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  • Hippocampal overexpression of mutant creb blocks long-term, but not short-term memory for a socially transmitted food preference. 15687228

    Phosphorylation of the transcription factor CREB on Ser133 is implicated in the establishment of long-term memory for hippocampus-dependent tasks, including spatial learning and contextual fear conditioning. We reported previously that training on a hippocampus-dependent social transmission of food preference (STFP) task increases CREB phosphorylation in the hippocampus of trained rats in comparisons with controls. In the current study, we tested the hypothesis that CREB function is necessary for long-term memory for STFP using herpes simplex viral (HSV) vector-mediated gene transfer. Rats received intrahippocampal infusions of HSV-mCREB (a mutant form of CREB, in which Ser133 has been replaced with Ala), HSV-LacZ, or saline, and were trained 3 d later. Rats were tested for food preference (demonstrated vs. novel foods) immediately (short-term test) and 11 d (long-term test) after training. Rats in all treatment groups showed a significant preference for the demonstrated food at the short-term memory test. At the long-term memory test, however, the percentage of demonstrated food eaten by mCREB-treated rats was significantly less than that eaten by the LacZ- or saline-treated rats. Quantitative Western blotting confirmed that mCREB-infused rats had significantly more hippocampal CREB protein than controls during training. The present results show that hippocampal CREB function is necessary for long-term, but not short-term memory for STFP.
    Document Type:
    Reference
    Product Catalog Number:
    06-863
    Product Catalog Name:
    Anti-CREB Antibody

  • Bedtime administration of NN2211, a long-acting GLP-1 derivative, substantially reduces fasting and postprandial glycemia in type 2 diabetes. 11812750

    Glucagon-like peptide 1 (GLP-1) is a potent glucose-lowering agent of potential interest for the treatment of type 2 diabetes. To evaluate actions of NN2211, a long-acting GLP-1 derivative, we examined 11 patients with type 2 diabetes, age 59 +/- 7 years (mean +/- SD), BMI 28.9 +/- 3.0 kg/m(2), HbA(1c) 6.5 +/- 0.6%, in a double-blind, placebo-controlled, crossover design. A single injection (10 microg/kg) of NN2211 was administered at 2300 h, and profiles of circulating insulin, C-peptide, glucose, and glucagon were monitored during the next 16.5 h. A standardized mixed meal was served at 1130 h. Efficacy analyses were performed for the fasting (7-8 h) and mealtime (1130-1530 h) periods. Insulin secretory rates (ISR) were estimated by C-peptide deconvolution analysis. Glucose pulse entrainment (6 mg x kg(-1) x min(-1) every 10 min) was evaluated by 1-min sampled measurements of insulin concentrations from 0930 to 1030 h and subsequent time series analysis of the insulin concentration profiles. All results are given as NN2211 versus placebo; statistical analyses were performed by analysis of variance. In the fasting state, plasma glucose was significantly reduced (6.9 +/- 1.0 vs. 8.1 +/- 1.0 mmol/l; P = 0.004), ISR was increased (179 +/- 70 vs. 163 +/- 66 pmol/min; P = 0.03), and plasma glucagon was unaltered (19 +/- 4 vs. 20 +/- 4 pg/ml; P = 0.17) by NN2211. Meal-related area under the curve (AUC)(1130-1530 h) for glucose was markedly reduced (30.6 +/- 2.4 vs. 39.9 +/- 7.3 mmol x l(-1) x h(-1); P 0.001), ISR AUC(1130-1530 h) was unchanged (118 +/- 32 vs. 106 +/- 27 nmol; P = 0.13), but the increment (relative to premeal values) was increased (65 +/- 22 vs. 45 +/- 11 nmol; P = 0.04). Glucagon AUC(1130-1530 h) was suppressed (77 +/- 18 vs. 82 +/- 17 pmol x l(-1) x h(-1); P = 0.04). Gastric emptying was significantly delayed as assessed by AUC(1130-1530 h) of 3-ortho-methylglucose (400 +/- 84 vs. 440 +/- 70 mg x l(-1) x h(-1); P = 0.02). During pulse entrainment, there was a tendency to increased high frequency regularity of insulin release as measured by a greater spectral power and autocorrelation coefficient (0.05 P 0.10). The pharmacokinetic profile of NN2211, as assessed by blood samplings for up to 63 h postdosing, was as follows: T(1/2) = 10.0 +/- 3.5 h and T(max) = 12.4 +/- 1.7 h. Two patients experienced gastrointestinal side effects on the day of active treatment. In conclusion, the long-acting GLP-1 derivative NN2211 effectively reduces fasting as well as meal-related (approximately 12 h postadministration) glycemia by modifying insulin secretion, delaying gastric emptying, and suppressing prandial glucagon secretion.
    Document Type:
    Reference
    Product Catalog Number:
    GL-32K
    Product Catalog Name:
    Glucagon RIA

  • Parkin expression profile in dopamine d3 receptor knock-out mice brains. 18612813

    Patients affected by autosomic recessive juvenile parkinsonism (ARJP) exhibit parkin gene mutations with brain decrease in dopamine D2/D3 binding sites. To date, there are no data indicating whether the reduction in dopamine D3 receptors (DRD3) may be associated with the expression of specific parkin variants. In the present study we investigated parkin expression profile in DRD3 knock-out mice brains. RT-PCR analysis was performed to assess qualitative changes in parkin isoforms' distribution pattern and in exons' expression both in wild type controls and dopamine D3 receptor's knock-out mice. Real-time PCR was performed to quantify single exons mRNA. Results demonstrated that exons 1, 2, 4, 6, 7, 8, were more expressed in wild type compared to dopamine D3 receptor KO mice brains while some other (3, 9, 10) were lower expressed. The expression levels of exons 5, 11 and 12 did not change in both animal groups. Our analysis was confirmed by western blot, which showed that parkin protein levels were influenced by the absence of DRD3.
    Document Type:
    Reference
    Product Catalog Number:
    AB5112
    Product Catalog Name:
    Anti-Parkin Antibody, a.a. 305-323

  • Cereal and nonfat milk support muscle recovery following exercise. 19442266

    ABSTRACT: BACKGROUND: This study compared the effects of ingesting cereal and nonfat milk (Cereal) and a carbohydrate-electrolyte sports drink (Drink) immediately following endurance exercise on muscle glycogen synthesis and the phosphorylation state of proteins controlling protein synthesis: Akt, mTOR, rpS6 and eIF4E. METHODS: Trained cyclists or triathletes (8 male: 28.0 +/- 1.6 yrs, 1.8 +/- 0.0 m, 75.4 +/- 3.2 kg, 61.0 +/- 1.6 ml O2*kg-1*min-1; 4 female: 25.3 +/- 1.7 yrs, 1.7 +/- 0.0 m, 66.9 +/- 4.6 kg, 46.4 +/- 1.2 mlO2*kg-1*min-1) completed two randomly-ordered trials serving as their own controls. After 2 hours of cycling at 60-65% VO2MAX, a biopsy from the vastus lateralis was obtained (Post0), then subjects consumed either Drink (78.5 g carbohydrate) or Cereal (77 g carbohydrate, 19.5 g protein and 2.7 g fat). Blood was drawn before and at the end of exercise, and at 15, 30 and 60 minutes after treatment. A second biopsy was taken 60 minutes after supplementation (Post60). Differences within and between treatments were tested using repeated measures ANOVA. RESULTS: At Post60, blood glucose was similar between treatments (Drink 6.1 +/- 0.3, Cereal 5.6 +/- 0.2 mmol/L, p .05), but after Cereal, plasma insulin was significantly higher (Drink 123.1 +/- 11.8, Cereal 191.0 +/- 12.3 pmol/L, p .05), and plasma lactate significantly lower (Drink 1.4 +/- 0.1, Cereal 1.00 +/- 0.1 mmol/L, p .05). Except for higher phosphorylation of mTOR after Cereal, glycogen and muscle proteins were not statistically different between treatments. Significant Post0 to Post60 changes occurred in glycogen (Drink 52.4 +/- 7.0 to 58.6 +/- 6.9, Cereal 58.7 +/- 9.6 to 66.0 +/- 10.0 mumol/g, p .05) and rpS6 (Drink 17.9 +/- 2.5 to 35.2 +/- 4.9, Cereal 18.6 +/- 2.2 to 35.4 +/- 4.4 %Std, p .05) for each treatment, but only Cereal significantly affected glycogen synthase (Drink 66.6 +/- 6.9 to 64.9 +/- 6.9, Cereal 61.1 +/- 8.0 to 54.2 +/- 7.2%Std, p .05), Akt (Drink 57.9 +/- 3.2 to 55.7 +/- 3.1, Cereal 53.2 +/- 4.1 to 60.5 +/- 3.7 %Std, p .05) and mTOR (Drink 28.7 +/- 4.4 to 35.4 +/- 4.5, Cereal 23.0 +/- 3.1 to 42.2 +/- 2.5 %Std, p .05). eIF4E was unchanged after both treatments. CONCLUSION: These results suggest that Cereal is as good as a commercially-available sports drink in initiating post-exercise muscle recovery.
    Document Type:
    Reference
    Product Catalog Number:
    05-736
    Product Catalog Name:
    Anti-phospho-Akt1/PKBα (Ser473) Antibody, clone SK703, rabbit monoclonal

  • Biologic significance of fascin expression in clear cell renal cell carcinoma: systematic analysis of primary and metastatic tumor tissues using a tissue microarray techn ... 16979727

    OBJECTIVES: To investigate the biologic significance of fascin, a globular actin cross-linking protein, involved in cell adhesion and motility, in primary and metastatic renal cell carcinoma (RCC). METHODS: A total of 136 primary clear cell RCCs and 54 clear cell RCC metastases were stained immunohistochemically using a tissue microarray technique. Distinct cytoplasmic staining was considered positive, and the staining results were associated with the pT stage, Fuhrman grade, tumor size, sarcomatoid morphology, and metastasis-free survival. For multivariate testing, Cox's proportional hazards regression model was used. RESULTS: Fascin expression was noted in 13 (10%) of 136 primary and 25 (46%) of 54 metastatic RCC specimens (P 0.001). Fascin expression was associated with high tumor stage (2 [3%] of 70 pT1 versus 11 [17%] of pT2/pT3; P = 0.008), high tumor grade (3 [3%] of 88 grade 1-2 versus 10 [21%] of 48 grade 3-4; P = 0.002), and large tumor size (P 0.001). In addition, 8 (62%) of 13 RCCs with sarcomatoid morphology expressed fascin compared with 5 (4%) of 123 RCCs without sarcomatoid transformation (P 0.001). Metastatic disease was noted in 10 (77%) of 13 patients with fascin-positive RCC compared with 26 (21%) of 121 patients with fascin-negative RCC (P 0.001). Multivariate analysis revealed pT Stage 1 or greater (P 0.001, risk ratio [RR] 8.6, 95% confidence interval [CI] 2.8 to 26.5), Fuhrman grade greater than 2 (P 0.001, RR 12.7, 95% CI 4.6 to 35.4), fascin expression (P 0.001, RR 7.2, 95% CI 3.0 to 17.4), and female gender (P = 0.02, RR 2.5, 95% CI 1.1 to 5.5) as independent predictors of metastatic disease. CONCLUSIONS: Fascin immunoreactivity in RCC proved to be an independent predictor of metastatic disease and was demonstrated in almost one half of RCC metastases. Thus, fascin may be a promising molecular target for future cancer therapy.
    Document Type:
    Reference
    Product Catalog Number:
    MAB3582
    Product Catalog Name:
    Anti-Fascin Antibody, clone 55K2

  • Bone metabolism in elite male rowers: adaptation to volume-extended training. 16583234

    We examined the effect of 6-month volume-extended training on bone metabolism in elite male rowers. Twelve elite male rowers (20.8+/-3.0 years; 192.9+/-4.7 cm; 91.9+/-5.3 kg; body fat 10.1+/-2.3%; VO2max 6.2+/-0.5 l min(-1)) participated in this study. Bone biochemical markers, hormones, bone mineral content (BMC), and bone mineral density (BMD) were assessed before and after training. Average weekly training volume was significantly higher (P0.05) during the 6 months of heavy training compared to relative rest (11.6+/-0.4 h week(-1) vs. 16.8+/-0.6 h week(-1)), while intensity remained the same. At the end of training, only arm BMD was significantly increased by 5.7%. Osteocalcin (16.6%), insulin-like growth factor-1 (IGF-1) (20.2%) and the bioavailability IGF-1 index (17.9%) were significantly increased. Before heavy training, relationships were observed between the whole body BMD and growth hormone (r=0.64; P or =0.02), lumbar spine BMD and 1.25(OH)2 vitamin D (r=0.69; P or =0.04), arm BMD and testosterone (r=0.59; P or =0.05), and arm BMD and adiponectin (r=0.59; P or =0.05). No relationship was found between BMC or BMD and blood biochemical measures 6 months later (r=0.56; P> or =0.05). In addition, osteocalcin was related to IGF-1 (r>0.58; P0.048) and bioavailability IGF-1 index (r>0.59; P or =0.055) before and after training. In summary, heavy training had a moderately favorable effect on BMD. Bone tissue at specific skeleton sites is sensitive to changes in training volume even in athletes with already high BMD values. Changes in BMD and bone formation may be caused by changes in specific hormones such as IGF-1 and adiponectin in male athletes.
    Document Type:
    Reference
    Product Catalog Number:
    HADP-61HK
    Product Catalog Name:
    Human Adiponectin RIA

  • Effects of estradiol and progesterone on gastric mucosal response to early Helicobacter pylori infection in female gerbils. 16579842

    BACKGROUND: Gender differences have been shown regarding the changes in the inflammatory response, gastrin secretion, and gastric acidity during Helicobacter pylori infection. Aim: To investigate the role of estradiol and progesterone in the changes of the gastric mucosa induced by H. pylori during the early stage of infection in female gerbils. MATERIALS AND METHODS: Thirty-three adult ovariectomized female gerbils were infected with H. pylori (SS1); 7 days after infection they were treated with low and high doses of estradiol (50 and 250 microg/60 days pellet), progesterone (15 and 50 mg/60 days pellet) and vehicle. Non-ovariectomized infected gerbils were used as control. Gerbils were euthanized after 6 weeks of infection. Histologic evaluation, immunohistochemical detection of proliferation cell nuclear antigen (PCNA), gastrin, and apoptosis by terminal deoxynucleotide nick end labeling (TUNEL) assay were performed. Positive cells for PCNA, TUNEL, and gastrin were counted in 10 oriented glands per animal. Two-sided p = .05 was considered significant. RESULTS: Estradiol-treated groups showed more intense and extended acute and follicular gastritis compared to the vehicle group, whereas progesterone-treated groups presented less gastritis than the other groups. Proliferation and apoptosis indexes were significantly lower in the vehicle group when compared with those of the control; both indexes were increased in the high-dose estradiol and progesterone groups as compared with those of the vehicle. Grade I nonmetaplastic atrophy was observed in the vehicle and progesterone groups. The high-dose progesterone group showed a significant reduction in the number of gastrin cells. CONCLUSIONS: Estradiol and progesterone participate in the gastric mucosal response to early H. pylori infection in gerbils.
    Document Type:
    Reference
    Product Catalog Number:
    S7101
    Product Catalog Name:
    ApopTag® Plus Peroxidase In Situ Apoptosis Kit

  • Synthesis and biodistribution of (11)C-GW7845, a positron-emitting agonist for peroxisome proliferator-activated receptor-{gamma}. 16204723

    The goal of this study was to synthesize and evaluate in vivo the peroxisome proliferator-activated receptor-gamma (PPARgamma) agonist (11)C-GW7845 ((S)-2-(1-carboxy-2-{4-[2-(5-methyl-2-phenyloxazol-4-yl)ethoxy]phenyl}ethylamino)benzoic acid methyl ester) ((11)C-compound 1). PPARgamma is a member of a family of nuclear receptors that plays a central role in the control of lipid and glucose metabolism. Compound 1 is an analog of tyrosine (inhibitor constant, 3.7 nmol/L), which is an inhibitor of experimental mammary carcinogenesis. METHODS: Protection of the carboxylic acid moiety of compound 1 was effected by treatment with N,N-dimethylformamide di-tert-butyl acetal to provide compound 2. Hydrolysis of the carbomethoxy group of compound 2 provided the benzoic acid (compound 3) that served as an immediate precursor to radiolabeling. Compound 3 underwent treatment with (11)C-methyl iodide followed by high-performance liquid chromatography to produce a radioactive peak sample that coeluted with a standard sample of compound 1. Analysis of biodistribution was undertaken by injecting male CD-1 mice via the tail vein with 6.03 MBq (163 microCi, 2.55 microg/kg) of (11)C-compound 1. To determine the tumor uptake of the radiotracer, 6 female SCID mice bearing MCF-7 xenografts were injected via the tail vein with 10.5 MBq (283 microCi, 0.235 microg/kg) of (11)C-compound 1. RESULTS: (11)C-Compound 1 was synthesized at an 8% radiochemical yield in 29 min with an average specific radioactivity of 1,222 GBq/micromol (33,024 mCi/micromol; n = 6) at the end of synthesis. Spleen (target)-to-muscle uptake and tumor-to-muscle uptake ratios were 3.1 and 1.5, respectively, but this uptake could not be blocked with unlabeled compound 1 at 2 mg/kg. CONCLUSION: Further structural modification, perhaps to generate a less lipophilic tyrosine analog, will be necessary to enable receptor-mediated PPARgamma imaging by this class of agents.
    Document Type:
    Reference
    Product Catalog Number:
    MAB3872
    Product Catalog Name:
    Anti-PPAR γ Antibody, isoform 1&2

  • BCAA catabolism in brown fat controls energy homeostasis through SLC25A44 31435015

    Branched-chain amino acid (BCAA; valine, leucine and isoleucine) supplementation is often beneficial to energy expenditure; however, increased circulating levels of BCAA are linked to obesity and diabetes. The mechanisms of this paradox remain unclear. Here we report that, on cold exposure, brown adipose tissue (BAT) actively utilizes BCAA in the mitochondria for thermogenesis and promotes systemic BCAA clearance in mice and humans. In turn, a BAT-specific defect in BCAA catabolism attenuates systemic BCAA clearance, BAT fuel oxidation and thermogenesis, leading to diet-induced obesity and glucose intolerance. Mechanistically, active BCAA catabolism in BAT is mediated by SLC25A44, which transports BCAAs into mitochondria. Our results suggest that BAT serves as a key metabolic filter that controls BCAA clearance via SLC25A44, thereby contributing to the improvement of metabolic health.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple
    Product Catalog Name:
    Multiple