MH_dev_211

Function of endogenous monoamine oxidase inhibitors ( tribulin ) . Recent research on tribulin [ low molecular weight endogenous inhibitory activity of monoamine oxidase ( MAO ) ] has confirmed that its level is increased in both human urine and rat tissues by stress or anxiety , and by anxiogenic drugs . However tribulin is now known to contain several different molecules . The raised inhibitory activity in rat tissues is selective for P21397 REA . There is a parallel decrease in P21397 REA activity ex vivo , suggesting a possible functional effect . Increase in endogenous MAO I may competitively inhibit the binding of irreversible MAO I drugs , and may also help to mediate some mood altering effects of other drugs , or procedures such as ECT . In human urine both P21397 REA I and P27338 REA I have been found to be increased in mild stress . Similar findings have been made with human saliva . Selective inhibitors of P21397 REA have been identified from human urine , and pig brain , but it is not yet clear to what extent they account for the P21397 REA I activity increased in stress . DB02095 is an endogenous selective inhibitor of P27338 REA ( P04264 REA approximately 3 microM ) . It has a distinct distribution in rat brain , with highest concentration in the hippocampus of 0.1 microgram / g . Its level is increased by pentylene tetrazole , and isatin is itself anxiogenic in rodent models . Its administration also increases monoamine levels in the brain . It is a potent antagonist of the P01160 REA receptor , and it may act to link the control of monoamine function and natriuresis .

Steroid hormone receptors and coregulators in endocrine-resistant and estrogen-independent breast cancer cells . Resistance to hormonal therapy is often a problem in the treatment of breast cancer patients . It has been suggested that resistance could be explained by altered nuclear hormone receptor or coregulator levels or inappropriately increased agonist activity of selective estrogen receptor modulator ( SERM ) . To test these hypotheses , we have established novel MCF - 7 cell line-derived in vitro models of anti-estrogen - and progestin-resistant and estrogen-independent breast cancer by long-term culture in the presence of toremifene and medroxyprogesterone acetate ( DB00603 SUB ) and in the absence of estradiol , respectively . Using cell growth and multiprobe ribonuclease protection assays , the expression of 5 nuclear hormone receptors and 9 coregulators as well as the alterations in the cell proliferation and target gene transcription in response to hormonal treatments were studied . P06401 REA ( PR ) expression was decreased and silencing mediator for retinoid acid and thyroid hormone receptors ( Q9Y618 REA ) and amplified in breast cancer - 1 ( Q9Y6Q9 REA ) expression increased in anti-estrogen-resistant cells . Estrogen caused PR and ERbeta upregulation in all cell lines , but we did not observe increased agonist activity of anti-estrogen measured by regulation of these estrogen target genes . Basal ERalpha levels and estrogenic growth response were decreased and p300 / CBP-associated factor ( pCAF ) and Q9Y6Q9 REA upregulated by estrogen in progestin-resistant cells , but coregulator levels were unchanged . Estrogen-independent cells were still estrogen-responsive and PR , nuclear receptor corepressor ( O75376 REA ) and Q9Y618 REA expression was increased whereas steroid receptor coactivator - 1 ( P12931 REA - 1a ) and CBP-related protein p300 ( p300 ) expression decreased . Their growth was inhibited by toremifene , but estradiol was able to abrogate this effect , which might have interesting clinical implications concerning the use of postmenopausal hormone replacement therapy .

Suppression of parathyroid hormone-related protein messenger RNA expression by medroxyprogesterone acetate in breast cancer tissues . The level of parathyroid hormone-related protein ( P12272 REA ) expressed in breast cancer tissue is closely related to the incidence of bone metastasis . We examined the P12272 REA mRNA expression in breast cancer tissues by coamplification polymerase chain reaction ( PCR ) in mole ratio to internal standard beta-actin mRNA . The P12272 REA expression was higher in premenopausal patients than in postmenopausal patients ( P < 0.05 ) . More pronounced difference by menopause found in estrogen receptor ( ER ) positive groups ( P < 0.001 ) indicated that the P12272 REA expression in breast cancer tissue is hormonally regulated and might be altered by endocrine agents . To clarify the changes of P12272 REA expression by endocrine therapy of breast cancer , we measured P12272 REA expression in the breast cancer tissue incubated for 24 h with 1 x 10 (-8 ) M of estradiol ( E2 ) , 1 x 10 ( - 6 ) M of tamoxifen ( TAM ) and 1 x 10 ( - 5 ) M of medroxyprogesterone acetate ( DB00603 SUB ) . The P12272 REA expression was decreased significantly by DB00603 SUB ( P < 0.005 ) , while E2 and TAM did not change the P12272 REA expression . P06401 REA ( PgR ) mRNA expression was also examined to confirm that the breast cancer tissue responds to E2 and TAM . The results were well compatible with the better therapeutic effect of DB00603 SUB reported for the treatment of breast cancer with bone metastases . As a potential candidate for the receptor that mediates the suppressive effect of DB00603 SUB , androgen receptor ( AR ) is suggested most probable . Present results also demonstrated that the clinical response of individual tumors is closely associated with the early in vitro changes of gene expression detected in the cancer specimen .

Role of chronic inhibition of dopamine-metabolizing enzymes in the regulation of renal sodium and phosphate excretion in the rat remnant kidney . BACKGROUND / AIMS : The present study examined the effects of chronic selective or combined inhibition of type A monoamine oxidase ( MAO ) and catechol-O-methyltransferase ( P21964 REA ) on daily urinary excretion of dopamine and metabolites and on natriuresis and phosphaturia in 3/4 nephrectomized ( 3/4 nx ) and Sham rats . METHODS : The 3/4 nx and Sham rats were placed in metabolic cages and received the P21397 REA - selective inhibitor Ro - 411049 ( 7.5 mg x kg ( - 1 ) bid ) and / or the P21964 REA - selective inhibitor DB03336 MEN 3-202 ( 30 mg x kg ( - 1 ) bid ) orally for 3 days during high sodium diet . RESULTS : Selective P21964 REA inhibition increased the urinary excretion of the deaminated metabolite ( 3,4- dihydroxyphenylacetic acid , DOPAC ) and decreased the urinary excretion of the methylated ( 3 - methoxytyramine , 3 - MT ) and deaminated plus methylated metabolite ( homovanillic acid , HVA ) in both groups . Selective P21397 REA inhibition increased the urinary excretion of 3 - MT and reduced the urinary excretion of both DOPAC and HVA in either 3/4 nx or Sham rats . Combined inhibition of P21397 REA and P21964 REA did not significantly change the urinary excretion of DOPAC and markedly decreased the urinary excretion of 3 - MT and HVA in both groups . Selective or combined inhibition of P21397 REA and P21964 REA did not alter the daily urinary excretion of dopamine , sodium or phosphate in either 3/4 nx or Sham rats . CONCLUSIONS : Chronic selective or combined inhibition of P21397 REA and P21964 REA is not of major importance in regulating the dopamine-dependent natriuresis and phosphaturia in either 3/4 nx or Sham rats .

Functional methionine synthase deficiency ( cblE and cblG ) : clinical and biochemical heterogeneity . Functional methionine synthase deficiency is generally characterized by homocystinuria and hypomethioninemia in the absence of methylmalonic aciduria . Patients are divided into two classes , cblE and cblG , on the basis of complementation analysis . Presentation has usually been in the first 2 years of life , but one patient came to medical attention at age 21 years with symptoms initially diagnosed as multiple sclerosis . Common findings among 11 patients ( 4 with cblE and 7 with cblG ) have included megaloblastic anemia ( all patients ) and various neurological deficits including developmental retardation ( 10 patients ) , cerebral atrophy ( 8 patients ) , hypotonia ( 7 patients ) , EEG abnormalities ( 6 patients ) , and nystagmus ( 5 patients ) . Hypertonia , seizures , blindness , and ataxia were less frequent . All patients have responded to therapy with cobalamin with resolution of anemia and biochemical abnormalities ; neurological deficits resolved more slowly and in some cases incompletely . DB00200 MEN has been more effective than cyanocobalamin . Fibroblasts from patients with cblE ( 5 patients ) and cblG ( 6 patients ) all showed decreased intracellular levels of methylcobalamin ( DB03614 ) and decreased incorporation of label from 5 - methyltetrahydrofolate into macromolecules , suggesting decreased activity of the DB03614 - dependent enzyme methionine synthase . Q99707 REA specific activity in extracts of all cblE fibroblasts was normal or near-normal under standard reducing conditions ; synthase specific activity in extracts of 5 cblG patients was low but was high in a 6th patient measured in another laboratory . Thus , there is heterogeneity among patients with functional methionine synthase deficiency both in clinical presentation and in the results of biochemical studies of cultured cells .

Q8NFH3 / Q8TCB0 mitogen-activated protein kinases inhibit atrial natriuretic peptide mRNA transcription in P40189 REA - mediated hypertrophic ventricular myocytes . OBJECTIVE : To understand the role of P01160 REA mRNA transcription regulation in P40189 REA - mediated cardiomyocyte hypertrophy , and the involved mitogen-activated protein kinase kinase ( MEK ) - extracellular signal-regulated kinase ( P29323 REA , also called Q8NFH3 / Q8TCB0 MAPK ) signaling pathway . METHODS : Isolated neonatal ventricular myocytes were treated with different concentrations of Q16619 REA ( 10 ( - 9 ) , 10 (-8 ) and 10 ( - 7 ) mol / L ) . MTT was used to analyze the viability and RT-PCR was used to detect P01160 REA mRNA levels in cardiomyocyte . To inhibit Q8NFH3 / Q8TCB0 MAPK activity in hypertrophic cardiomyocytes , the cells were pretreated with a specific Q02750 REA inhibitor . RESULTS : Q16619 REA significantly induced P01160 REA mRNA expression and the viability of cardiomyocytes in a dose - and time-dependent manner . Furthermore , blocking Q8NFH3 / Q8TCB0 MAPK activity by the special Q02750 REA inhibitor upregulated the P01160 REA mRNA . CONCLUSIONS : Q8NFH3 / Q8TCB0 MAPK have an important role in suppressing P01160 REA mRNA transcription and cell activity in P40189 REA - mediated hypertrophic ventricular myocytes .

DB09337 cleaning in P01160 REA experiments at the ETR and Q99707 REA .

Comparison of bevacizumab , ranibizumab , and pegaptanib in vitro : efficiency and possible additional pathways . PURPOSE : Vascular endothelial growth factor ( P15692 REA ) antagonists are the therapy of choice for age-related macular degeneration . DB01270 MEN and pegaptanib have been approved by the United States Food and Drug Administration , whereas bevacizumab is used off label . In this study , the authors compare these P15692 REA inhibitors directly regarding their efficiency to neutralize P15692 REA in a quantifiable in vitro system . METHODS : Porcine retina-retinal pigment epithelium-choroid organ culture and Q96AT9 cell culture were prepared from fresh eyes , cultivated in a perfusion chamber , and treated with clinically relevant concentrations of bevacizumab , ranibizumab and pegaptanib . P15692 REA content of the supernatant was analyzed with ELISA . Additionally , the influence of bevacizumab and ranibizumab on intracellular P15692 REA was analyzed with Western blot . RESULTS : At clinically significant doses , bevacizumab ( 0.25 mg / mL ) and ranibizumab ( 0.125 mg / mL ) neutralized P15692 REA completely for 6 hours , whereas pegaptanib ( 0.08 mg / mL ) showed no effect . DB00112 and ranibizumab neutralized P15692 REA significantly up to 16 hours . When diluted , bevacizumab lost its inhibiting properties at a concentration of 975 ng / mL , and ranibizumab neutralized P15692 REA up to a concentration of 120 ng / mL . Both substances significantly diminished P15692 REA expression in Western blot . CONCLUSIONS : At clinical doses , bevacizumab and ranibizumab are equally potent in neutralizing P15692 REA . To neutralize P15692 REA completely in this system , a fraction of the clinical dose is needed . DB01270 MEN is more efficient at neutralizing P15692 REA when diluted . DB04895 showed no effect in this system , which might help explain the clinical experience regarding this drug . A direct effect of ranibizumab and bevacizumab on P15692 REA protein expression indicates additional pathways of P15692 REA inhibitors .

Isolation and characterization of P62333 REA . A novel ATPase family component of the yeast 26 S proteasome . Using a genetic strategy designed to find proteins involved in the function of the Saccharomyces cerevisiae transcriptional activator GAL 4 , we isolated mutants in two genes which rescue a class of gal 4 activation domain mutants . One of these genes , P62195 REA , encodes a member of a large family of putative ATPases , the Conserved ATPase containing Domain ( CAD ) proteins ( also known as AAA proteins ) that are involved in a wide variety of cellular functions . Subsequently , P62195 REA was identified as a subunit of the 26 S proteasome . We have now cloned the gene defined by the second complementation group . P62333 REA encodes an essential 49 - kDa protein that is also a member of the CAD family and is 43 % identical to P62195 REA . The mutation in sug 2-1 , like that in sug 1-1 , is found in the CAD near the highly conserved ATPase motif . We present biochemical and genetic evidence that P62333 REA is associated in vivo with P62195 REA and is a novel P27708 REA subunit of the 26 S proteasome . With its highly conserved mammalian homologs , human Q8NFH3 and ground squirrel CADp 44 , P62333 REA defines a new class of proteasomal CAD proteins .

Modulation of the folate receptor alpha gene by the estrogen receptor : mechanism and implications in tumor targeting . The folate receptor ( FR ) type alpha is a promising target for diagnostic imaging agents and therapeutic intervention in major subtypes of gynecological malignancies ; however , the receptor levels in the tumors are variable and are generally relatively low in estrogen receptor ( ER ) - positive tumors . Here we report that the P15328 REA gene promoter is repressed in the presence of 17beta - estradiol and derepressed by the antiestrogens tamoxifen and ICI 182780 in a promoter-specific and P03372 REA - dependent manner in carcinoma cell lines including HeLa ( cervical carcinoma ) , BG - 1 ( ovarian carcinoma ) , and IGROV - 1 ( ovarian carcinoma ) . The ligand and ER dose response of the P15328 REA promoter and its time course paralleled those of a classical estrogen response element-mediated effect . Antiestrogens produced an ER-dependent increase of up to 36 - fold in the expression of the endogenous P15328 REA gene . Deletion analysis and P15328 REA / SV40 promoter chimeras showed that the ER effect is mediated exclusively within the G / C-rich region in the TATA-less P4 promoter of P15328 REA ; electrophoretic mobility shift analysis demonstrated interaction of ER at only one of three G / C-rich elements . Q92731 REA only modestly affected P15328 REA promoter activity but did not diminish the P03372 REA - mediated effects . The ER corepressor , Q9Y618 REA , enhanced the repression by 17beta - estradiol / ER , but ER coactivators , including P12931 REA family members , did not appreciably impact the ER ligand response . The results suggest that in ER + tumors , P15328 REA expression is directly and actively suppressed and predict that a brief treatment with antiestrogens will boost P15328 REA expression by passive derepression , enhancing the efficacy of FR-targeted diagnostic and therapeutic applications . They also reveal novel aspects of gene repression by ER .

Down-regulation of human osteoblast PTH / P12272 REA receptor mRNA in end-stage renal failure . BACKGROUND : Resistance to the action of parathyroid hormone ( PTH ) has been demonstrated in end-stage renal failure and is considered to be important in the pathogenesis of secondary hyperparathyroidism . The mechanism of resistance is unknown . However , altered regulation of cellular PTH / PTH-related protein ( PTH / P12272 REA ) receptor ( Q03431 REA ) has been assumed to be important . METHODS : We have used in situ hybridization to examine Q03431 REA mRNA expression by osteoblasts in human bone and have compared the expression in high - and low-turnover renal bone disease , high-turnover nonrenal bone disease ( healing fracture callus and Pagetic bone ) , and normal bone . Bone biopsies were formalin fixed , ethylenediaminetetraacetic acid decalcified , and paraffin wax embedded . A 1.8 kb Q03431 REA cDNA probe , labeled with 35S , was used , and the hybridization signal was revealed by autoradiography . The density of signal over osteoblasts was quantitated using a semiautomated Leica image analysis software package . RESULTS : The mean density of Q03431 REA mRNA signal over osteoblasts in renal high-turnover bone was only 36 % of that found in nonrenal high-turnover bone ( P < 0.05 ) and 51 % of that found in normal bone ( P < 0.05 ) . Osteoblast Q03431 REA mRNA signal in adynamic bone from individuals with diabetes mellitus was 28 % of normal bone ( P < 0.05 ) and 54 % of that found in renal high-turnover bone ( P < 0.05 ) . CONCLUSIONS : These results demonstrate a down-regulation of osteoblast Q03431 REA mRNA in end-stage renal failure in comparison to normal and high-turnover bone from otherwise healthy individuals , and provide an insight into the mechanisms of " skeletal resistance " to the actions of PTH .

Estrogenic chemicals at puberty change ERalpha in the hypothalamus of male and female rats . The effects of two environmental endocrine disruptors , the synthetic pharmaceutical estrogen DB00977 ( EE ) and bisphenol-A ( Q03001 REA ) , were analysed in male and female rats in a very sensitive developmental period , puberty . Immunohistochemistry was used to evaluate changes in the number of cells expressing estrogen receptors ( P03372 REA ) in the arcuate nucleus ( ARC ) , ventromedial nucleus ( VMH ) and medial preoptic area ( DB00603 SUB ) of the hypothalamus . Animals were treated during early puberty , from P01160 REA 23 to P01160 REA 30 , with EE and Q03001 REA given orally every day . They were then sacrificed and perfused on P01160 REA 37 or P01160 REA 90 , and blood and brains were collected for hormonal determination ( testosterone and estradiol ) and immunohistochemistry ( estrogen receptors , ER ) . At P01160 REA 37 , ER-labelled neurons were higher in males than in females in the ARC and DB00603 SUB . EE and Q03001 REA increased ER-labelled neurons in the ARC and DB00603 SUB . At P01160 REA 90 , females showed higher ER-labelled neurons in the VMH . EE and Q03001 REA increased ER-labelled neurons in the DB00603 SUB in females . EE increased testosterone in males at P01160 REA 37 and estradiol in females at P01160 REA 90 . These results indicate the ability of estrogenic chemicals to change the reproductive neural circuits during puberty in male and female rats .

DB05595 MEN , an anti-folate receptor α antibody , does not block binding of folate or anti-folates to receptor nor does it alter the potency of anti-folates in vitro . PURPOSE : DB00158 is a cofactor in the synthesis of purines and pyrimidines ; folate analogs are potent cytotoxic drugs . P15328 REA ( FRα ) , a protein-mediating cellular accumulation of folate ( and anti-folates ) , has limited expression in normal tissues and is overexpressed by numerous carcinomas . Limited distribution and high affinity for folic acid have resulted in the development of antibodies or the use of folic acid coupled to toxins or radionuclides as therapeutic and imaging agents . DB05595 MEN is an anti-FRα antibody in clinical trials for ovarian and non-small cell lung cancers . Our goal was to evaluate the effect of farletuzumab on binding and uptake of folates and anti-folates and the potency of anti-folates in vitro . METHODS : Direct binding and uptake of radiolabeled folates and anti-folates and the assessments of drug concentration of drug that inhibited cell growth 50 % ( IC ( 50 ) ) in vitro in the presence or absence of antibody . RESULTS : DB05595 MEN did not block membrane binding of radiolabeled folic acid , 5 - methyltetrahydrofolate , pemetrexed , and other anti-folates ; folic acid blocked > 95 % . DB05595 MEN had a minimal effect on the cytoplasmic accumulation of 5 - methyltetrahydrofolate or pemetrexed ; folic acid had a considerable but variable effect on the different cell lines . As a single agent , farletuzumab did not affect cell viability or the IC ( 50 ) of pemetrexed and other anti-folates in vitro . CONCLUSIONS : DB05595 MEN does not block FRα binding of folates and anti-folates , minimally retards folate delivery via FRα-mediated transport , and minimally retards the growth of cells in vitro . Concomitant use of farletuzumab and pemetrexed is not contraindicated .

DB00741 response to stress is associated with myocardial remodeling in salmonid fishes . Cardiac disease is frequently reported in farmed animals , and stress has been implicated as a factor for myocardial dysfunction in commercial fish rearing . DB00741 is a major stress hormone in teleosts , and this hormone has adverse effects on the myocardium . Strains of rainbow trout ( Oncorhynchus mykiss ) selected for divergent post-stress cortisol levels [ high responsive ( HR ) and low responsive ( LR ) ] have been established as a comparative model to examine how fish with contrasting stress-coping styles differ in their physiological and behavioral profiles . We show that the mean cardiosomatic index ( CSI ) of adult HR fish was 34 % higher than in LR fish , mainly because of hypertrophy of the compact myocardium . To characterize the hypertrophy as physiological or pathological , we investigated specific cardiac markers at the transcriptional level . HR hearts had higher mRNA levels of cortisol receptors ( MR , GR1 and GR2 ) , increased P53805 levels [ suggesting enhanced pro-hypertrophic nuclear factor of activated T-cell ( NFAT ) signaling ] and increased P15692 REA gene expression ( reflecting increased angiogenesis ) . Elevated collagen ( Col 1a2 ) expression and deposition in HR hearts supported enhanced fibrosis , whereas the heart failure markers P01160 REA and DB04899 were not upregulated in HR hearts . To confirm our results outside the selection model , we investigated the effect of acute confinement stress in wild-type European brown trout , Salmo trutta . A positive correlation between post-stress cortisol levels and CSI was observed , supporting an association between enhanced cortisol response and myocardial remodeling . In conclusion , post-stress cortisol production correlates with myocardial remodeling , and coincides with several indicators of heart pathology , well-known from mammalian cardiology .

P28562 REA regulates bone mass , osteoblast gene expression , and responsiveness to parathyroid hormone . Parathyroid hormone ( PTH ) signaling via PTH 1 receptor ( Q03431 REA ) involves mitogen-activated protein kinase ( MAPK ) pathways . MAPK phosphatase 1 ( P28562 REA ) dephosphorylates and inactivates MAPKs in osteoblasts , the bone-forming cells . We previously showed that Q03431 REA activation in differentiated osteoblasts upregulates P28562 REA and downregulates pERK 1/2- MAPK and cyclin D1 . In this study , we evaluated the skeletal phenotype of Mkp 1 knockout ( KO ) mice and the effects of PTH in vivo and in vitro . Microcomputed tomography analysis of proximal tibiae and distal femora from 12 - week-old Mkp 1 KO female mice revealed osteopenic phenotype with significant reduction ( 8-46 % ) in bone parameters compared with wild-type ( WT ) controls . Histomorphometric analysis showed decreased trabecular bone area in KO females . Levels of serum osteocalcin ( OCN ) were lower and serum tartrate-resistant acid phosphatase 5b ( TRAP 5b ) was higher in KO animals . Treatment of neonatal mice with DB05829 MEN ( 1-34 ) for 3 weeks showed attenuated anabolic responses in the distal femora of KO mice compared with WT mice . Primary osteoblasts derived from KO mice displayed delayed differentiation determined by alkaline phosphatase activity , and reduced expressions of Ocn and Runx 2 genes associated with osteoblast maturation and function . Cells from KO females exhibited attenuated PTH response in mineralized nodule formation in vitro . Remarkably , this observation was correlated with decreased PTH response of matrix Gla protein expression . Expressions of pERK 1/2 and cyclin D1 were inhibited dramatically by PTH in differentiated osteoblasts from WT mice but much less in osteoblasts from Mkp 1 KO mice . In conclusion , P28562 REA is important for bone homeostasis , osteoblast differentiation and skeletal responsiveness to PTH .

Potent and selective inhibition of human nitric oxide synthases . Inhibition by non-amino acid isothioureas . DB02539 MEN was a potent competitive inhibitor of human nitric oxide synthase ( NOS ) , with Ki values of 17 , 36 , and 29 nM for the inducible ( i ) , endothelial ( e ) , and neuronal ( n ) isozymes , respectively . Unlike some potent inhibitors of NOS , no time dependence was observed . DB02539 MEN was not a detectable substrate for P29474 REA . DB02539 MEN was also a potent inhibitor of mouse P35228 REA ( Ki value of 5.2 nM ) , and its binding perturbed the spectrum of P35228 REA consistent with its altering the environment of the bound heme . The optimum binding of S-ethyl - and S-isopropylisothiourea relative to 70 other analogs suggested that these alkyl substitutions fit into a small hydrophobic pocket . Most isothioureas were 2-6- fold selective for the human P35228 REA ( Ki for P35228 REA versus Ki for P29474 REA ) , with one being 19 - fold selective . The cyclized mimics of S-ethylisothiourea , 2 - NH2 - thiazoline , and 2 - NH2 - thiazole , were also competitive inhibitors of human NOS . A third structural class of inhibitors , bisisothioureas , were , in general , the most selective in their inhibition of human P35228 REA . S , S ' - ( 1,3- Phenylenebis ( 1,2- ethanediyl ) ) bisisothiourea was 190 - fold selective ( Ki value of 0.047 microM against P35228 REA versus 9.0 microM against P29474 REA ) . These results demonstrate that potent and selective inhibition of human NOS isozymes is achievable .

Altered regulation of renal nitric oxide , atrial natriuretic peptide and cyclooxygenase systems in aldosterone escape in rats . The present study was aimed to determine whether there is an altered role of local nitric oxide ( NO ) , atrial natriuretic peptide ( P01160 REA ) and cyclooxygenase ( P36551 REA ) systems in the kidney in association with the aldosterone escape . Male Sprague-Dawley rats were used . DB04630 ( 200 microg / day ) was infused through entire time course . The control group was kept on a low sodium diet ( 0.02 mEq / day ) , and the experimental group was supplied with a higher sodium diet ( 2 . / day ) . Four days after beginning the regimen , the kidneys were taken . The protein expression of NO synthase ( NOS ) and P36551 REA isoforms was determined by semiquantitative immunoblotting . The mRNA expression of components of P01160 REA system was determined by real-time polymerase chain reaction . The activities of soluble and particulate guanylyl cyclases were determined by the amount of cGMP generated in responses to sodium nitroprusside and P01160 REA , respectively . There developed aldosterone escape in the experimental group . Accordingly , the renal content and the urinary excretion of NO increased . The expression of P29475 REA was increased in the inner medulla . Neither the expression of P29474 REA nor that of P35228 REA was changed . The expression and the catalytic activity of soluble guanylyl cyclase remained unaltered . The mRNA expression of P01160 REA was increased . Neither the expression of P16066 REA or P17342 nor the activity of particulate guanylyl cyclase was altered in the papilla . The protein expression of P35354 REA was increased in the inner medulla , while that of P23219 REA remained unchanged . In conclusion , the upregulation of P29475 REA , P01160 REA , and P35354 REA may be causally related with the aldosterone escape .

Tp53 - associated growth arrest and DNA damage repair gene expression is attenuated in mammary epithelial cells of rats fed whey proteins . Dietary protection from mammary cancer is likely coordinated through multiple signaling pathways , based on the known heterogeneity of the disease and the distinct origins of mammary tumor cells . The present study examined the modulatory effects of dietary intake of whey protein hydrolysate ( WPH ) relative to casein ( CAS ) , on mammary epithelial cell resistance to endogenous DNA damage using Tp53 gene expression and signaling as a read-out , and on systemic proapoptotic and immune surveillance activity , in young adult female Sprague-Dawley rats . Rats were fed AIN - 93G diets made with CAS or WPH as the sole protein source beginning at gestation d 4 . At postnatal day ( P01160 REA ) 50 , mammary glands of rats fed WPH had lower levels of activated Tp53 and p38 mitogen-activated protein kinase proteins , and reduced transcript levels for Tp53 - associated DNA damage repair , growth arrest , and proapoptotic genes than those of CAS-fed rats . Serum from WPH-fed rats had greater apoptotic activity in MCF - 7 tumor cells than that from rats fed CAS . Serum levels of monocyte chemoattractant protein ( MCP ) - 1 were higher in WPH - than in CAS-fed rats . MCF - 7 cells treated with CAS serum + recombinant rat P13500 REA had apoptotic activity and Tp53 and P38936 REA gene expression levels comparable to those treated with WPH serum or recombinant P13500 REA . Results indicate that mammary glands of rats fed a WPH diet are more protected from endogenous DNA damage than are those of CAS-fed rats , and identify P13500 REA as a potential serum biomarker for the positive effects of healthy diets .

P04150 REA and sequential P04637 REA activation by dexamethasone mediates apoptosis and cell cycle arrest of osteoblastic MC3T3 - E1 cells . Glucocorticoids play a pivotal role in the proliferation of osteoblasts , but the underlying mechanism has not been successfully elucidated . In this report , we have investigated the molecular mechanism which elucidates the inhibitory effects of dexamethasone on murine osteoblastic MC3T3 - E1 cells . It was found that the inhibitory effects were largely attributed to apoptosis and P55008 phase arrest . Both the cell cycle arrest and apoptosis were dependent on glucocorticoid receptor ( GR ) , as they were abolished by GR blocker DB00834 MENMAX DB00834 MEN pre-treatment and GR interference . P55008 phase arrest and apoptosis were accompanied with a p53 - dependent up-regulation of P38936 REA and pro-apoptotic genes Q13794 REA and PUMA . We also proved that dexamethasone ca n't induce apoptosis and cell cycle arrest when p53 was inhibited by p53 RNA interference . These data demonstrate that proliferation of MC3T3 - E1 cell was significantly and directly inhibited by dexamethasone treatment via aberrant GR activation and subsequently P04637 REA activation .

Effects of nimesulide , a preferential cyclooxygenase - 2 inhibitor , on carrageenan-induced pleurisy and stress-induced gastric lesions in rats . Intrapleural injection of carrageenan in rats increased prostaglandin E2 ( DB00917 ) production and induced newly synthesized cyclooxygenase - 2 ( P35354 REA ) in pleural exudate cells without affecting P23219 REA levels . DB04743 MEN , a preferential inhibitor of P35354 REA , reduced pleural DB00917 production and was almost as active as indomethacin and 10 times more active than ibuprofen . Only P23219 REA , and nc P35354 REA , was detected in gastric mucosal cells , and DB00917 concentration of gastric mucosa was significantly decreased by indomethacin and ibuprofen . The decrease in gastric DB00917 production induced by indomethacin and ibuprofen was enhanced in stressed rats , resulting in aggravation of stress-induced gastric lesions at anti-inflammatory doses . However , nimesulide did not produce stress-induced gastric lesions even at 30 times the anti-inflammatory dose . This supports the hypothesis that inhibition of P23219 REA causes unwanted side effects and inhibition of P35354 REA produces anti-inflammatory effects .

Analysis of CAD gene amplification using a combined approach of molecular genetics and cytogenetics . CAD is a multifunctional protein which catalyzes the first three steps of de novo uridine biosynthesis . Rodent cells resistant to DB03459 MEN , a specific inhibitor of the ATCase activity of CAD , overproduce the P27708 REA and CAD mRNA as a direct result of the amplification of the CAD gene . In order to study the mechanism of CAD gene amplification , a functional Syrian hamster CAD gene was inserted into a cosmid vector using molecular cloning techniques . The cloned genes were assayed for biological function by fusing CAD-deficient Chinese hamster ovary ( CHO ) cell mutants with protoplasts of E . coli containing the CAD cosmids . Two clones with functional CAD genes were isolated and shown to contain inserts 40 and 45 kb long . The cloned genes could also be introduced into wild type CHO cells by selecting for cells which became resistant to high DB03459 MEN concentrations in a single step . Transformations of mutant and wild type CHO cells contained multiple active copies of the donated Syrian hamster CAD genes in addition to their endogenous CHO CAD genes . The cloned genes in all transformants analyzed are integrated into host cell chromosomes at single locations defined by in situ hybridization . Independently isolated transformants contain the donated genes in different chromosomes . Co-transformation of CHO cells with two different genes by protoplast fusion is also shown to be possible .