MH_dev_11

Evaluation of pharmacological profile of meloxicam as an anti-inflammatory agent , with particular reference to its relative selectivity for cyclooxygenase - 2 over cyclooxygenase - 1 . We studied the anti-inflammatory activity of meloxicam on rat carrageenin-induced pleurisy and its toxicity for rat gastric mucosa , relative to its in vitro inhibitory potency against partially purified cyclooxygenase ( P36551 REA ) - 1 and P35354 REA preparations in order to clarify the pharmacological profile of the compound as an anti-inflammatory agent . In rat carrageenin-induced pleurisy , the plasma exudation rate peaked at 5 h , at which time P35354 REA was detectable in cells from the pleural exudate . Meloxicam and piroxicam ( 1 and 3 mg / kg ) and NS - 398 ( 3 mg / kg ) showed almost equal anti-inflammatory potency against 5 - hour pleurisy . A single oral administration of the compounds caused a dose-dependent increase in the number of rats with gastric mucosal erosion . The ED50 value for meloxicam ( 5.92 mg / kg ) was significantly higher than that for piroxicam ( 1.76 mg / kg ) , indicating that meloxicam is safer . DB00328 MENMAX DB00328 MEN showed intermediate safety ( 2.59 mg / kg ) . In in vitro experiments , indometacin inhibited P23219 REA about 1.7 times more potently than P35354 REA . NS - 398 inhibited P35354 REA with an IC50 of 0.32 microM , but never affected P23219 REA activity , even at 100 microM . In the same assay system , meloxicam inhibited P35354 REA about 12 times more selectively than P23219 REA . Piroxicam , however , inhibited both isoforms almost equally . These results indicate that meloxicam is a potent anti-inflammatory agent with low gastric toxicity . One reason for its in vivo pharmacological profile may be related to its relative selectivity for P35354 REA over P23219 REA . Thus , meloxicam may belong to a group of P35354 REA selective anti-inflammatory agents with a better safety profile than conventional P23219 REA and P35354 REA nonselective anti-inflammatory agents .

Anti-Parkinson ' s disease drugs and pharmacogenetic considerations . INTRODUCTION : The development of pharmacogenetic-based clinical practice guidelines for the use of anti-Parkinson ' s disease drugs requires , as a pre-requisite , the identification and validation of genetic biomarkers . These biomarkers are then used as surrogate endpoints . This review analyzes potential genetic biomarkers which can be used to improve anti-Parkinson ' s disease therapy . AREAS COVERED : The authors present an overview of current knowledge of pharmacogenetic implications of anti-Parkinson ' s disease drugs , including genes coding for the corresponding drug-metabolizing enzymes and drug targets . The gene / drug pairings with the strongest potential for pharmacogenetic recommendations include : P33261 REA / benztropine , P21964 REA / levodopa and entacapone , P20813 REA / selegiline , P22309 REA / entacapone , P14416 REA / ropinirole , pramipexole and cabergoline , and P35462 REA / ropinirole and pramipexole . Evidence supporting the effect of substrates , inhibitor or inducers for drug specific metabolizing enzymes in anti-Parkinson ' s disease drug response includes P05177 REA in the response to ropinirole and rasagiline , and P08684 REA in the response to bromocriptine , lisuride , pergolide and cabergoline . The authors present and discuss the current information on gene variations according to the 1000 genomes catalog and other databases with regards to anti-Parkinson ' s disease drugs . They also review and discuss the clinical implications of these variations . EXPERT OPINION : The goal of pharmacogenomic testing for anti-Parkinson ' s disease drugs should be conservative and aimed at selecting determined drugs for determined patients . However , much additional research is still needed to obtain reliable pre-prescription tests .

A case of a novel mutant vasopressin receptor-dependent nephrogenic diabetes insipidus with bilateral non-obstructive hydronephrosis in a middle aged man : differentiation from aquaporin-dependent nephrogenic diabetes insipidus by response of factor VII and P04275 REA to 1 - diamino - 8 - arginine vasopressin administration . We describe a case of a novel mutant vasopressin 2 receptor ( P30518 REA ) - dependent nephrogenic diabetes insipidus ( NDI ) with bilateral non-obstructive hydronephrosis in a middle aged man . This could be distinguished from aquaporin 2 ( P41181 REA ) - dependent NDI by the response of factor VIII and P04275 REA ( P04275 REA ) to DB00035 SUB ( DB00035 SUB ) administration . A 47 - year-old man was admitted to hospital because of polyuria , which had been present from infancy and was suspected of causing non-obstructive hydronephrosis . DB00117 mother ' s father , the older brother of his mother and his second daughter also all had polyuria . Sodium concentration , osmolality and vasopressin in blood were high , while sodium concentration and osmolality in urine were low . There were no changes in urine osmolality , factor VIII and P04275 REA in response to DB00035 SUB infusion . Neither was heart rate , diastolic blood pressure nor facial flushing affected . These findings suggested this case was P30518 REA - dependent NDI rather than P41181 REA - dependent NDI . Molecular genetic analysis demonstrated that the patient had a P30518 REA missense mutation involving a substitution of cysteine for arginine at position 104 ( R104C ) located in the first extracellular loop of the P30518 REA . It was also found that the patient ' s mother and his second daughter were heterozygous for this R104C mutation .

Protective effect of treatment with low-dose gliclazide in a model of middle cerebral artery occlusion and reperfusion in rats . The aim of this study was to explore the expression of sulfonylurea receptor 1 ( Q09428 REA ) , the regulatory subunit of the NCCa - DB00171 channel , and to investigate the protective effects of gliclazide following middle cerebral artery occlusion ( MCAO ) / reperfusion in male Wistar rats . Adult rats underwent 2h of the left MCAO using the intraluminal thread technique before reperfusion . The core areas of the infarct at different reperfusion time points were examined for the mRNA level and protein expression of Q09428 REA using reverse transcription-polymerase chain reaction ( RT-PCR ) and western blotting respectively . DB01120 MEN was administered intravenously into the right jugular vein for 12h simultaneously with the reperfusion . The number of apoptotic cells was determined using the TUNEL assay . The neurological functional deficits were evaluated using Bederson ׳ s test , and the cerebral infarction volume was visualized with TTC staining . We found up-regulation of Q09428 REA mRNA and protein levels in ischemic infarct tissues after reperfusion following MCAO , and Q09428 REA mRNA and protein were maximally upregulated 8-12 h after a 2 - hour ischemia . The treatment with low-dose of gliclazide reduced the total number of TUNEL-positive cells , the neurological functional deficits and the brain infarct volume . These results suggest that the Q09428 REA - regulated NCCa - DB00171 channel may be associated with MCAO / reperfusion injury and the infarct-reducing effects of intravenous treatment with gliclazide may be due , in part , to the blocked upregulation of Q09428 REA expression , the decreased infarct size and the reduced apoptosis in the ischemia-reperfusion brain .

Antidiabetic sulfonylurea enhances secretagogue-induced adrenocorticotropin secretion and proopiomelanocortin gene expression in vitro . The presence of high-affinity binding sites for antidiabetic sulfonylureas ( SUs ) and the expression of SU receptor ( Q09428 REA ) messenger RNA in the adenohypophyseal cells have recently been reported . In this study , we examined the effects of SU on P01189 REA gene expression and DB01285 secretion using the AtT 20PL cell line , a subclone of AtT 20 in which the rat P01189 REA 5 ' - promoter-luciferase fusion gene was stably incorporated . A representative SU glibenclamide inhibited the basal P01189 REA 5 ' - promoter activity . In contrast , glibenclamide enhanced forskolin - or P06850 REA - induced P01189 REA expression in a dose-dependent manner . Interestingly , the latter effect was not observed under treatment with DB07954 , a nonselective phosphodiesterase inhibitor . Furthermore , diazoxide , an opener of the DB00171 - sensitive K + channel , only antagonized the suppressive effect of glibenclamide . Lastly , RT-PCR analysis showed that mouse Q09428 REA ( but not SUR 2 ) messenger RNA was expressed in this cell line . These results suggest that , in AtT 20PL cells , SU has dual effects , i . e . a suppressive effect on basal P01189 REA expression through diazoxide-sensitive ( DB00171 - sensitive ) K + - channel-mediated mechanism , and an enhancing effect on DB02527 / protein kinase A-stimulated P01189 REA expression through a different mechanism ( probably mediated by phosphodiesterase ) . To our knowledge , this is the first report showing the effect of SU on the expression of the anterior pituitary hormone gene .

P14416 REA desensitization by dopamine or corticotropin releasing factor in ventral tegmental area neurons is associated with increased glutamate release . Neurons of the ventral tegmental area ( VTA ) are the source of dopaminergic ( DAergic ) input to important brain regions related to addiction . Prolonged exposure of these VTA neurons to moderate concentrations of dopamine ( DA ) causes a time-dependent decrease in DA-induced inhibition , a complex desensitization called DA inhibition reversal ( P30518 REA ) . P30518 REA is mediated by conventional protein kinase C ( cPKC ) through concurrent stimulation of D2 and D1 - like DA receptors , or by D2 stimulation concurrent with activation of some Gq-linked receptors . DB01285 releasing factor ( CRF ) acts via Gq , and can modulate glutamater neurotransmission in the VTA . In the present study , we used brain slice electrophysiology to characterize the interaction of DA , glutamate antagonists , and CRF agonists in the induction and maintenance of P30518 REA in the VTA . Glutamate receptor antagonists blocked induction but not maintenance of P30518 REA . Putative blockers of neurotransmitter release and store-operated calcium channels blocked and reversed P30518 REA . CRF and the CRF agonist urocortin reversed inhibition produced by the D2 agonist quinpirole , consistent with our earlier work indicating that Gq activation reverses quinpirole-mediated inhibition . In whole cell recordings , the combination of urocortin and quinpirole , but not either agent alone , increased spontaneous excitatory postsynaptic currents ( sEPSCs ) in VTA neurons . Likewise , the combination of a D1 - like receptor agonist and quinpirole , but not either agent alone , increased sEPSCs in VTA neurons . In summary , desensitization of D2 receptors induced by dopamine or CRF on DAergic VTA neurons is associated with increased glutamatergic signaling in the VTA .

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 .

DB00067 - induced P04275 REA secretion from endothelial cells involves V2 receptors and DB02527 . DB00067 and its analogue DB00035 SUB ( DB00035 SUB ) are known to raise plasma P04275 REA ( P04275 REA ) levels . DB00035 SUB is used as a hemostatic agent for the treatment of von Willebrand ' s disease . However , its cellular mechanisms of action have not been elucidated . DB00035 SUB , a specific agonist for the vasopressin V2 receptor ( P30518 REA ) , exerts its antidiuretic effect via a rise in DB02527 in kidney collecting ducts . We tested the hypothesis that DB00035 SUB induces P04275 REA secretion by binding to P30518 REA and activating DB02527 - mediated signaling in endothelial cells . P04275 REA secretion from human umbilical vein endothelial cells ( HUVECs ) can be mediated by DB02527 , but DB00035 SUB is ineffective , presumably due to the absence of P30518 REA . We report that DB00035 SUB stimulates P04275 REA secretion in a DB02527 - dependent manner in HUVECs after transfection of the P30518 REA . In addition , vasopressin and DB00035 SUB induce P04275 REA secretion in human lung microvascular endothelial cells ( HMVEC-L ) . These cells ( but not HUVECs ) express endogenous P30518 REA , as shown by RT-PCR . DB00067 - induced P04275 REA secretion is mimicked by DB00035 SUB and inhibited by the selective P30518 REA antagonist SR121463B . It is mediated by DB02527 , since it is inhibited by the protein kinase A inhibitor Rp - 8CPT - cAMPS . These results indicate that vasopressin induces DB02527 - mediated P04275 REA secretion by a direct effect on endothelial cells . They also demonstrate functional expression of P30518 REA in endothelial cells , and provide a cellular mechanism for the hemostatic effects of DB00035 SUB .

Protective effects of metallothionein on isoniazid and rifampicin-induced hepatotoxicity in mice . Isoniazid ( DB00951 MEN ) and DB01045 ( RFP ) are widely used in the world for the treatment of tuberculosis , but the hepatotoxicity is a major concern during clinical therapy . Previous studies showed that these drugs induced oxidative stress in liver , and several antioxidants abated this effect . Metallothionein ( MT ) , a member of cysteine-rich protein , has been proposed as a potent antioxidant . This study attempts to determine whether endogenous expression of MT protects against DB00951 MEN and RFP-induced hepatic oxidative stress in mice . Wild type ( MT + / + ) and MT-null ( MT - / - ) mice were treated intragastrically with DB00951 MEN ( 150 mg / kg ) , RFP ( 300 mg / kg ) , or the combination ( 150 mg / kg DB00951 MEN + 300 mg / kg RFP ) for 21 days . The results showed that MT - / - mice were more sensitive than MT + / + mice to DB00951 MEN and RFP-induced hepatic injuries as evidenced by hepatic histopathological alterations , increased serum Q9NRA2 levels and liver index , and hepatic oxidative stress as evidenced by the increase of MDA production and the change of liver antioxidant status . Furthermore , DB00951 MEN increased the protein expression of hepatic P05181 REA and DB00951 MEN / RFP ( alone or in combination ) decreased the expression of hepatic P05177 REA . These findings clearly demonstrate that basal MT provides protection against DB00951 MEN and RFP-induced toxicity in hepatocytes . The P05181 REA and P05177 REA were involved in the pathogenesis of DB00951 MEN and RFP-induced hepatotoxicity .

Effects of Luteolin on Liver , Kidney and Brain in Pentylentetrazol-Induced Seizures : Involvement of Metalloproteinases and NOS Activities . OBJECTIVE : Flavonoids are an important group of recognized antioxidants in plants . Luteolin ( LUT ) is a natural flavonoid in the plant kingdom . This study was aimed to investigate the effects of the LUT in the liver , kidney and brain of pentylentetrazol ( PTZ ) - induced seizure and the relationship between nitric oxide synthases ( P35228 REA , P29474 REA ) and matrix metalloproteinases ( P08253 REA , P14780 REA ) . MATERIALS AND METHODS : LUT ( 10 mg / kg ) was given intraperitoneally during two weeks prior to seizure induction . A single dose PTZ 80 mg / kg i . p . was administered and seizures were observed and evaluated with regard to latency , frequency and stage for one hour . RESULTS : Seizure frequen cy after PTZ administration was significantly decreased in LUT pretreated rats ( p < 0.05 ) . An increase of immunhistochemical reactions of P35228 REA and P08253 REA , but a decrease of P29474 REA activity , were observed in rat hippocampus and peripheral tissues during the PTZ induced seizures . LUT pretreatment reversed the P35228 REA and P08253 REA activity to the control levels and significantly increased the P29474 REA activity ( p < 0.001 ) . CONCLUSION : LUT seems to have an effective role in reducing the seizure frequency and a protective role on peripheral organ injury in animal models of seizure . The protective effect of LUT in seizures and the seizure induced peripheral tissue damage warrant further investigations .

The protective effect of rebamipide on paracellular permeability of rat gastric epithelial cells . BACKGROUND : Barrier function in gastric epithelial cells is essential for the gastric defence mechanism against acid back-diffusion into the mucosal layer . Our previous study indicated that trans-epithelial resistance ( Q9NZ01 ) of rat gastric epithelial cells was rapidly increased when the cells were exposed to acid . This response to acid was diminished by indometacin . AIM : Evaluate the effects of a mucoprotective agent , rebamipide , on the nonsteroidal anti-inflammatory drug ( NSAID ) - induced increase of gastric epithelial permeability . METHODS : Rat gastric epithelial cells were plated on tissue culture inserts . Cells were exposed to a NSAID ( indometacin , 10-7 M ) . Trans-epithelial permeability was measured by Q9NZ01 and diffusion rate of 14C - mannitol . The effect of rebamipide was evaluated by measuring Q9NZ01 . Endogenous prostaglandin E2 ( DB00917 ) production in culture medium was also measured . RESULTS : DB00328 MEN gradually and significantly decreased Q9NZ01 and increased 14C - manitol permeability . Rebamipide reversed the indometacin-induced changes in epithelial permeability and induced DB00917 synthesis . This induction was blocked by either indometacin or a Cyclooxygenase ( P36551 REA ) - 2 specific inhibitor . CONCLUSIONS : P36551 REA inhibitors such as indometacin inhibit regulation of epithelial permeability by reducing DB00917 . P23219 REA has an important role in the gastric defense mechanism . Rebamipide suppressed an indometacin-induced increase in gastric epithelial permeability by increasing DB00917 levels in a P35354 REA dependent manner .

Synthesis , biological activity and HPLC validation of 1,2 , 3,4- tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 MEN ) and 4 - fluorobenzoic acid ( 4 - FBA ) possessing activity towards acetylcholinesterase ( P22303 REA ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4 - FBA and diamino derivatives of 1,2 , 3,4- tetrahydroacridine . The compounds P13671 REA - 2KW / HCl , P13671 REA - 4KW / HCl and P13671 REA - 3KW / HCl have four-fold higher antiacetylcholinesterase activity than DB00382 MEN . All of the acquired compounds present higher selectivity towards P22303 REA than DB00382 MEN and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 REA - 2KW / HCl , P13671 REA - 3KW / HCl and P13671 REA - 4KW / HCl . DB00382 MEN and 4 - FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile / buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v / v ) ( overall pH 4 ) . A 1.5 ml / min flow rate and a 247 nm wavelength were chosen for this method . P13671 REA - 2KW / HCl , P13671 REA - 3KW / HCl and P13671 REA - 4KW / HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 ° C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg / ml and 6 μg / ml for P13671 REA - 2KW / HCl , P13671 REA - 3KW / HCl and P13671 REA - 4KW / HCl , 0.04 μg / ml and 0.12 μg / ml for DB00382 MEN , 0.42 μg / ml and 1.41 μg / ml for 4 - FBA , respectively .

A novel mutation in P30518 REA causing congenital nephrogenic diabetes insipidus with complete resistance to antidiuretic hormone . A 6 - month-old male infant presented with failure to thrive . Hypernatraemia and elevated serum osmolality in the presence of low urine sodium and osmolality led to the diagnosis of diabetes insipidus . Administration of DB00035 SUB ( dDAVP ) neither decreased urine volume nor increased urine osmolality indicating congenital nephrogenic diabetes insipidus . Molecular analysis in the arginine-vasopressin receptor - 2 gene ( P30518 REA ) located on chromosome Xq28 demonstrated a novel 5 - base pair deletion ( c . 962-966 delACCCC ; g . 1429-1433 delACCCC ) leading to a shift of the reading frame ( p . Asn 321fs ) and a premature termination codon implying an absent or non-functional protein . Treatment with hydrochlorothiazide , amiloride and indomethacin led to a favourable clinical course .

DB00067 triggers senescence in K-ras transformed cells via RhoA-dependent downregulation of cyclin D1 . DB00067 ( AVP ) , a vasoactive peptide hormone that binds to three G-protein coupled receptors ( V1R , P30518 REA , and V3R ) , has long been known to activate V1R and elicit mitogenesis in several cell types , including adrenal glomerulosa cells . However , in the mouse Q03519 REA adrenocortical malignant cell line , AVP triggers not only a canonical mitogenic response but also novel RhoA-GTP-dependent mechanisms which downregulate cyclin D1 , irreversibly inhibiting K-ras oncogene-driven proliferation . In Q03519 REA cells , AVP blocks cyclin D1 expression , induces senescence-associated beta-galactosidase ( SAbeta-Gal ) and inhibits proliferation . However , ectopic expression of cyclin D1 renders Q03519 REA cells resistant to both SAbeta-Gal induction and proliferation inhibition by AVP . In addition , ectopic expression of the dominant negative RhoAN 19 mutant blocks RhoA activation , yielding Q03519 REA cell sub-lines which are no longer susceptible to cyclin D1 downregulation , SAbeta-Gal induction , or proliferation inhibition by AVP . Furthermore , inhibiting RhoA with P01024 REA exoenzyme protects Q03519 REA cells from AVP proliferation inhibition and SAbeta-Gal induction . On the other hand , AVP treatment does not activate caspases 3 and 7 , and the caspase inhibitor Ac-DEVD-CMK does not protect Q03519 REA cells from proliferation inhibition by AVP , implying that AVP does not trigger apoptosis . These results underline a pivotal survival activity of cyclin D1 that protects K-ras oncogene-dependent malignant cells from senescence .

Ectopic DB01285 syndrome caused by bronchial carcinoid tumor indistinguishable from Cushing ' s disease . A 75 - year-old woman was admitted to our hospital because of a poor glycemic control . She was found to have Cushingoid feature and dynamic endocrine tests showed elevated plasma DB01285 and cortisol levels , lack of their circadian rhythm , non-suppressibility to high-dose dexamethasone , responsiveness to P06850 REA , but not to DB00035 SUB , and suppression to octreotide . Pituitary Q9BWK5 showed an equivocal small lesion . CT scan of the chest showed two nodular lesions in the right lung ( S5 , S7 ) , while a mild uptake was noted only in S5 lesion by DB09150 - PET , but positive uptake was only in S7 lesion by somatostatin receptor scintigraphy ( SRS ) . Inferior petrosal sinus sampling revealed a gradient of plasma DB01285 after P06850 REA stimulation , consistent with the diagnosis of Cushing ' s disease . She underwent middle and inferior lobectomy of the right lung . The resected tumor in S7 was consistent with the diagnosis of a bronchial carcinoid tumor with positive DB01285 immunoreactivity , while that of S5 was cryptococcal granuloma . RT-PCR revealed abundant expressions of P01189 REA and SSTR ( - 1 , - 2 , - 5 ) , but not of P34998 REA and P47901 REA . Postoperatively , abnormal endocrine data were normalized along with improvement of hypertension and diabetes . This was a diagnostic challenging case with ectopic DB01285 syndrome indistinguishable from Cushing ' s disease by various endocrine and imaging tests , among which SRS successfully localized the tumor responsible for ectopic DB01285 secretion .

Determination of cobimetinib in human plasma using protein precipitation extraction and high-performance liquid chromatography coupled to mass spectrometry . Inhibition of Q96HU1 / P29323 REA kinase ( MEK ) is a promising strategy to control the growth of tumors that are dependent on aberrant signaling in the MEK pathway . DB05239 MEN ( P16260 REA - 0973 ) ( S ) - [ 3,4- Difluoro - 2 - ( 2 - fluoro - 4 - iodo-phenylamino ) - phenyl ] - ( ( S ) - 3 - hydroxy - 3 - piperidin - 2 - yl-azetidin - 1 - yl ) - methanone ) inhibits proliferation of a variety of human tumor cell lines by inhibiting Q02750 REA and P36507 REA . A specific high performance liquid chromatography-mass spectrometric assay was developed and validated for the determination of cobimetinib in human plasma . The overall mean recovery using protein precipitation extraction with acetonitrile was found to be 54.1 % . The calibration curve was ranged from 0.20 to 100ng / mL . The LLOQ was sensitive enough to detect terminal phase concentrations of the drug . The intra - and inter-assay precision ( % CV ) was within 10.3 % and 9.5 % for cobimetinib . The assay accuracy ( % RE ) was within ± 13.7 % of the nominal concentration values for cobimetinib with the normal analytical QCs . The developed assay was successfully used to analyze the human plasma samples ( for pharmacokinetic analysis ) from clinical trials .

Differential actions of vasopeptidase inhibition versus angiotensin-converting enzyme inhibition on diuretic therapy in experimental congestive heart failure . BACKGROUND : DB00886 MEN ( OMA ) , a vasopeptidase inhibitor , simultaneously inhibits angiotensin-converting enzyme ( P12821 REA ) and neutral endopeptidase , which degrades vasodilatory factors ( eg , P35318 REA ) and natriuretic peptides . Based on the beneficial cardiorenal and humoral properties of the natriuretic peptides , we hypothesized that an acute vasopeptidase inhibitor with or without diuretic would result in more favorable cardiorenal and hormonal actions than P12821 REA inhibition plus diuretic ( ACEI + D ) in congestive heart failure . METHODS AND RESULTS : We compared the actions of OMA alone and with diuretic ( OMA + D ) to ACEI + D in a model of pacing-induced congestive heart failure . OMA + D decreased pulmonary arterial and pulmonary capillary wedge pressures to a greater level than OMA alone or ACEI + D . Glomerular filtration rate was lower with ACEI + D than with either OMA group . Plasma renin activity and aldosterone immediately increased with ACEI + D , whereas OMA + D resulted in higher plasma renin activity and a delayed increase in aldosterone . OMA alone did not increase plasma renin activity and aldosterone , but resulted in a sustained increase in plasma adrenomedullin , with higher urinary atrial natriuretic peptide , adrenomedullin , and cGMP excretions than with ACEI + D . CONCLUSIONS : Acute administration of OMA with or without diuretic results in more favorable cardiorenal and humoral responses in experimental congestive heart failure than does ACEI + D . There is no acute activation of renin and aldosterone with OMA alone such as occurs with ACEI + D and OMA + D . Thus , OMA with or without a diuretic possesses beneficial cardiorenal and humoral actions comparable to those observed with ACEI + D that can be explained by potentiation of natriuretic peptides .

Desmopressin ( DB00035 SUB ) induces NO production in human endothelial cells via V2 receptor - and DB02527 - mediated signaling . The hemostatic agent desmopressin ( DB00035 SUB ) also has strong vasodilatory effects . DB00035 SUB is a selective agonist for the vasopressin V2 receptor ( P30518 REA ) , which is coupled to DB02527 - dependent signaling . DB00035 SUB - induced vasodilation may be due to endothelial NO synthase ( P29474 REA ) activation . This hypothesis implies DB02527 - mediated P29474 REA activation . It also implies wide extrarenal , endothelial P30518 REA expression . We show that in human umbilical vein endothelial cells ( HUVECs ) the DB02527 - raising agents forskolin and epinephrine increase NO production , as measured by a l-NMMA-inhibitable rise in cellular cGMP content . They also increase P29474 REA enzymatic activity , in a partly calcium-independent manner . DB02527 - mediated P29474 REA activation is associated with phosphorylation of residue Ser 1177 , in a phosphatidyl inositol 3 - kinase ( PI3K ) - independent manner . HUVECs do not express P30518 REA . However , after heterologous P30518 REA expression , DB00035 SUB induces DB02527 - dependent P29474 REA activation via Ser 1177 phosphorylation . We have previously found P30518 REA expression in cultured lung endothelial cells . By real time quantitative RT-PCR , we now find a wide P30518 REA distribution notably in heart , lung and skeletal muscle . These results indicate that DB00035 SUB and other DB02527 - raising agents can activate P29474 REA via PI3K - independent Ser 1177 phosphorylation in human endothelial cells . This mechanism most likely accounts for DB00035 SUB - induced vasodilation .

Correlation between tumor volume response to radiotherapy and expression of biological markers in patients with cervical squamous cell carcinoma . OBJECTIVE : To determine the factors associated with tumor volume response to radiotherapy ( RT ) in cervical cancer patients , and the relationship between the tumor volume response and alteration of the expression of biological markers during RT . METHODS : Twenty consecutive patients with cervical squamous cell carcinoma who received definitive RT were enrolled . Tumor volumes were calculated by Q9BWK5 examinations performed at the start of RT ( pre-RT ) , at the fourth week of RT ( mid-RT ) , and 1 month after RT completion ( post-RT ) . Two serial punch biopsies were performed at pre - and mid-RT , and immunohistochemical staining was performed for cyclooxygenase ( P36551 REA ) - 2 and epidermal growth factor receptor ( P00533 REA ) . RESULTS : For the pre-RT evaluation , fourteen ( 70 % ) and eleven ( 55 % ) patients showed positive immunoreactivity for P35354 REA and P00533 REA , respectively . Among the seven patients whose median percentage residual tumor at mid-RT ( P30518 REA ) was greater than 0.5 , seven ( 100 % , p= 0.0515 ) and five ( 71.4 % , p= 0.3742 ) patients showed positive immunoreactivity for P35354 REA and P00533 REA , respectively . The logistic regression analysis showed that positive immunoreactivity for both P35354 REA and P00533 REA at pre-RT were associated with P30518 REA ( p= 0.0782 ) . For the mid-RT evaluation , eight cases showed an interval increase in the distribution of immunoreactivity for P35354 REA , and six out of the eight patients had a P30518 REA greater than 0.5 ( p= 0.2222 ) . CONCLUSION : The poor mid-RT tumor response was associated with the coexpression of P35354 REA and P00533 REA .

Preparation of phosphorylated starch by dry-heating in the presence of pyrophosphate and its calcium-phosphate solubilizing ability . Starch was phosphorylated through dry-heating in the presence of pyrophosphate at various conditions , and the characteristics of phosphorylated starch ( PS ) were examined . Starch phosphorylation increases as the pH increases from 3 to 6 , but diminishes at pH 7 . Increased temperatures enhance phosphorylation . Data from ( 31 ) P NMR suggests that starch phosphorylation occurs mainly at the P01024 REA - OH and P13671 REA - OH of the glucose residue . The phosphate linkage is mainly due to monostarch monophosphate . Although starch had almost no calcium phosphate-solubilising capacity , this capacity was markedly enhanced by phosphorylation . X-ray diffraction analysis indicates that the crystal structure of hydroxyapatite was not present in the calcium phosphate-PS complex .

Loss of Jak 2 impairs endothelial function by attenuating P04049 REA / Q02750 REA / Sp - 1 signaling along with altered P29474 REA activities . A number of inhibitors have been used to dissect the functional relevance of Jak 2 in endothelial homeostasis , with disparate results . Given that Jak 2 deficiency leads to embryonic lethality , the exact role of Jak 2 in the regulation of postnatal endothelial function is yet to be fully elucidated . We generated a model in which Jak 2 deficiency can be induced by tamoxifen in adult mice . Loss of Jak 2 significantly impaired endothelium-dependent response capacity for vasodilators . Matrigel plug assays indicated a notable decrease in endothelial angiogenic function in Jak 2 - deficient mice . Studies in a hindlimb ischemic model indicated that Jak 2 activity is likely to be a prerequisite for prompt perfusion recovery , based on the concordance of temporal changes in Jak 2 expression during the course of ischemic injury and perfusion recovery . A remarkable delay in perfusion recovery , along with reduced capillary and arteriole formation , was observed in Jak 2 - deficient mice . Antibody array studies indicated that loss of Jak 2 led to repressed P29474 REA expression . In mechanistic studies , Jak 2 deficiency attenuated P04049 REA / Q02750 REA signaling , which then reduced activity of Sp - 1 , an essential transcription factor responsible for P29474 REA expression . These data are important not only for understanding the exact role that Jak 2 plays in endothelial homeostasis , but also for assessing Jak 2 - based therapeutic strategies in a variety of clinical settings .

Mechanisms for epigallocatechin gallate induced inhibition of drug metabolizing enzymes in rat liver microsomes . DB03823 gallate ( EGCG ) inhibits drug metabolizing enzymes by unknown mechanisms . Here we examined if the inhibition is due to covalent-binding of EGCG to the enzymes or formation of protein aggregates . EGCG was incubated with rat liver microsomes at 1-100 μM for 30min . The EGCG-binding proteins were affinity purified using m-aminophenylboronic acid agarose and probed with antibodies against glyceraldehyde - 3 - phosphate dehydrogenase ( P04406 REA ) , actin , cytochrome P450 ( CYP ) 1A1 , P05177 REA , CYP 2B1 / 2 , P05181 REA , CYP 3A , catechol-O-methyltransferase ( P21964 REA ) and microsomal glutathione transferase 1 ( P10620 REA ) . All but actin and soluble P21964 REA were positively detected at ≥ 1μM EGCG , indicating EGCG selectively bound to a subset of proteins including membrane-bound P21964 REA . The binding correlated well with inhibition of CYP activities , except for P05181 REA whose activity was unaffected despite evident binding . The antioxidant enzyme P10620 REA , but not cytosolic GSTs , was remarkably inhibited , providing novel evidence supporting the pro-oxidative effects of EGCG . When microsomes incubated with EGCG were probed on Western blots , all but the actin and P05181 REA antibodies showed a significant reduction in binding at ≥ 1μM EGCG , suggesting that a fraction of the indicated proteins formed aggregates that likely contributed to the inhibitory effects of EGCG but were not recognizable by antibodies against the intact proteins . This raised the possibility that previous reports on EGCG regulating protein expression using P04406 REA as a reference should be revisited for accuracy . Remarkable protein aggregate formation in EGCG-treated microsomes was also observed by analyzing Coomassie Blue-stained SDS-PAGE gels . EGCG effects were partially abolished in the presence of 1mM glutathione , suggesting they are particularly relevant to the in vivo conditions when glutathione is depleted by toxicant insults .

Absolute bioavailability and effect of formulation change , food , or elevated pH with rabeprazole on cobimetinib absorption in healthy subjects . DB05239 MEN is a potent and highly selective inhibitor of Q02750 REA / 2 . Since cobimetinib exhibited absorption variability in cancer patients , a series of single-dose studies in healthy subjects were conducted to determine absolute bioavailability and elucidate potential effects of formulation , food , and elevated gastric pH on cobimetinib bioavailability . Three crossover trials were performed with a 20 mg cobimetinib oral dose : absolute bioavailability using a 2 mg intravenous infusion ( n = 13 ) , relative bioavailability of tablets versus capsules and food effect ( n = 20 ) , and drug interaction with a proton pump inhibitor ( 20 mg of rabeprazole daily for 5 days prior to cobimetinib administration ; n = 20 ) . Absolute bioavailability of cobimetinib was 46.2 % ( 24.2 , CV % ) , likely due to metabolism rather than incomplete absorption . The mean systemic clearance of cobimetinib was low ( 11.7 L / h [ 28.2 , CV % ] ) . Administration of cobimetinib tablets with a high-fat meal delayed drug absorption ( prolonged tmax ) but had no statistically significant effect on cobimetinib exposure ( Cmax and AUC 0 - ∞ ) . Tablet and capsule formulations of cobimetinib showed comparable exposures . DB05239 MEN exhibited delayed absorption ( tmax ) in the presence of rabeprazole , with no statistically significant effects on drug exposure ( Cmax and AUC 0 - ∞ ) in the fasted state . In conclusion , cobimetinib oral absorption was not affected by change in formulation , food , or elevated gastric pH .

Effects of retinol binding protein - 4 on vascular endothelial cells . The study was designed to investigate the effect of retinol binding protein ( P02753 REA ) - 4 on the phosphatidylinositol 3 - kinase ( PI3K ) and mitogen-activated protein kinase ( MAPK ) pathways , which mediate the effects of insulin in vascular endothelial cells . The effects of P02753 REA on nitric oxide ( NO ) and insulin-stimulated endothelin - 1 ( ET - 1 ) secretion and on phosphorylation ( p ) of Akt , endothelial NO synthetase ( P29474 REA ) , and extracellular signal-regulated kinase ( P29323 REA ) 1/2 were investigated in bovine vascular aortic endothelial cells ( BAECs ) . P02753 REA showed an acute vasodilatatory effect on aortic rings of rats within a few minutes . In BAECs , P02753 REA - treatment for 5min significantly increased NO production , but inhibited insulin-stimulated ET - 1 secretion . P02753 REA - induced NO production was not inhibited by tetraacetoxymethylester ( BAPTA-AM ) , an intracellular calcium chelator , but was completely abolished by wortmannin , a PI3K inhibitor . P02753 REA significantly increased p-Akt and p - P29474 REA production , and significantly inhibited p - P27361 REA / 2 production . Triciribine , an Akt inhibitor , and wortmannin significantly inhibited P02753 REA - induced p-Akt and p - P29474 REA production . Inhibition of Akt 1 by small interfering RNA decreased p - P29474 REA production enhanced by P02753 REA in human umbilical vein endothelial cells . In conclusion , P02753 REA has a robust acute effect of enhancement of NO production via stimulation of part of the PI3K / Akt / P29474 REA pathway and inhibition of P27361 REA / 2 phosphorylation and insulin-induced ET - 1 secretion , probably in the MAPK pathway , which results in vasodilatation .

Permanent neonatal diabetes mellitus in China . BACKGROUND : Permanent neonatal diabetes mellitus ( PNDM ) is a rare disease , which is defined as the onset of diabetes before the age of 6 months with persistence through life . Infants with Q14654 REA or Q09428 REA genetic mutations may respond to oral sulfonylurea therapy . Currently , there are limited studies about the genetic analysis and long-term follow-up of PNDM . CASE PRESENTATION : We report four cases of PNDM . None of the infants or their parents had P01308 REA , Q14654 REA , or Q09428 REA genetic mutations . One infant underwent continuous subcutaneous insulin infusion ( CSII ) and the other infants underwent multiple injections of insulin ( MII ) . In these infants , PNDM persisted from 35 months to 60 months of follow-up . Three infants maintained fairly stable blood sugar levels , and one infant had poor sugar control . CONCLUSIONS : We suggest that all of the infants with PNDM should undergo genetic evaluation . For infants without Q14654 REA and Q09428 REA genetic mutations , oral sulfonylurea should not be considered as treatment . CSII is a useful method for overcoming the difficulties of diabetes , and it may also improve the quality of life of both infants and their parents .

Functional characterization of vasopressin receptor 2 mutations causing partial and complete congenital nephrogenic diabetes insipidus in Thai families . BACKGROUND : P30518 REA mutations cause most cases of nephrogenic diabetes insipidus ( NDI ) ; 211 P30518 REA mutations have been described , but only 7 are described causing partial NDI . METHODS : Two unrelated Thai boys had polyuria and polydipsia in infancy but had normal electrolytes and serum osmolality at 2 years of age . Patient 1 could not concentrate his urine in response to water deprivation or 1 - desamino - 8-D - arginine vasopressin ( DB00035 SUB ) ; patient 2 could concentrate to approximately 600 mosm / l . The patients ' P30518 REA genes were sequenced and the identified mutations were re-created in P30518 REA cDNA expression vectors . P30518 REA activities were measured by stimulating transfected HEK 293T cells with arginine vasopressin ( AVP ) or DB00035 SUB , and assessing the resulting DB02527 production by the activation of a luciferase reporter . RESULTS : Patient 1 carried the previously described missense mutation R181C ; patient 2 carried the novel missense mutation M311V . When transiently transfected into HEK 293T cells , 6.8 x 10 ( - 12 ) M AVP induced the half-maximal response ( EC50 ) of the wild-type , whereas the EC50 value for R181C was 5.9 x 10 ( - 9 ) M and for M311V was 2.6 x 10 ( - 10 ) M . Responses to DB00035 SUB were qualitatively similar but required 10 - fold higher concentrations . CONCLUSION : The novel P30518 REA mutation M311V retains partial activity and results in a milder form of NDI .

Lipoxygenase pathway mediates increases of airway resistance and lung inflation induced by exposure to nanotitanium dioxide in rats . Nanotitanium dioxide particle ( nTiO 2 ) inhalation has been reported to induce lung parenchymal injury . After inhalation of nTiO 2 , we monitored changes in P09917 REA , endothelial nitric oxide synthase ( P29474 REA ) , and inducible nitric oxide synthase ( P35228 REA ) mRNA in rat lung tissue . Lung function parameters include specific airway resistance ( SRaw ) , peak expiratory flow rate ( PEF ) , functional residual capacity ( FRC ) , and lung compliance ( Cchord ) ; blood white blood cell count ( WBC ) , nitric oxide ( NO ) , hydrogen peroxide , and lactic dehydrogenase ( LDH ) ; and lung lavage leukotriene C4 , interleukin 6 ( P05231 REA ) , tumor necrotic factor α ( TNFα ) , hydroxyl radicals , and NO . Leukotriene receptor antagonist MK571 and P09917 REA inhibitor MK886 were used for pharmacologic intervention . Compared to control , nTiO 2 exposure induced near 5 - fold increase in P09917 REA mRNA expression in lung tissue . P35228 REA mRNA increased while P29474 REA mRNA decreased . Lavage leukotriene C4 ; P05231 REA ; TNFα ; NO ; hydroxyl radicals ; and blood WBC , NO , hydrogen peroxide , and LDH levels rose . Obstructive ventilatory insufficiency was observed . MK571 and MK886 both attenuated the systemic inflammation and lung function changes . We conclude that inhaled nTiO 2 induces systemic inflammation , cytokine release , and oxidative and nitrosative stress in the lung . The lipoxygenase pathway products , mediated by oxygen radicals and WBC , play a critical role in the obstructive ventilatory insufficiency induced by nTiO 2 .

Mapping of an ordered set of 14 cosmids to human chromosome 12p by two-color in situ hybridization . To map human chromosome 12p aberrations by fluorescence in situ hybridization ( Q5TCZ1 ) , cosmids were isolated or obtained for 14 known 12p loci ( D12S119 , D12S158 , D12S178 , P36941 REA , D12S380E , P01023 REA , Q13936 REA , P10767 REA , P04406 REA , P01116 REA , P04280 , P20742 , P60174 REA , and P04275 REA ) . Using two-color Q5TCZ1 with three labeled probes to interphase nuclei , and to prometaphase chromosomes where possible , the order of these loci was sequentially determined to be pter-D 12S158 - D12S380E - Q13936 REA - P10767 REA - P36941 REA - P04275 REA - P04406 REA - P60174 REA - P01023 REA - P20742 - P04280 - D12S 178 - D12S119 - P01116 REA - cen . Two cell lines were analyzed with this set of cosmids . The EBV-transformed cell line TA carries a der ( 12 ) with a deletion of bands 12p13 . 1 --> p11 . 2 . D12S178 , D12S119 , and P01116 REA were absent in the der ( 12 ) , whereas the other loci were present . The second cell line , GM01203A , exhibits a balanced t ( 4 ; 12 ) ( 4q25 ; 12p13 . 3 ) with a breakpoint between P10767 REA and P36941 REA .

Antiproliferative effect of DB00035 SUB analogs on human breast cancer cells . BACKGROUND : Desmopressin ( dDAVP ) , a synthetic nonapeptide derivative of arginine vasopressin , is a safe antidiuretic and hemostatic compound that acts as a selective agonist for the vasopressin V2 membrane receptor ( P30518 REA ) . It is known that dDAVP can inhibit progression of residual metastatic cells in preclinical models . Among other mechanisms , the compound induces an agonist effect on P30518 REA present in tumor cells . RESULTS / DISCUSSION : Looking for novel analogs with improved anti-tumor activity , positions 4 and 5 , at the conformational peptide loop , were substituted . The analog [ V ( 4 ) Q ( 5 ) ] dDAVP ( [ 4 - valine 5 - glutamine ] desmopressin ) exhibited a significantly higher antiproliferative effect than dDAVP in cultures of MCF - 7 , a P30518 REA - expressing human breast carcinoma cell line . The chiral isomer of this analog and tetrapeptide fragments corresponding to the loop region were also assessed . CONCLUSION : Preclinical evaluation of the anti-tumor activity of [ V ( 4 ) Q ( 5 ) ] dDAVP in animal models is warranted .

Mass spectrometry and hydrogen / deuterium exchange measurements of alcohol-induced structural changes in cellular retinol-binding protein type I . To bind and release its ligand , cellular retinol-binding protein type I ( P09455 REA ) needs to undergo conformational and dynamic changes to connect the inner , solvent-shielded cavity , where retinol is found to bind , and the outside medium . DB00162 dissociation in vitro is favoured by water / alcohol mixtures whose moderately low dielectric constants mimic a property characteristic of the membrane microenvironment where this process occurs in vivo . Apo - and holo - P09455 REA , in either water / methanol or water / trifluoroethanol ( TFE ) mixtures , were analyzed at equilibrium by electrospray ionization with orthogonal quadrupole time-of-flight mass spectrometry ( P19957 REA - Q-TOFMS ) to identify the alcohol-induced species . The questions were asked whether the presence of alcohols affects protein dynamics , as reflected by hydrogen / deuterium ( H / D ) exchange monitored by continuous-labelling experiments , and to which extent retinol dissociation influences the process . With increasing methanol , at pH near neutrality , apo - P09455 REA exhibits a progressively more compact conformation , resulting in reduced H / D exchange with respect to the native protein in water . DB00162 dissociation from the holo-protein did not promote hydrogen replacement . Similarly , in the presence of the low TFE concentration sufficient to cause retinol dissociation , the hydrogen exchange of the resulting apo-protein was not exalted . However , in contrast with the alkanol , higher TFE concentrations induced a transition of apo - P09455 REA to a new alpha-helix conformation capable of exchanging all available hydrogen atoms .

Therapy with a synthetic retinoid - - ( Ro 10-1670 ) etretin - - increases the cellular retinoic acid-binding protein in nonlesional psoriatic skin . Cellular retinol ( P09455 REA ) - and retinoic acid ( CRABP ) - binding proteins were determined in samples of lesional and nonlesional skin of psoriatic patients , before and during oral administration of a synthetic retinoid , DB00459 MEN ( Ro 10-1670 ) . A 200 % increase in CRABP levels , measured by the ability of the protein to bind retinoic acid , was observed in the normal skin during treatment . The P09455 REA levels were not altered during therapy . The results show that P09455 REA and CRABP are independently regulated in human skin and suggest that synthetic retinoids may exert their pharmacologic effects by interfering with the regulation of natural retinoic acid receptors .

Acute diabetes insipidus mediated by vasopressinase after placental abruption . CONTEXT : Postpartum , diabetes insipidus ( DI ) can be part of Sheehan ' s syndrome or lymphocytic hypophysitis in combination with anterior pituitary hormone deficiencies . In contrast , acute onset of isolated DI in the postpartum period is unusual . CASE PRESENTATION : This patient presented at 33 weeks gestation with placental abruption , prompting a cesarean delivery of twins . Immediately after delivery , she developed severe DI . The DI could be controlled with the vasopressinase-resistant DB00035 SUB ( DB00035 SUB ) , but not with arginine vasopressin ( AVP ) , and it resolved within a few weeks . OBJECTIVE : The aim of this study was to demonstrate that the postpartum DI in this patient was caused by the release of placental vasopressinase into the maternal bloodstream . METHODS AND RESULTS : Cells were transiently transfected with the AVP receptor 2 ( P30518 REA ) and treated with either AVP or DB00035 SUB in the presence of the patient ' s serum collected postpartum or 10 weeks after delivery . The response to the different treatments was evaluated by measuring the activity of a DB02527 - responsive firefly luciferase reporter construct . The in vitro studies demonstrate that the patient ' s postpartum serum disrupts activation of the P30518 REA by AVP , but not by the vasopressinase-resistant DB00035 SUB . CONCLUSIONS : Placental abruption can rarely be associated with acute postpartum DI caused by release of placental vasopressinase into the bloodstream . This clinical entity must be considered in patients with placental abruption and when evaluating patients presenting with DI after delivery .

Identification , characterization and rescue of a novel vasopressin - 2 receptor mutation causing nephrogenic diabetes insipidus . OBJECTIVE : X-linked nephrogenic diabetes insipidus ( XNDI ) , caused by mutations in the V2 vasopressin receptor ( P30518 REA ) , is clinically distinguished from central diabetes insipidus ( CDI ) by elevated serum vasopressin ( AVP ) levels and unresponsiveness to 1 - desamino - 8-d - arginine vasopressin ( DB00035 SUB ) . We report two infants with XNDI , and present the characterization and functional rescue of a novel P30518 REA mutation . PATIENTS : Two male infants presented with poor growth and hypernatraemia . Both patients had measurable pretreatment serum AVP and polyuria that did not respond to DB00035 SUB , suggesting NDI . However , both also had absent posterior pituitary bright spot on Q9BWK5 , which is a finding more typical of CDI . METHODS : The P30518 REA gene encoding P30518 REA was sequenced . The identified novel missense mutation was re-created by site-directed mutagenesis and expressed in HEK 293 cells . P30518 REA activity was assessed by the ability of transfected cells to produce DB02527 in response to stimulation with DB00035 SUB . Membrane localization of P30518 REA was assessed by fluorescence microscopy . RESULTS : Patient 1 had a deletion of P30518 REA ; patient 2 had the novel mutation L57R . In transiently transfected HEK 293 cells , DB00035 SUB induced detectable but severely impaired L57R P30518 REA activity compared to cells expressing wild-type P30518 REA . Fluorescence microscopy showed that myc-tagged wild-type P30518 REA localized to the cell membrane while L57R P30518 REA remained intracellular . A nonpeptide P30518 REA chaperone , SR121463 , partially rescued L57R P30518 REA function by allowing it to reach the cell membrane . CONCLUSIONS : L57R P30518 REA has impaired in vitro activity that can be partially improved by treatment with a P30518 REA chaperone . The posterior pituitary hyperintensity may be absent in infants with XNDI .

DB01017 MEN inhibits P09917 REA activation and brain inflammation after focal cerebral ischemia in rats . AIM : To determine whether the anti-inflammatory effect of minocycline on postischemic brain injury is mediated by the inhibition of P09917 REA ( 5 - P28300 REA ) expression and enzymatic activation in rats . METHODS : Focal cerebral ischemia was induced for 30 min with middle cerebral artery occlusion , followed by reperfusion . The ischemic injuries , endogenous IgG exudation , the accumulation of neutrophils and macrophage / microglia , and 5 - P28300 REA mRNA expression were determined 72 h after reperfusion . 5 - P28300 REA metabolites ( leukotriene B4 and cysteinyl leukotrienes ) were measured 3 h after reperfusion . RESULTS : DB01017 MEN ( 22.5 and 45 mg / kg , ip , for 3 d ) attenuated ischemic injuries , IgG exudation , and the accumulation of neutrophils and macrophage / microglia 72 h after reperfusion . It also inhibited 5 - P28300 REA expression 72 h after reperfusion and the production of leukotrienes 3 h after reperfusion . CONCLUSION : DB01017 MEN inhibited postischemic brain inflammation , which might be partly mediated by the inhibition of 5 - P28300 REA expression and enzymatic activation .

Urinary pheromones promote P29323 REA / Akt phosphorylation , regeneration and survival of vomeronasal ( P30518 REA ) neurons . The G protein-coupled pheromone receptor neurons ( V1R and P30518 REA ) of the vomeronasal organ ( VNO ) are continually replaced throughout the lifetime of the mouse . Moreover , active signalling of V2Rs via the transient receptor potential 2 ( TRPC 2 ) channel is necessary for regeneration of receptors , as the TRPC 2 null mutant mouse showed a 75 % reduction of V2Rs by the age of two months . Here we describe P30518 REA mediated signalling in a neuronal line established from vomeronasal stem cells taken from postnatal female mice . Cells were immunoreactive for Galpha ( o ) and P30518 REA , whereas V1R and Galpha ( i ) immunoreactivity could not be detected . Biological ligands ( dilute urine and its protein fractions ) were found to increase proliferation and survival of these neurons . Dilute mouse urine but not artificial urine also induced P29323 REA , Akt and CREB signalling in a dose dependent way . The volatile fraction of male mouse urine alone was without effect while the fraction containing peptides ( > 5 kDa ) also stimulated P29323 REA and Akt phosphorylation . The P29323 REA , Akt and CREB phosphorylation response was sensitive to pertussis toxin , confirming the involvement of P30518 REA linked Galpha ( o ) . Dilute mouse urine or its high molecular weight protein fraction increased survival and proliferation of these neurons . Hence , urinary pheromones , which signal important social information via mature neurons , also promote survival and proliferation of their regenerating precursors . These data show that regenerating V2Rs respond to urine and the urinary peptides by activation of the Ras - P29323 REA and P19957 REA - Akt pathways , which appear to be important for vomeronasal neural survival and proliferation .

Expression of cytosolic retinoid-binding protein genes in human skin biopsies and cultured keratinocytes and fibroblasts . Using reverse transcription coupled to polymerase chain reaction we have studied the mRNA expression of serum retinol-binding protein and cytosolic receptors for retinol and retinoic acid in skin biopsies , and in cultured epidermal keratinocytes and dermal fibroblasts . Transcripts for cellular retinol-binding protein ( P09455 REA ) I and cellular retinoic-acid-binding protein ( CRABP ) I were found in normal skin , keratinocytes , and fibroblasts . CRABP II transcripts were detected in skin and keratinocytes . A decreased mRNA expression of CRABP I and an increased mRNA expression of CRABP II were found in lesional psoriatic skin compared with uninvolved skin . mRNA transcripts for serum retinol-binding protein ( s - P02753 REA ) were detected in all tissues and cells . The biological importance of s - P02753 REA expression in keratinocytes and fibroblasts is not known , but hypothetically this protein may be involved in the intracellular shuttling of retinol and retinoic acid , or in the retransportation of cellular retinoids into the extracellular space .

Adrenomedullin - O60895 REA system is crucially involved in retinal angiogenesis . Adrenomedullin ( P35318 REA ) is an endogenous peptide first identified as a strong vasodilating molecule . We previously showed that in mice , homozygous knockout of P35318 REA ( P35318 REA ( - / - ) ) or its receptor regulating protein , O60895 REA ( O60895 REA ( - / - ) ) , is embryonically lethal due to abnormal vascular development , thereby demonstrating the importance of P35318 REA and its receptor signaling to vascular development . P35318 REA expression in the retina is strongly induced by ischemia ; however , its role in retinal pathophysiology remains unknown . Here , we analyzed oxygen-induced retinopathy ( OIR ) using heterozygous P35318 REA and O60895 REA knockout mice models ( P35318 REA ( + / - ) or O60895 REA ( + / - ) , respectively ) . In addition , we analyzed the role of the P35318 REA - O60895 REA system during earlier stages of retinal angiogenesis using an inducible endothelial cell-specific O60895 REA knockout mouse line ( DI-E - O60895 REA ( - / - ) ) . Finally , we assessed the ability of antibody-induced P35318 REA blockade to control pathological retinal angiogenesis in OIR . In OIR , neovascular tufts , avascular zones , and hypoxic areas were all smaller in P35318 REA ( + / - ) retinas compared with wild-type mice . P35318 REA ( + / - ) retinas also exhibited reduced levels of P15692 REA and P29474 REA expression . DI-E - O60895 REA ( - / - ) showed abnormal retinal vascular patterns in the early stages of development . However , P35318 REA enhanced the proliferation and migration of retinal endothelial cells . Finally , we found intravitreal injection of anti - P35318 REA antibody reduced pathological retinal angiogenesis . In conclusion , the P35318 REA - O60895 REA system is crucially involved in retinal angiogenesis . P35318 REA and its receptor system are potential therapeutic targets for controlling pathological retinal angiogenesis .

The Drosophila insulin receptor activates multiple signaling pathways but requires insulin receptor substrate proteins for DNA synthesis . The Drosophila insulin receptor ( P30518 REA ) contains a 368 - amino-acid COOH-terminal extension that contains several tyrosine phosphorylation sites in YXXM motifs . This extension is absent from the human insulin receptor but resembles a region in insulin receptor substrate ( P41252 REA ) proteins which binds to the phosphatidylinositol ( PI ) 3 - kinase and mediates mitogenesis . The function of a chimeric P30518 REA containing the human insulin receptor binding domain ( hDIR ) was investigated in 32D cells , which contain few insulin receptors and no P41252 REA proteins . P01308 REA stimulated tyrosine autophosphorylation of the human insulin receptor and hDIR , and both receptors mediated tyrosine phosphorylation of Shc and activated mitogen-activated protein kinase . P35568 REA was required by the human insulin receptor to activate PI 3 - kinase and p70s6k , whereas hDIR associated with PI 3 - kinase and activated p70s6k without P35568 REA . However , both receptors required P35568 REA to mediate insulin-stimulated mitogenesis . These data demonstrate that the P30518 REA possesses additional signaling capabilities compared with its mammalian counterpart but still requires P35568 REA for the complete insulin response in mammalian cells .

Hypoxia-induced neuroinflammatory white-matter injury reduced by minocycline in SHR / SP . Hypertensive small vessel disease is a major cause of vascular cognitive impairment ( VCI ) . Spontaneously hypertensive / stroke prone rats ( SHR / SP ) with unilateral carotid artery occlusion ( UCAO ) and a Japanese permissive diet ( JPD ) have white-matter ( WM ) damage similar to that seen in VCI . We hypothesized that WM injury was due to hypoxia-mediated , blood-brain barrier ( BBB ) disruption . Twelve-week-old SHR / SP had UCAO / JPD and were studied with immunohistochemistry , biochemistry , multimodal magnetic resonance imaging ( Q9BWK5 ) , and Morris water maze ( MWM ) testing . One week after UCAO / JPD , WM showed a significant increase in hypoxia inducible factor - 1α ( HIF - 1α ) , which increased further by 3 weeks . Prolyl hydroxylase - 2 ( Q9GZT9 ) expression decreased at 1 and 3 weeks . Infiltrating T cells and neutrophils appeared around endothelial cells from 1 to 3 weeks after UCAO / JPD , and matrix metalloproteinase - 9 ( P14780 REA ) colocalized with inflammatory cells . At 3 weeks , WM immunostained for IgG , indicating BBB leakage . DB01017 MEN ( 50 mg / kg intraperitoneally ) was given every other day from weeks 12 to 20 . Multimodal Q9BWK5 showed that treatment with minocycline significantly reduced lesion size and improved cerebral blood flow . DB01017 MEN improved performance in the MWM and prolonged survival . We propose that BBB disruption occurred secondary to hypoxia , which induced an P14780 REA - mediated infiltration of leukocytes . DB01017 MEN significantly reduced WM damage , improved behavior , and prolonged life .

The P08908 REA - receptor agonist flibanserin reduces drug-induced dyskinesia in O75916 REA - deficient mice . Drug-induced dyskinesia is a major complication of dopamine replacement therapy in advanced Parkinson ' s disease consisting of dystonia , chorea and athetosis . Agonists at P08908 REA - receptors attenuate levodopa-induced motor complications in non-human primates . Mice with increased dopamine D2 receptor ( P14416 REA ) signalling due to the lack of expression of the regulator of G-protein signalling 9 ( O75916 REA ) also develop dyskinesia following levodopa treatment . We investigated whether the P08908 REA - receptor agonist flibanserin compared with buspirone reduces motor abnormalities induced by levodopa or quinelorane , a selective dopamine D2 - receptor agonist . Following dopamine depletion via reserpine , 40 mice ( 20 wild-type and 20 O75916 REA knock-out ) were treated with flibanserin or buspirone in combination with levodopa or quinelorane . Motor behaviour was analysed using open field analysis . O75916 REA knock-out mice displayed significantly more drug-induced dystonia ( p < 0.04 ; t test ) than wild type . In quinelorane-treated wild-type mice flibanserin as well as buspirone significantly reduced dystonia ( p < 0.05 ) . In O75916 REA knock-out animals again both reduced quinelorane-induced dystonia . However , flibanserin was significantly more effective ( p = 0.003 ) . Following reserpine pretreatment and administration of levodopa wild-type and Q99697 REA 9 knock-out mice showed mild to moderate dystonia . Surprisingly , 10 mg / kg buspirone increased dystonia in both animal groups , whereas it was decreased by 10 mg / kg flibanserin . However , compared with levodopa alone only the increase of dystonia by buspirone was significant ( p < 0.04 ) . DB04908 MEN showed promising antidyskinetic effects in a model of drug-induced dyskinesia . Our data underline the possible benefit of P08908 REA agonists in drug-induced dyskinesia .

Pharmacodynamics and pharmacokinetics of omapatrilat in heart failure . The purpose of this study was to determine the pharmacodynamics and pharmacokinetics of omapatrilat , administered orally ( 25 mg ) or intravenously ( 10 mg ) in 19 New York Heart Association class II and class III congestive heart failure ( CHF ) patients versus 17 healthy controls matched for age , race , gender , and weight . The plasma concentrations of atrial natriuretic peptide ( P01160 REA ) increased by approximately 20 % and 30 % in CHF and control subjects , respectively , at 4 hours after intravenous or oral omapatrilat administration . Similar elevation in the cyclic guanosine monophosphate concentration ( 25 % to 35 % ) and P01160 REA urinary excretion ( 21 ng / 24 h to 22 ng / 24 h ) was seen in all treatment groups after omapatrilat administration . P12821 REA activity was > 90 % inhibited at 4 hours after dosing and remained approximately 60 % to 70 % inhibited at 24 hours after dosing . The levels of endothelin - 1 and endothelin - 2 remained unchanged after oral or intravenous administration of omapatrilat . The maximal reduction in seated blood pressure compared with baseline was similarfor CHF and control subjects . Clinical pharmacokinetic parameters were similar in both groups after intravenous dosing , but maximum concentration and area under the concentration-time curve were elevated in CHF patients compared with controls after oral dosing . DB00886 MEN was well tolerated ; differences in systemic exposure and metabolism between CHF patients and controls did not appear to be clinically significant .