MH_dev_125

DB01296 MEN sulfate suppresses the expressions of urokinase plasminogen activator and inhibitor and gelatinases during the early stage of osteoarthritis . BACKGROUND : DB01296 MEN sulfate may have an ex vivo inhibitory effect on the plasminogen activator ( PA ) / plasmin system and gelatinases expression during the early development of osteoarthritis ( OA ) . METHODS : We compared the levels of urokinase-type PA ( u-PA ) , PA inhibitor - 1 ( P05121 REA ) and gelatinases ( matrix metalloproteinase - 2 and - 9 [ P08253 REA and - 9 ] ) in a series of chondral , meniscal , and synovial cultures of early OA after treatment with or without glucosamine sulfate . RESULTS : Gelatin zymography revealed that glucosamine sulfate could suppress P08253 REA secretion in chondral , meniscal and synovial cultures and also decrease P14780 REA production in synovial and meniscal cultures . ELISA data also showed the suppressive effects of glucosamine sulfate on u-PA and P05121 REA production in synovial cultures at 48 h . CONCLUSIONS : Our data suggest that one of the therapeutic effects of glucosamine sulfate is to down-regulate the expressions of u-PA , P05121 REA , P08253 REA and P14780 REA that underlie the destruction of articular cartilage in the early stage of OA , and therefore to delay the joint failure .

DB01017 SUB protects against permanent cerebral ischemia in wild type but not in matrix metalloprotease - 9 - deficient mice . DB01017 SUB is protective in models of transient middle cerebral artery occlusion ( MCAO ) . We studied whether minocycline and doxycycline , another tetracycline derivative , provide protection in permanent MCAO . Because minocycline inhibits matrix metalloprotease - 9 ( P14780 REA ) , we also compared minocycline ' s protective effect in wild type ( wt ) and P14780 REA knock-out ( ko ) mice . Wt FVB / N , Balb / C , and two lines of P14780 REA ko and their wt C57Bl / 6 control mice were subjected to 24 - or 72 - hour permanent MCAO . Drug administration was started either 12 hours before or 2 hours after the onset of MCAO . Infarct size was determined by triphenyltetrazolium staining or P24752 REA - weighted Q9BWK5 . Zymography was used to study the expression of MMPs . In wt strains , tetracycline treatments started before MCAO reduced the infarct size by 25 % to 50 % , whereas the treatment started after MCAO was not protective . DB01017 SUB inhibited ischemia-provoked pro - P14780 REA induction in wt mice , but was not protective in P14780 REA ko mice . Pro - P08253 REA was induced by MCAO in wt and P14780 REA ko mice . MCAO-induced pro - P08253 REA was downregulated by minocycline treatment in wt mice but remained in P14780 REA ko mice at the same level as in saline-treated wt mice . Tetracyclines are protective in permanent MCAO when the treatment is started before the insult . DB01017 SUB may provide protection by interfering with MMPs .

DB01017 SUB effects on cerebral edema : relations with inflammatory and oxidative stress markers following traumatic brain injury in mice . One of the severe complications following traumatic brain injury ( TBI ) is cerebral edema and its effective treatment is of great interest to prevent further brain damage . This study investigated the effects of minocycline , known for its anti-inflammatory properties , on cerebral edema and its respective inflammatory markers by comparing different dose regimens , on oxidative stress and on neurological dysfunction following TBI . The weight drop model was used to induce TBI in mice . The brain water content was measured to evaluate cerebral edema . Inflammatory markers were detected by ELISA ( IL - 1beta ) , zymography and Western blot ( P14780 REA ) . The oxidative stress marker ( glutathione levels ) and neurological function were measured by Griffith technique and string test , respectively . DB01017 SUB was administered i . p . once ( 5 min ) , twice ( 5 min and 3 h ) or triple ( 5 min , 3 h and 9 h ) following TBI . The first dose of minocycline only varied ( 45 or 90 mg / kg ) , whereas the following doses were all at 45 mg / kg . The single and double administrations of minocycline reduced the increase of inflammatory markers at 6 h post-TBI . DB01017 SUB also reduced cerebral edema at this time point , only after double administration and at the high dose regimen , although with no effect on the TBI-induced oxidized glutathione increase . The anti-edematous effect of minocycline persisted up to 24 h , upon a triple administration , and accompanied by a neurological recovery . In conclusion , we reported an anti-edematous effect of minocycline after TBI in mice according to a specific treatment regimen . These findings emphasize that the beneficial effects of minocycline depend on the treatment regimen following a brain injury .

Evidence of in vitro differential secretion of 72 and 92 kDa type IV collagenases after selective exposure to lipopolysaccharide in human fetal membranes . Premature rupture of chorioamniotic membranes complicated with intrauterine infection has been associated to degradation of extracellular matrix ( Q13201 REA ) , which could explain local morphological changes . We used a culture system in which the chorioamniotic membranes form two independent chambers , allowing for the selective stimulation of either the amnion ( Q9BXJ7 ) and / or the choriodecidua ( Q8NE62 ) regions . Lipopolysaccharide ( 500 ng / ml ) was added to the Q9BXJ7 and / or the Q8NE62 ; secretions and gelatinolytic activity of matrix metalloproteinase ( MMP ) - 2 and P14780 REA were measured in both compartments by enzyme-linked immunosorbent assay ( ELISA ) and zymography . Secretions of P01033 REA , P16035 REA and Q99727 were measured by ELISA . Both metalloproteinases were immunolocalized in tissue sections . All stimulation modalities induced a similar proMMP - 2 and proMMP - 9 secretion pattern in the Q8NE62 with concentrations of 2.49 ng / ml and 90.91 pg / ml , respectively ; the Q9BXJ7 showed no significant changes . The active forms of both enzymes did not change with any stimulation modality . P01033 REA , P16035 REA and Q99727 secretions remained without significant changes ( P = 0.41 ) . Q13201 REA degradation and structural disarrangement were evident after stimulation . Secretion of proMMP - 2 and proMMP - 9 mainly in the Q8NE62 , presence of active forms associated to the tissue and minor changes in TIMPs secretion could favor Q13201 REA degradation and explain the weakening and thinning associated with the pathological rupture of chorioamniotic membranes .

Experimental models of neuroprotection relevant to multiple sclerosis . Activated T cells , particularly those of the T-helper ( Th ) 1 subset , have the capacity to kill neurons . Strategies for preventing such damage may include deviation of activated T cells into the Th2 subset ( e . g . , via use of glatiramer acetate ) , alteration of functional properties of Th1 cells ( e . g . , through use of interferon [ P27352 REA ] - beta or IV immunoglobulin ) , and inhibition of activated cell migration into the CNS ( e . g . , by employing P27352 REA - beta or natalizumab ) . P14780 REA ( P14780 REA ) also causes neuron death in neurotoxicity models , and examination of medications with MMP inhibitory activity indicates that minocycline is capable of preventing such damage . DB01017 SUB also has other properties relevant to conferring neuroprotection , such as inhibition of microglial activity and apoptosis pathways . In a small pilot study in patients with relapsing-remitting multiple sclerosis , minocycline treatment produced favorable outcomes in terms of gadolinium-enhancing lesions and clinical course . Further studies are needed to establish whether experimental neuroprotection strategies involving these mechanisms may be translated into preventing neurodegeneration in multiple sclerosis .

Preterm Cervical Ripening in humans . Preterm birth ( PTB ) is the leading cause of neonatal mortality and morbidity . Despite the current treatment procedures , the incidence of PTB has not changed in the past thirty years . Incomplete understanding of the biological and patophysiological mechanisms underlying preterm delivery is the major obstacle to prevent PTB . Cervical ripening is necessary for vaginal delivery and understanding of preterm cervical ripening is required for developing new treatment strategies . Several important substances such as P09429 REA and its receptors , P06850 REA and its receptors and numerous cytokines are localized in the cervix and undergo distinct changes in labour . Other important - molecules , such as P06850 REA , P24387 REA , P34998 REA , Q13324 REA , P09429 REA , O60603 REA , O00206 REA , P22301 REA , IL - 12 , are localized in the cervical epithelium , also indicating their role in the process of cervical ripening during labour . Furthermore , P06850 REA stimulates P10145 REA secretion from both preterm and term cervical fibroblasts . Recent studies from our group show that major - inflammatory changes occur in the cervix at labour irrespective of gestational age . This indicates that cervical ripening at both term and preterm is an inflammatory process even if no infection is present . However , preterm cervical ripening still entails some differences from term cervical ripening , for example in the down-regulation of mRNA expression of Toll-like receptors ( TLR - 2 and TLR - 4 ) and IL - 12 , higher levels of P22301 REA in cervical epithelium , and presents different secretion patterns of cervical fibroblasts . Moreover , preterm cervical ripening , like preterm delivery itself , is a multifactorial disorder with pathways which are partly different from those involved in PPROM and infected preterm labour .

Identification of new candidate therapeutic target genes in triple-negative breast cancer . Triple-negative breast cancer ( TNBC ) is a subgroup of breast cancer that is negative for estrogen and progesterone receptor and P04626 REA protein expression . It is characterized by its aggressive behavior and by the lack of targeted therapies . To identify new therapeutic targets in TNBC , we used real-time quantitative RT-PCR to analyze 63 TNBC samples in terms of their mRNA expression of 26 genes coding for the major proteins currently targeted by drugs used to treat other cancers or undergoing clinical trials in breast cancer . Six of the 26 genes tested ( P15692 REA , P12931 REA , P09874 REA , Q05397 REA , P04049 REA , and P22607 REA ) were significantly upregulated in 13 % to 46 % of the TNBCs . None of the 6 genes was specifically upregulated in the TNBCs compared with 3 other classical breast tumor subtypes . No association was observed between overexpression of these 6 genes ( except for P22607 REA ) and P42336 REA mutation status . These results confirm the interest of targeting P15692 REA and P09874 REA in ongoing clinical trials in TNBC patients and also identify new target genes ( P12931 REA , Q05397 REA , P04049 REA , and P22607 REA ) . Clinical trials could be initiated easily with existing drugs . Our results also suggest that these target genes might serve as predictive biomarkers of the TNBC treatment response .

Metabolism of risperidone to 9 - hydroxyrisperidone by human cytochromes P450 2D6 and 3A4 . DB00734 is a relatively new antipsychotic drug that has been reported to improve both the positive and the negative symptoms of schizophrenia and produces relatively few extrapyramidal side effects at low doses . Formation of 9 - hydroxyrisperidone , an active metabolite , is the most important metabolic pathway of risperidone in human . In the present study , in vitro metabolism of risperidone ( 100 microM ) was investigated using the recombinant human cytochrome P450 ( CYP ) enzymes P04798 REA , P05177 REA , P10632 REA , P11712 REA - arg 144 , P11712 REA - cys 144 , P33261 REA , P10635 REA , P08684 REA and P20815 REA supplemented with an NADPH-generating system . DB01267 MENMAX DB01267 MEN was determined by a new HPLC method with an Hypersil CN column and a UV detector . Of these enzymes , CYPs 2D6 , 3A4 and 3A5 were found to be the ones capable of metabolising risperidone to 9 - hydroxyrisperidone , with activities of 7.5 , 0.4 and 0.2 pmol pmol ( - 1 ) CYP min ( - 1 ) , respectively . A correlation study using a panel of human liver microsomes showed that the formation of 9 - hydroxyrisperidone is highly correlated with P10635 REA and 3A activities . Thus , both P10635 REA and 3A4 are involved in the 9 - hydroxylation of risperidone at the concentration of risperidone used in this study . This observation is confirmed by the findings that both quinidine ( inhibitor of P10635 REA ) and ketoconazole ( inhibitor of P08684 REA ) can inhibit the formation of 9 - hydroxyrisperidone . Furthermore , inducers of CYP can significantly increase the formation of 9 - hydroxyrisperidone in rat . The formation of 9 - hydroxyrisperidone is highly correlated with testosterone 6beta - hydroxylase activities , suggesting that inducible CYP 3A contributes significantly to the metabolism of risperidone in rat .

Gene expression profiling in chronic myeloid leukemia patients treated with hydroxyurea . Using array technology that allows the simultaneous detection of gene expression of hundreds of genes , four patients with chronic myeloid leukemia ( CML ) were investigated at diagnosis and after starting administration of hydroxyurea . To detect the gene expression of peripheral blood mononuclears and granulocytes Human Cancer cDNA Array ( CLONTECH ) with 588 gene probes was used . Gene expression mononuclear and granulocyte profiles of patients at diagnosis were compared with the control profiles . The significant expression changes observed in most patients seemed to be important . Increased expression of c-jun N-terminal kinase 2 ( P45984 REA ) , integrin alpha E , P22894 REA , P14780 REA was detected in both fractions of most patients . In some samples P12004 REA , P51858 REA , MAPK p38 , P13987 REA increased expressions were found . Significant down-regulation of expression in patients was detected in genes P11802 REA inhibitor A , Q00577 , notch 1 in mononuclears ; P52630 REA , P42229 REA , P10276 REA , Q8WXI8 - 1 , junB , caspase 4 in granulocytes ; Q00534 REA , P35638 REA , P00533 REA - 3 , cadherin 5 in both fractions . Expression profiles detected in patients at diagnosis did not differ markedly from those after one-week treatment with hydroxyurea . Only in a few genes were significant changes after hydroxyurea administration observed and inter-individual expression differences were rather common .

SNPs in DNA repair genes associated to meningitis and host immune response . In vitro and in animal models , P27695 REA , O15527 REA , and P09874 REA have been proposed as being involved with inflammatory response . In this work , we have investigated if the SNPs P27695 REA Asn 148Glu , O15527 REA Ser 326Cys , and P09874 REA Val 762Ala are associated to meningitis . The patient genotypes were investigated by PIRA-PCR or PCR-RFLP . DNA damages were detected in genomic DNA by Fpg treatment . IgG and IgA were measured from plasma and the cytokines and chemokines were measured from cerebrospinal fluid samples using Bio-Plex assays . A higher frequency ( P < 0.05 ) of P27695 REA DB00142 allele in bacterial meningitis ( BM ) and aseptic meningitis ( AM ) patients was observed . The genotypes DB00174 / DB00174 in control group and DB00174 / DB00142 in BM group was also higher . For the SNP O15527 REA Ser 326Cys , the genotype DB00151 / DB00151 was more frequent ( P < 0.05 ) in BM group . The frequency of P09874 REA DB00161 / DB00161 genotype was higher in control group ( P < 0.05 ) . The occurrence of combined SNPs is significantly higher in BM patients , indicating that these SNPs may be associated to the disease . Increasing in sensitive sites to Fpg was observed in carriers of P27695 REA DB00142 allele or O15527 REA DB00151 allele , suggesting that SNPs affect DNA repair activity . Alterations in IgG production were observed in the presence of SNPs P27695 REA Asn 148Glu , O15527 REA Ser 326Cys or P09874 REA Val 762Ala . Moreover , reduction in the levels of P05231 REA , IL - 1Ra , P13500 REA / P13500 REA and P10145 REA / P10145 REA was observed in the presence of P27695 REA DB00142 allele in BM patients . In conclusion , we obtained indications of an effect of SNPs in DNA repair genes on the regulation of immune response in meningitis .

Large-scale association study for structural soundness and leg locomotion traits in the pig . BACKGROUND : Identification and culling of replacement gilts with poor skeletal conformation and feet and leg ( FL ) unsoundness is an approach used to reduce sow culling and mortality rates in breeding stock . Few candidate genes related to soundness traits have been identified in the pig . METHODS : In this study , 2066 commercial females were scored for 17 traits describing body conformation and FL structure , and were used for association analyses . Genotyping of 121 SNPs derived from 95 genes was implemented using Sequenom ' s MassARRAY system . RESULTS : Based on the association results from single trait and principal components using mixed linear model analyses and false discovery rate testing , it was observed that P02649 REA , P34820 , P30988 REA , P08123 REA , P20849 REA , DKFZ , P35555 REA and VDBP were very highly significantly ( P < 0.001 ) associated with body conformation traits . The genes P09917 REA , P34820 , P30988 REA , O00300 REA , P30559 REA and Q9UBV4 were very highly significantly ( P < 0.001 ) associated with FL structures , and P02649 REA , P30988 REA , P08123 REA , P30968 REA , Q14623 REA , P42898 REA and Q9UBV4 were highly significantly ( P < 0.01 ) associated with overall leg action . Strong linkage disequilibrium between P30988 REA and P08123 REA on SSC 9 was detected , and haplotype - ACGACC - was highly significantly ( P < 0.01 ) associated with overall leg action and several important FL soundness traits . CONCLUSION : The present findings provide a comprehensive list of candidate genes for further use in fine mapping and biological functional analyses .

DB01017 SUB attenuates hypoxia-inducible factor - 1α expression correlated with modulation of p53 and AKT / P42345 REA / p70S6K / Q13541 REA pathway in ovarian cancer : in vitro and in vivo studies . Hypoxia-inducible factor ( HIF ) - 1α is the key cellular survival protein under hypoxia , and is associated with tumor progression and angiogenesis . We have recently shown the inhibitory effects of minocycline on ovarian tumor growth correlated with attenuation of vascular endothelial growth factor ( P15692 REA ) and herein report a companion laboratory study to test if these effects were the result of HIF - 1α inhibition . In vitro , human ovarian carcinoma cell lines ( A2780 , OVCAR - 3 and SKOV - 3 ) were utilized to examine the effect of minocycline on Q9BYW2 and its upstream pathway components to elucidate the underlying mechanism of action of minocycline . Mice harboring OVCAR - 3 xenografts were treated with minocycline to assess the in vivo efficacy of minocycline in the context of Q9BYW2 . DB01017 SUB negatively regulated HIF - 1α protein levels in a concentration-dependent manner and induced its degradation by a mechanism that is independent of prolyl-hydroxylation . The inhibition of HIF - 1α was found to be associated with up-regulation of endogenous p53 , a tumor suppressor with confirmed role in HIF - 1α degradation . Further studies demonstrated that the effect of minocycline was not restricted to proteasomal degradation and that it also caused down-regulation of HIF - 1α translation by suppressing the AKT / P42345 REA / p70S6K / Q13541 REA signaling pathway . DB01017 SUB treatment of mice bearing established ovarian tumors , led to suppression of HIF - 1α accompanied by up-regulation of p53 protein levels and inactivation of AKT / P42345 REA / p70S6K / Q13541 REA pathway . These data reveal the therapeutic potential of minocycline in ovarian cancer as an agent that targets the pro-oncogenic factor HIF - 1α through multiple mechanisms .

DB01017 SUB accelerates hypoxia-inducible factor - 1 alpha degradation and inhibits hypoxia-induced neovasculogenesis through prolyl hydroxylase , von Hippel-Lindau-dependent pathway . Hypoxia-mediated stress responses are important in tumor progression , especially when tumor growth causes the tumor to become deprived of its blood supply . The oxygen-labile transcription factor hypoxia-inducible factor - 1 alpha ( HIF - 1α ) plays a critical role in regulating hypoxia stress-related gene expression and is considered a novel therapeutic target . Lung adenocarcinoma cell lines were exposed to minocycline , followed by incubation at hypoxic condition for 3-6 h . Here , we show that minocycline , a second-generation tetracycline , can induce HIF - 1α proteasomal degradation under hypoxia by increasing the expression prolyl hydroxylase - 2 and HIF - 1α / von Hippel-Lindau protein interaction , thereby overcoming hypoxia-induced HIF - 1α stabilization . Neither repression of hypoxia-induced phosphatidylinositol - 3 kinase / Akt / mammalian target of rapamycin pathway nor inhibition of Hsp 90 was required for minocycline-induced HIF - 1α degradation . The HIF - 1α degradation-enhancing effect of minocycline was evident in both cancerous and primary cells . DB01017 SUB - pretreated , hypoxia-conditioned cells showed a clear reduction in hypoxia response element reporter expression and amelioration of vascular endothelial growth factor C / D ( P49767 REA / D ) , matrix metalloproteinase 2 , and glucose transporter 1 expression . By decreasing P15692 REA secretion of cancerous cells , minocycline could suppress endothelial cell neovasculogenesis . These findings suggest a novel application of minocycline in the treatment of tumor angiogenesis as well as hypoxia-related diseases .

Clinical and pathogenic aspects of candidate genes for lithium prophylactic efficacy . A number of candidate genes for lithium prophylactic efficacy have been proposed , some of them being also associated with a predisposition to bipolar illness . The aim of the present study was to investigate a possible association between polymorphisms of 14 common genes with the quality of prophylactic lithium response in patients with bipolar mood disorder , in relation to the putative role of these genes in the pathogenesis of this disorder . Some association with lithium prophylactic efficacy was found for the polymorphisms of P31645 REA , P21728 REA , P21964 REA , P23560 REA and P06241 REA genes , but not for 5HT2A , 5HT2C , P14416 REA , P35462 REA , P21917 REA , GSK - 3 , Q16620 REA , Q13224 REA and P14780 REA . Possible aspects of these genes with regard to the mechanism of lithium activity and pathogenesis of bipolar mood disorder are discussed .

Matrix metalloproteinase ( MMP ) - 9 , but not P08253 REA , is involved in the development and progression of C protein-induced myocarditis and subsequent dilated cardiomyopathy . Repeated or continuous inflammation of the heart is one of the initiation factors for dilated cardiomyopathy ( DCM ) . In previous studies , we established a DCM animal model by immunizing rats with cardiac C protein . In the present study , we analyze the role of matrix metalloproteinases ( MMPs ) in experimental autoimmune carditis ( EAC ) and subsequent DCM to elucidate the pathomechanisms of this disease . In this model , inflammation begins approximately 9 days after immunization . At that time , MMP activities were detected by in situ zymography . Real-time PCR analysis revealed continuous up-regulation of P08253 REA mRNA from 2 wk and thereafter . P14780 REA mRNA , however , had only a transient increase at 2 wk . Double staining with in situ zymography and cell markers demonstrated that gelatinase ( P08253 REA and P14780 REA ) - expressing cells are infiltrating macrophages during the early stage and cardiomyocytes at later stages . DB01017 SUB , which inhibits P14780 REA activities more strongly than P08253 REA , significantly suppressed EAC , but an P08253 REA - specific inhibitor , TISAM , did not affect the course of the disease . Furthermore , immunohistochemical examination revealed that minocycline treatment suppressed T cell and macrophage infiltration strongly , whereas TISAM did not . These findings indicate that P14780 REA , but not P08253 REA , is involved in the pathogenesis of the acute phase of EAC , and further suggest that P14780 REA inhibitors , minocycline and its derivatives , may be useful therapies for EAC and DCM .

DB01296 MEN sulfate inhibits P01375 REA and P01579 REA - induced production of P05362 REA in human retinal pigment epithelial cells in vitro . PURPOSE : DB01296 MEN sulfate ( GS ) is a naturally occurring sugar that possesses some immunosuppressive effects in vitro and in vivo , but its mechanism is unknown . We investigated whether GS could modulate the proinflammatory cytokine-induced expression of the gene for intercellular adhesion molecule ( ICAM ) - 1 , an inflammatory protein in human retinal pigment epithelial ( Q96AT9 ) cells . METHODS : ARPE - 19 cells were used as a model to determine the effects of GS on the expression of the P05362 REA gene upregulated by P01375 REA or P01579 REA , by Western blot analysis and semiquantitative reverse transcription polymerase chain reaction ( RT-PCR ) . The activation and nuclear translocation of the nuclear factors NF-kappaB and P42224 REA were evaluated by immunocytochemistry , Western blot analysis , and electrophoretic mobility shift assay ( EMSA ) . RESULTS : Both P01375 REA and P01579 REA increased the expression of P05362 REA at the mRNA and protein levels in a time - and dose-dependent manner in ARPE - 19 cells . GS effectively downregulated the P01375 REA - or P01579 REA - induced expression of P05362 REA in the protein and mRNA level in a dose-dependent manner . GS further inhibited the nuclear translocation of p65 proteins in P01375 REA and phosphorylated P42224 REA in P01579 REA - stimulated ARPE - 19 cells . CONCLUSIONS : GS inhibits the expression of the P05362 REA gene in ARPE - 19 cell stimulated with P01375 REA or P01579 REA through blockade of NF-kappaB subunit p65 and nuclear translocation of P42224 REA . This study has demonstrated a potentially important property of GS in reducing P05362 REA mediated inflammatory mechanisms in the eye .

PPARγ and RXR ligands disrupt the inflammatory cross-talk in the hypoxic breast cancer stem cells niche . Cancer stem cells ( CSCs ) are affected by the local micro-environment , the niche , in which inflammatory stimuli and hypoxia act as steering factors . Here , two nuclear receptors ( NRs ) agonists , i . e . pioglitazone ( PGZ ) , a ligand of peroxisome proliferator activated receptor-γ , and 6 - OH - 11 - O-hydroxyphenanthrene ( IIF ) , a ligand of retinoid X receptors , were investigated for their capability to interference with the cross-talk between breast CSCs and the niche compartment . We found that IIF potentiates the ability of PGZ to hamper the mammospheres-forming capability of human breast tumours and MCF 7 cancer cells , reducing the expression of CSCs regulatory genes ( Notch 3 , P78504 REA , O43623 REA , P05231 REA , P02649 REA , Hypoxia inducible factor - 1α and Q16790 REA ) . Notably , these effects are not observed in normal-MS obtained from human breast tissue . Importantly , NRs agonists abolish the capability of hypoxic MCF 7 derived exosomes to induce a pro-inflammatory phenotype in mammary glands fibroblasts . Moreover , NRs agonist also directly acts on breast tumour associated fibroblasts to downregulate nuclear factor-κB pathway and metalloproteinases ( P08253 REA and P14780 REA ) expression and activity . In conclusion , NRs agonists disrupt the inflammatory cross-talk of the hypoxic breast CSCs niche .

P06850 REA enhances the invasiveness and migration of Ishikawa cells , possibly by increasing matrix metalloproteinase - 2 and matrix metalloproteinase - 9 . P06850 REA ( P06850 REA ) , synthesized in the hypothalamus , is also produced at several extrahypothalamic sites and in normal endometrial cells . P06850 REA exerts antiproliferative activity on oestrogen-dependent tumour cell lines ( Ishikawa cells and breast cancer cells ) via the P06850 REA receptor - 1 . This study investigated the potential role of P06850 REA as a factor affecting endometrial migration and invasion in Ishikawa cells , and the possible mechanisms involved in this process . Increasing concentrations of P06850 REA ( 1 , 10 and 100 nM ) significantly reduced the proliferation of Ishikawa cells but increased the invasiveness these cells compared with the control group . All three concentrations of P06850 REA significantly increased matrix metalloproteinase ( MMP ) - 2 and P14780 REA levels in Ishikawa cells . In conclusion , P06850 REA inhibited the growth of Ishikawa cells but enhanced their invasiveness , possibly by increasing P08253 REA and P14780 REA levels . These findings suggest that P06850 REA might induce invasion and migration by upregulating P08253 REA and P14780 REA in endometrial cancer .

DB01017 SUB improves functional outcomes , memory deficits , and histopathology after endovascular perforation-induced subarachnoid hemorrhage in rats . Subarachnoid hemorrhage ( Q53FZ2 ) results in significant long-lasting cognitive dysfunction . Therefore , evaluating acute and long-term outcomes after therapeutic intervention is important for clinical translation . The aim of this study was to use minocycline , a known neuroprotectant agent , to evaluate the long-term benefits in terms of neurobehavior and neuropathology after experimental Q53FZ2 in rats , and to determine which neurobehavioral test would be effective for long-term evaluation . Q53FZ2 was induced by endovascular perforation in adult male Sprague-Dawley rats ( n = 118 ) . The animals were treated with intraperitoneal injection of minocycline ( 45 mg / kg or 135 mg / kg ) or vehicle 1 h after Q53FZ2 induction . In the short-term , animals were euthanized at 24 and 72 h for evaluation of neurobehavior , brain water content , and matrix metalloproteinase ( MMP ) activity . In the long-term , neurobehavior was evaluated at days 21-28 post - Q53FZ2 , and histopathological analysis was done at day 28 . High-dose but not low-dose minocycline reduced brain water content at 24 h , and therefore only the high-dose regimen was used for further evaluation , which reduced P14780 REA activity at 24 h . Further , high-dose minocycline improved spatial memory and attenuated neuronal loss in the hippocampus and cortex . The rotarod , T-maze , and water maze tests , but not the inclined plane test , detected neurobehavioral deficits in Q53FZ2 rats at days 21-28 . This study demonstrates that minocycline attenuates long-term functional and morphological outcomes after endovascular perforation-induced Q53FZ2 . Long-term neurobehavioral assessments using the rotarod , T-maze , and water maze tests could be useful to evaluate the efficacy of therapeutic intervention after experimental Q53FZ2 .

DB01017 SUB with aspirin : an approach to attenuate diabetic nephropathy in rats . Degradation of extracellular matrix ( Q13201 REA ) by enhanced production of matrix metalloproteinase - 2 ( P08253 REA ) and matrix metalloproteinase - 9 ( P14780 REA ) in diabetes leads to nephropathy . Cyclooxygenases ( P36551 REA ) further increase levels of these MMPs . The objective of present study was to inhibit P08253 REA and P14780 REA by combination of minocycline and aspirin to treat diabetic nephropathy . Diabetes was induced in male Wistar rats by streptozotocin ( Q11206 REA , 55 mg / kg i . p . ) . Four weeks after diabetes induction , the rats were treated with minocycline ( 50 mg / kg , p . o . ) , aspirin ( 50 mg / kg , p . o . ) , or minocycline ( 50 mg / kg , p . o . ) plus aspirin ( 50 mg / kg , p . o . ) for a period of 4 weeks . At the end of eighth week fluid input , urine output , and renal function tests were carried out for diagnosis of diabetic nephropathy . Renal hypertrophy was measured and histopathology was done to evaluate renal damage . Diabetes produced significant loss of body weight , polyuria , polydipsia , hyperglycemia , and increase in blood pressure . Serum creatinine , urea , and blood urea nitrogen levels were found to be increased significantly in the Q11206 REA group diabetic rats . Treatment with combination of minocycline and aspirin significantly prevented the rise in creatinine , urea , and blood urea nitrogen levels and increased creatinine clearance . Image analysis of kidneys revealed that collagen level was significantly decreased in combined treated group when compared with control . Results of present study suggest that P08253 REA and P14780 REA inhibition in presence of P36551 REA inhibitor prevents the development of experimental diabetic nephropathy in rats and can be a potential approach for the treatment .

Delivering minocycline into brain endothelial cells with liposome-based technology . DB01017 SUB has been proposed as a way to blunt neurovascular injury from matrix metalloproteinases ( MMPs ) during stroke . However , recent clinical trials suggest that high levels of minocycline may have deleterious side-effects . Here , we showed that very high minocycline concentrations damage endothelial cells via calpain / caspase pathways . To alleviate this potential cytotoxicity , we encapsulated minocycline in liposomes . Low concentrations of minocycline could not reduce tumor necrosis factor α ( TNFα ) - induced P14780 REA release from endothelial cells . But low concentrations of minocycline-loaded liposomes significantly reduced TNFα-induced P14780 REA release . This study provides proof-of-concept that liposomes may be used to deliver lower levels of minocycline for targeting MMPs in cerebral endothelium .

DB01017 SUB suppresses experimental autoimmune encephalomyelitis by increasing tissue inhibitors of metalloproteinases . Matrix metalloproteinases ( MMPs ) that are secreted by activated T cells play a significant role in degradation of the extracellular matrix around the blood vessels and facilitate autoimmune neuroinflammation ; however , it remains unclear how MMPs act in lesion formation and whether MMP-targeted therapies are effective in disease suppression . In the present study , we attempted to treat experimental autoimmune encephalomyelitis ( EAE ) by administration of small interfering RNAs ( siRNAs ) for P08253 REA , P14780 REA , and minocycline , all of which have MMP-inhibiting functions . DB01017 SUB , but not siRNAs , significantly suppressed disease development . In situ zymography revealed that gelatinase activities were almost completely suppressed in the spinal cords of minocycline-treated animals , while significant gelatinase activities were measured in the EAE lesions of control animals . However , P08253 REA and P14780 REA mRNAs and proteins in the spinal cords of treated rats were unexpectedly upregulated . At the same time , mRNA for tissue inhibitors of MMPs ( P01033 REA ) - 1 and - 2 were also upregulated . The EnzChek Gelatinase / Collagenase assay using tissue containing native MMPs and TIMPs demonstrated that gelatinase activity levels in the spinal cords of treated rats were suppressed to the same level as those in normal spinal cord tissues . Finally , double immunofluorescent staining demonstrated that P14780 REA immunoreactivities of treated rats were almost the same as those of control rats and that P14780 REA and P01033 REA immunoreactivities were colocalized in the spinal cord . These findings suggest that minocycline administration does not suppress MMPs at mRNA and protein levels but that it suppresses gelatinase activities by upregulating TIMPs . Thus , MMP-targeted therapies should be designed after the mechanisms of candidate drugs have been considered .

Q16552 REA promotes p38 MAPK-dependent endothelial activation enhancing neutrophil recruitment to sites of inflammation . Neutrophilic inflammation plays an important role in lung tissue destruction occurring in many chronic pulmonary diseases . Neutrophils can be recruited to sites of inflammation via the action of the cytokine Q16552 REA . In this study , we report that Q96F46 REA and Q8NAC3 mRNA expression is significantly increased in asthmatic bronchoscopic biopsies and that these receptors are not only expressed on epithelial and inflammatory cells but also on endothelial cells . Q16552 REA potently stimulates lung microvascular endothelial cells to produce chemoattractants ( P10145 REA and derivatives of the P09917 REA pathway ) that selectively drive neutrophil but not lymphocyte chemotaxis . Moreover , Q16552 REA promotes endothelial activation by inducing the expression of endothelial adhesion markers ( P16581 REA , P19320 REA , and P05362 REA ) in a p38 MAPK-dependent manner . This increased expression of adhesion molecules stimulates the trans-endothelial migration of neutrophils , as well as the transmigration of HT - 29 colon carcinoma cells , suggesting a further role in promoting lung metastasis . Finally , Q16552 REA increased neutrophil adhesion to the endothelium in vivo as determined by intravital microscopy of mice cremaster muscle . Overall , our results demonstrate that Q16552 REA is a potent activator of the endothelium in vivo leading to neutrophil infiltration . Therefore , preventing neutrophil recruitment by blocking the action of Q16552 REA on endothelial cells may prove to be highly beneficial in diseases in which neutrophilic inflammation plays a key role .

PEG minocycline-liposomes ameliorate CNS autoimmune disease . BACKGROUND : DB01017 SUB is an oral tetracycline derivative with good bioavailability in the central nervous system ( CNS ) . DB01017 SUB , a potent inhibitor of matrix metalloproteinase ( MMP ) - 9 , attenuates disease activity in experimental autoimmune encephalomyelitis ( EAE ) , an animal model of multiple sclerosis ( MS ) . Potential adverse effects associated with long-term daily minocycline therapy in human patients are concerning . Here , we investigated whether less frequent treatment with long-circulating polyethylene glycol ( PEG ) minocycline liposomes are effective in treating EAE . FINDINGS : Performing in vitro time kinetic studies of PEG minocycline-liposomes in human peripheral blood mononuclear cells ( PBMCs ) , we determined that PEG minocycline-liposome preparations stabilized with CaCl ( 2 ) are effective in diminishing P14780 REA activity . Intravenous injections of PEG minocycline-liposomes every five days were as effective in ameliorating clinical EAE as daily intraperitoneal injections of minocycline . Treatment of animals with PEG minocycline-liposomes significantly reduced the number of CNS-infiltrating leukocytes , and the overall expression of P14780 REA in the CNS . There was also a significant suppression of P14780 REA expression and proteolytic activity in splenocytes of treated animals , but not in CNS-infiltrating leukocytes . Thus , leukocytes gaining access to the brain and spinal cord require the same absolute amount of P14780 REA in all treatment groups , but minocycline decreases the absolute cell number . CONCLUSIONS : Our data indicate that less frequent injections of PEG minocycline-liposomes are an effective alternative pharmacotherapy to daily minocycline injections for the treatment of CNS autoimmune diseases . Also , inhibition of P14780 REA remains a promising treatment target in EAE and patients with MS .

DB01017 SUB modulates cytokine and gene expression profiles in the brain after whole-body exposure to radiation . An effective countermeasure against radiation damage to normal tissues is urgently needed . The major goal of the present study was to determine if minocycline could modify the immunomodulatory effects of radiation on the brain . C57BL / 6 mice were treated with minocycline intraperitoneally for 5 days beginning immediately before total-body exposure to 0 , 1 , 2 and 3 Gray ( Gy ) ( 60 ) Co γ-rays . Brains were collected on days 4 and 32 post-irradiation for cytokine and gene analyses . DB01017 SUB treatment significantly increased the levels of interleukin ( IL ) - 10 , P40933 REA and vascular endothelial growth factor ( P15692 REA ) in the brain on day 4 in one or more irradiated groups compared to radiation-alone ( p < 0.05 ) . P22301 REA is anti-inflammatory , P40933 REA can prevent apoptosis and P15692 REA is nuroprotective . On day 32 , the drug decreased IL - 1β in the 2 - Gy group ( p < 0.05 vs . 2 - Gy alone ) ; this cytokine is implicated in immune-related central nervous system pathologies . Microarray analysis of brains on day 32 showed that while radiation increased expression of inflammatory genes such as Il1f10 , Il17 , Tnfrsf 11b , Tnfsf 12 , Il12b and Il1f8 , these were no longer up-regulated in the minocycline-treated groups . Similarly , the pro-apoptotic gene Bik and nitric oxide synthase producer ( Q8IVI9 ) were no longer up-regulated in the drug-treated groups . Pathway analysis based on gene data suggested that catenin-β 1 and tumor suppressor-related transcription regulation were significantly activated by radiation and / or minocycline ( activation z-score > 2.0 ) . Overall , the data warrant further testing of minocycline as a potential neuroprotectant against radiation-induced damage .

O60315 mediates multiple pathways regulating cell proliferation , migration , invasion , and apoptosis in glioma . BACKGROUND : The aim of the present study was to analyze the expression of Zinc finger E-box Binding homeobox 2 ( O60315 ) in glioma and to explore the molecular mechanisms of O60315 that regulate cell proliferation , migration , invasion , and apoptosis . METHODOLOGY / PRINCIPAL FINDINGS : Expression of O60315 in 90 clinicopathologically characterized glioma patients was analyzed by immunohistochemistry . Furthermore , siRNA targeting O60315 was transfected into U251 and U87 glioma cell lines in vitro and proliferation , migration , invasion , and apoptosis were examined separately by MTT assay , Transwell chamber assay , flow cytometry , and western blot . RESULTS : The expression level of O60315 protein was significantly increased in glioma tissues compared to normal brain tissues ( P < 0.001 ) . In addition , high levels of O60315 protein were positively correlated with pathology grade classification ( P = 0.024 ) of glioma patients . Knockdown of O60315 by siRNA suppressed cell proliferation , migration and invasion , as well as induced cell apoptosis in glioma cells . Furthermore , O60315 downregulation was accompanied by decreased expression of P11802 REA / 6 , P12004 REA D1 , P12004 REA E , Q01094 REA , and c-myc , while p15 and P38936 REA were upregulated . Lowered expression of O60315 enhanced P12830 REA levels but also inhibited β-Catenin , P08670 REA , P19022 REA , and Snail expression . Several apoptosis-related regulators such as P42574 REA , P55212 REA , P55211 REA , and Cleaved-PARP were activated while PARP was inhibited after O60315 siRNA treatment . CONCLUSION : Overexpression of O60315 is an unfavorable factor that may facilitate glioma progression . Knockdown O60315 expression by siRNA suppressed cell proliferation , migration , invasion and promoted cell apoptosis in glioma cells .

DB01017 SUB protects PC12 cells against DB01221 - induced injury via inhibiting P09917 REA activation . Recently , we have reported that minocycline , a semi-synthetic tetracycline with neuroprotective effects , inhibits the in vitro ischemic-like injury and P09917 REA ( 5 - P28300 REA ) activation in PC12 cells . In the present study , we further determined whether minocycline protects PC12 cells from excitotoxicity via inhibiting 5 - P28300 REA activation . We used N-methyl-d-aspartate ( DB01221 , 200 microM ) to induce early ( exposure for 6 h ) and delayed ( exposure for 6 h followed by 24 h recovery ) injuries . We found that DB01221 receptor antagonist ketamine , 5 - P28300 REA inhibitor caffeic acid and minocycline concentration dependently attenuated DB01221 - induced early and delayed cell injuries ( viability reduction and cell death ) . However , only ketamine ( 1 microM ) inhibited DB01221 - evoked elevation of intracellular calcium . In addition , immunohistochemical analysis showed that DB01221 induced 5 - P28300 REA translocation to the nuclear membrane after 1 - to 6 - h exposure which was confirmed by Western blotting , indicating that 5 - P28300 REA was activated . DB01221 , caffeic acid and minocycline ( each at 1 microM ) inhibited 5 - P28300 REA translocation after early injury . After delayed injury , PC12 cells were shrunk , and 5 - P28300 REA was translocated to the nuclei and nuclear membrane ; ketamine , caffeic acid and minocycline inhibited both cell shrinking and 5 - P28300 REA translocation . As a control , P16050 REA inhibitor baicalein showed a weak effect on cell viability and death , but no effect on 5 - P28300 REA translocation . Therefore , we conclude that the protective effect of minocycline on DB01221 - induced injury is partly mediated by inhibiting 5 - P28300 REA activation .

DB01017 SUB partially inhibits caspase - 3 activation and photoreceptor degeneration after photic injury . PURPOSE : To evaluate the possible role of caspase - 3 in retinal photic injury , and to investigate whether minocycline can ameliorate light-induced photoreceptor degeneration . METHODS : Retinal photic injury was induced in rats by exposure to intense light . Expression of caspase - 3 was studied using Western blot analysis , immunohistochemical staining and enzyme activity assay . Apoptotic photoreceptor cells were detected by the TdT-dUTP terminal nick-end labeling ( TUNEL ) method . DB01017 SUB ( 15 , 30 or 45 mg / kg ) was administered before or after photic injury in rats randomly assigned to pretreatment and posttreatment groups . DB01017 SUB and vehicle-treated retinas subjected to photic injury were compared with respect to Western blotting , enzyme activity assay , quantitative counts of TUNEL stains , morphometry of the outer nuclear layer ( ONL ) thickness and histopathological examination . RESULTS : After light exposure , active caspase - 3 and poly-adenosine diphosphate-ribose-polymerase were upregulated in the retinas and increased caspase - 3 immunoreactivity was observed in the ONL . P42574 REA enzyme activity increased in the retinas that underwent photic injury , and this increase was significantly reduced in minocycline pretreated ( 30 and 45 mg / kg ) and posttreated ( 45 mg / kg ) groups . Intraperitoneal administration of minocycline before or after photic injury in rats also resulted in less TUNEL-positive photoreceptors , as assessed by the quantitative TUNEL counts . The degree of retinal degeneration , measured by the ONL thickness 14 days after photic injury , was significantly improved in minocycline pretreatment ( 45 mg / kg ) rats . CONCLUSIONS : We demonstrate that increased caspase - 3 activities localize specifically within the ONL after photic injury , and that minocycline partially inhibits caspase - 3 activation and photoreceptor degeneration in this animal model .

Effects of baicalein on apoptosis , cell cycle arrest , migration and invasion of osteosarcoma cells . Baicalein is a bioactive flavonoid that is widely used in ancient China . However , its effects on the most common primary malignant bone tumor , osteosarcoma , remain unknown . In the present study , we investigated the effects of baicalein in osteosarcoma cells . Our results indicate baicalein might be an efficacious anti-osteosarcoma drug . We found that baicalein could inhibit cell proliferation in a time - and dose-dependent manner . Additionally , we demonstrated that baicalein promotes osteosarcoma cell apoptosis , and our mechanistic studies suggest that this is mediated by caspase activation , especially caspase - 3 . We also showed that the down-regulation of Bcl - 2 and concurrent increase in Bax and Bim levels contribute to the apoptosis induced by baicalein . In addition , we observed that baicalein induces P55008 cell cycle arrest by decreasing cyclin D1 and cyclin-dependent kinase 4 ( P11802 REA ) . Furthermore , our data verifies that baicalein can reduce osteosarcoma cell adhesion , migration and invasion in vitro , which indicates its potential to inhibit osteosarcoma metastasis . The decrease in expression of matrix metalloproteinases ( MMP ) - 2 and P14780 REA may contribute to the effects of baicalein . Taken together , our results provide evidence that baicalein plays important roles in anti-osteosarcoma therapy , and thus may serve as a novel and efficient candidate agent for osteosarcoma treatment .

Poly ( DB02059 ) polymerase - 1 signalling of the DNA damage induced by P11387 REA poison in D54 ( p53wt ) and U251 ( p53mut ) glioblastoma cell lines . Glioblastomas are widely characterised by the mutation of the p53 gene and p53 disruption sensitizes glioblastoma cells to P11387 REA ( TOPO I ) inhibitor-mediated apoptosis . We investigated the effects of combined treatments with the P11387 REA inhibitor DB01030 MEN and the poly ( DB02059 ) polymerase - 1 inhibitor DB02690 in D54 ( p53wt ) and U251 ( p53mut ) glioblastoma cell lines . Analysis of cell growth and cell cycle kinetics showed a synergistic anti-proliferative effect of 10 nM TPT and 10 microM DB02690 and a G2 / M block of the cell cycle . We also evaluated , the influence of TPT + / - DB02690 treatment on P09874 REA and p53 activity . We got evidences of a TPT-dependent increase of P09874 REA auto-modification level in both the cells . Moreover , in the D54 ( p53wt ) cells we found that in co-treatments DB02690 incremented the TPT-dependent stimulation of p53 transcriptional activity and increased the P38936 REA nuclear amount . Conversely , in U251 ( p53mut ) cells we found that DB02690 incremented the TPT-dependent apoptosis characterised by P09874 REA proteolysis . Our findings suggest that the modulation of P09874 REA can be considered a strategy in the potentiation of the chemotherapeutic action of TOPO I poisons in glioblastoma cells apart from their p53 status .

The effects of minocycline and tetracycline on the mitotic response of human peripheral blood-lymphocytes . The effects of minocycline and tetracycline on the mitotic response of human peripheral blood lymphocytes was investigated in vitro . The effects of the antibiotics on the mitotic response of purified lymphocytes stimulated with P01584 REA varied according to the individual from whom the lymphocytes were obtained . At concentrations above those reported to be present in serum during conventional therapy ( 2-8 mg / l ) , there was a tendency for both minocycline and tetracycline to suppress the mitotic response . DB01017 SUB was superior to tetracycline in this respect . However , at physiological concentrations the antibiotics either had no significant effect , suppressed the mitotic response ( minocycline at 2 mg / l with one of six donors ) , or enhanced the mitotic response ( tetracycline at 2 and 8 mg / l with four of six donors ) . The stimulatory effect of tetracycline was not demonstrated when lymphocytes were cultured in whole blood for up to seven days with the antibiotic alone . Similar effects of the antibiotics were observed when mononuclear cell fractions isolated from six donors were stimulated with an optimal concentration of phytohaemagglutinin ( PHA ) . Stimulation of lymphocytes in whole blood cultures with PHA in the presence of minocycline and tetracycline revealed that , under these culture conditions , the antibiotics could suppress the mitotic response of lymphocytes at physiological doses with cells from a majority of donors .

DB01076 MEN inhibits RhoC function and limits head and neck cancer metastasis . OBJECTIVE : RhoC oncogene is a well characterized marker of metastasis in a majority of invasive cancers , including HNSCC . Elevated RhoC expression has been found to be associated with distant metastasis . Statins are a class of drugs that are used to reduce cholesterol levels by inhibiting P04035 REA activity which in turns prevents mevalonate synthesis , which is a precursor for synthesis of cholesterol and prenylation . Interestingly , the proper function of Rho proteins depends on prenylation . Significantly , it has been reported that metastasis in human melanoma can be reduced by atorvastatin which inhibits RhoC activity by preventing its geranylgeranylation . Given that RhoC is a key oncogene involved in metastasis , we hypothesized DB01076 MEN can reduce head and neck metastasis by inhibiting RhoC activity . METHODS : In vitro and in vivo studies were carried out to evaluate the ability of DB01076 MEN to inhibit RhoC function and HNSCC metastasis . Cell motility , proliferation , cell invasion , and colony formation assays were performed according to the standard protocols . RESULTS : DB01076 MEN treatment significantly reduced the active form of RhoC in vitro and diminished cell motility , invasion , proliferation and colony formation . Importantly , we observed a significant decrease in p - P27361 REA / 2 and p - P40763 REA in DB01076 MEN treated cell lines . In vivo experiments revealed inhibition of angiogenesis and lung metastases with DB01076 MEN therapy . CONCLUSIONS : This study is the first of its kind to establish a potential role of DB01076 MEN in head and neck cancer therapy . These findings suggest that DB01076 MEN can be a potential low risk adjuvant therapy to minimize metastases in aggressive forms of HNSCC .

Genetic factors influencing outcome from neurotrauma . PURPOSE OF REVIEW : Clinical outcome after neurotrauma is considerably variable and can only partly be explained by known prognostic factors . There is converging evidence from genetic research that a number of genetic variants may contribute to this variability . This review provides recent data from human studies , published in the previous year , on genetic factors influencing outcome after neurotrauma . The bibliographic databases MEDLINE , EMBASE and PsycINFO were searched to identify relevant studies . RECENT FINDINGS : Genetic susceptibility to various aspects of clinical outcome after neurotrauma was reported in recent clinical studies . Genetic loci investigated include polymorphisms in P02649 REA , P21397 REA , P23560 REA , NOS 3 , P05231 REA , P12036 , P31645 REA , P21964 REA , P48454 REA and Q8IX03 genes . The importance of these findings and future directions are discussed . SUMMARY : Recent genetic studies have revealed emerging aspects and extended the existing knowledge regarding the pathogenesis of neurotrauma and the genetic influence on phenotypic diversity . A better understanding of the underlying biological pathways and molecular mechanisms of an individual ' s response to neurotrauma may hold the promise of novel treatment strategies and improved clinical outcome .

Intrahypothalamic perfusion with interleukin - 1 - beta stimulates the local release of corticotropin-releasing hormone and arginine vasopressin and the plasma adrenocorticotropin in freely moving rats : a comparative perfusion of the paraventricular nucleus and the median eminence . It is almost generally accepted that an acute-phase DB01285 response induced by interleukin ( IL ) - 1 is mediated principally by P06850 REA release from the hypothalamus . However , the precise cellular site of action of IL - 1 in activating the P06850 REA neuronal system remains to be determined . Two likely candidates comprise the paraventricular nucleus ( PVN ) where P06850 REA neuronal cell bodies are located , and the median eminence ( ME ) where their nerve endings are terminated . Therefore , in this study we performed a comparative perfusion of the ME and the PVN with increasing concentrations of recombinant human P01584 REA utilizing the push-pull perfusion technique in freely moving rats . We measured the plasma DB01285 and ME and PVN levels of P06850 REA , and also of AVP , because AVP , another secretagogue of DB01285 , has its cell body in the PVN and axon terminals partly in the ME . In control groups , the ME or the PVN was perfused with artificial cerebrospinal fluid between 12:00 and 15:00 h , and perfusates and blood samples were collected every 20 min . In the other groups , either the ME or the PVN was perfused with three increasing concentrations ( 0.1 , 1.0 , and 10 nM ) of recombinant human P01584 REA dissolved in artificial cerebrospinal fluid only between 13:00 and 14:00 h with all the other procedures run in the same way as in the controls . In the control perfusions , the hypothalamic release of P06850 REA and AVP and the plasma DB01285 did not change significantly during the entire period of observation . ( ABSTRACT TRUNCATED AT 250 WORDS )

Sense p16 and antisense Q03405 REA bicistronic construct inhibits angiogenesis and induces glioma cell death . High-grade gliomas comprise the most malignant type of primary brain tumor and are relatively frequent in adults . Recent studies have indicated that the loss of p16 , an inhibitor of P11802 REA , promotes the acquisition of malignant characteristics in gliomas . A correlation between overexpression of urokinase-type plasminogen activator receptor ( Q03405 REA ) and glioblastoma invasion has also been established . Moreover , Q03405 REA / integrin binding has been shown to initiate or potentiate integrin signaling through focal adhesion kinase and / or src kinases . Our previous studies demonstrated that downregulation of Q03405 REA expression and restoration of p16 regress glioma growth in nude mice and downregulate alphavbeta 3 integrin receptor expression . Here , we show the effect of a bicistronic construct on alphavbeta 5 integrin receptor expression , angiogenesis and the biochemical pathway that causes glioma cell death . The U251 glioblastoma and a glioblastoma xenograft cell line transduced with a recombinant replication-defective adenovirus vector containing the cDNA of wild-type p16 and antisense RNA of Q03405 REA significantly inhibited human mammary epithelial cell capillary formation and vascular endothelial growth factor ( P15692 REA ) expression . Inactivation of anti-apoptotic molecules such as Akt , PARP , activation of caspases and accumulation of heteroduplex chromosomal DNA in pre - P55008 phase of the cell cycle was demonstrated by Western blotting , caspase activity assay and FACS analysis . Nuclear DNA fragmentation upon induction of apoptosis was scored using the TUNEL assay . Significant downregulation of alphavbeta 5 integrin receptor expression was also confirmed by FACS analysis , immunoprecipitation and RT-PCR . Taken together , the results demonstrate that the sense p16 and anti-sense Q03405 REA bicistronic construct significantly inhibits angiogenesis , induces apoptosis by deregulation of the PI3K - Akt pathway and downregulates alphavbeta 5 integrin receptor expression .

Analysis of breast cancer related gene expression using natural splines and the Cox proportional hazard model to identify prognostic associations . Many studies correlating gene expression data to clinical parameters assume a linear increase or decrease of the clinical parameter under investigation with the expression of a gene . We have studied genes encoding important breast cancer-related proteins using a model for survival-type data that is based on natural splines and the Cox proportional hazard model , thereby removing the linearity assumption . Expression data of 16 genes were studied in relation to metastasis-free probability in a cohort of 295 consecutive breast cancer patients treated at The Netherlands Cancer Institute . The independent predictive power for disease outcome of the 16 individual genes was tested in a multivariable model with known clinical and pathological risk factors . There is a linear relationship between increasing expression and a higher or lower hazard for distant metastasis for P03372 REA , Q15303 REA , P15692 REA , O96020 REA , Q15910 REA , and Q96NZ9 ; for P04626 REA , P21860 REA , P24385 REA , P24864 REA , O75530 REA , P61073 REA , P32248 REA , P48061 REA , and P05121 REA there is no clear increase or decrease ; and for P00533 REA there seems to be a non-linear relation . Multivariable analysis showed that the 70 - gene prognosis profile outperforms all the other variables in the model ( hazard-rate 5.4 , 95 % CI 2.5- 11.7 ; P = 0.000018 ) . P00533 REA - expression seems to have a non-linear relation with disease outcome , indicating that lower but also higher expression of P00533 REA are associated with worse outcome compared to intermediate expression levels ; the other genes show no or a linear relation .

Cardiac uptake of minocycline and mechanisms for in vivo cardioprotection . OBJECTIVES : The ability of minocycline to be transported into cardiac cells , concentrate in normal and ischemic myocardium , and act as a cardioprotector in vivo was examined . We also determined minocycline ' s capacity to act as a reducer of myocardial oxidative stress and matrix metalloproteinase ( MMP ) activity . BACKGROUND : The identification of compounds with the potential to reduce myocardial ischemic injury is of great interest . Tetracyclines are antibiotics with pleiotropic cytoprotective properties that accumulate in normal and diseased tissues . DB01017 SUB is highly lipophilic and has shown promise as a possible cardioprotector . However , minocycline ' s potential as an in vivo cardioprotector as well as the means by which this action is attained are not well understood . METHODS : Rats were subjected to 45 min of ischemia and 48 h of reperfusion . Animals were treated 48 h before and 48 h after thoracotomy with either vehicle or 50 mg / kg / day minocycline . Tissue samples were used for biochemical assays and cultured cardiac cells for minocycline uptake experiments . RESULTS : DB01017 SUB significantly reduced infarct size ( approximately 33 % ) , tissue P14780 REA activity , and oxidative stress . DB01017 SUB was concentrated approximately 24 - fold in normal ( 0.5 mmol / l ) and approximately 50 - fold in ischemic regions ( 1.1 mmol / l ) versus blood . Neonatal rat cardiac fibroblasts , myocytes , and adult fibroblasts demonstrated a time - and temperature-dependent uptake of minocycline to levels that approximate those of normal myocardium . CONCLUSIONS : Given the high intracellular levels observed and results from the assessment of in vitro antioxidant and MMP inhibitor capacities , it is likely that minocycline acts to limit myocardial ischemic injury via mass action effects .

The potential role of PD0332991 ( DB09073 MEN ) in the treatment of multiple myeloma . INTRODUCTION : Multiple myeloma ( MM ) remains an incurable malignancy indicating a need for continued investigation of novel therapies . Recent studies have highlighted the role of cyclin-dependent kinases ( CDK ) in the pathogenesis of MM . PD0332991 ( DB09073 MEN ) is an orally bioavailable , highly selective inhibitor of the P11802 REA / 6 - cyclin complex and downstream retinoblastoma protein ( Rb ) activation pathway that induces cell cycle arrest in the P55008 phase . AREAS COVERED : In this review , the authors summarize the role of the P11802 REA / 6 signaling pathway in MM . They also summarize the development of PD0332991 as a specific inhibitor of P11802 REA / 6 , and the reported preclinical and clinical data supporting the potential role of PD0332991 in MM . EXPERT OPINION : While PD0332991 is essentially cytostatic , inducing prolonged P55008 arrest , it enhances the cytotoxic effect of other agents effective in MM , including bortezomib and lenalidomide , as confirmed in early phase clinical trials . However , with a plethora of other drugs of different classes being tested in MM , further development of PD0332991 will depend on defining the most efficacious combination with least toxicity . An unexplored opportunity remains the potential protective effect of PD0332991 against lytic bone lesions of MM . The next few years are likely to better define the place of PD0332991 in the treatment of MM .

Expression of vitamin D3 receptor and retinoid receptors in human breast cancer : identification of potential heterodimeric receptors . DB00169 ( VD ) and all-trans-retinoic acid ( DB00755 MEN ) have been postulated as a novel treatment option for breast carcinoma . Since the combined effects of retinoids and VD derivatives are attributed to heterodimeric interactions between members of the nuclear receptor family , the expression patterns of the heterodimers formed by vitamin D3 receptor ( P11473 REA ) and the retinoid receptors RARs ( P10276 REA , P10826 REA and P13631 REA ) and RXRs ( RXR-alpha , RXR-beta and RXR-gamma ) have been studied by immunohistochemistry in benign and malignant breast tissues . Present results revealed that immunoexpressions to all receptor types studied were higher in both in situ and infiltrative carcinomas than in benign breast diseases . In a variable number of cases of infiltrative carcinoma , immunostaining appeared in the nucleus , whereas in the other two disorders immunostaining was only cytoplasmic . The correlation established between P11473 REA and the different isoforms of retinoid receptors revealed that P11473 REA seems to select mainly P10276 REA to form heterodimers and to exert their properties as transcription factor . The results of this study suggest that this heterodimer plays a critical role in cancer malignancy , and its presence indicates those patient groups presenting a better response to adjuvant therapies based on the combination of vitamin D and DB00755 MEN .

Spin trapping agent phenyl-N-tert-butylnitrone prevents diisopropylphosphorofluoridate-induced excitotoxicity in skeletal muscle of the rat . Indirect evidence suggests that reactive oxygen species ( ROS ) may mediate muscle fiber necrosis following muscle hyperactivity induced by the anticholinesterase diisopropylphosphorofluoridate ( DB00677 MEN ) . Pronounced muscle fasciculations and muscle fiber necrosis were seen when acetylcholinesterase ( P22303 REA ) activity was reduced to less than 30 % of control . The spin trapping agent phenyl-N-tert-butylnitrone ( PBN ) was used in vivo to directly assess the formation of ROS during DB00677 MEN ( 1.75 mg / kg , s . c . ) induced muscle hyperactivity . Pretreatment with PBN ( 300 mg / kg , i . p . ) , the concentration necessary for in vivo spin trapping , prevented muscle hyperactivity as well as necrosis and attenuated the DB00677 MEN induced P22303 REA inhibition otherwise seen in DB00677 MEN only treated rats . PBN had no effect when given after fasciculations were established . Muscle extracts from PBN and DB00677 MEN treated rats subjected to electron spin resonance ( P03372 REA ) spectroscopy tested negative for ROS . While the role of PBN as an antioxidant is well established , its prophylactic effect against excitotoxity induced by an P22303 REA inhibitor are due to its protection of P22303 REA , an unexpected non-antioxidant action .

Allele frequencies of single nucleotide polymorphisms ( SNPs ) in 40 candidate genes for gene-environment studies on cancer : data from population-based Japanese random samples . Knowledge of genetic polymorphisms in gene-environment studies may contribute to more accurate identification of avoidable risks and to developing tailor-made preventative measures . The aim of this study was to describe the allele frequencies of single nucleotide polymorphisms ( SNPs ) of select genes , which may be included in future gene-environment studies on cancer in Japan . SNP typing was performed on middle-aged Japanese men randomly selected from the general population in five areas of Japan . We genotyped and calculated allele frequencies of 153 SNPs located on 40 genes : P04798 REA , Q16678 REA , P11712 REA , P33261 REA , P05181 REA , P05093 REA , P11511 REA , P35869 REA , P03372 REA , Q92731 REA , ERRRG , P06401 REA , P07099 REA , P34913 REA , P37059 REA , P37058 REA , P28161 REA , P21266 REA , GSTT 2 , P09211 REA , NAT 1 , NAT 2 , P21964 REA , P07327 REA , P00325 REA , P00326 REA , P05091 REA , P35228 REA , NOS 3 , P01583 REA , P01584 REA , O15527 REA , P36639 REA [ P36639 REA ] , P14416 REA , P35462 REA , P21917 REA , P31645 REA , P04150 REA [ GCCR ] , P42898 REA , and P15559 REA . In the present study , the Japanese allele frequencies were verified by using nationwide population samples .

DB01017 SUB exerts multiple inhibitory effects on vascular endothelial growth factor-induced smooth muscle cell migration : the role of P27361 REA / 2 , PI3K , and matrix metalloproteinases . Widely used tetracycline antibiotics affect many cellular functions relevant to human vascular disease including cell proliferation , migration , and matrix remodeling . We examined whether minocycline inhibited human aortic smooth muscle cell ( HASMC ) migration induced by vascular endothelial growth factor ( P15692 REA ) . After the establishment of an optimal dose , minocycline treated HASMC were exposed to P15692 REA . HASMC migration , matrix metalloproteinase ( MMP ) - 2 and P14780 REA activities , mitogen-activated protein kinase ( MAPK ) , and phosphatidylinositol 3 - kinase ( PI3K ) phosphorylation were determined by smooth muscle cell ( SMC ) invasion assay , real-time polymerase chain reaction , zymograms , and Western blot analysis , respectively . We demonstrated that P15692 REA and platelet-derived growth factor ( PDGF ) - induced SMC migration in a dose-dependent manner . P14780 REA , but not P08253 REA , mRNA was increased during P15692 REA stimulation . P14780 REA activity was increased from 1.5- to 2.5- fold in a dose-dependent manner ( P < 0.05 ) . Both P27361 REA / 2 and PI3K / AKt pathways were activated during P15692 REA - induced HASMCs migration . We then demonstrated that minocycline can inhibit P15692 REA - induced HASMC migration ( P < 0.05 ) . The effects may be through the inhibition of P14780 REA mRNA transcription , protein activities and downregulation of P27361 REA / 2 and PI3K / Akt pathway phosphorylation . Our results indicated that minocycline exerts multiple effects on P15692 REA - induced SMC migration , including inhibition of P14780 REA mRNA transcription and protein activities and downregulating P27361 REA / 2 and PI3K signal pathways , suggesting minocycline may be a potentially therapeutic approach to inhibit disease process induced angiogenesis .

DB00104 MEN and the novel multireceptor ligand somatostatin receptor agonist pasireotide ( DB06663 ) block the adrenalectomy-induced increase in mitotic activity in male rat anterior pituitary . The novel somatostatin receptor agonist pasireotide binds with high affinity to somatostatin receptors P30872 REA , 2 , 3 , and 5 . Acting principally through the latter , it inhibits basal and P06850 REA - stimulated DB01285 secretion from the AtT 20 corticotroph cell line and DB01285 release from a proportion of human corticotroph adenomas both in vitro and in vivo . Data supporting an additional antiproliferative effect has led to pasireotide being explored as a potential therapy for patients with Cushing ' s disease . We have compared the effects of pasireotide and octreotide on adrenalectomy-induced mitotic and apoptotic activity in the male rat anterior pituitary . Adrenalectomized rats were treated with daily sc injections of vehicle , pasireotide , or octreotide . Changes in proliferation and apoptosis were determined 2-6 d postoperatively . DB06663 and octreotide had no effect on baseline pituitary cell turnover and no measurable effects on apoptosis . However , the wave of increased mitotic activity normally seen in the pituitary after adrenalectomy was completely abolished . Nevertheless , pasireotide and octreotide did not diminish the increase in DB01285 - immunopositive cell index after adrenalectomy , indicating that cell division and differentiation of hormonally null cells in the pituitary are under independent control . In conclusion , basal cell turnover in the pituitary is not inhibited by pasireotide or octreotide . Bilateral adrenalectomy stimulates differentiation of preexisting null cells into DB01285 - positive cells . Cell division after bilateral adrenalectomy occurs in a specific subpopulation of hormonally null cells that are equally sensitive to the antiproliferative effects of pasireotide and octreotide , implicating P30874 REA receptors in this antimitotic response .

The role of atorvastatin in regulating the immune response leading to vascular damage in a model of Kawasaki disease . Superantigens have been implicated in a number of diseases including Kawasaki disease ( KD ) , a multi-system vasculitis resulting in coronary artery aneurysms . We have characterized a murine disease model in which coronary arteritis is induced by a novel superantigen found in Lactobacillus casei cell wall extract ( LCWE ) . Using this animal model of KD , we have identified three pathogenic steps leading to coronary artery aneurysm formation . These steps include T cell activation and proliferation , production of the proinflammatory cytokine tumour necrosis factor ( P01375 REA ) - α and up-regulation of matrix metalloproteinase 9 ( P14780 REA ) , an elastolytic protease . In addition to their cholesterol-lowering effects , 3 - hydroxy - 3 - methylglutaryl ( HMG ) coenzyme A ( DB01992 ) reductase inhibitors ( statins ) have pleotropic immunomodulatory properties . Thus , we examined the effect of atorvastatin in modulating each of these three critical pathogenic processes leading to aneurysm formation in the disease model . DB01076 MEN inhibited lymphocyte proliferation in response to superantigen stimulation in a dose-dependent manner . This inhibition was also observed for production of soluble mediators of inflammation including interleukin ( IL ) - 2 and P01375 REA - α . The inhibitory effect on proliferation was rescued completely by mevalonic acid , confirming that the mechanism responsible for this inhibitory activity on immune activation was inhibition of P04035 REA . Similarly , P01375 REA - α-induced P14780 REA production was reduced in a dose-dependent manner in response to atorvastatin . Inhibition of extracellular-regulated kinase ( P29323 REA ) phosphorylation appears to be the mechanism responsible for inhibition of P14780 REA production . In conclusion , atorvastatin is able to inhibit critical steps known to be important in the development of coronary aneurysms , suggesting that statins may have therapeutic benefit in patients with KD .

Understanding the molecular mechanism of blood-brain barrier damage in an experimental model of Japanese encephalitis : correlation with minocycline administration as a therapeutic agent . The blood-brain barrier ( BBB ) serves to protect the central nervous system ( CNS ) from damage by exogenous molecules . Japanese encephalitis ( JE ) , caused by a neurotropic flavivirus , leads to inflammation in the CNS , neuronal death and also compromises the structural and functional integrity of the BBB . DB01017 SUB , a semisynthetic tetracycline , has been found to be broadly protective in neurological disease models featuring inflammation and cell death and at present , is being evaluated in clinical trials . In the present study , we propose that the neuroprotective role of minocycline in experimental models of JE extends also to the protection of the BBB . Damage to the BBB was assessed by Evan ' s blue dye exclusion test after minocycline treatment following Japanese encephalitis virus ( JEV ) infection . A breakdown of the BBB occurred in mice inoculated intravenously with JEV . This resulted in leakage of protein-bound Evan ' s blue dye into the brain tissue . Semi-quantitative RT-PCR revealed an up-regulation of chemokine receptors and adhesion molecules following JEV infection . Immunostaining showed leukocyte and neutrophil infiltration following JEV infection . Intraperitoneal injection of minocycline , beginning 24h post-JEV infection , abrogated the effects by reducing BBB damage , decreasing expression of P35228 REA , Cox - 2 , P15692 REA and also by reducing the elevated level of transcript of chemokine receptors and adhesion molecules in the brain . Matrix metalloproteinases ( MMPs ) are known to disrupt the BBB and minocycline was found to significantly decrease the activity of P14780 REA in brain tissue homogenates . Thus , minocycline , administered at a clinically relevant time , appears to maintain blood-brain barrier integrity following JEV infection .

DB01017 SUB inhibits malignant ascites of ovarian cancer through targeting multiple signaling pathways . OBJECTIVES : To evaluate the effect of minocycline on the expression of cytokines and growth factors responsible for malignant ascite formation . METHODS : In vitro , cells obtained from malignant ascites were pre-treated with minocycline ( 0-100 μmol / L ) and exposed briefly to hypoxia . In vivo , female nude mice bearing OVCAR - 3 tumors were treated orally in drinking water with minocycline for 4 weeks . Plasma , ascites , and tumors were analyzed . RESULTS : DB01017 SUB blocked hypoxia-induced surge in interleukin - 6 ( P05231 REA ) , its soluble receptor ( sIL - 6R ) and vascular endothelial growth factor ( P15692 REA ) levels in concentration-dependent manner . In mice , orally administered minocycline led to dramatic reduction in tumor weight and malignant ascite volume . P05231 REA , sIL 6R and in particular P15692 REA levels were highly suppressed in plasma , ascite fluid and tumor tissue by minocycline . In addition , tumors from minocycline treated mice expressed profoundly lower levels of phosphorylated extracellular regulated kinases ( p-Erk 1/2 ) and p-Akt . DB01017 SUB was also effective at suppressing transforming growth factor beta ( TGF-β 1 ) and increasing vascular endothelial cadherin ( P33151 REA ) expression thereby providing molecular confirmation for its effects on malignant ascite formation . CONCLUSION : Orally administered minocycline is highly effective in suppressing ovarian cancer-induced malignant ascites by targeting cytokines and growth factors essential for tumor growth and malignant ascite formation .

[ Moclobemide ( DB01171 MEN ) , the first P21397 REA - inhibitor : really something new ? ] .

Associations between single nucleotide polymorphisms in cellular viral receptors and attachment factor-related genes and humoral immunity to rubella vaccination . BACKGROUND : Viral attachment and cell entry host factors are important for viral replication , pathogenesis , and the generation and sustenance of immune responses after infection and / or vaccination , and are plausible genetic regulators of vaccine-induced immunity . METHODS : Using a tag-SNP approach in candidate gene study , we assessed the role of selected cell surface receptor genes , attachment factor-related genes , along with other immune genes in the genetic control of immune response variations after live rubella vaccination in two independent study cohorts . RESULTS : Our analysis revealed evidence for multiple associations between genetic variants in the P15151 REA , Q92692 REA , Q9NNX6 / Q9NNX6 , P10826 REA , Q16653 , P05231 REA and other immune function-related genes and rubella-specific neutralizing antibodies after vaccination ( meta p-value < 0.05 ) . CONCLUSION : Our results indicate that multiple SNPs from genes involved in cell adhesion , viral attachment , and viral entry , as well as others in genes involved in signaling and / or immune response regulation , play a role in modulating humoral immune responses following live rubella vaccination .

Angiotensin II promotes poly ( ADP-ribosyl ) ation of c-Jun / c-Fos in cardiac fibroblasts . Although c-Jun / c-Fos ( activator protein 1 , P05412 REA ) contributes importantly to Ang II-induced cardiac fibrosis through induction of extracellular matrix protein over-expression in cardiac fibroblasts , the mechanism by which Ang II promotes c-Jun / c-Fos transactivation remains unclear . In this study , we demonstrated that c-Fos and c-Jun were poly ( ADP-ribosyl ) ated in cultured cardiac fibroblasts . Southwestern blot and EMSA assays showed that incubation of nuclear extracts with NAD ( + ) and active DNA increased the basal DNA binding activities of c-Jun ( 31.0+ / -1.0 % , P < 0.01 ) and P05412 REA ( 14.2+ / -3.1 % , P < 0.01 ) ; incubation of recombinant c-Fos or / and c-Jun with P09874 REA , NAD ( + ) and active DNA increased the basal DNA binding activities of c-Jun ( 48.3+ / -4.2 % , P < 0.01 ) and P05412 REA ( 21.2+ / -1.5 % , P < 0.01 ) . Treatment with Ang II promoted P09874 REA activation and enhanced poly ( ADP-ribosyl ) ation of c-Fos ( 14.1+ 1.1 % , P < 0.01 ) and c-Jun ( 15.5+ / -5.6 % , P < 0.01 ) . Ang II also increased the basal DNA binding activities of c-Jun ( 13.5+ / -2.4 % , P < 0.01 ) and P05412 REA ( 18.7+ / -3.5 % , P < 0.01 ) in cultured cells . Inhibition of P09874 REA by PJ34 or siRNA effectively prevented Ang II-induced increases in the DNA binding of c-Jun and P05412 REA , and decreased P05412 REA - driven transcription ( including collagen Ialpha 1 and IIIalpha 1 , P14780 REA and P01033 REA ) . This study illustrated that c-Jun and c-Fos were poly ( ADP-ribosyl ) ated by P09874 REA , and poly ( ADP-ribosyl ) ation enhanced the DNA binding of P05412 REA . Ang II promoted poly ( ADP-ribosyl ) ation of c-Jun and c-Fos through activation of P09874 REA and , subsequently , enhanced P05412 REA - driven transcription in cardiac fibroblasts .

DB01017 SUB protects Schwann cells from ischemia-like injury and promotes axonal outgrowth in bioartificial nerve grafts lacking Wallerian degeneration . DB01017 SUB , a broad-spectrum antimicrobial tetracycline , acts neuroprotectively in ischemia . Recently , however , minocycline has been revealed to have ambiguous effects on nerve regeneration . Thus its effects in a rat sciatic nerve transplantation model and on cultivated Schwann cells stressed by oxygen glucose deprivation ( OGD ) were studied . The negative effect of minocycline on Wallerian degeneration , the essential initial phase of degeneration / regeneration after nerve injury , that was recently demonstrated , was excluded by using predegenerated nerve and Schwann cell-enriched muscle grafts , both free of Wallerian degeneration . They were compared with common nerve grafts . The principle findings were that in vitro minocycline provided protective effects against OGD-induced death of Schwann cells by preventing permeability of the mitochondrial membrane . It suppressed the OGD-mediated induction of HIF - 1alpha and Q07812 REA , and stabilized / induced BCL - 2 . P99999 REA release and cleavage of procaspase - 3 were diminished ; release and translocation of O95831 and cytotoxic cleavage of actin into fractin were stopped . In common nerve grafts , minocycline , besides its direct anti-ischemic effect , hampered revascularization by down-regulation of P14780 REA and P15692 REA prolonging ischemia and impeding macrophage recruitment . In bioartificial nerve grafts that were free of Wallerian degeneration and revealed lower immunogenicity , minocycline aided the regeneration process . Here , the direct anti-ischemic effect of minocycline on Schwann cells , which are mandatory for successful peripheral nerve regeneration , dominated the systemic anti-angiogenic / pro-ischemic effects . In common nerve grafts , however , where Wallerian degeneration is a prerequisite , the anti-angiogenic and macrophage-depressing effect is an obstacle for regeneration .

Analysis of biological and technical variability in gene expression assays from formalin-fixed paraffin-embedded classical Hodgkin lymphomas . DB03843 - fixed paraffin-embedded ( FFPE ) tissues are invaluable sources of biological material for research and diagnostic purposes . In this study , we aimed to identify biological and technical variability in RT-qPCR TaqMan ® assays performed with FFPE-RNA from lymph nodes of classical Hodgkin lymphoma samples . An Q9UNW9 - nested 6 - level design was employed to evaluate P10415 REA , P42574 REA , Q15306 REA , P61626 REA and P42224 REA gene expression . The most variable genes were P42574 REA ( low expression ) and P61626 REA ( high expression ) . Total variability decreased after normalization for all genes , except by P61626 REA . Genes with moderate and low expression were identified and suffered more the effects of the technical manipulation than high-expression genes . Pre-amplification was shown to introduce significant technical variability , which was partially alleviated by lowering to a half the amount of input RNA . Ct and Cy0 quantification methods , based on cycle-threshold and the kinetic of amplification curves , respectively , were compared . Cy0 method resulted in higher quantification values , leading to the decrease of total variability in P42574 REA and P61626 REA genes . The mean individual noise was 0.45 ( 0.31 to 0.61 SD ) , indicating a variation of gene expression over ~ 1.5 folds from one case to another . We showed that total variability in RT-qPCR from FFPE-RNA is not higher than that reported for fresh complex tissues , and identified gene - , and expression level-sources of biological and technical variability , which can allow better strategies for designing RT-qPCR assays from highly degraded and inhibited samples .

DB01017 SUB reduces renal microvascular leakage in a rat model of ischemic renal injury . Tetracyclines exhibit significant anti-inflammatory properties , inhibit matrix metalloproteinases ( MMPs ) , and are protective in models of ischemia-reperfusion injury ( IRI ) . Both inflammatory cascades and MMP activation have been demonstrated to modulate microvascular permeability . Because increased microvascular permeability occurs during IRI in a variety of organ systems including the kidney , we hypothesized that minocycline , a semisynthetic tetracycline , would diminish microvascular leakage during renal IRI . To test this hypothesis , we used intravital 2 - photon microscopy to examine leakage of fluorescent dextrans from the vasculature in a rodent model of IRI . DB01017 SUB significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Although minocycline diminished leukocyte accumulation in the kidney following ischemia , areas of leukocyte accumulation did not correlate with areas of microvascular permeability in either the saline - or minocycline-pretreated animals . DB01017 SUB diminished the perivascular increase in P08253 REA and P14780 REA , as well as the increase in P08253 REA activity 24 h after ischemia . ABT - 518 , a specific inhibitor of P08253 REA and P14780 REA , also significantly reduced the extent of dextran ( 500 kDa ) leakage from the renal microvasculature 24 h after ischemia . Our results indicate that minocycline mitigates the renal microvascular permeability defect following IRI . This effect is spatially distinct from the effect of minocycline on leukocyte accumulation and may be related to diminished activity of MMPs on the integrity of the perivascular matrix .

DB01017 SUB prevents dynorphin-induced neurotoxicity during neuropathic pain in rats . Despite many advances , our understanding of the involvement of prodynorphin systems in the development of neuropathic pain is not fully understood . Recent studies suggest an important role of neuro-glial interactions in the dynorphin effects associated with neuropathic pain conditions . Our studies show that minocycline reduced prodynorphin mRNA levels that were previously elevated in the spinal and / or dorsal root ganglia ( Q86YR7 ) following sciatic nerve injury . The repeated intrathecal administration of minocycline enhanced the analgesic effects of low-dose dynorphin ( 0.15 nmol ) and U50 , 488H ( 25-100 nmol ) and prevented the development of flaccid paralysis following high-dose dynorphin administration ( 15 nmol ) , suggesting a neuroprotective effect . DB01017 SUB reverts the expression of IL - 1β and P05231 REA mRNA within the spinal cord and IL - 1β mRNA in Q86YR7 , which was elevated following intrathecal administration of dynorphin ( 15 nmol ) . These results suggest an important role of these proinflammatory cytokines in the development of the neurotoxic effects of dynorphin . Similar to minocycline , a selective inhibitor of P14780 REA ( P14780 REA levels are reduced by minocycline ) exerts an analgesic effect in behavioral studies , and its administration prevents the occurrence of flaccid paralysis caused by high-dose dynorphin administration ( 15 nmol ) . In conclusion , our results underline the importance of neuro-glial interactions as evidenced by the involvement of IL - 1β and P05231 REA and the minocycline effect in dynorphin-induced toxicity , which suggests that drugs that alter the prodynorphin system could be used to better control neuropathic pain .

Statin Modulation of Human T-Cell Proliferation , IL - 1β and Q16552 REA Production , and IFN-γ T Cell Expression : Synergy with Conventional Immunosuppressive Agents . P04035 REA inhibitors ( statins ) have been demonstrated to be immunomodulatory for human immune-mediated disease and in experimental models . The aim of this study was to compare statin-mediated immunosuppressive effects on human T-cell responses in vitro with those of conventional immunosuppressives ( dexamethasone , cyclosporin A ( DB00091 ) , mycophenolate , and rapamycin ) . Statins ( atorvastatin , lovastatin , and simvastatin ) were investigated for their modulatory effects on human PBMC viability , cytokine profiles , and T-cell proliferation . At concentrations that inhibited anti-CD 3/28- stimulated T-cell proliferation ( P < 0.01 ) , simvastatin significantly decreased intracellular P01730 REA ( + ) T-cell expression of IFN-γ ( P < 0.01 ) to levels similar to those induced by conventional immunosuppressives . DB01076 MEN and lovastatin also decreased IFN-γ expression , although to a lesser degree ( P < 0.05 ) . All three statins reduced levels of Q16552 REA production ( P < 0.01 ) . However , in response to anti-CD 3/28 stimulation , simvastatin significantly upregulated IL - 1β production ( P < 0.05 ) . The profile of cytokines produced in response to anti-CD 3/28 stimulation was similar when both atorvastatin and dexamethasone were added as compared with dexamethasone alone , suggesting that atorvastatin can synergise with dexamethasone with respect to immunomodulation of cytokines . This data supports the hypothesis of selective statin-mediated immunomodulatory effects on human immune cells .

Chemotactic and cytotoxic effects of minocycline on human retinal pigment epithelial cells . PURPOSE : To reveal the effects of minocycline , an anti-inflammatory and neuroprotective agent , on the viability and physiological properties of retinal pigment epithelial ( Q96AT9 ) cells and to compare the effects with those of triamcinolone acetonide . METHODS : The proliferation of human Q96AT9 cells in vitro was investigated with a bromodeoxyuridine immunoassay ; chemotaxis was examined with a Boyden chamber assay . Cell viability was determined by trypan blue exclusion . The gene expression of growth factors and P14780 REA was determined with real-time RT-PCR , and the secretion of P15692 REA was examined with ELISA . The phosphorylation of p38 MAPK and P27361 REA / 2 proteins was determined with Western blot analysis . RESULTS : DB01017 SUB at low concentrations ( 50 nM - 20 microM ) stimulated chemotaxis and decreased the proliferation of Q96AT9 cells . DB01017 SUB at high concentrations ( above 5 microM ) decreased the viability of Q96AT9 cells through the induction of cell necrosis . The chemotactic effect of minocycline was mediated by the stimulation of autocrine PDGF signaling and the activation of p38 MAPK . DB01017 SUB promoted the expression of PDGF-B , P14210 REA , P15692 REA , and P14780 REA and increased the amounts of phosphorylated p38 and P27361 REA / 2 proteins in Q96AT9 cells . DB00620 reduced PDGF-evoked chemotaxis and P15692 REA expression and secretion and had no significant effects on cell viability and proliferation . DB00620 did not reverse the effects of minocycline on cell proliferation , chemotaxis , or viability or the expression of P15692 REA . CONCLUSIONS : Low-dose minocycline induces the activation of Q96AT9 cells , as indicated by the activation of p38 and P27361 REA / 2 and by enhanced chemotaxis mediated by autocrine PDGF signaling . High-dose minocycline induces Q96AT9 cell degeneration .

DB01017 SUB increases phosphorylation and membrane insertion of neuronal GluR 1 receptors . The tetracycline antibiotic minocycline beneficially affects neuronal functioning and also inhibits the enzyme P09917 REA ( 5 - P28300 REA ) . We hypothesized that similar to 5 - P28300 REA inhibitors , minocycline may increase phosphorylation and membrane insertion of the glutamate receptor GluR 1 . The experiments were performed in primary cultures of mouse striatal neurons and in the prefrontal cortex and striatum of minocycline-treated mice . In vitro , low micromolar minocycline concentrations increased GluR 1 phosphorylation at Ser 845 and Ser 831 and increased the surface content of GluR 1 . DB01017 SUB also increased GluR 1 phosphorylation in vivo . Increased GluR 1 phosphorylation and minocycline treatment have been associated with antidepressant and memory-enhancing activities . Direct consequences of minocycline-increased GluR 1 phosphorylation are yet to be established .

Effects of phenytoin , ketamine , and atropine methyl nitrate in preventing neuromuscular toxicity of acetylcholinesterase inhibitors soman and diisopropylphosphorofluoridate . Toxic manifestations of acetylcholinesterase inhibitors ( P22303 REA - I ) include muscle twitching and muscle fiber necrosis , in addition to muscarinic manifestations of acetylcholine excess . The P22303 REA - Is pinacolyl methylphosphonofluoridate ( soman ) or diisopropylphosphorofluoridate ( DB00677 MEN ) were administered to rats to produce spontaneous muscle fiber discharges . Soman produced discharges that arose primarily from the central nervous system ( CNS ) , while those due to DB00677 MEN were generated from the peripheral nerves as well as the CNS . Three drugs were tested for their potential to reduce muscle fiber discharges : atropine methyl nitrate ( Q9BXJ7 ) , ketamine , and phenytoin . DB01221 caused a significant decrease in discharges of CNS origin , while Q9BXJ7 and phenytoin had no effect . For muscle fiber discharges of peripheral origin , all three drugs produced a significant drop in muscle fiber discharges , but phenytoin showed slightly more efficacy than the others . P22303 REA - I-induced muscle hyperactivity arises from actions on the CNS and on the peripheral nerve in varying proportions for different P22303 REA - Is . Treatment for the toxicity of P22303 REA - Is on muscle may be accomplished by administering drugs with distinctive pharmacological actions at target sites in the CNS and peripheral nervous system ( PNS ) where P22303 REA - Is exert their effects . By attenuating the effects of P22303 REA - Is at specific CNS or PNS sites , the neuromuscular toxicity can be reduced in a manner specific to the characteristic sites of toxicity of each P22303 REA - I .

MicroRNA -384-5 p regulates ischemia-induced cardioprotection by targeting phosphatidylinositol -4,5- bisphosphate 3 - kinase , catalytic subunit delta ( PI3K p110δ ) . MicroRNAs ( miRNAs ) are a novel class of powerful , endogenous regulators of gene expression . This study identified 16 differentially expressed miRNAs in ischemic myocardium of rats using TaqMan Low Density Array . In addition , bioinformatics analyses , such as Gene ontology and Pathway assays , were applied to determine the apoptosis pathway , only regulated by miR -384-5 p , and all the associated target genes ( O00329 REA , Q08209 REA , P16298 REA , P63098 REA , P42574 REA and P01583 REA ) . These target genes , besides P42338 REA , were shown to be significantly up-regulated by qRT-PCR assay , which further suggested that O00329 REA , Q08209 REA , P63098 REA , P42574 REA , P01583 REA could be regulated by miR -384-5 p . MTT , Western blot , qRT-PCR and luciferase assays were used to investigate the role of miR -384-5 p in myocardial ischemia . We found that cleaved caspase 3 expression was up-regulated by miR -384-5 p and down-regulated by miR -384-5 p inhibitor suggesting that apoptosis pathway was regulated by miR -384-5 p . We also found that miR -384-5 p suppressed cell viability while miR -384-5 p inhibitor improved it , confirming H9c2 cell survival was affected by miR -384-5 p . In addition , the O00329 REA protein level in H9c2 cells was up-regulated by miR -384-5 p inhibitor . We found that miR -384-5 p expression level decreased and O00329 REA protein level increased in both ischemic myocardium of rats and hypoxic H9c2 cells , and that miR -384-5 p suppress O00329 REA expression through a miR -384-5 p binding site within the 3 ' untranslational region of O00329 REA . These results show that miR -384-5 p , an important protecting factor , plays a significant role in cardioprotection by regulating O00329 REA in myocardial ischemia .

Glioma-associated microglial P14780 REA expression is upregulated by O60603 REA signaling and sensitive to minocycline . The invasiveness of malignant gliomas is one of the major obstacles in glioma therapy and the reason for the poor survival of patients . Glioma cells infiltrate into the brain parenchyma and thereby escape surgical resection . Glioma associated microglia / macrophages support glioma infiltration into the brain parenchyma by increased expression and activation of extracellular matrix degrading proteases such as matrix metalloprotease ( MMP ) 2 , P14780 REA and membrane-type 1 MMP . In this work we demonstrate that , P14780 REA is predominantly expressed by glioma associated microglia / macrophages in mouse and human glioma tissue but not by the glioma cells . Supernatant from glioma cells induced the expression of P14780 REA in cultured microglial cells . Using mice deficient for different Toll-like receptors we identified O60603 REA / 6 as the signaling pathway for the glioma induced upregulation of microglial P14780 REA . Also in an experimental mouse glioma model , O60603 REA deficiency attenuated the upregulation of microglial P14780 REA . Moreover , glioma supernatant triggered an upregulation of O60603 REA expression in microglia . Both , the upregulation of P14780 REA and O60603 REA were attenuated by the antibiotic minocycline and a p38 mitogen-activated protein kinase antagonist in vitro . DB01017 SUB also extended the survival rate of glioma bearing mice when given to the drinking water . Thus glioma cells change the phenotype of glioma associated microglia / macrophages in a complex fashion using O60603 REA as an important signaling pathway and minocycline further proved to be a potential candidate for adjuvant glioma therapy .