interacts_with DB00382?
Effects of repeated administration of tetrahydroaminoacridine ( DB00382 ) on muscarinic receptor subtypes in the rat brain . The effects of a chronic treatment ( 21 days ) with the acetylcholinesterase ( P22303 ) inhibitor tetrahydroaminoacridine ( DB00382 ) on muscarinic receptors subtypes were investigated at various times after the last administration of the drug , in various brain areas including cortex , striatum , hippocampus and cerebellum . Forty eight hours after the end of chronic DB00382 treatment , the number of muscarinic receptors , labelled with [(3)H] Q5H8A3 , was significantly lowered in the cortex and the striatum but not in the hippocampus or cerebellum . High affinity pirenzepine binding sites ( M(1) receptors ) , directly assayed using [(3)H]pirenzepine saturation assays or estimated by pirenzepine [(3)H] Q5H8A3 competition , were lowered only in the cortex and in the striatum of DB00382 -treated rats . In contrast , the number of low affinity pirenzepine sites ( M(2) receptors ) , was not significantly modified . At shorter wash-out period ( 18 h ) , the density of M(1) receptors decreased by 26 , 46 and 52 % in the hippocampus , cerebral cortex and striatum , respectively . In all cases , K(d) values remained unchanged suggesting that the loss of M(1) sites was not due to a modification of radioligand affinity for the receptors . Although DB00382 displayed a micromolar affinity for M(1) and M(2) receptors in vitro , this P22303 inhibitor did not interfere with the receptor assays since no trace of residual free DB00382 was detected in rat brain at 48 h post-treatment . These results suggest that chronic treatment with DB00382 produced a selective down-regulation of M(1) receptors ; they also indicate that these receptors may be regulated differently in cortical , striatal , hippocampal or cerebellar regions . TATA-driven transcriptional initiation and regulation of the rat serotonin P08908 receptor gene . The transcriptional initiation and regulation of the rat serotonin P08908 receptor gene were characterized . By three types of analyses , a single brain-specific site of transcriptional initiation was localized to -967 bp upstream of the translation initiation codon that is utilized both in hippocampus and in the rat raphe RN46A cell line . This major site of transcriptional initiation was located 58 bp downstream from a consensus TATA element , suggesting TATA-driven transcription of the rat P08908 receptor . To identify the promoter activity of the receptor gene , progressive 5' deletions of the -2,719/-117-bp fragment of the P08908 promoter linked to luciferase gene were transfected into P08908 -negative ( pituitary GH4C1 , Q9BTT4 myoblast , and P13671 glioma ) and P08908 -positive ( septal SN-48 and raphe RN46A ) cell lines . Enhancer regions were identified within a fragment between nucleotides -426 and -117 that selectively enhanced transcription in P08908 -positive cells . A nonselective enhancer/promoter that mediated expression in all cell lines was located upstream between -1,519 and -426 bp in a DNA segment containing consensus TATA , CCAAT , SP-1 , and AP-1 elements as well as a poly-GT26 dinucleotide repeat . Strong repression of transcription in all cell lines was conferred by the region upstream of -1,519 bp that contains a 152-bp DNA segment with > 80 % identity to RANTES , tumor necrosis factor-beta , and other immune system genes . Our results indicate that TATA-driven expression of the P08908 receptor is regulated by a novel proximal tissue-specific enhancer region , a nonselective promoter , and an upstream repressor region that is distinct from previously identified neuron-specific repressors . A fully human recombinant IgG-like bispecific antibody to both the epidermal growth factor receptor and the insulin-like growth factor receptor for enhanced antitumor activity . Both the epidermal growth factor receptor ( P00533 ) and the insulin-like growth factor receptor ( IGFR ) have been implicated in the tumorigenesis of a variety of cancers . Here we propose that simultaneous targeting of both receptors with a bispecific antibody would lead to enhanced antitumor activity . To this end , we produced a recombinant human IgG-like bispecific antibody , a Di-diabody , using the variable regions from two antagonistic antibodies : DB05774 to P00533 and DB05759 to IGFR . The Di-diabody binds to both P00533 and IGFR and effectively blocked both P01133 - and IGF-stimulated receptor activation and tumor cell proliferation . The Di-diabody also inherited the biological properties from both of its parent antibodies ; it triggers rapid and significant IGFR internalization and degradation and mediates effective antibody-dependent cellular cytotoxicity in a variety of tumor cells . Finally , the Di-diabody strongly inhibited the growth of two different human tumor xenografts in vivo . Our results underscore the benefits of simultaneous targeting of two tumor targets with bispecific antibodies . A Nile blue based infrared fluorescent probe : imaging tumors that over-express cyclooxygenase-2 . The first Golgi-localized cyclooxygenase-2 ( P35354 ) -specific near-infrared ( Q9Y3T9 ) fluorescent probe , Niblue- P13671 -IMC , able to detect cancer cells , was designed . Importantly , Niblue- P13671 -IMC preferentially labeled the tumors in a mouse tumor model with deep tissue penetration capacity . It may be a promising molecular tool for guiding tumor resection during surgery . Short-term biomarker modulation prevention study of anastrozole in women at increased risk for second primary breast cancer . The selective estrogen receptor modulators ( SERM ) , Tamoxifen and raloxifen reduce risk breast cancer . Patient acceptance of SERMs for breast cancer prevention is low due to toxicities . New agents with a better toxicity profile are needed . P11511 inhibitors ( AI ) reduce the risk of contralateral breast cancer and risk of new breast cancer in high risk women . However , the mechanism by which AIs reduce breast risk is not known . Surrogate biomarkers are needed to evaluate the effect of preventive agents . The objective of this prospective short-term prevention study was to evaluate the effect of anastrozole on biomarkers in breast tissue and serum of women at increased risk for developing a contralateral breast cancer . Women with a history of stage I , II breast cancer who started anastrozole for standard adjuvant treatment were eligible . Patients underwent baseline fine needle aspiration of the unaffected breast and serum collection for biomarker analysis before starting anastrozole at 1 mg per oral/day and again at 6 months . Biomarkers included changes in cytology , insulin-like growth factor 1 ( DB01277 ) , P08833 ( P08833 ) , and P17936 . Thirty-seven patients were enrolled . There was a significant modulation in serum P08833 levels between pre- and postsamples ( P = 0.02 ) . No change was observed in DB01277 , P17936 , and breast cytology.We showed a significant modulation of P08833 levels with six months anastrozole . DB01217 is currently being studied as a prevention agent in a large phase III trial and our results provide support for continued evaluation of P08833 as a surrogate endpoint biomarker in prospective breast chemoprevention studies . NFκB inhibitors induce cell death in glioblastomas . Identification of novel target pathways in glioblastoma ( GBM ) remains critical due to poor prognosis , inefficient therapies and recurrence associated with these tumors . In this work , we evaluated the role of nuclear-factor-kappa-B ( NFκB ) in the growth of GBM cells , and the potential of NFκB inhibitors as antiglioma agents . NFκB pathway was found overstimulated in GBM cell lines and in tumor specimens compared to normal astrocytes and healthy brain tissues , respectively . Treatment of a panel of established GBM cell lines ( U138MG , U87 , U373 and P13671 ) with pharmacological NFκB inhibitors ( BAY117082 , parthenolide , MG132 , curcumin and arsenic trioxide ) and NFκB-p65 siRNA markedly decreased the viability of GBMs as compared to inhibitors of other signaling pathways such as MAPKs ( P29323 , JNK and p38 ) , PKC , P00533 and PI3K/Akt . In addition , NFκB inhibitors presented a low toxicity to normal astrocytes , indicating selectivity to cancerous cells . In GBMs , mitochondrial dysfunction ( membrane depolarization , bcl-xL downregulation and cytochrome c release ) and arrest in the G2/M phase were observed at the early steps of NFκB inhibitors treatment . These events preceded sub- P55008 detection , apoptotic body formation and caspase-3 activation . Also , NFκB was found overstimulated in cisplatin-resistant P13671 cells , and treatment of GBMs with NFκB inhibitors overcame cisplatin resistance besides potentiating the effects of the chemotherapeutics , cisplatin and doxorubicin . These findings support NFκB as a potential target to cell death induction in GBMs , and that the NFκB inhibitors may be considered for in vivo testing on animal models and possibly on GBM therapy . Synthesis , biological activity and HPLC validation of 1,2,3,4-tetrahydroacridine derivatives as acetylcholinesterase inhibitors . The synthesis and biochemical evaluation of new hybrids of tacrine ( DB00382 ) and 4-fluorobenzoic acid ( 4-FBA ) possessing activity towards acetylcholinesterase ( P22303 ) and butyrylcholinesterase ( BuChE ) inhibition are presented . The compounds of interest were obtained from the reaction of activated 4-FBA and diamino derivatives of 1,2,3,4-tetrahydroacridine . The compounds P13671 -2KW/HCl , P13671 -4KW/HCl and P13671 -3KW/HCl have four-fold higher antiacetylcholinesterase activity than DB00382 . All of the acquired compounds present higher selectivity towards P22303 than DB00382 and lower selectivity towards BuChE . In addition , a rapid , selective and stability-indicating HPLC method was developed and validated for the determination of P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl . DB00382 and 4-FBA were found to be the main impurities . Chromatographic separation was achieved isocratically on a Waters Symmetry C18 150 × 3.9 mm , 4 μm column with a mobile phase of acetonitrile/buffer ( 17 mM sodium dodecyl sulphate and 8.3 mM sodium dihydrogen phosphate , 50:50 v/v ) ( overall pH 4 ) . A 1.5 ml/min flow rate and a 247 nm wavelength were chosen for this method . P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl were subjected to acidic and basic hydrolysis , chemical oxidation , thermal exposition at 60 °C and intense UV light . The limits of detection ( LOD ) and quantification ( LOQ ) were less than 2 μg/ml and 6 μg/ml for P13671 -2KW/HCl , P13671 -3KW/HCl and P13671 -4KW/HCl , 0.04 μg/ml and 0.12 μg/ml for DB00382 , 0.42 μg/ml and 1.41 μg/ml for 4-FBA , respectively . miR-17 family of microRNAs controls O15520 -mediated embryonic lung epithelial branching morphogenesis through Q16539 and P40763 regulation of E-Cadherin distribution . The miR-17 family of microRNAs has recently been recognized for its importance during lung development . The transgenic overexpression of the entire miR-17-92 cluster in the lung epithelium led to elevated cellular proliferation and inhibition of differentiation , while targeted deletion of miR-17-92 and miR-106b-25 clusters showed embryonic or early post-natal lethality . Herein we demonstrate that miR-17 and its paralogs , miR-20a , and miR-106b , are highly expressed during the pseudoglandular stage and identify their critical functional role during embryonic lung development . Simultaneous downregulation of these three miRNAs in explants of isolated lung epithelium altered O15520 induced budding morphogenesis , an effect that was rescued by synthetic miR-17 . E-Cadherin levels were reduced , and its distribution was altered by miR-17 , miR-20a and miR-106b downregulation , while conversely , beta-catenin activity was augmented , and expression of its downstream targets , including Bmp4 as well as Fgfr2b , increased . Finally , we identified Stat3 and Mapk14 as key direct targets of miR-17 , miR-20a , and miR-106b and showed that simultaneous overexpression of Stat3 and Mapk14 mimics the alteration of E-Cadherin distribution observed after miR-17 , miR-20a , and miR-106b downregulation . We conclude that the mir-17 family of miRNA modulates O15520 -FGFR2b downstream signaling by specifically targeting Stat3 and Mapk14 , hence regulating E-Cadherin expression , which in turn modulates epithelial bud morphogenesis in response to O15520 signaling . Preparation of phosphorylated starch by dry-heating in the presence of pyrophosphate and its calcium-phosphate solubilizing ability . Starch was phosphorylated through dry-heating in the presence of pyrophosphate at various conditions , and the characteristics of phosphorylated starch ( PS ) were examined . Starch phosphorylation increases as the pH increases from 3 to 6 , but diminishes at pH 7 . Increased temperatures enhance phosphorylation . Data from (31)P NMR suggests that starch phosphorylation occurs mainly at the P01024 -OH and P13671 -OH of the glucose residue . The phosphate linkage is mainly due to monostarch monophosphate . Although starch had almost no calcium phosphate-solubilising capacity , this capacity was markedly enhanced by phosphorylation . X-ray diffraction analysis indicates that the crystal structure of hydroxyapatite was not present in the calcium phosphate-PS complex . The promotion of iron-induced generation of reactive oxygen species in nerve tissue by aluminum . DB01370 is suspected to play a role in several neurological disorders . Reactive oxygen species ( ROS ) lead to oxidative stress , which is thought to be a possible mechanism for neurological damage . Interactions between aluminum and iron , a known promoter of prooxidant events , were studied in cerebral tissues using a fluorescent probe to measure rates of generation of ROS . Al2(SO4)3 alone failed to stimulate ROS production over a wide range of concentrations ( 50-1000 microM ) . The aluminum-deferrioxamine chelate in the absence of iron could also not potentiate ROS formation . However , Al2(SO4)3 potentiated FeSO4-induced ROS , with a maximal effect at 10 microM Fe and 500 microM Al . DB01575 , a hydrated aluminum silicate , did not potentiate iron-induced ROS formation . Ferritin had a minor stimulatory effect on ROS generation , but this was not potentiated by the concurrent presence of Al2(SO4)3 . P02787 had no effect on basal rates of ROS generation , but when Al2(SO4)3 was also present , ROS production was enhanced . It is concluded that : 1 . There is a potentiation of iron-induced ROS by aluminum salts ; 2 . Free or complexed aluminum alone is not a key producer of ROS ; and 3 . High rates of ROS production are unlikely to be owing to the displacement by aluminum iron from its biologically sequestered locations . P10275 abnormalities in identical twins with oligospermia . Clinical and biochemical studies . Identical twin brothers presented with oligospermia , small testes , normal male phenotypes , elevated serum luteinizing hormone levels , and normal or elevated serum testosterone levels . Both men had low to low-normal cytosol androgen receptor binding capacity in cultured fibroblasts from pubic skin biopsy specimens . Qualitative abnormalities of cellular androgen receptors were suggested by low-normal or low nuclear androgen uptake in fibroblasts from both brothers as well as abnormal thermolability and subnormal molybdate stabilization of androgen receptors from one brother . In vivo androgen sensitivity was assessed in one twin following administration of testosterone or the non-aromatizable androgen fluoxymesterone . DB01185 suppressed serum luteinizing hormone and serum testosterone/estradiol-binding globulin , and although testosterone suppressed both serum luteinizing hormone and serum follicle-stimulating hormone , the suppression of serum luteinizing hormone by testosterone was subnormal . Both subjects showed marked exaggeration of the serum 17-hydroxyprogesterone increase after administration of human chorionic gonadotropin , despite normal serum testosterone increases , suggesting a block in testicular 17,20-desmolase , which converts 17-hydroxyprogesterone to testosterone . These studies suggest that oligospermia and block of the enzyme 17,20-desmolase may be the earliest manifestations of androgen resistance , and the finding of the syndrome of oligospermia , normal male phenotype , and androgen receptor abnormalities in identical twins indicates a genetic etiology of this disorder . Pharmacology of the calcium sensing receptor . DB01373 sensing receptor ( P41180 ) is a G-protein couple receptor which plays a key role in calcium homeostasis in vertebrates . Its extracellular domain is sensitive to divalent cations , aminoacids and polyamines . In parathyroid glands , P41180 activation causes parathyroid hormone ( PTH ) reduction and subsequently a decrease in blood calcium concentration . In PTH-dependent disorders , e.g. primary and secondary hyperparathyroidism ( Q9HD23 ) , the need for therapeutic options other than surgery led to the synthesis of various allosteric P41180 agonists ( calcimimetics ) , such as cinacalcet . DB01012 is the only calcimimetic approved for Q9HD23 secondary to chronic kidney disease ( CDK ) , parathyroid carcinoma , and , in some countries , primary Q9HD23 . Clinical trials showed that cinacalcet reduced PTH and calcemia both in CDK and primary Q9HD23 , lowering the risk of bone fractures , surgery , and cardiovascular complications in the former patients . Long-term safety and pharmacoeconomics have to be fully tested yet . Few both in vitro and in vivo studies showed an association between Arg990Gly- P41180 polymorphism and cinacalcet sensitivity , though in patients with severe P41180 inactivating mutations the drug substantially retained its positive clinical effects . Recently , a new class of allosteric antagonists of P41180 , i.e. calcilytics , has been synthesized . Calcilytics are structurally similar to calcimimetics , but exert their effects acting on a different allosteric site . Infusion of calcilytics was followed by transient rise in PTH and calcium . One of these compounds , DB05255 , was able to increase femur BMD in post menopausal women , but with induction of mild hyperparathyroidism . In the future , calcilytics may contribute to the osteoporosis treatment choice . Identification of a novel extracellular cation-sensing G-protein-coupled receptor . The C family G-protein-coupled receptors contain members that sense amino acid and extracellular cations , of which calcium-sensing receptor ( P41180 ) is the prototypic extracellular calcium-sensing receptor . Some cells , such as osteoblasts in bone , retain responsiveness to extracellular calcium in P41180 -deficient mice , consistent with the existence of another calcium-sensing receptor . We examined the calcium-sensing properties of Q5T6X5 , a newly identified member of this family . Alignment of Q5T6X5 with P41180 revealed conservation of both calcium and calcimimetic binding sites . In addition , calcium , magnesium , strontium , aluminum , gadolinium , and the calcimimetic P0C0P6 568 resulted in a dose-dependent stimulation of Q5T6X5 overexpressed in human embryonic kidney cells 293 cells . Also , osteocalcin , a calcium-binding protein highly expressed in bone , dose-dependently stimulated Q5T6X5 activity in the presence of calcium but inhibited the calcium-dependent activation of P41180 . Coexpression of beta-arrestins 1 and 2 , regulators of G-protein signaling P41220 or P49798 , the RhoA inhibitor P01024 toxin , the dominant negative Galpha(q)-(305-359) minigene , and pretreatment with pertussis toxin inhibited activation of Q5T6X5 by extracellular cations . Reverse transcription-PCR analyses showed that mouse Q5T6X5 is widely expressed in mouse tissues , including bone , calvaria , and the osteoblastic cell line MC3T3-E1 . These data suggest that in addition to sensing amino acids , Q5T6X5 is a cation- , calcimimetic- , and osteocalcin-sensing receptor and a candidate for mediating extracellular calcium-sensing responses in osteoblasts and possibly other tissues . [ Preliminary study on the mechanism of connexin 43 gene transfection in the control of glioma cell proliferation ] . OBJECTIVE : To study the mechanism involved in the control of glioma cell proliferation with transfection of connexin ( Cx ) 43 gene . METHODS : P13671 rat glioma and TJ905 human glioblastoma cell lines without P17302 gene expression were transfected with Cx43cDNA mediated by lipofectamine . Northern blot , in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation , TUNEL method for determination of cell apoptosis , scrape loading dye transfer ( SLDT ) for GJIC , Western blot and immunohistochemical technology for P09038 , PDGF , P00533 , P05019 and P17936 expression . RESULTS : Cx 43 gene transfected glioma cells showed decreased proliferation , restored GJIC and decreased P09038 , PDGF , P17936 , except P00533 expression and cell apoptosis which showed no change . CONCLUSION : The mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors . Immunohistochemical analysis of brain lesions using P04271 and glial fibrillary acidic protein antibodies in arundic acid- ( DB05343 ) treated stroke-prone spontaneously hypertensive rats . Stroke-prone spontaneously hypertensive rats ( SHRSP ) used as a model of essential hypertension cause a high incidence of brain stroke on the course of hypertension . Incidences and sizes of brain lesions are known to relate to the astrocyte activities . Therefore , relation between brain damage and the expression profile of the astrocytes was investigated with morphometric and immunohistochemical analyses using astrocyte marker antibodies of P04271 and glial fibrillary acidic protein ( P14136 ) with or without arundic acid administration , a suppressor on the activation of astrocytes . Arundic acid extended the average life span of SHRSP . An increase in brain tissue weight was inhibited concomitant with a lower rate of gliosis/hemosiderin deposit/scarring in brain lesions . P04271 - or P14136 -positive dot and filamentous structures were decreased in arundic acid-treated SHRSP , and this effect was most pronounced in the cerebral cortex , white matter , and pons , and less so in the hippocampus , diencephalon , midbrain , and cerebellum . Blood pressure decreased after administration of arundic acid in the high-dose group ( 100 mg/kg/day arundic acid ) , but not in the low-dose group ( 30 mg/kg/day ) . These data indicate that arundic acid can prevent hypertension-induced stroke , and may inhibit the enlargement of the stroke lesion by preventing the inflammatory changes caused by overproduction of the P04271 protein in the astrocytes . Tanshinone IIA inhibits constitutive P40763 activation , suppresses proliferation , and induces apoptosis in rat P13671 glioma cells . P40763 ( P40763 ) is usually constitutively activated in a variety of malignancies . Thus , P40763 may be a promising target for treatment of tumor cells . Recently , Tanshinone IIA ( Tan IIA ) , a major active constituent from the root of Salvia miltiorrhiza Bunge , was reported to have apoptosis inducing effects on a large variety of cancer cells . In this study , we evaluate the anti-proliferation and apoptosis inducing effects of Tan IIA on P13671 glioma cells . Cell growth and proliferation were measured by MTT assay , cell apoptosis was observed by flow cytometry and DNA-fragmentation analysis . Further more , we investigated inhibitory effects of Tan IIA on P40763 activity and its downstream targets : Bcl-XL , cyclin D1 . Alteration of P40763 activity was examined by measuring their DNA binding activity and tyrosine phosphorylation . Changes in the expression levels of Bcl-XL and cyclin D1 were examined by Western blot analysis . We found that the cellular growth were inhibited and cell apoptosis were observed after the treatment with Tan IIA . The P40763 activity was significantly reduced by Tan IIA parallel with a significant attenuation of expression of Bcl-XL and cyclin D1 . These results suggest that Tan IIA may serve as an effective adjunctive reagent in the treatment of glioma for its targeting of constitutive P40763 signaling . Neuroendocrine response to clonidine and 8-OH-DPAT in rats following chronic administration of desipramine or sertraline . 1. Rats were administered either desipramine ( DB01151 ) or sertraline daily at doses 7.5 mg kg-1 or 10 mg kg-1 , i.p. , respectively and the effects on the functional state of hypothalamic neuroendocrine control mechanisms assessed by measurements of plasma hormones following acute drug challenge . The effects of treatment on gross behaviour and brain adrenoceptor density were also determined . 2 . Both DB01151 and sertraline caused significant reduction in activity measured as ambulation and rearing at 14 days of treatment . 3 . All animals were chronically cannulated after 14 days of treatment and tested for neuroendocrine response to acute i.v. clonidine ( 50 micrograms kg-1 ) or 8-hydroxy-2-(di-n-propylamino)tetralin ( 8-OH-DPAT , 250 micrograms kg-1 ) after 21 or more days of treatment . 4 . Rats treated with DB01151 but not sertraline showed a virtually complete suppression of the growth hormone ( GH ) secretion elicited by clonidine in controls , while the secretion of corticosterone was augmented . 5 . Treatment with DB01151 but not sertraline led to a significantly greater 8-OH-DPAT-induced secretion of prolactin than in the control rats , while the plasma concentrations of corticosterone following 8-OH-DPAT were not influenced by either DB01151 or sertraline treatment . 6 . The density ( but not the affinity ) of cerebral cortical binding of [ 3H ] -dihydroalprenolol was significantly reduced by DB01151 treatment . 7 . These results show that DB01151 treatment blunted the sensitivity of post-synaptic alpha 2-adrenoceptors , accompanied by complex interactions manifested as increased responsiveness of alpha 1-adrenoceptors and P08908 receptors . Sertraline had no significant neurendocrine effects at a dose which significantly reduced gross activity . DB00624 potentiates the hypoxic ventilatory response of adult male rats subjected to neonatal stress . Neonatal stress disrupts development of homeostatic systems . During adulthood , male rats subjected to neonatal maternal separation ( Q5H8A3 ) are hypertensive and show a larger hypoxic ventilatory response ( HVR ) , with greater respiratory instability during sleep . Neonatal stress also affects sex hormone secretion ; hypoxia increases circulating testosterone of Q5H8A3 ( but not control ) male rats . Given that these effects of Q5H8A3 are not observed in females , we tested the hypothesis that testosterone elevation is necessary for the stress-related increase of the HVR in adult male rats . Pups subjected to Q5H8A3 were placed in an incubator for 3 h per day from postnatal day 3 to 12 . Control pups remained undisturbed . Rats were reared until adulthood , and the HVR was measured by plethysmography ( fractional inspired O2 = 0.12 , for 20 min ) . We used gonadectomy to evaluate the effects of reducing testosterone on the HVR . Gonadectomy had no effect on the HVR of control animals but reduced that of Q5H8A3 animals below control levels . Immunohistochemistry was used to quantify androgen receptors in brainstem areas involved in the HVR . P10275 expression was generally greater in Q5H8A3 rats than in control rats ; the most significant increase was noted in the caudal region of the nucleus tractus solitarii . We conclude that the abnormal regulation of testosterone is important in stress-related augmentation of the HVR . The greater number of androgen receptors within the brainstem may explain why Q5H8A3 rats are more sensitive to testosterone withdrawal . Based on the similarities of the cardiorespiratory phenotype of Q5H8A3 rats and patients suffering from sleep-disordered breathing , these results provide new insight into its pathophysiology , especially sex-based differences in its prevalence . PP2Cdelta ( Ppm1d , O15297 ) , an endogenous inhibitor of p38 MAPK , is regulated along with Trp53 and Cdkn2a following p38 MAPK inhibition during mouse preimplantation development . Preimplantation embryos utilize mitogen-activated protein kinase signaling ( MAPK ) pathways to relay signals from the external environment to prepare appropriate responses and adaptations to a changing milieu . It is therefore important to investigate how MAPK pathways are regulated during preimplantation development . This study was conducted to investigate whether PP2Cdelta ( Ppm1d , O15297 ) is expressed during mouse preimplantation development and to determine the influences of p38 MAPK inhibition on expression of Trp53 ( p53 ) , Ppm1d , ( O15297 ) , and Cdkn2a ( p16 ) during mouse preimplantation development . Our results indicate that Trp53 , Ppm1d , and Cdkn2a mRNAs and TRP53 and PP2Cdelta proteins are expressed throughout mouse preimplantation development . Treatment of 2-cell embryos with DB04338 ( potent inhibitor of p38 MAPK alpha/beta/ Q16539 /11 ) significantly increased Trp53 , Ppm1d and Cdkn2a and Mapk14 mRNA levels at 12 and 24 hr . Treatment of 8-cell embryos with DB04338 for 12 hr increased Trp53 , Ppm1d , and Cdkn2a mRNA levels , but not Mapk14 mRNA levels . Treatment of 8-cell embryos for 24 hr increased Trp53 , and Ppm1d mRNA levels , but decreased Cdkn2a and Mapk14 mRNA levels . Therefore , blockade of p38 MAPK activity is associated with embryo stage specific influences on Trp53 , Ppm1d , Cdkn2a , and Mapk14 expression during mouse preimplantation development . These results define downstream targets of p38 MAPK during preimplantation development and indicate that the p38 MAPK pathway regulates Trp53 , Ppm1d , and Cdkn2a expression . This study increases our understanding of the mechanisms controlling preimplantation development and of the interactions between preimplantation embryos and their culture environments . DB00151 oxidation in the mitogenic P04271 protein leads to changes in phosphorylation by catalytic CKII-alpha subunit . The glial-derived calcium-binding protein P04271 can be secreted to act as a neurotrophic factor or a mitogen , stimulating proliferation of glial cells . The extracellular P04271 activities rely on the oxidation of the protein cysteine residues ( Kligman , D. , and Marshak , D. R. ( 1985 ) Proc. Natl. Acad. Sci. U. S. A. 82 , 7136-7139 ; Winningham-Major , F. , Staecker , J. L. , Barger , S. W. , Coats , S. , and Van Eldik , L. J. ( 1989 ) J. Cell Biol. 109 , 3063-3071 ) . Here we show that oxidation of the P04271 cysteine residues , DB00151 -68 and DB00151 -84 , induces a conformational change in the protein structure , unmasking a canonical CKII phosphorylation site located within the typical EF-hand calcium-binding site IIbeta . Intrasubunit disulfide-bridged P04271 monomer and disulfide-bonded P04271 dimer are phosphorylated by the catalytic CKII-alpha subunit on DB00133 -62 with a Km of 0.5 microM and a Vmax of 10 pmol/min/100 pmol of P04271 . Oxidized P04271 is the best in vitro CKII-alpha substrate identified so far . Next we show that intrasubunit disulfide-bridged P04271 monomer is the most potent P04271 species to stimulate [3H]thymidine uptake by P13671 glial cells in culture . In addition , the phosphorylated intrasubunit disulfide-bridged P04271 monomer retains apparent mitogenic activity toward P13671 glial cells , and hence , 32P-labeled P04271 should be a useful probe for characterizing the mechanisms by which extracellular oxidized P04271 functions . Finally , we show that formation of intrasubunit disulfide-bridged P04271 monomer is stimulated by peroxynitrite anion , suggesting that production of mitogenic P04271 species could be enhanced in neuropathology associated with peroxynitrite anion production . Embryo transfer induces a subclinical endometritis in recipient mares which can be prevented by treatment with non-steroid anti-inflammatory drugs . We tested the hypothesis that subclinical endometritis occurs after embryo transfer ( ET ) in the horse . Recipient mares were treated with meclofenamic acid ( M ) or flunixin meglumin ( F ) after ET or were left untreated ( n=9 per group ) . Embryos were re-collected 4 days after transfer . Endometrial biopsies were taken for histology and analysis of cyclooxygenase-2 ( P35354 ) by immunohistochemistry and for PCR . Bacteriological swabs were collected from the uterus and lavage fluid of donor and recipient mares . Progesterone and prostaglandin F(2alpha) release was analysed in recipient mares after ET . Four days after ET , four embryos were recovered from group M and three from group F and untreated mares , each . The number of polymorph nuclear neutrophils was reduced in treated mares ( p < 0.05 ) . Expression of mRNA for inflammatory cytokines did not differ between groups . In group M , expression of endometrial prostaglandin-E-synthase was higher than in group F ( p < 0.05 ) . Three out of nine control mares underwent preterm luteolysis ( p < 0.05 vs. treatment groups ) , prostaglandin release ( p < 0.05 ) and the number of P35354 positive cells ( p < 0.01 ) were significantly higher than in treated mares . Only few bacteriological swabs were positive . In conclusion , treatment of embryo recipient mares with non-steroid anti-inflammatory drugs inhibits the inflammatory response of the endometrium after ET . DB00939 may have advantages in comparison to flunixin meglumin due to a different influence on prostaglandin synthesis that may not result in inhibition of embryonic mobility . P02787 subtypes and variants in Germany ; further evidence for a Tf null allele . Isoelectric focusing ( IEF ) with carrier ampholytes was used for the determination of transferrin C subtypes and transferrin B and D variants in a sample of 1125 unrelated individuals from Southern Germany . The observed TfC allele frequencies were Tf*C1 = 0.7872 , Tf* P06681 = 0.1365 , and Tf* P01024 = 0.0675 . The rare C subtype P13671 was observed twice . A new C subtype , called Q99622 , was observed and identified by IEF with immobilized pH gradients . The rare C subtypes C4 and Q99618 were also studied by this method . TfB and TfD variants were found with a heterozygous frequency of 1.53 % . One new TfD was found which is located between D1 and D2 and therefore named D1-2 . Evidence for a Tf null allele was obtained in a child and the putative father ; they were considered to be heterozygous for an allele Tf0 . The theoretical exclusion rate for paternity examinations was calculated for the Tf system and found to be 17.95 % .