MH_dev_322

Effects of external calcium on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . DB01373 is a known signalling molecule in eukaryotic cells and plays a central role in the regulation of many cellular processes . In the following study , we report on the effect of external calcium treatments on the biotransformation of DB06749 to ginsenoside Rd by Paecilomyces bainier 229-7 . We observed that the intracellular calcium content of P . bainier 229-7 mycelia was increased in response to exposure to high external Ca ( 2 + ) concentrations . Both ginsenoside Rd biotransformation and β-glucosidase activity were both found to be dependent on the external calcium concentration . At an optimal Ca ( 2 + ) concentration of 45 mM , maximal ginsenoside Rd bioconversion rate of 92.44 % was observed and maximal β-glucosidase activity of 0.1778 U was reached in a 72 - h biotransformation . The Ca ( 2 + ) channel blocker Verapamil blocked the trans-membrane influx of calcium and decreased ginsenoside Rd biotransformatiom . In addition , β-glucosidase activity and ginsenoside Rd content decreased by 36.0 and 29.2 % respectively after a 72 - h incubation in the presence of 0.05 mM P62158 ( P62158 ) antagonist DB00850 MENMAX DB00850 MEN . These results suggest that both Ca ( 2 + ) channels and P62158 are involved in ginsenoside Rd biotransformation via regulation of β-glucosidase activity . This is the first report regarding the effects of calcium signal transduction on biotransformation and enzyme activity in fungi .

DB00472 SUB - induced proliferation and differentiation of neural progenitor cells isolated from rat postnatal cerebellum . Previous studies have shown that the serotonin-reuptake inhibitor ( SSRI ) fluoxetine affects neural progenitors derived from postnatal cerebellum or adult hippocampus and stimulates their proliferation . In the human cerebellum , the proliferation of cerebellar granule cells ( CGC ) continues until the 11th postnatal month and could be influenced in infants by breastfeeding-delivered SSRIs . Current information about fluoxetine effects on postnatal cerebellar neural progenitors is limited . Here we report the characterization of fluoxetine actions on rat postnatal cerebellar neural progenitors . RT-PCR and immunostaining revealed the expression of serotonin transporter ( P31645 REA ) , 5HT ( 1A ) receptors , tryptophan hydroxylase ( P17752 REA ) , and serotonin ( 5HT ) . Protracted in vitro fluoxetine treatment increased cell proliferation and differentiation . The proliferative effects of fluoxetine , 5HT , and the selective agonist of 5HT ( 1A ) receptors trans - 8 - hydroxy - 2 - ( N-n-propyl-N - 3 ' - iodo - 2 ' - propenyl ) aminotetralin ( 8 - OH-PIPAT ) were abolished by the selective antagonist of 5HT ( 1A ) receptors , N - [ 2 - [ 4 - ( 2 - methoxyphenyl ) - 1 - piperazinyl ] ethyl ] - N - ( 2 - pyridinyl ) cyclohexanecarboxamide trihydrochloride ( WAY - 100635 ) . Furthermore , fluoxetine-induced activation of both the DB02527 - response element-binding ( CREB ) protein and extracellular signal-regulated protein kinases ( P27361 REA / 2 ) , which was abolished by the selective inhibitor of Q96HU1 kinase kinase ( MEK ) 1,4- diamino -2,3- dicyano -1,4- bis ( 2 - aminophenylthio ) butadiene ( U0126 ) , and increased cyclin D1 expression . All these effects were prevented by WAY - 100635 . Collectively , our results demonstrate that rat postnatal cerebellum contains neural progenitors capable of proliferating and differentiating in response to fluoxetine exposure , possibly through the activation of 5HT ( 1A ) receptors . The relevance of these findings for possible SSRI effects on the developing postnatal / infant human cerebellum should be explored .

DB00472 SUB pharmacogenetics in child and adult populations . Although fluoxetine is useful in the treatment of major depression , 30-40 % of the patients do not respond to therapy . The response seems to be influenced by certain genes which are involved in the drug ' s pharmacodynamics and pharmacokinetics . The present study reviews the literature on genetic contributions to fluoxetine response in children and adults , and concludes that the different polymorphisms of P10635 REA and P11712 REA may influence the blood concentrations of fluoxetine . If the childhood dose is adjusted for weight , differences between children and adults are unlikely . As regards the genes that influence the drug ' s pharmacodynamics , polymorphisms of P31645 REA , P08908 REA and P21397 REA seem to be involved in the response to fluoxetine , while the genes P21964 REA , P34998 REA , PDEA 1 , PDEA 11 P49841 REA and serpin - 1 also seem to play a role . Comparison of different studies reveals that the results are not always consistent , probably due to methodological differences . Other factors such as gender or ethnicity may also influence treatment response .

Acyl substitution at the ortho position of anilides enhances oral bioavailability of thiophene sulfonamides : TBC 3214 , an P25101 REA selective endothelin antagonist . DB06268 MEN ( 3 , TBC 11251 ) ( Wu et al . J . Med . Chem . 1997 , 40 , 1690 ) is an orally active ET ( A ) selective endothelin antagonist that attenuates pulmonary vascular hypertension and cardiac hypertrophy in rats ( Tilton et al . Pulm . Pharmacol . Ther . 2000 , 13 , 87 ) . It has demonstrated efficacy in a phase II clinical trial for congestive heart failure ( Givertz et al . Circulation 2000 , 101 , 2922 ) . During the discovery of 3 , we observed several structure-oral bioavailability relationships . To investigate whether there is any generality in these trends , we synthesized some similar pairs of compounds in the latest series ( Wu et al . J . Med . Chem . 1999 , 42 , 4485 ) and evaluated their oral properties . In both series , an acyl group at the 2 - position of the anilide of these thiophene sulfonamides improved oral bioavailability . As a result of this exercise , TBC 3214 ( 17 ) was identified as a sitaxsentan follow-on candidate . It is very potent ( IC ( 50 ) for ET ( A ) = 40 pM ) and highly selective for ET ( A ) vs ET ( B ) receptors ( 400 000 - fold ) , with a half-life of > 4 h and oral bioavailability of 25 % in rats , 42 % in cats , and 70 % in dogs .

Molecular events caused by mechanical stress in bone . The shape of bone changes as a result of bone remodeling corresponding to physical circumstances such as mechanical stress . The tissue which receives the loaded mechanical stress most efficiently is bone matrix . Recent studies revealed the function of osteocytes as mechanosensors in the early stage of bone remodeling . Loaded mechanical stress is converted to a series of biochemical reactions , and finally activates osteoclasts and osteoblasts to cause bone resorption and formation . Biochemical and molecular biological studies have recently resulted in the identification of the gene of which expression level is changed by mechanical stress . DB00435 ( NO ) and DB02527 is secreted in response to mechanical stress in the immediate early stage . Genes encoding enzymes such as glutamate / aspartate transporter ( P43003 REA ) , nitric oxide synthetase ( NOS ) and prostaglandin G / H synthetase ( P35354 REA ) are identified as mechanical stress-responsive . The expression level of P05019 REA is enhanced under the control of PTH / P12272 REA . The expression of c-fos is increased by loading of mechanical stress . P05412 REA , a heterodimer of c - P01100 REA / c - P05412 REA , functions as a transcription factor of downstream gene ( s ) . Elements including P05412 REA sites , cyclic AMP response elements ( CRE ) and shear stress response elements ( SSRE ) are found in the promoter region of mechanical stress-response genes . The enhanced expression of osteopontin ( P10451 REA ) in the osteocytes of bone resorption sites was demonstrated by in situ hybridization and immunohistochemistry and transdifferentiation of chondrocytes with the abundant expression of P12643 REA and - 4 in the process of distraction osteogenesis was observed .

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 .

Arctic mutant Aβ40 aggregates on α7 nicotinic acetylcholine receptors and inhibits their functions . Amyloid β protein ( Aβ ) plays a central role in the pathogenesis of Alzheimer ' s disease ( AD ) . Point mutations within the Aβ sequence associated with familial AD ( DB03147 ) are clustered around the central hydrophobic core of Aβ . Several types of mutations within the Aβ sequence have been identified , and the ' Arctic ' mutation ( E22G ) has a purely cognitive phenotype typical of AD . Previous studies have shown that the primary result of the ' Arctic ' mutation is increased formation of Aβ protofibrils . However , the molecular mechanism underlying this effect remains unknown . Aβ42 binds to a neuronal nicotinic acetylcholine receptor subunit , neuronal acetylcholine receptor subunit alpha - 7 ( P36544 REA ) , with high affinity and , thus , may be involved in the pathogenesis of AD . Therefore , to clarify the molecular mechanism of Arctic mutation-mediated DB03147 , we focused on P36544 REA as a target molecule of Arctic Aβ . We performed an in vitro binding assay using purified P36544 REA and synthetic Arctic Aβ40 , and demonstrated that Arctic Aβ40 specifically bound to P36544 REA . The aggregation of Arctic Aβ40 was enhanced with the addition of P36544 REA . Furthermore , the function of P36544 REA was detected by measuring Ca ( 2 + ) flux and phospho - Q8TCB0 / 42 MAPK ( P27361 REA / 2 ) activation . Our results indicated that Arctic Aβ40 aggregation was enhanced by the addition of P36544 REA , which destabilized the function of P36544 REA via inhibition of Ca ( 2 + ) responses and activation of P27361 REA / 2 . These findings indicate that Arctic Aβ mutation may be involved in the mechanism underlying DB03147 . This mechanism may involve binding and aggregation , leading to the inhibition of P36544 REA functions .

Taming psoriatic keratinocytes - - PTHs ' uses go up another notch . The native parathyroid hormone ( PTH ) and several of its N-terminal adenylyl cyclase-activating fragments and their analogs have become the star stimulators of bone growth for treating osteoporosis , accelerating fracture healing , and strengthening the anchorage of prosthetic bone implants and one of them ( Lilly ' s Forteo - - recombinant DB05829 MEN - ( 1-34 ) has recently arrived in the clinic . But something entirely different has been lurking in the background-the ability of the adenylyl cyclase stimulating DB05829 MEN - ( 1-34 ) to calm hyperproliferating keratinocytes and reduce psoriatic lesions . By contrast PTH - ( 7-34 ) which can not stimulate adenylyl cyclase actually stimulates keratinocyte proliferation . Normal keratinocytes make P12272 REA after they lift off the basal lamina and have stopped cycling . But they have an unconventional PTH / P12272 REA receptor which is not coupled to adenylyl cyclase . Psoriatic keratinocytes do not make P12272 REA and have only a broken-down , proliferation-limiting terminal differentiation-driving Notch-Notch ligand mechanism . Putting these and other facts together produces a possible picture of an exogenously applied adenylyl cyclase-activating PTH pinch hitting for the missing P12272 REA and restoring normal keratinocyte proliferative activity epidermal structure by stimulating dermal fibroblasts which do have the conventional adenylyl cyclase-linked Q03431 REA and in response directly or indirectly restore the overlying basal keratinocytes ' Notch-Notch ligand terminal differentiation-driving mechanism and consequently a normal epidermal structure .

Growth-inhibitory effects of vitamin D analogues and retinoids on human pancreatic cancer cells . Retinoids and vitamin D are important factors that regulate cellular growth and differentiation . An additive growth-inhibitory effect of retinoids and vitamin D analogues has been demonstrated for human myeloma , leukaemic and breast cancer cells . We set out to study the effects of the vitamin D analogue EB1089 and the retinoids all-trans - and 9 - cis-retinoic acid on the human pancreatic adenocarcinoma cell lines Capan 1 and Capan 2 and the undifferentiated pancreatic carcinoma cell line Hs766T . The cell lines investigated expressed vitamin D receptor , retinoic acid receptor ( RAR ) - alpha and gamma as determined by polymerase chain reaction after reverse transcription . P10826 REA was expressed only in Hs766T cells . Addition of all-trans-retinoic acid increased the amount of P10276 REA mRNA in the three cell lines and induced P10826 REA mRNA in Capan 1 and Capan 2 cells . All-trans-retinoic acid at a concentration of 10 nM inhibited the growth of Capan 1 and Capan 2 cells by 40 % relative to controls . DB00523 MEN was less effective . Neither all-trans-retinoic acid nor 9 - cis-retinoic acid affected the growth of Hs766T cells . EB1089 , if added alone to the cells , did not significantly inhibit growth . However , the combination of 1 nM EB1089 with 10 nM all-trans-retinoic acid exerted a growth-inhibitory effect of 90 % in Capan 1 cells and of 70 % in Capan 2 cells . Our data suggest that vitamin D analogues together with retinoids inhibit the growth of human pancreatic cancer cells . However , in vivo studies are necessary to examine the potential use of retinoids and vitamin D analogues on pancreatic cancer .

Evaluation of an aldose reductase inhibitor on lens metabolism , ATPases and antioxidative defense in streptozotocin-diabetic rats : an intervention study . AIMS / HYPOTHESIS : P15121 REA inhibitors ( ARIs ) prevent biochemical abnormalities associated with diabetic complications . We evaluated whether a short-term intervention with an adequate dose of Q9Y4X5 REA , introduced at the very early , precataractous stage , reversed diabetes-induced metabolic imbalances , down-regulation of ATPases and oxidative stress in the lens . Methods . The groups included mature control and streptozotocin-diabetic rats treated with or without Q9Y4X5 REA sorbinil ( 65 mg x kg ( - 1 ) x day ( - 1 ) , in the diet , for 2 weeks after 4 weeks of untreated diabetes ) . Free cytosolic NAD + : DB00157 and NADP + : NADPH ratios were calculated from the lactate dehydrogenase and malic enzyme systems . Concentrations of metabolites and adenine nucleotides , Na + / K + - ATPase , H + - ATPase and Ca + + - independent Mg + + - ATPase activities and variables of oxidative stress were measured in individual lenses . Results . DB02712 MEN treatment essentially corrected diabetes-induced sorbitol and fructose accumulation , myo-inositol depletion , decrease in free cytosolic NAD + : DB00157 ratio and energy deficiency . Malondialdehyde accumulation , reduced glutathione depletion and the increase in oxidized glutathione : reduced glutathione ratio were partially corrected . Free cytosolic NADP + : NADPH ratio and 4 - hydroxyalkenal concentrations were similarly increased in diabetic rats treated with or without Q9Y4X5 REA . DB02712 MEN did not counteract diabetes-induced down-regulation of the three ATPase activities . CONCLUSION / INTERPRETATION : All biochemical changes assessed in our study are known to be prevented by ARIs . Despite the essential normalization of the sorbitol pathway activity , only part of them were , however , reversed by the Q9Y4X5 REA treatment introduced at the very early , i . e . precataractous , stage of diabetes . Therefore , intervention studies can easily underestimate the importance of aldose reductase in the pathogenesis of diabetic complications and should be interpreted with caution .

CP -101,606 , a potent neuroprotectant selective for forebrain neurons . The neuroprotective activity of ( 1S , 2S ) - 1 - ( 4 - hydroxyphenyl ) - 2 - ( 4 - hydroxy - 4 - phenylpiperidino ) - 1 - propanol ( CP -101,606 ) , an N-methyl-D-aspartate ( DB01221 ) receptor antagonist structurally similar to ( ( + / - ) - ( R * , S * ) - alpha - ( 4 - hydroxyphenyl ) - beta-methyl - 4 - ( phenylmethyl ) - 1 - + + + piperidineethanol ( ifenprodil ) , was investigated in neurons in primary culture . CP -101,606 potently and efficaciously protected hippocampal neurons from glutamate toxicity but was > 900 - fold less effective for cerebellar granule neurons . The neuroprotective activity in the hippocampal neurons is mediated through a high affinity binding site distinct from the agonist and thienylcyclohexylpiperidine ( DB01520 MEN ) binding sites of the DB01221 receptor . Autoradiography indicates the CP -101,606 binding site is localized in forebrain , most notably in hippocampus and the outer layers of cortex . The functional selectivity for hippocampal neurons , forebrain localization of binding sites , and structural relation to ifenprodil suggest that CP -101,606 is an DB01221 antagonist highly selective for Q13224 REA subunit containing receptors .

DB00472 SUB induces preventive and complex effects against colon cancer development in epithelial and stromal areas in rats . DB00472 SUB ( 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 .

P22303 REA antagonist potentiated insulin action in fed but not fasted state . The glucose disposal effect of insulin is doubled in response to a meal . This meal-induced insulin sensitization results from insulin acting on the liver , in the presence of a permissive hepatic parasympathetic feeding signal and elevated hepatic glutathione ( DB00143 ) , to release hepatic insulin-sensitizing substance ( HISS ) , a hormone that acts selectively on skeletal muscle to stimulate insulin-mediated glucose uptake . Blockade of the parasympathetic feeding signal to the liver , either through surgical denervation or atropine-mediated antagonism of hepatic muscarinic receptors , eliminates the HISS response , resulting in HISS-dependent insulin resistance ( HDIR ) and decreasing the response to insulin by approximately 55 % in the fed state . P01308 REA action in Sprague-Dawley rats , as determined with a rapidly sampled , transient euglycemic clamp in response to insulin ( 50 mU / kg ) , is decreased in a dose-dependent manner by atropine . In this study , we have used the ED75 atropine-induced model of HDIR . After a submaximal dose of atropine , potentiation of the remaining parasympathetic effect with the acetylcholinesterase antagonist neostigmine significantly restored postprandial insulin sensitization in a dose-dependent manner with peak effect at 0.1 microg / kg / min . DB01400 MEN reversed the insulin resistance induced by partial fasting and partial muscarinic inhibition ( hepatic DB00143 levels are at fed levels ) , but not that induced by surgical hepatic denervation ( DB00143 normal , no nerve signal ) or 24 - h fasting ( low DB00143 ) . No potentiation of the response to insulin by neostigmine occurred in normal , fed rats . The data suggest the use of either direct or indirectly acting cholinergic agonists for the treatment of impaired postprandial insulin sensitization .

O15554 REA channels participate in the EMT induced by O75365 in colorectal cancer . Studies have shown that phosphatase of regenerating liver - 3 ( O75365 ) promotes the invasion , migration , and metastasis of human tumor cells by facilitating an epithelial-mesenchymal transition ( EMT ) . However , the mechanism by which O75365 induces tumor cell EMT is unknown . Our previous research revealed that O75365 promotes LoVo cell proliferation by up-regulating O15554 REA channels . In the current study , we explored the mechanism by which O75365 mediates EMT . We demonstrated that O75365 induced the expression of O15554 REA channels , leading to EMT and the down-regulation of P12830 REA . Further studies revealed that O15554 REA channels increased intracellular calcium levels and activated components of cell signaling downstream of calcium , including P62158 - kinase II and glycogen synthase kinase - 3 beta ( P49841 REA ) , which increased Snail expression . Inhibiting O15554 REA with siRNA and TRAM - 34 , a specific inhibitor , restored P12830 REA expression and inhibited Snail expression . These results implicated the up-regulation of O15554 REA channels in the O75365 - mediated induction of EMT and promotion of cancer metastasis .

Induction of retinoic acid receptor-beta suppresses cyclooxygenase - 2 expression in esophageal cancer cells . Since retinoic acid receptor ( RAR ) - beta mRNA is frequently lost during esophageal carcinogenesis and esophageal cancer cells that do not express P10826 REA are resistant to retinoic acid ( RA ) , we stably transfected P10826 REA expression vector into an esophageal cancer cell line TE - 8 and an antisense P10826 REA into TE - 3 cells . Transfection of P10826 REA decreased cell growth and colony formation and induced apoptosis in TE - 8 cells . Antisense P10826 REA - transfected TE - 3 cells had a shorter doubling time and became resistant to RA . Induction of P10826 REA decreased P35354 REA expression in P10826 REA transfected TE - 8 cells , whereas antisense P10826 REA transfected TE - 3 cells increased P35354 REA expression . The inhibitory effect of P10826 REA on P35354 REA expression was further enhanced in the presence of RA , which was blocked by an RAR antagonist . The synthetic retinoid N - ( 4 - hydroxyphenyl ) retinamide , which does not bind effectively to P10826 REA , had no effect on P35354 REA suppression . Furthermore , RA blocked bile acid-induced P35354 REA expression and prostaglandin E ( 2 ) production only in the P10826 REA positive cells . Our data demonstrated that anticancer effect of P10826 REA may be related to its ability to suppress P35354 REA expression and support that the loss of P10826 REA expression may contribute to esophageal carcinogenesis .

Enhanced killing of cancer cells by poly ( ADP-ribose ) polymerase inhibitors and topoisomerase I inhibitors reflects poisoning of both enzymes . Poly ( ADP-ribose ) polymerase - 1 ( P09874 REA ) plays critical roles in the regulation of DNA repair . Accordingly , small molecule inhibitors of PARP are being developed as agents that could modulate the activity of genotoxic chemotherapy , such as topoisomerase I poisons . In this study we evaluated the ability of the PARP inhibitor veliparib to enhance the cytotoxicity of the topoisomerase I poisons topotecan and camptothecin ( CPT ) . DB07232 MEN increased the cell cycle and cytotoxic effects of topotecan in multiple cell line models . Importantly , this sensitization occurred at veliparib concentrations far below those required to substantially inhibit poly ( ADP-ribose ) polymer synthesis and at least an order of magnitude lower than those involved in selective killing of homologous recombination-deficient cells . Further studies demonstrated that veliparib enhanced the effects of CPT in wild-type mouse embryonic fibroblasts ( MEFs ) but not Parp 1 ( - / - ) MEFs , confirming that P09874 REA is the critical target for this sensitization . Importantly , parental and Parp 1 ( - / - ) MEFs had indistinguishable CPT sensitivities , ruling out models in which P09874 REA catalytic activity plays a role in protecting cells from topoisomerase I poisons . To the contrary , cells were sensitized to CPT in a veliparib-independent manner upon transfection with P09874 REA E988K , which lacks catalytic activity , or the isolated P09874 REA DNA binding domain . These results are consistent with a model in which small molecule inhibitors convert P09874 REA into a protein that potentiates the effects of topoisomerase I poisons by binding to damaged DNA and preventing its normal repair .

Prenatal activation of maternal O15455 REA receptors by viral-mimetic poly ( I : C ) modifies Q13224 REA expression in embryos and sonic hedgehog in offspring in the absence of kynurenine pathway activation . Activation of the immune system during pregnancy is believed to lead to psychiatric and neurological disorders in the offspring , but the molecular changes responsible are unknown . Polyinosinic :p olycytidylic acid ( poly ( I : C ) ) is a viral-mimetic double-stranded RNA complex which activates Toll-Like-Receptor - 3 and can activate the metabolism of tryptophan through the oxidative kynurenine pathway to compounds that modulate activity of glutamate receptors . The aim was to determine whether prenatal administration of poly ( I : C ) affects the expression of neurodevelopmental proteins in the offspring and whether such effects were mediated via the kynurenine pathway . Pregnant rats were treated with poly ( I : C ) during late gestation and the offspring were allowed to develop to postnatal day 21 ( P21 ) . Immunoblotting of the brains at P21 showed decreased expression of sonic hedgehog , a key protein in dopaminergic neuronal maturation . Expression of α-synuclein was decreased , while tyrosine hydroxylase was increased . Disrupted in Schizophrenia - 1 ( DISC - 1 ) and P28335 REA receptor levels were unaffected , as were the dependence receptors Unc 5H1 , Unc 5H3 and Deleted in Colorectal Cancer ( P43146 REA ) , the inflammation-related transcription factor NFkB and the inducible oxidative enzyme cyclo-oxygenase - 2 ( P35354 REA ) . An examination of embryo brains 5 h after maternal poly ( I : C ) showed increased expression of Q13224 REA , with reduced doublecortin and P43146 REA but no change in NFkB . Despite altered protein expression , there were no changes in the kynurenine pathway . The results show that maternal exposure to poly ( I : C ) alters the expression of proteins in the embryos and offspring which may affect the development of dopaminergic function . The oxidation of tryptophan along the kynurenine pathway is not involved in these effects .

Pathogenesis of spinally mediated hyperalgesia in diabetes . Hyperalgesia to noxious stimuli is accompanied by increased spinal cyclooxygenase ( P36551 REA ) - 2 protein in diabetic rats . The present studies were initiated to establish causality between increased spinal P35354 REA activity and hyperalgesia during diabetes and to assess the potential involvement of polyol pathway activity in the pathogenesis of spinally mediated hyperalgesia . Rats with 1 , 2 , or 4 weeks of streptozotocin-induced diabetes exhibited significantly increased levels of spinal P35354 REA protein and activity , along with exaggerated paw flinching in response to 0.5 % paw formalin injection . Increased flinching of diabetic rats was attenuated by intrathecal pretreatment with a selective P35354 REA inhibitor immediately before formalin injection , confirming the involvement of P35354 REA activity in diabetic hyperalgesia . Chronic treatment with insulin or ICI 222155 , an aldose reductase inhibitor ( Q9Y4X5 REA ) previously shown to prevent spinal polyol accumulation and formalin-evoked hyperalgesia in diabetic rats , prevented elevated spinal P35354 REA protein and activity in diabetic rats . In contrast , the Q9Y4X5 REA IDD 676 had no effect on spinal polyol accumulation , elevated spinal P35354 REA , or hyperalgesia to paw formalin injection . In the spinal cord , aldose reductase immunoreactivity was present solely in oligodendrocytes , which also contained P35354 REA immunoreactivity . Polyol pathway flux in spinal oligodendrocytes provides a pathogenic mechanism linking hyperglycemia to hyperalgesia in diabetic rats .

Differential involvement of Galpha 12 and Galpha 13 in receptor-mediated stress fiber formation . The ubiquitously expressed heterotrimeric guanine nucleotide-binding proteins ( G-proteins ) G12 and Q99941 have been shown to activate the small GTPase Rho . Rho stimulation leads to a rapid remodeling of the actin cytoskeleton and subsequent stress fiber formation . We investigated the involvement of G12 or Q99941 in stress fiber formation induced through a variety of Gq / P49842 - coupled receptors . Using fibroblast cell lines derived from wild-type and Galphaq / Galpha 11 - deficient mice , we show that agonist-dependent activation of the endogenous receptors for thrombin or lysophosphatidic acid and of the heterologously expressed bradykinin B2 , vasopressin V1A , endothelin P25101 REA , and serotonin P28335 REA receptors induced stress fiber formation in either the presence or absence of Galphaq / Galpha 11 . Stress fiber assembly induced through the muscarinic M1 and the metabotropic glutamate subtype 1alpha receptors was dependent on Gq / P49842 proteins . The activation of the Gq / P49842 - coupled endothelin ETB and angiotensin AT1A receptors failed to induce stress fiber formation . Lysophosphatidic acid , B2 , and P28335 REA receptor-mediated stress fiber formation was dependent on Galpha 13 and involved epidermal growth factor ( P01133 REA ) receptors , whereas thrombin , P25101 REA , and V1A receptors induced stress fiber accumulation via Galpha 12 in an P01133 REA receptor-independent manner . Our data demonstrate that many Gq / P49842 - coupled receptors induce stress fiber assembly in the absence of Galphaq and Galpha 11 and that this involves either a Galpha 12 or a Galpha 13 / P01133 REA receptor-mediated pathway .

Nuclear factor kappa B inhibition improves conductance artery function in type 2 diabetic mice . BACKGROUND : We previously reported that enhanced nuclear factor kappa B ( NFκB ) activity is responsible for resistance arteries dysfunction in type 2 diabetic mice . METHODS : In this study , we aimed to determine whether augmented NFκB activity also impairs conductance artery ( thoracic aorta ) function in type 2 diabetic mice . We treated type 2 diabetic ( db ( - ) / db ( - ) ) and control ( db ( - ) / db ( + ) ) mice with two NFκB inhibitors ( dehydroxymethylepoxyquinomicin , 6 mg / kg , twice a week and IKK-NBD peptide , 500 µg / kg / day ) for 4 weeks . RESULTS : As expected , the NFκB inhibition did not affect blood glucose level and body weight . Thoracic aorta vascular endothelium-dependent relaxation ( EDR ) , determined by the wire myograph , was impaired in diabetic mice compared with control and was significantly improved after NFκB inhibition . Interestingly , thoracic EDR was also rescued in db ( - ) / db ( - p50NFκB - / - ) and db ( - ) / db ( - P09874 REA - / - ) double knockout mice compared with db ( - ) / db ( - ) mice . Similarly , the acute in vitro down regulation of NFκB-p 65 using p65 shRNA lentiviral particles in arteries from db ( - ) / db ( - ) mice also improved thoracic aorta EDR . Western blot analysis showed that the p65NFκB phosphorylation , cleaved P09874 REA and P35354 REA expression were increased in thoracic aorta from diabetic mice , which were restored after NFκB inhibition and in db ( - ) / db ( - p - 50NFκB - / - ) and db ( - ) / db ( - P09874 REA - / - ) mice . CONCLUSIONS : The present results indicate that in male type 2 diabetic mice , the augmented NFκB activity also impairs conductance artery function through P09874 REA and P35354 REA - dependent mechanisms .

Effects of an alpha 7 - nicotinic agonist on default network activity in schizophrenia . BACKGROUND : 3 - ( 2,4- dimethoxybenzylidene ) - anabaseine ( DB05708 MEN ) is a partial agonist at α7 nicotinic acetylcholine receptors that has been evaluated clinically for treatment of schizophrenia . This study examined the effects of DB05708 MEN on default network activity as a biomarker for drug effects on pathologic brain function associated with schizophrenia . METHODS : Placebo and two doses of DB05708 MEN were administered in a random , double-blind crossover design during a Phase 2 study of DB05708 MEN . Functional magnetic resonance imaging was performed on 16 nonsmoking patients with schizophrenia while they performed a simple eye movement task . Independent component analysis was used to identify the default network component . Default network changes were evaluated in the context of a polymorphism in P36544 REA , the α7 - nicotinic acetylcholine receptor subunit gene , which was previously found to be associated with schizophrenia . RESULTS : Compared with placebo , both 150 and 75 mg twice daily DB05708 MEN altered default network activity , including a reduction in posterior cingulate , inferior parietal cortex , and medial frontal gyrus activity and an increase in precuneus activity . The most robust difference , posterior cingulate activity reduction , was affected by P36544 REA genotype . CONCLUSIONS : The observed DB05708 MEN - related changes are consistent with improved default network function in schizophrenia . Pharmacogenetic analysis indicates mediation of the effect through the α7 - nicotinic receptor . These results further implicate nicotinic cholinergic dysfunction in the disease and suggest that default network activity may be a useful indicator of biological effects of novel therapeutic agents .