality of life and accounts for 20-30 % of all cancer-related deaths . Activation of the renin-angiotensin system causes skeletal muscle wasting and weakness . We tested the hypothesis that treatment with the angiotensin converting enzyme ( P12821 REA ) inhibitor , perindopril , would enhance whole body and skeletal muscle function in cachectic mice bearing Colon - 26 ( C - 26 ) tumors . CD2F1 mice received a subcutaneous injection of phosphate buffered saline or C - 26 tumor cells inducing either a mild or severe cachexia . The following day , one cohort of C - 26 mice began receiving perindopril in their drinking water ( 4 mg kg ( - 1 ) day ( - 1 ) ) for 21 days . In mild and severe cachexia , perindopril increased measures of whole body function ( grip strength and rotarod ) and reduced fatigue in isolated contracting diaphragm muscle strips ( p < 0.05 ) . In severely cachectic mice , perindopril reduced tumor growth , improved locomotor activity and reduced fatigue of tibialis anterior muscles in situ ( p < 0.05 ) , which was associated with increased oxidative enzyme capacity ( succinate deyhydrogenase , p < 0.05 ) . DB00790 SUB attenuated the increase in Q969Q1 REA and P05231 REA mRNA expression and enhanced Akt phosphorylation in severely cachectic mice but neither body nor muscle mass was increased . These findings support the therapeutic potential of P12821 REA inhibition for enhancing whole body function and reducing fatigue of respiratory muscles in early and late stage cancer cachexia and should be confirmed in future clinical trials . Since P12821 REA inhibition alone did not enhance body or muscle mass , co-treatment with an anabolic agent may be required to address these aspects of cancer cachexia .

[ An effect of perindopril on the level of tumor necrosis factor-alpha and matrix metalloproteinase - 9 in peripheral blood in the acute period of atherothrombotic ischemic stroke and myocardial infarction ] . Twenty-nine patients with acute atherothrombotic ischemic stroke and 36 patients with acute Q-wave myocardial infarction have been studied . Each group has been stratified into 2 subgroups : patients of subgroups A received an P12821 REA inhibitor perindopril in the complex therapy from the 1st day of disease . Patients of subgroups B were not assigned to this drug . Along with routine tests , the level of tumor necrosis factor-alpha and matrix metalloproteinase - 9 ( P14780 REA ) measured with ELISA using test-systems ( Q02223 REA Diagnostics , USA ) and reagents ( R & D , England ) have been determined . The administration of perindopril did not cause side-effects , including arterial hypotonia after the first dosage , in patients in the acute period of atherothrombotic ischemic stroke and myocardial infarction . DB00790 SUB may decrease the activity of P14780 REA in these patients and produces an anticytokine effect . Some similar mechanisms of ischemic lesions of the heart and the brain and a commonness of biochemical " response " to the same medical intervention ( the administration of an P12821 REA inhibitor perindopril ) in patients of both groups were found . The results support the pathogenetic validity of perindopril therapy in the secondary prevention of ischemic stroke and myocardial infarction .

Dual effect of angiotensin-converting enzyme inhibition on angiogenesis in type 1 diabetic mice . OBJECTIVE : We analyzed the beneficial therapeutic effect of angiotensin converting enzyme inhibitor ( ACEI ) on both retinal and hind limb neovascularization in diabetic mice . METHODS AND RESULTS : Diabetic mice ( streptozotocin , 40 mg / kg ) were treated with or without ACEI ( DB00790 SUB , 3 mg / kg per day ) or AT1 receptor blocker ( DB00796 , 20 mg / kg ) for 4 months . Hind limb ischemia was then induced by right femoral artery ligature for 1 additional month . In the ischemic leg , angiographic score , capillary density , and foot perfusion were increased by 2.7 , 2.0- fold , and 1.6- fold , respectively , in ACEI-treated diabetic mice compared with untreated diabetic animals ( P < 0.01 ) . ACEI also raised vascular endothelial growth factor ( P15692 REA ) protein level by 1.4- fold in ischemic diabetic leg . This ACEI pro-angiogenic effect was totally blunted in diabetic bradykinin B2 receptor-deficient animals , suggesting that it was mediated by the bradykinin pathway . In the diabetic retina , angiotensinogen and P12821 REA mRNA levels were increased by 2.8- fold and 4.1- fold , respectively ( P < 0.01 versus nondiabetic mice ) , highlighting a local activation of renin-angiotensin system . Diabetes also raised P15692 REA protein level by 1.5- fold ( P < 0.05 versus nondiabetic mice ) . Treatments with ACEI and AT1 receptor blocker hampered diabetes-induced P15692 REA upregulation and retinal neovascularization . CONCLUSIONS : P12821 REA inhibition improved neovascularization in the diabetic ischemic leg through activation of bradykinin signaling , whereas it reduced vessel growth in the diabetic retina through inhibition of overacting Ang II pathway .

Comparison of the novel antipsychotic ziprasidone with clozapine and olanzapine : inhibition of dorsal raphe cell firing and the role of P08908 REA receptor activation . Ziprasidone is a novel antipsychotic agent which binds with high affinity to P08908 REA receptors ( Ki = 3.4 nM ) , in addition to P28221 REA , 5 - HT2 , and D2 sites . While it is an antagonist at these latter receptors , ziprasidone behaves as a P08908 REA agonist in vitro in adenylate cyclase measurements . The goal of the present study was to examine the P08908 REA properties of ziprasidone in vivo using as a marker of central P08908 REA activity the inhibition of firing of serotonin-containing neurons in the dorsal raphe nucleus . In anesthetized rats , ziprasidone dose-dependently slowed raphe unit activity ( ED50 = 300 micrograms / kg i . v . ) as did the atypical antipsychotics clozapine ( ED50 = 250 micrograms / kg i . v . ) and olanzapine ( ED50 = 1000 micrograms / kg i . v . ) . Pretreatment with the P08908 REA antagonist WAY -100,635 ( 10 micrograms / kg i . v . ) prevented the ziprasidone-induced inhibition ; the same dose of WAY -100,635 had little effect on the inhibition produced by clozapine and olanzapine . Because all three agents also bind to alpha 1 receptors , antagonists of which inhibit serotonin neuronal firing , this aspect of their pharmacology was assessed with desipramine ( DB01151 MEN ) , a NE re-uptake blocker previously shown to reverse the effects of alpha 1 antagonists on raphe unit activity . DB01151 MEN ( 5 mg / kg i . v . ) failed to reverse the inhibitory effect of ziprasidone but produced nearly complete reversal of that of clozapine and olanzapine . These profiles suggest a mechanism of action for each agent , P08908 REA agonism for ziprasidone and alpha 1 antagonism for clozapine and olanzapine . The P08908 REA agonist activity reported here clearly distinguishes ziprasidone from currently available antipsychotic agents and suggests that this property may play a significant role in its pharmacologic actions .

In vivo imaging visualizes discoid platelet aggregations without endothelium disruption and implicates contribution of inflammatory cytokine and integrin signaling . The mechanism by which thrombotic vessel occlusion occurs independently of plaque development or endothelial cell ( EC ) disruption remains unclear , largely because of an inability to visualize the formation of thrombus , especially at the single-platelet level in real time . Here we demonstrate that rapidly developing thrombi composed of discoid platelets can be induced in the mesenteric capillaries , arterioles , and large-sized arteries of living mice , enabling characterization of the kinetics of thrombosis initiation and the multicellular interrelationships during thrombus development . Platelet aggregation without EC disruption was triggered by reactive oxygen species ( ROS ) photochemically induced by moderate power laser irradiation . The inflammatory cytokines P01375 REA - α and IL - 1 could be key components of the EC response , acting through regulation of P04275 REA mobilization to the cell surface . Thrombus formation was then initiated by the binding of platelet GPIbα to endothelial P04275 REA in our model , and this effect was inhibited by the ROS scavenger DB06151 . Actin linker talin-dependent activation of alphaIIb-beta 3 integrin or Rac 1 in platelets was required for late-phase thrombus stability . Our novel imaging technology illustrates the molecular mechanism underlying inflammation-based thrombus formation by discoid platelets on undisrupted ECs and suggests control of ROS could be a useful therapeutic target for the prevention of thrombotic diseases .

Quantitative trait locus mapping of susceptibilities to butylated hydroxytoluene-induced lung tumor promotion and pulmonary inflammation in CXB mice . We have reported previously [ Bauer , A . K . et al . ( 2001 ) Exp . Lung Res . , 27 , 197-216 ] that the 13 CXB recombinant inbred mouse strains derived from BALB / cByJ and C57BL / 6J progenitors vary in their responsiveness to both lung tumor promotion and pulmonary inflammation induced by chronic administration of butylated hydroxytoluene ( BHT ) . Herein we have applied these data , along with markers known to be polymorphic among these strains , to conduct linkage analysis of these susceptibilities . This enabled us to assign provisional quantitative trait loci ( QTL ) that govern these strain variations in susceptibility as a genetic approach to assessing the influence of inflammation on tumorigenesis . A Chr 15 ( Q04695 REA -55.6 cM ) QTL regulated susceptibility to two-stage carcinogenesis , a protocol in which chronic BHT exposure followed a single urethane injection ; a similar QTL on Chr 15 ( 46.7- 61.7 cM ) influenced BHT induction of cyclooxygenase - 2 ( P35354 REA ) expression . A Chr 18 ( 37-41 cM ) QTL modulated both the number of lung tumors induced by 3 - methylcholanthrene ( MCA ) injection with subsequent treatment with BHT as well as BHT-induced ingress of macrophages into airways . Other chromosomal sites that affected either the degree of BHT-elicited macrophage infiltration , Chr 9 ( 48-61 cM ) , or P35354 REA induction , Chr 10 ( 59-65 cM ) , were reported to influence susceptibility to lung tumorigenesis in other strains . The fact that common chromosomal locations regulate both inflammation and carcinogenesis suggests a pathogenic role of inflammatory mediators in tumor development that may be exploited for chemoprevention of lung cancer .

17 DB00783 overcomes a P55008 block induced by P04035 REA inhibitors and fosters cell cycle progression without inducing P27361 REA and - 2 Q96HU1 kinases activation . P04035 REA inhibitors , such as DB00227 MEN and Simvastatin , cause cell cycle arrest by interfering with the mitogenic activity of mitogens present in culture media . Cells are induced to pause in P55008 and can readily resume growth upon removal of the enzymatic block . DB00286 , acting via their nuclear receptor , are mitogens for different normal and transformed cell types , where they foster cell cycle progression and cell division . In estrogen-responsive MCF - 7 human breast cancer cells , but not in non responsive cells , 17 beta-estradiol ( E2 ) induces cells arrested with DB00227 MEN or Simvastatin to proliferate in the presence of inhibitor , without restoring P04035 REA activity or affecting the protein prenylation pattern . Mitogenic stimulation of P55008 - arrested MCF - 7 cells with E2 includes primary transcriptional activation of c-fos , accompanied by transient binding in vivo of the estrogen receptor and / or other factors to the ERE and the estrogen-responsive DNA region of this proto-oncogene , as detected by dimethylsulphate genomic footprinting analysis . Mitogenic stimulation of growth-arrested MCF - 7 cells by E2 occurs , under these conditions , without evident activation of P27361 REA and - 2 kinases , and thus independently from the mitogen-responsive signal transduction pathways that converge on these enzymes .

Modeling of Q14654 REA and inhibition mechanism of the natural ligand , ellagic acid , using molecular docking . Diabetes mellitus is a disorder in which blood sugar ( glucose ) levels are abnormally high because the body does not produce enough insulin to meet its needs . Post-prandial hyperglycemia ( PPHG ) is an independent risk factor for the development of macro vascular complications . It is now recognized that normalizing post-prandial blood glucose is more difficult than normalizing fasting glucose . DB01345 channels are the most widely distributed type of ion channel and are found in virtually all living organisms . The function of KATP channels is best understood in pancreatic beta cells , the membrane potential of which is responsive to external glucose concentration . Beta cells show a remarkably complex electrical bursting behavior in response to an increase in glucose level . DB00731 and DB00222 MEN are a class of insulin secretagog agents that lowers blood glucose levels by stimulating insulin secretion from the pancreas . These compounds interact with the DB00171 - sensitive potassium ( K + DB00171 ) channel in pancreatic beta cells . However , the side effects of these drugs overpass their uses , and the need to identify compounds with less adverse effects is exigent . In our research study , we used the natural compound ellagic acid , which is an already proven anti-carcinogen , anti-mutagen , and anticancer initiator , for its anti-diabetic activity in comparison to the two commercial drugs ( DB00731 and DB00222 MEN ) . The drugs and the compounds were docked to the DB00171 - dependent potassium channel and their energy value showed that the compound had higher binding value than the commercial drugs . Then an ADME / Tox analysis for the compound was carried out which showed that ellagic can be a possible lead molecule .

Different effects of perindopril and enalapril on monocyte cytokine release in coronary artery disease patients with normal blood pressure . BACKGROUND : Favorable effects of angiotensin-converting enzyme ( P12821 REA ) inhibitor treatment on the incidence of cardiovascular and cerebrovascular mortality and morbidity are not limited to patients with elevated blood pressure . As suggested by our previous results , the physicochemical and pharmacokinetic differences between drugs may markedly contribute to the strength of pleiotropic effects of P12821 REA inhibitors . METHODS : The present study was aimed at comparing the effects of serum - and tissue-type P12821 REA inhibitors on monocyte release of proinflammatory cytokines in normotensive patients with stable coronary artery disease . The participants were randomized to 90 - day treatment with enalapril ( 20 mg daily , n = 29 ) , perindopril ( 4 mg daily , n = 27 ) or placebo ( n = 28 ) . Plasma levels of lipids , glucose , insulin and high sensitivity P02741 REA ( hsCRP ) , as well as monocyte release of proinflammatory cytokines were determined before and after 30 days of therapy , and at the end of the treatment . RESULTS : Lipopolysaccharide-stimulated monocytes from normotensive patients with stable coronary artery disease released significantly more P01375 REA - α , interleukin - 1β and monocyte chemoattractant protein - 1 in comparison with monocytes from 23 matched control subjects . Their baseline hsCRP levels were also higher . DB00790 SUB reversed the disease-induced changes in cytokine release and reduced plasma hsCRP , while the effect of enalapril was much more limited . The effect on both drugs on cytokine release was stronger in insulin-resistant than insulin-sensitive subjects . CONCLUSIONS : Our results indicate that perindopril is superior to enalapril in producing monocyte-suppressing and systemic anti-inflammatory effects in normotensive patients with coronary artery disease . This action may contribute to the clinical effectiveness of tissue P12821 REA inhibitors in the therapy of atherosclerosis-related disorders , particularly in insulin-resistant subjects .

DB00790 SUB : possible use in cancer therapy . Since angiogenesis is essential for the growth of any solid tumor , emerging efforts are being made to develop antiangiogenic therapy . To date , however , no antiangiogenic agent has become widely available for the clinical setting . Angiotensin I-converting enzyme ( P12821 REA ) inhibitors are commonly used as antihypertensive agents and it has recently been suggested that they decrease the risk of cancer . Studies have found that an P12821 REA inhibitor , perindopril , is a potent inhibitor of experimental tumor development and angiogenesis at a clinically comparable dose . The potent angiogenic factor , vascular endothelial growth factor ( P15692 REA ) , is significantly suppressed by perindopril and also inhibits P15692 REA - induced tumor growth . In vitro studies showed that perindopril is not cytotoxic to either tumor cells or endothelial cells . Since perindopril is already in widespread clinical use without serious side effects , it may represent a potential new strategy for anticancer therapy .

P62158 interacts with angiotensin-converting enzyme - 2 ( Q9BYF1 ) and inhibits shedding of its ectodomain . P12821 REA - 2 ( Q9BYF1 ) is a regulatory protein of the renin-angiotensin system ( DB01367 ) and a receptor for the causative agent of severe-acute respiratory syndrome ( P49591 REA ) , the P49591 REA - coronavirus . We have previously shown that Q9BYF1 can be shed from the cell surface in response to phorbol esters by a process involving P01375 REA converting enzyme ( P78536 REA ; P78536 REA ) . In this study , we demonstrate that inhibitors of calmodulin also stimulate shedding of the Q9BYF1 ectodomain , a process at least partially mediated by a metalloproteinase . We also show that calmodulin associates with Q9BYF1 and that this interaction is decreased by calmodulin inhibitors .

Ds-echinoside A , a new triterpene glycoside derived from sea cucumber , exhibits antimetastatic activity via the inhibition of NF-κB-dependent P14780 REA and P15692 REA expressions . Ds-echinoside A ( DSEA ) , a non-sulfated triterpene glycoside , was isolated from the sea cucumber Pearsonothuria graeffei . In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion , migration , invasion , and angiogenesis . In this study , we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2 , with a half-maximal inhibitory concentration ( IC₅₀ ) of 2.65 μmol / L , and suppressed Hep G2 cell adhesion , migration , and invasion in a dose-dependent manner . DSEA also reduced tube formation of human endothelial cells ECV - 304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane ( P62158 ) assay in vivo . Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase - 9 ( P14780 REA ) , which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis . DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase - 1 ( P01033 REA ) , an important regulator of P14780 REA activation . From the results of Western blotting , the expressions of nuclear factor-kappa B ( NF-κB ) and vascular endothelial growth factor ( P15692 REA ) were found to be remarkably reduced by DSEA . These findings suggest that DSEA exhibits a significant anti-metastatic activity through the specific inhibition of NF-κB-dependent P14780 REA and P15692 REA expressions .

[ Network pharmacology study on major active compounds of Fufang Danshen formula ] . OBJECTIVE : To investigate the correlations between multi-compounds of Fufang Danshen formula and their multi targets and multi diseases . METHOD : Literature knowledge of nine major active compounds from Fufang Danshen formula , including tanshinone II ( A ) , salvianolic acid B , protocatechuic aldehyde , danshensu , cryptotanshinone , notoginsenoside Q96GN5 , ginsenoside Rg1 , DB06749 and borneol were collected from PubMed . Combined with cardiovascular related diseases and genes from OMIM database , the corresponding multi-compound-multi - target-multi-disease network was constructed and visualized by Cytoscape software . RESULT : AND CONCLUSION : Network analysis showed that the 9 compounds could modulate 42 cardiovascular associated genes ( e . g . P37231 REA , P12821 REA , Q14654 REA , P51787 REA , Q09428 REA , et al ) , which related to 30 cardiovascular associated diseases including non-insulin-dependent diabetes mellitus , hyperinsulinemic hypoglycemia , hypertension , and coronary heart disease . These results suggested new potential indications of Fufang Danshen formula .

P12821 REA activity is involved in the mechanism of increased endogenous nitric oxide synthase inhibitor in patients with type 2 diabetes mellitus . The renin-angiotensin system plays an important role in the elevation of asymmetric dimethylarginine ( DB01686 ) , an endogenous inhibitor of nitric oxide synthase , in hypertensive patients , so the present study was designed to examine whether angiotensin-converting enzyme ( P12821 REA ) activity is also involved in the mechanism of DB01686 elevation in type 2 diabetes mellitus ( NIDDM ) . A crossover study was performed to determine if P12821 REA inhibition with perindopril ( 4 mg / day ) for 4 weeks decreases serum DB01686 concentration and plasma P04275 REA ( P04275 REA ) level ( a marker of endothelial injury ) in 11 patients with NIDDM . None of the patients was treated with insulin or oral hypoglycemic drugs , and none had major diabetic complications . Before the protocol began , serum DB01686 and plasma P04275 REA were significantly higher in the 11 NIDDM patients , when compared with 8 control subjects without diabetes . DB00790 SUB did not affect blood pressure or glucose metabolism , but did significantly decrease serum DB01686 and plasma P04275 REA . These results suggest that endothelial injury associated with DB01686 elevation may be present even in patients with non-complicated NIDDM , and that increased activity of P12821 REA may be involved in such endothelial dysfunction .

Differential selectivity of insulin secretagogues : mechanisms , clinical implications , and drug interactions . The sulphonylurea receptor ( Q09428 REA ) subunits of K ( DB00171 ) channels are the targets for several classes of therapeutic drugs . Sulphonylureas close K ( DB00171 ) channels in pancreatic beta-cells and are used to stimulate insulin release in type 2 diabetes , whereas the K ( DB00171 ) channel opener nicorandil acts as an antianginal agent by opening K ( DB00171 ) channels in cardiac and vascular smooth muscle . The predominant type of Q09428 REA varies between tissues : Q09428 REA in beta-cells , SUR 2A in cardiac muscle , and SUR 2B in smooth muscle . Sulphonylureas and related drugs exhibit differences in tissue specificity , as the drugs interact to varying degrees with different types of Q09428 REA . DB01120 and tolbutamide are beta-cell selective and reversible . DB00222 MEN , glibenclamide , and repaglinide , however , inhibit cardiac and smooth muscle K ( DB00171 ) channels in addition to those in beta-cells and are only slowly reversible . Similar properties have been observed by recording K ( DB00171 ) channel activity in intact cells and in Xenopus oocytes expressing cloned K ( DB00171 ) channel subunits . While K ( DB00171 ) channels in cardiac and smooth muscle are largely closed under physiological conditions ( but open during ischaemia ) , they are activated by antianginal agents such as nicorandil . Under these conditions , they may be inhibited by sulphonylureas that block SUR 2 - type K ( DB00171 ) channels ( e . g . , glibenclamide ) . Care should , therefore , be taken when choosing a sulphonylurea if potential interactions with cardiac and smooth muscle K ( DB00171 ) channels are to be avoided .

Association of sixty-one non-synonymous polymorphisms in forty-one hypertension candidate genes with blood pressure variation and hypertension . We previously selected a group of hypertension candidate genes by a key word search using the OMIM database of NCBI and validated 525 coding single nucleotide polymorphisms ( SNPs ) in 179 hypertension candidate genes by DNA sequencing in a Japanese population . In the present study , we examined the association between 61 non-synonymous SNPs and blood pressure variations and hypertension . We used DNA samples taken from 1,880 subjects in the Suita study , a population-based study using randomly selected subjects . Analyses of covariance adjusting for age , body mass index , hyperlipidemia , diabetes , smoking , drinking , and antihypertensive medication revealed that 17 polymorphisms in 16 genes ( P04114 REA , CAST , P51801 REA , O60931 REA , P10912 REA , P13807 REA , P08603 REA , O95163 , Q14654 REA , P11150 REA , P06858 REA , P41231 REA , Q15165 REA , P02730 REA , TRH , P04275 REA ) were significantly associated with blood pressure variations . Multivariate logistic regression analysis with adjustment for the same factors revealed that 11 polymorphisms in 11 genes ( CAST , P16410 REA , P12259 REA , GC , P10912 REA , P11150 REA , Q13093 REA , P02730 REA , SLCI 8A1 , TRH , P04275 REA ) showed significant associations with hypertension . Five polymorphisms in five genes , CAST ( calpastatin ) , P11150 REA ( hepatic lipase ) , P02730 REA ( band 3 anion transporter ) , TRH ( thyrotropin-releasing hormone ) , and P04275 REA ( P04275 REA ) , were significantly associated with both blood pressure variation and hypertension . Thus , our study suggests that these five genes were susceptibility genes for essential hypertension in this Japanese population .

P10275 REA YAC transgenic mice recapitulate SBMA motor neuronopathy and implicate VEGF 164 in the motor neuron degeneration . X-linked spinal and bulbar muscular atrophy ( SBMA ) is an inherited neuromuscular disorder characterized by lower motor neuron degeneration . SBMA is caused by polyglutamine repeat expansions in the androgen receptor ( AR ) . To determine the basis of AR polyglutamine neurotoxicity , we introduced human AR yeast artificial chromosomes carrying either 20 or 100 CAGs into mouse embryonic stem cells . The AR100 transgenic mice developed a late-onset , gradually progressive neuromuscular phenotype accompanied by motor neuron degeneration , indicating striking recapitulation of the human disease . We then tested the hypothesis that polyglutamine-expanded AR interferes with CREB binding protein ( CBP ) - mediated transcription of vascular endothelial growth factor ( P15692 REA ) and observed altered CBP-AR binding and P15692 REA reduction in AR100 mice . We found that mutant AR-induced death of motor neuron-like cells could be rescued by P15692 REA . Our results suggest that SBMA motor neuronopathy involves altered expression of P15692 REA , consistent with a role for P15692 REA as a neurotrophic / survival factor in motor neuron disease .

P28335 REA receptor involvement in female rat lordosis behavior . Adult , hormone-primed , ovariectomized rats ( P05231 REA - 344 ) with bilateral implants within the ventromedial nucleus of the hypothalamus ( VMN ) , were injected with 0.5 microgram estradiol benzoate followed 48 h later with 500 microgram progesterone . This priming produced rats with 2 different levels of sexual receptivity . Rats with a lordosis to mount ratio ( L / M ) > /= 0.5 were used to examine the potential lordosis-inhibiting effects of the 5 - Q13049 REA receptor antagonist , R ( + ) - a - ( 2 , 3 - dimethoxyphenyl ) - 1 - [ 2 ( 4 - fluoro-phenylethyl ) ] - 4 - piperidine-methanol ( MDL 100,907 ) , and the P28335 REA receptor antagonist , 5 - methyl - 1 - ( 3 - pyridylcarbamoyl ) -1,2 , 3,5- tetrahydropyrrolo [ 2 , 3 - f ] indole ( SB 206553 ) . Rats with low sexual receptivity ( L / M < 0.5 ) were bilaterally infused with the 5 - Q13049 REA / 2C receptor agonist , ( + / - ) - 1 - ( 2,5- dimethoxy - 4 - iodophenyl ) - 2 - aminopropane HCl ( DOI ) , or DOI plus either MDL 100,907 or SB 206553 to determine if either drug would attenuate the lordosis-facilitating effects of DOI . The P28335 REA receptor antagonist , but not the 5 - Q13049 REA receptor antagonist , effectively inhibited lordosis behavior . Similarly , SB 206553 was more effective than MDL 100,907 in reducing the DOI-induced increase in lordosis responding . However , both drugs limited the duration of lordosis responding initiated by DOI . These results are consistent with prior suggestions that 5 - Q13049 REA / 2C receptors within the VMN are involved in the modulation of lordosis behavior and lead to the suggestion that P28335 REA , rather than 5 - Q13049 REA , receptors are primarily responsible for the effects of 5 - HT2 receptor-active drugs on lordosis behavior .

Investigation of mechanisms mediating 8 - OH-DPAT-induced impairment of spatial memory : involvement of P08908 REA receptors in the dorsal hippocampus in rats . The purpose of this study was to identify mechanisms that mediate the impairment of spatial memory induced by 8 - hydroxy - 2 - ( di-n-propylamino ) tetralin ( 8 - OH-DPAT ) , a P08908 REA / P34969 REA receptor agonist , in the eight-arm radial maze in rats . WAY - 100635 and NAN - 190 , P08908 REA receptor antagonists , reversed the impairment of spatial memory induced by systemic injection of 8 - OH-DPAT ( 1 mg / kg , i . p . ) . On the other hand , the alpha 1 - adrenoceptor antagonist prazosin and a selective P34969 REA receptor antagonist SB269970 had no effect on 8 - OH-DPAT-induced impairment of spatial memory . Bilateral microinjection of 8 - OH-DPAT ( 4 microg / side ) impaired spatial memory when injected into the dorsal hippocampus ( DH ) . Contrastingly , spatial memory was unaffected by microinjections of 8 - OH-DPAT into the other six areas examined : ventral hippocampus ( VH ) , central amygdaloid nucleus ( P12821 REA ) , lateral hypothalamus ( LH ) , nucleus accumbens ( NAc ) , and dorsal ( DR ) and median ( MR ) raphe nucleus . Furthermore , NAN - 190 significantly reversed the impairment of spatial memory induced by intra-DH injection of 8 - OH-DPAT . These findings suggest that P08908 REA receptors in the DH play an important role in the mechanisms underlying the 8 - OH-DPAT-induced impairment of spatial memory in rats .

Implantation of P15692 REA transfected preadipocytes improves vascularization of fibrin implants on the cylinder chorioallantoic membrane ( P62158 ) model . The successful substitution or augmentation of soft tissues by implantation of three dimensional cell constructs , consisting of human preadipocytes and fibrin glue as a carrier matrix , requires a rapid and homogeneous vascularization of the whole implant in order to provide a sufficient blood supply of centrally situated cells . Previous investigations have shown that under in vivo conditions primary human preadipocytes induce vascularization of fibrin matrices by secretion of several growth factors , such as P15692 REA and P09038 REA . The current study investigates whether vascularization of implants can be improved by transplantation of preadipocytes following transfection with a P15692 REA - vector . Transfection was performed by electroporation with an pCMX-GFP and pCMX-VEGF 165 vector . Transfection efficiency ( GFP expression ) and P15692 REA expression were determined in vitro by FACS analysis and P15692 REA immunoassay , respectively . In vivo investigations to determine the vascularization of the implants were performed on the cylinder chorioallantoic membrane ( P62158 ) . Four million P15692 REA transfected cells were transferred within a fibrin matrix onto the P62158 on the 7 ( th ) day of incubation and after 8 days the vascularization of the implant was histologically examined and evaluated by means of a computer-assisted image analysis program . Transfection of preadipocytes with the GFP vector by electroporation yielded transfection efficiencies between 12 % and 41 % of surviving cells . Results of the P15692 REA immunoassay demonstrated that P15692 REA expression was significantly higher following transfection . Investigations on the P62158 outlined a significantly higher rate of vascularization in the transfected vs . control population . Our investigations demonstrate that primary human preadipocytes can be successfully transfected by electroporation with a P15692 REA vector . The enhanced P15692 REA expression on transfected cells results in an increase of vascularization of the cell constructs on the P62158 .

Regulation of P01160 REA secretion from isolated atria by prostaglandins and cyclooxygenase - 2 . Cyclooxygenase ( P36551 REA ) is a key enzyme regulating the production of various prostaglandins ( PGs ) from arachidonic acid . Angiotensin II has been reported to be an important inflammatory mediator , which increases P35354 REA . The aim of this study was to determine the role of various PGs and P35354 REA in the regulation of atrial natriuretic peptide ( P01160 REA ) secretion . PGF 2alpha and PGD 2 caused dose-dependent increases in P01160 REA release and intra-atrial pressure . The potency for the stimulation of P01160 REA secretion by PGF 2alpha was higher than that by PGD 2 . In contrast , DB00917 , DB01240 , PGJ 2 , and thromboxane A2 did not show any significant effects . The increases in intra-atrial pressure and P01160 REA secretion induced by PGF 2alpha and PGD 2 were significantly attenuated by the pretreatment with an inhibitor of PGF 2alpha receptor . By the pretreatment with an inhibitor for phospholipase C ( P98160 REA ) , inositol 3 - phosphate ( IP3 ) receptor , protein kinase C ( PKC ) , or myosin light chain kinase ( MLCK ) , PGF 2alpha - mediated increase in P01160 REA secretion and positive inotropy were attenuated . Inhibitor for P23219 REA or P35354 REA did not cause any significant effects on atrial parameters . In hypertrophied rat atria , PGF 2alpha - induced positive inotropy and P01160 REA secretion were markedly attenuated whereas P35354 REA inhibitor stimulated P01160 REA secretion . The expression of P35354 REA increased and the expression of PGF 2alpha receptor mRNA decreased in hypertrophied rat atria . These results suggest that PGF 2alpha increased the P01160 REA secretion and positive inotropy through P98160 REA - IP3 - PKC-MLCK pathway , and the modulation of P01160 REA secretion by P35354 REA inhibitor and PGF 2alpha may partly relate to the development of renal hypertension .

The use of microcalorimetry and HPLC for the determination of degradation kinetics and thermodynamic parameters of DB00790 SUB Erbumine in aqueous solutions . DB00790 SUB Erbumine ( O15534 REA ) is one of the newly used angiotensin-converting enzyme inhibitors ( P12821 REA inhibitors ) and is used for the treatment of patients with hypertension and symptomatic heart failure . It has two main degradation pathways , i . e . the degradation by hydrolysis and the degradation by cyclization . An isothermal heat conduction microcalorimetry ( MC ) and high pressure liquid chromatography ( HPLC ) were used for the characterization of aqueous solutions of O15534 REA and its stability properties . The rates of heat evolved during degradation of perindopril were measured by MC as a function of temperature and pH and from these data rate constant and change in enthalpy of the reactions were determined . With the HPLC method the concentration of perindopril and its degradation products were measured as a function of time in aqueous solutions of different pH that were stored at different temperatures . We demonstrated that reactions of degradation of perindopril at observed conditions follow the first order kinetics . The Arrhenius equation for each pH was determined . At pH 6.8 only one degradation pathway is present , i . e . the degradation by hydrolysis . Degradation constants for this pathway calculated from MC data are in good agreement with those obtained from HPLC . MC as a non-specific technique was shown to be useful in studies of O15534 REA when one reaction was present in the sample and also when more chemical and physical processes were simultaneously running .

P10275 REA promotes esophageal cancer cell migration and proliferation via matrix metalloproteinase 2 . Esophageal squamous cell carcinoma ( ESCC ) is one of the most common malignancies worldwide . P10275 REA ( AR ) plays an important role in many kinds of cancers . However , the molecular mechanisms of AR in ESCC are poorly characterized . In the present study , Western blot analysis and real-time quantitative PCR were performed to identify differentially expressed AR in 40 ESCC tissue samples , which revealed that the messenger RNA ( mRNA ) and protein expression of AR is upregulated in the ESCC tissue samples . AR overexpression induced increases in ESCC cell invasion and proliferation in vitro . Silencing of AR inhibited the proliferation of KYSE 450 cells which have a relatively high level of AR , and the invasion of KYSE 450 cells was distinctly suppressed . Furthermore , AR knockdown led to substantial reductions in matrix metalloproteinase 2 ( P08253 REA ) and p-AKT levels in ESCC cell lines , but no significant change in AKT and P14780 REA expression . These results suggest that AR is involved in tumor progression , and thus , AR could represent selective targets for the molecularly targeted treatments of ESCC .

REV-ERBα inhibits the P35354 REA expression in bovine uterus endometrium stromal and epithelial cells exposed to ovarian steroids . The nuclear receptor REV-ERBα ( encoded by P20393 REA ) has a critical role in metabolism and physiology as well as circadian rhythm . Here , we investigated the possible contribution of clock genes including P20393 REA to the secretion of prostaglandin F2α ( PGF 2α ) from bovine uterine stromal ( USCs ) and epithelial cells ( UECs ) by modulating the expression of P35354 REA . The circadian oscillation of clock genes in the cells was weak compared with that reported in rodents , but the expression of O00327 REA , O15534 REA , and P20393 REA was changed temporally by treatment with ovarian steroids . Significant expression of clock genes including P20393 REA was detected in USCs exposed to progesterone . P20393 REA was also significantly expressed in UECs exposed to estradiol . The expression of P35354 REA was suppressed in USCs exposed to progesterone , while the expression was initially suppressed in UECs exposed to estradiol and then increased after long-term exposure to estradiol . O00327 REA knockdown with specific siRNA caused a significant decrease in the transcript levels of P20393 REA and P35354 REA in USCs , but not in UECs . The production of PGF 2α also decreased in USCs after O00327 REA knockdown , while its level did not significantly change in UECs . The transcript level of P35354 REA was increased by treatment with the antagonist of REV-ERBα in both cell types , but the agonist was ineffective . In these two cell types treated with the agonist or antagonist , the PGF 2α production coincided well with the P35354 REA expression . Collectively , these results indicate that REV-ERBα plays an inhibitory role in the expression of P35354 REA in both bovine USCs and UECs treated with ovarian steroids .