MH_dev_238

The efficacy of activated protein C in murine endotoxemia is dependent on integrin CD11b . DB00055 MEN ( P25054 REA ) , the only FDA-approved biotherapeutic drug for sepsis , possesses anticoagulant , antiinflammatory , and barrier-protective activities . However , the mechanisms underlying its anti-inflammatory functions are not well defined . Here , we report that the antiinflammatory activity of P25054 REA on macrophages is dependent on integrin CD11b / P05107 REA , but not on endothelial protein C receptor ( Q9UNN8 ) . We showed that CD11b / P05107 REA bound P25054 REA within specialized membrane microdomains / lipid rafts and facilitated P25054 REA cleavage and activation of protease-activated receptor - 1 ( PAR 1 ) , leading to enhanced production of sphingosine - 1 - phosphate ( Q14703 REA ) and suppression of the proinflammatory response of activated macrophages . Deletion of the gamma-carboxyglutamic acid domain of P25054 REA , a region critical for its anticoagulant activity and Q9UNN8 - dependent barrier protection , had no effect on its antiinflammatory function . Genetic inactivation of CD11b , PAR 1 , or sphingosine kinase - 1 , but not Q9UNN8 , abolished the ability of P25054 REA to suppress the macrophage inflammatory response in vitro . Using an LPS-induced mouse model of lethal endotoxemia , we showed that P25054 REA administration reduced the mortality of wild-type mice , but not CD11b - deficient mice . These data establish what we believe to be a novel mechanism underlying the antiinflammatory activity of P25054 REA in the setting of endotoxemia and provide clear evidence that the antiinflammatory function of P25054 REA is distinct from its barrier-protective function and anticoagulant activities .

Involvement of sphingosine kinase in P01375 REA - stimulated tetrahydrobiopterin biosynthesis in P13671 REA glioma cells . In P13671 REA glioma cells , the sphingolipid second messenger ceramide potentiates expression of inducible nitric-oxide synthase ( P35228 REA ) induced by tumor necrosis factor alpha ( P01375 REA ) without affecting P30793 REA ( GTPCH ) , the rate-limiting enzyme in the biosynthesis of 6 ( R ) - DB00360 ( BH ( 4 ) ) , a cofactor required for P35228 REA activity . P01375 REA also stimulates sphingosine kinase , the enzyme that phosphorylates sphingosine to form sphingosine - 1 - phosphate ( Q8TCT9 ) , a further metabolite of ceramide . Several clones of P13671 REA cells , expressing widely varying levels of sphingosine kinase , were used to examine the role of Q8TCT9 in regulation of GTPCH and BH ( 4 ) biosynthesis . Overexpression of sphingosine kinase , with concomitant increased endogenous Q8TCT9 levels , potentiated the effect of P01375 REA on GTPCH expression and activity and BH ( 4 ) biosynthesis . In contrast , enforced expression of sphingosine kinase had no effect on P35228 REA expression or NO formation . Furthermore , N , N-dimethylsphingosine , a potent sphingosine kinase inhibitor , completely eliminated the increased GTPCH activity and expression induced by P01375 REA . Surprisingly , we found that , although P13671 REA cells can secrete Q8TCT9 , which is enhanced by P01375 REA , treatment of P13671 REA cells with exogenous Q8TCT9 or dihydro - Q8TCT9 had no affect on BH ( 4 ) biosynthesis . However , both Q8TCT9 and dihydro - Q8TCT9 markedly stimulated P29323 REA 1/2 in P13671 REA cells , which express cell surface Q8TCT9 receptors . Interestingly , although this P29323 REA activation was blocked by PD98059 , which also reduced cellular proliferation induced by enforced expression of sphingosine kinase , PD98059 had no effect on GTPCH activity . Collectively , these results suggest that only intracellularly generated Q8TCT9 plays a role in regulation of GTPCH and BH ( 4 ) levels .

Somatostatin preserved blood brain barrier against cytokine induced alterations : possible role in multiple sclerosis . Multiple sclerosis ( MS ) is an inflammatory neurological disorder associated with demyelination , impaired blood brain barrier ( BBB ) , axonal damage and neuronal loss . In the present study , we measured somatostatin ( P61278 REA ) and tumor necrosis factor-α ( P01375 REA - α ) like immunoreactivity in P04141 REA samples from MS and non-MS patients . We also examined the role of P61278 REA in cytokines and lipopolysaccharide ( LPS ) - induced damage to the BBB using human brain endothelial cells in culture . Most of the cerebrospinal fluid ( P04141 REA ) samples studied from definite MS patients exhibited lower somatostatin ( P61278 REA ) - like immunoreactivity and higher expression of P01375 REA - α in comparison to non-MS patients . Treatment of cells with cytokines and LPS blocked P61278 REA secretion and decreased P61278 REA expression . Human brain endothelial cells expressed all five somatostatin receptors ( SSTRs ) with increased expression of P30874 REA and 4 upon treatment with cytokines and LPS . Cytokines and LPS-induced disruption of the tight junction proteins Zonula occludens ( ZO - 1 ) organization was restored in presence of P61278 REA , P30874 REA or P31391 REA selective agonists . Furthermore , inflammation induced changes in extracellular signal-regulated kinases ( P27361 REA / 2 and Q13164 REA ) signaling and altered expression of endothelial and inducible nitric oxide synthase are modulated in presence of P61278 REA . These data indicate that decreased levels of P61278 REA contribute to failure of the BBB in MS .

Glutamate modulators as potential therapeutic drugs in schizophrenia and affective disorders . Severe psychiatric disorders such as schizophrenia are related to cognitive and negative symptoms , which often are resistant to current treatment approaches . The glutamatergic system has been implicated in the pathophysiology of schizophrenia and affective disorders . A key component is the dysfunction of the glutamatergic N-methyl-D-aspartate ( DB01221 ) receptor . Substances regulating activation / inhibition of the DB01221 receptor have been investigated in schizophrenia and major depression and are promising in therapeutic approaches of negative symptoms , cognition , and mood . In schizophrenia , add-on treatments with glycine , D-serine , D-alanine , D-cycloserine , D-amino acid oxidase inhibitors , glycine transporter - 1 ( P48067 REA ) inhibitors ( e . g . , sarcosine , bitopertin ) and agonists ( e . g . , DB05096 MEN ) or positive allosteric modulator ( e . g . , ADX 71149 ) of group II metabotropic glutamate receptors ( mGluRs ) have been studied . In major depression , the DB01221 receptor antagonists ( e . g . , ketamine , AZD 6765 ) , Q13224 REA subtype antagonists ( e . g . , traxoprodil , MK - 0657 ) , and partial agonists ( e . g . , D-cycloserine , GLYX - 13 ) at the glycine site of the DB01221 receptor have been proven to be effective in animal studies and first clinical trials . In addition , clinical studies of Q14416 REA / 3 antagonist BCI - 838 ( a prodrug of BCI - 632 ( MGS 0039 ) ) , Q14416 REA / 3 - negative allosteric modulators ( NMAs ) ( e . g . , RO499819 , RO4432717 ) , and P41594 REA NAMs ( e . g . , AZD 2066 , RO4917523 ) are in progress . Future investigations should include effects on brain structure and activation to elucidate neural mechanisms underlying efficacy of these drugs .

Activation of spinal alpha - 7 nicotinic acetylcholine receptor attenuates remifentanil-induced postoperative hyperalgesia . The activation of alpha - 7 nicotinic acetylcholine receptors ( α7 - nAchRs ) are currently being considered as novel therapeutic approaches for managing hyperalgesia in inflammation and chronic neuropathic pain , but the role of a7 - nAChRs on opioids induced hyperalgesia remain unknown . The present study investigated the effects of α7 - nAChRs selective agonists PHA - 543613 and type II positive allosteric modulators ( PAMs ) PNU - 120596 in remifentanil induced postoperative hyperalgesia . As the results shown , intrathecal treatment with both α7 - nAChRs agonists and type II PAMs could attenuate remifentanil induced hyperalgesia by increasing paw withdrawal mechanical threshold ( PWMT ) and paw withdrawal thermal latency ( PWTL ) . Furthermore , we also investigated the protein level of proinflammatory cytokines and phosphorylation N-methyl-d-aspartate receptor 2B subunit ( p - Q13224 REA ) in the spinal cord . Our data indicated that activation of α7 - nAchRs decreased the proinflammatory cytokines ( P01375 REA - α , P05231 REA ) and p - Q13224 REA protein level in the spinal cord . The depression of the increased levels of proinflammatory cytokines and p - Q13224 REA after remifentanil treatment may contribute to the anti-hyperalgesia effects of PHA - 543613and PNU - 120596 via α7 - nAChRs . Therefore , our findings demonstrated that α7 - nAChRs may be potential candidates for treating opioids induced hyperalgesia .

Novel synthesis of various orthogonally protected Cα-methyllysine analogues and biological evaluation of a vapreotide analogue containing ( S ) - α-methyllysine . Prochiral malonic diesters containing a quaternary carbon center have been successfully transformed into a diverse set of ( t ) Boc-Fmoc-α ( 2,2 ) - methyllysine-OH analogues through chiral malonic half-ester intermediates obtained via enzymatic ( Pig Liver Esterase , PLE ) hydrolysis . The variety of chiral half-ester intermediates , which vary from 1 to 6 methylene units in the side chain , are achieved in moderate to high optical purity and in good yields . The PLE hydrolysis of malonic diesters with various side chain lengths appears to obey the Jones ' s PLE model according to the stereochemical configurations of the resulting chiral half-esters . The established synthetic strategy allows the construction of both enantiomers of α ( 2,2 ) - methyllysine analogues , and a ( S ) - β ( 2,2 ) - methyllysine analogue from a common synthon by straightforward manipulation of protecting groups . Two different straightforward and cost effective synthetic strategies are described for the synthesis of α ( 2,2 ) - methyllysine analogues . The described strategies should find significant usefulness in preparing novel peptide libraries with unnatural lysine analogues . A DB04894 MEN analogue incorporating ( S ) - α ( 2,2 ) - methyllysine was prepared . However , the DB04894 MEN analogue with ( S ) - α-methyl-α-lysine is found to lose its specific binding to somatostatin receptor subtype 2 ( P30874 REA ) .

[ Measurement of rifampicin and clarithromycin in serum by high-performance liquid chromatography with electrochemical detection ] . DB01045 ( RFP ) induces hepatic drug-metabolizing enzymes , making drug interactions a very important clinical problem . DB01211 MENMAX DB01211 MEN ( P62158 ) metabolism is reportedly enhanced by induction of hepatic drug-metabolizing enzymes ( P08684 REA ) by RFP , so that the blood lend of P62158 decreases when RFP is administered concurrently . We connected an electrochemical detector to a high-performance liquid chromatograph ( HPLC ) for simple , rapid , easy measurement of blood concentrations of RFP and P62158 . Using samples of patient serum , normal serum , and reference standards , we compared HPLC by an external laboratory and the results of LC / MS / MS analysis with those of this new assay . A strong correlation was seen between our HPLC results and those of the external laboratory in RFP levels ( r = 0.975 , p < 0.01 ) . A strong correlation was also seen between results we obtained for P62158 with the electrochemical detector in this assay and values measured by LC / MS / MS analysis ( r = 0.995 , p < 0.01 ) . Our method enabled simple , rapid measurement of RFP and P62158 by connecting the HPLC and electrochemical detector in tandem . This system was used to modulate dosage during combined therapy with RFP and P62158 . The therapeutic effect for nontuberculous mycobacteriosis is expected to improve , and our HPLC is expected to be useful for simple , rapid , easy measurement of blood concentrations .

Differential interleukin - 10 ( P22301 REA ) and IL - 23 production by human blood monocytes and dendritic cells in response to commensal enteric bacteria . Human peripheral blood contains antigen-presenting cells ( P25054 REA ) , including dendritic cells ( DC ) and monocytes , that may encounter microbes that have translocated from the intestine to the periphery in disease states like HIV - 1 infection and inflammatory bowel disease . We investigated the response of DC and monocytes in peripheral blood mononuclear cells ( PBMC ) to a panel of representative commensal enteric bacteria , including Escherichia coli , Enterococcus sp . , and Bacteroides fragilis . All three bacteria induced significant upregulation of the maturation and activation markers P25942 REA and Q01151 on myeloid dendritic cells ( mDC ) and plasmacytoid dendritic cells ( pDC ) . However , only mDC produced cytokines , including interleukin - 10 ( P22301 REA ) , IL - 12p40 / 70 , and tumor necrosis factor alpha ( P01375 REA - α ) , in response to bacterial stimulation . Cytokine profiles in whole PBMC differed depending on the stimulating bacterial species : B . fragilis induced production of IL - 23 , IL - 12p70 , and P22301 REA , whereas E . coli and Enterococcus induced an P22301 REA - predominant response . mDC and monocyte depletion experiments indicated that these cell types differentially produced P22301 REA and IL - 23 in response to E . coli and B . fragilis . Bacteroides thetaiotaomicron did not induce levels of IL - 23 similar to those of B . fragilis , suggesting that B . fragilis may have unique proinflammatory properties among Bacteroides species . The addition of recombinant human P22301 REA to PBMC cultures stimulated with commensal bacteria abrogated the IL - 23 response , whereas blocking P22301 REA significantly enhanced IL - 23 production , suggesting that P22301 REA controls the levels of IL - 23 produced . These results indicate that blood mDC and monocytes respond differentially to innate stimulation with whole commensal bacteria and that P22301 REA may play a role in controlling the proinflammatory response to translocated microbes .

P30047 REA - dependent and - independent inhibitors of P30793 REA . P30047 REA ( P30047 REA ) mediates the feedback inhibition of P30793 REA activity by ( 6R ) - L-erythro - DB00360 ( BH4 ) through protein complex formation . Since guanine and BH4 have a common pyrimidine ring structure , we examined the inhibitory effect of guanine and its analogs on the enzyme activity . DB02377 MEN , 8 - hydroxyguanine , 8 - methylguanine , and 8 - bromoguanine inhibited the enzyme activity in a P30047 REA - dependent and pH-dependent manner and induced complex formation between P30793 REA and P30047 REA . The type of inhibition by this group is a mixed type . All these properties were shared with BH4 . In striking contrast , inhibition by DB01667 and 8 - mercaptoguanine was P30047 REA - independent and pH-independent . The type of inhibition by DB01667 and 8 - mercaptoguanine was a competitive type . The two compounds did not induce complex formation between the enzyme and P30047 REA . These results demonstrate that guanine compounds of the first group bind to the BH4 - binding site of the P30793 REA / P30047 REA complex , whereas DB01667 and 8 - mercaptoguanine bind to the active site of the enzyme . Finally , the possible implications in Lesch-Nyhan syndrome and Parkinson diseases of the inhibition of P30793 REA by guanine and 8 - hydroxyguanine are discussed .

Expression of human all-trans-retinoic acid receptor beta and its ligand-binding domain in Escherichia coli . DB00755 , one of the hormonally active derivatives of vitamin A , occurs physiologically in plasma at a concentration below 10 nmol / l . The methods currently used for its quantification are based on HPLC , need about 1 ml of serum , are relatively laborious and thus not well suited for mass analysis . The affinity and specificity of retinoic acid receptors for all-trans-retinoic acid encouraged us to express both the entire human retinoic acid receptor beta ( P10826 REA ) and two versions of its retinoic acid-binding domain in Escherichia coli in the hope that these recombinant proteins might be used as binders in a ligand-binding assay for all-trans-retinoic acid . The recombinant receptors , the whole receptor [ P10826 REA - ( Q93033 REA - Q448 ) ] , corresponding to domains A-F , and the ligand-binding domain [ P10826 REA - ( E149 - Q448 ) ] , corresponding to domains D-F , were expressed in the vector pET 3d / BL21 ( DE3 ) as inclusion bodies , solubilized with guanidinium chloride , renatured and purified by ion-exchange chromatography . P10826 REA - ( P193 - Q448 ) , corresponding to domains E-F , was expressed in the vector pET 3d / BL21 ( DE3 ) pLysS , and purified by reversed-phase chromatography . Under non-denaturing conditions , the expressed whole receptor [ P10826 REA - ( Q93033 REA - Q448 ) ] and the D-F construct ( P10826 REA - ( E149 - Q448 ) ] behaved chromatographically as monomeric proteins whereas the E-F construct [ P10826 REA - ( P193 - Q448 ) ] had a strong tendency to aggregate . P10826 REA - ( Q93033 REA - Q448 ) and P10826 REA - ( E149 - Q448 ) had similar Kd values for all-trans-retinoic acid ( 1.4 and 0.6 nmol / l respectively ) whereas P10826 REA - ( P193 - Q448 ) bound all-trans-retinoic acid less avidly ( Kd 9.6 nmol / l ) . DB00523 MEN bound to P10826 REA - ( E149 - Q448 ) and P10826 REA - ( Q93033 REA - Q448 ) as avidly as all-trans-retinoic acid . Competition experiments showed weak or no binding of 4 - oxo-all-trans-retinoic acid , 4 - oxo - 13 - cis-retinoic acid , 13 - cis-retinoic acid , acitretin and retinol by P10826 REA - ( E149 - Q448 ) .

Molecular systematics of armadillos ( Xenarthra , Dasypodidae ) : contribution of maximum likelihood and Bayesian analyses of mitochondrial and nuclear genes . The 30 living species of armadillos , anteaters , and sloths ( Mammalia : Xenarthra ) represent one of the three major clades of placentals . Armadillos ( Cingulata : Dasypodidae ) are the earliest and most speciose xenarthran lineage with 21 described species . The question of their tricky phylogeny was here studied by adding two mitochondrial genes ( P03886 REA [ P03886 REA ] and 12S ribosomal RNA [ 12S rRNA ] ) to the three protein-coding nuclear genes ( alpha 2B adrenergic receptor [ P18089 REA ] , breast cancer susceptibility exon 11 [ P38398 REA ] , and P04275 REA exon 28 [ P04275 REA ] ) yielding a total of 6869 aligned nucleotide sites for thirteen xenarthran species . The two mitochondrial genes were characterized by marked excesses of transitions over transversions-with a strong bias toward CT transitions for the 12S rRNA-and exhibited two - to fivefold faster evolutionary rates than the fastest nuclear gene ( P18089 REA ) . Maximum likelihood and Bayesian phylogenetic analyses supported the monophyly of Dasypodinae , Tolypeutinae , and Euphractinae , with the latter two armadillo subfamilies strongly clustering together . Conflicting branching points between individual genes involved relationships within the subfamilies Tolypeutinae and Euphractinae . Owing to a greater number of informative sites , the overall concatenation favored the mitochondrial topology with the classical grouping of Cabassous and Priodontes within Tolypeutinae , and a close relationship between Euphractus and Chaetophractus within Euphractinae . However , low statistical support values associated with almost equal distributions of apomorphies among alternatives suggested that two parallel events of rapid speciation occurred within these two armadillo subfamilies .

The Retinoic Acid Receptor-alpha mediates human T-cell activation and Th2 cytokine and chemokine production . BACKGROUND : We have recently demonstrated that all-trans-retinoic acid ( DB00755 ) and 9 - cis-retinoic acid ( 9 - cis RA ) promote P05112 REA , P05113 REA and P35225 REA synthesis , while decreasing P01579 REA and P01375 REA expression by activated human T cells and reduces the synthesis of IL - 12p70 from accessory cells . Here , we have demonstrated that the observed effects using DB00755 and 9 - cis RA are shared with the clinically useful RAR ligand , 13 - cis retinoic acid ( 13 - cis RA ) , and the retinoic acid receptor-alpha ( P10276 REA ) - selective agonist , AM580 but not with the P10826 REA / gamma ligand , 4 - hydroxyphenylretinamide ( DB05076 ) . RESULTS : The increase in type 2 cytokine production by these retinoids correlated with the expression of the T cell activation markers , Q07108 and P28907 REA . The P10276 REA - selective agonist , AM580 recapitulated all of the T cell activation and type 2 cytokine-inducing effects of DB00755 and 9 - DB00982 , while the P10276 REA - selective antagonist , RO 41-5253 , inhibited these effects . CONCLUSION : These results strongly support a role for P10276 REA engagement in the regulation of genes and proteins involved with human T cell activation and type 2 cytokine production .

P18089 REA genotype differentially modulates stress-induced neural activity in the amygdala and hippocampus during emotional memory retrieval . RATIONALE : DB00368 MEN interacts with stress hormones in the amygdala and hippocampus to enhance emotional memory consolidation , but the noradrenergic-glucocorticoid interaction at retrieval , where stress impairs memory , is less understood . OBJECTIVES : We used a genetic neuroimaging approach to investigate whether a genetic variation of the noradrenergic system impacts stress-induced neural activity in amygdala and hippocampus during recognition of emotional memory . METHODS : This study is based on genotype-dependent reanalysis of data from our previous publication ( Li et al . Brain Imaging Behav 2014 ) . Twenty-two healthy male volunteers were genotyped for the P18089 REA gene encoding the α2B - adrenergic receptor . Ten deletion carriers and 12 noncarriers performed an emotional face recognition task , while their brain activity was measured with fMRI . During encoding , 50 fearful and 50 neutral faces were presented . One hour later , they underwent either an acute stress ( Trier Social Stress Test ) or a control procedure which was followed immediately by the retrieval session , where participants had to discriminate between 100 old and 50 new faces . RESULTS : A genotype-dependent modulation of neural activity at retrieval was found in the bilateral amygdala and right hippocampus . Deletion carriers showed decreased neural activity in the amygdala when recognizing emotional faces in control condition and increased amygdala activity under stress . Noncarriers showed no differences in emotional modulated amygdala activation under stress or control . Instead , stress-induced increases during recognition of emotional faces were present in the right hippocampus . CONCLUSION : The genotype-dependent effects of acute stress on neural activity in amygdala and hippocampus provide evidence for noradrenergic-glucocorticoid interaction in emotional memory retrieval .

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

The role of selectins and P05107 REA in leukotriene B4 - mediated white blood cell emigration in human skin grafts transplanted on SCID mice . The purpose of this study was to examine the role of selectins and P05107 REA cell adhesion molecules ( CAMs ) in inflammation induced by injection of leukotriene B4 ( LTB 4 ) into human skin . To accomplish this , the expression of CAMs and the ability of specific antibodies against CAMs to block white blood cell ( WBC ) transmigration were studied in an in vivo model consisting of human skin transplanted onto mice with the severe combined immune deficiency ( SCID ) mutation . The results indicate that LTB 4 - induced WBC transmigration in the human / SCID model is rapid and pronounced ; however , it is not accompanied by a significant upregulation of the baseline expression of endothelial P16109 REA , P16581 REA , P05362 REA or P19320 REA . An anti-murine P05107 REA mAb markedly inhibited white cell infiltration ( 89 % inhibition ) confirming the importance of beta 2 integrins in the process . The role of selectins was also examined . P61006 REA - 14 , a bioactive antibody against murine P14151 REA inhibited transmigration by 66 % . A significant , but smaller , effect ( 39 % inhibition ) was observed by blocking P16581 REA function . These results indicate that LTB 4 - induced inflammation does not require upregulation of endothelial P62158 expression and , in contrast to P01375 REA alpha-induced transmigration , is only partially blocked by anti - P16581 REA antibodies .

P05231 REA , P01579 REA and P01375 REA production by liver-associated T cells and acute liver injury in rats administered concanavalin A . The relationship between the development of acute hepatitis and the production of P01375 REA P01579 REA and P05231 REA by liver-associated T lymphocytes following intravenous injection of concanavalin A ( Con A ) was studied in rats . Following a single injection of Con A , there was a dose and time-dependent correlation in the serum levels of serum alanine aminotransferase ( ALT ) , P05231 REA , P01579 REA and P01375 REA . These increases correlated with an increase in the numbers of P01730 REA + , CD8 + and CD25 + T cells in blood and P01730 REA + and CD25 + T cells in the liver perfusate , but not with CD8 + T cells in liver perfusate . Increased levels of P05231 REA , P01579 REA and P01375 REA were constitutively produced by liver-associated P01730 REA + T cells when cultured . In Con A-stimulated cultures , liver-associated P01730 REA + T cells secreted increasing levels of P01375 REA in a time-dependent manner following Con A injection , but P01375 REA production by peripheral blood lymphocytes was transient with peak levels detected at 1 h which then declined over 24 h . Histological examination of the liver revealed fatty change , hepatocyte degeneration and necrosis , with an associated cell infiltrate of neutrophils and P01730 REA + T cells both in the portal areas and around the central veins . These results support the hypothesis that Con A-induced liver damage is mediated by P01730 REA + T cells acting within the liver , at least in part through the secretion of P01375 REA , P01579 REA and P05231 REA .

Changes in circulating lymphocyte subpopulations following administration of the leucocyte function-associated antigen - 3 ( LFA - 3 ) / IgG 1 fusion protein alefacept . DB00092 MEN , a recombinant leucocyte function-associated antigen - 3 ( LFA - 3 ) / IgG 1 fusion protein approved for the treatment of psoriasis , is reported to reduce selectively the numbers of circulating P01730 REA ( + ) CD45RO ( + ) and CD8 ( + ) CD45RO ( + ) T cells , while sparing the naive cells . The purpose of the present study was to elucidate further the effect of alefacept on various circulating lymphocyte subsets . Sixteen patients , 12 with chronic plaque psoriasis and four with pustular psoriasis , received alefacept 7.5 mg once weekly for 12 weeks . Blood samples collected at study entry and after 12 weeks of treatment were analysed by four-colour flow cytometry . There were statistically significant reductions in the total number of conventional memory ( CD45RA ( - ) P26842 REA ( + ) ) and effector ( CD45RA ( - ) P26842 REA ( - ) or CD45RA ( + ) P26842 REA ( - ) ) T cells , including P01730 REA ( + ) and CD8 ( + ) T cells expressing CD161 and CD8 ( + ) T cells expressing cutaneous lymphocyte-associated antigen ( DB01211 MEN ) . Natural killer ( NK ) T cells were also reduced significantly , while no statistically significant changes were seen in NK cells and P01730 REA ( + ) CD25 ( high ) cells . The affected subpopulations were all characterized by a high expression of P06729 REA . However , P01730 REA ( + ) CD25 ( low ) , and P01730 REA ( + ) DB01211 MEN ( + ) cells , which also expressed relative high levels of P06729 REA , were not reduced significantly . Our results suggest a heterogeneous effect of alefacept on the circulating memory T cell population , indicating that high expression of P06729 REA may not , by itself , be sufficient to explain the reduction in cell count for a specific subpopulation .

Genetic dissection of atypical antipsychotic-induced weight gain : novel preliminary data on the pharmacogenetic puzzle . Atypical antipsychotics such as clozapine represent a significant improvement over typical antipsychotics in the treatment of schizophrenia , particularly regarding extrapyramidal symptoms . Despite their benefits , use is limited by the occurrence of adverse reactions such as sedation and weight gain . This article provides a comprehensive review and discussion of obesity-related pathways and integrates these with the known mechanisms of atypical antipsychotic action to identify candidate molecules that may be disrupted during antipsychotic treatment . Novel preliminary data are presented to genetically dissect these obesity pathways and elucidate the genetic contribution of these candidate molecules to clozapine-induced weight gain . There is considerable variability among individuals with respect to the ability of clozapine to induce weight gain . Genetic predisposition to clozapine-induced weight gain has been suggested . Therefore , genetic variation in these candidate molecules may predict patient susceptibility to clozapine-induced weight gain . This hypothesis was tested for 10 genetic polymorphisms across 9 candidate genes , including the serotonin 2C , 2A , and 1A receptor genes ( P28335 REA / 2A / 1A ) ; the histamine H1 and H2 receptor genes ( P35367 REA / P25021 REA ) ; the cytochrome P450 1A2 gene ( P05177 REA ) ; the beta 3 and alpha , alpha-adrenergic receptor genes ( P13945 REA / ADRAIA ) ; and tumor necrosis factor alpha ( P01375 REA ) . Prospective weight gain data were obtained for 80 patients with schizophrenia who completed a structured clozapine trial . Trends were observed for P13945 REA , ADRA 1A , P01375 REA , and P28335 REA ; however , replication in larger , independent samples is required . Although in its infancy , psychiatric pharmacogenetics will in the future aid clinical practice in the prediction of response and side effects , such as antipsychotic-induced weight gain , and minimize the current " trial and error " approach to prescribing .

P25021 REA mediated relaxation of buffalo ( Bubalus bubalus ) ureter . On the buffalo ureter , histamine did not elicit any direct effect . However , it caused concentration-dependent relaxation of the tissues precontracted by carbachol , phenylephrine , norepinephrine , KCI or BaCl 2 and also inhibited the contractile effect of carbachol . DB08805 MEN selectively antagonised the relaxation and inhibition of contractile response but mepyramine did not show this effect . Isoprenaline , dobutamine , salbutamol , verapamil and papaverine neither produced any direct effect nor relaxed the carbachol-contracted tissues ; norepinephrine and epinephrine had contractile effects . Hence , the histamine-induced relaxation was mediated through the activation of H2 receptors and not through adrenergic mechanisms or blockade of Ca ( 2 + ) - channels or inhibition of cyclic nucleotide phosphodiesterase .

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

The P04035 REA inhibitors , simvastatin , lovastatin and mevastatin inhibit proliferation and invasion of melanoma cells . BACKGROUND : A number of recent studies have suggested that cancer incidence rates may be lower in patients receiving statin treatment for hypercholesterolemia . We examined the effects of statin drugs on in vitro proliferation , migration and invasion of melanoma cells . METHODS : The ability of lovastatin , mevastatin and simvastatin to inhibit the melanoma cell proliferation was examined using cytotoxicity and apoptosis assays . Effects on cell migration and invasion were assessed using transwell invasion and migration chambers . Hypothesis testing was performed using 1 - way Q9UNW9 , and Student ' s t-test . RESULTS : DB00227 SUB , mevastatin and simvastatin inhibited the growth , cell migration and invasion of HT144 , M14 and SK - P61006 REA - 28 melanoma cells . The concentrations required to inhibit proliferation of melanoma cells ( 0.8- 2.1 microave previously been achieved in a phase I clinical trial of lovastatin in patients with solid tumours , ( 45 mg / kg / day resulted in peak plasma concentrations of approximately 3.9 micro CONCLUSION : Our results suggest that statin treatment is unlikely to prevent melanoma development at standard doses . However , higher doses of statins may have a role to play in adjuvant therapy by inhibiting growth and invasion of melanoma cells .