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Schematic representation of key components of the 6HNic (6HNic)-responsive transgene regulation system (NICE). As a binary transcription-control system, NICE consists of an artificial 6HNic-dependent transactivator (NT), assembled by fusing the A.nicotinovorans pAO1 6HNic oxidase repressor HdnoR to functional mammalian transactivation domains (T; e.g. H.simplex VP16, p65 of human NF-κB, a domain of human E2F4) and a chimeric promoter engineered by placing HdnoR-specific operator modules (ONIC) adjacent to a minimal version of the human cytomegalovirus immediate early promoter (PhCMVmin). In the absence of 6HNic (â6HNic), NT binds to PNIC via direct HdnoR-ONIC interaction and induces PhCMVmin-mediated transcription of the gene of interest (goi). However, 6HNic modifies NT Values are mean ± SD of six rats aP < 0.001, bP < 0.01. Hyperlipidemic control (Group 2) was compared with control (Group 1). Hyperlipidemic + A. indicus treated (Group 3) and hyperlipidemic + gemfibrozil (Group 4) was compared with hyperlipidemic control (Group 2). Schematic drawing of the setup used in experiment 1 (side view). While foraging from the nest to the feeder the ants passed a narrowed section of the channel, within which they were recorded from above (x-y plane, see arrow). The inclination of this section could be altered from level ground (Ï = 0°) to almost vertical (Ï = 90°). Improved capture of known glycoproteins. Identified proteins were cross-referenced to the UniProt Knowledgebase for known glycoproteins. The percent of total proteins that were matched to known glycoproteins is shown, in comparison to that typically determined by global proteomic approaches. Induction of apoptosis in mouse PBMC by DMF or MMF for 48h as determined with PI (A) and Annexin (B) staining by flow cytometry.For controls PBMC were treated with 1.0% methanol or with medium alone (CO). Results are shown as means ± SEM. Significant post hoc effects versus controls (treated with methanol) are indicated by asterisks (**p<0.01, ***p<0.001).
Impact of competence in the stationary phase type microtiter biofilm model. In this model, biofilm formation was evaluated by both crystal violet staining and analysis at the spectrophotometer. The FP23 strain (non-capsulated TIGR4) was compared with its isogenic mutants in comD (FP231) and comC (FP259). The comC mutant FP259 was also assayed with addition of synthetic CSP to the medium (striped bars). The experiment was performed in BHI and read after 24 (panel A) or 48 hours (panel B) of incubation at 37°C. The differences in biofilm formations between the wt and the comC and comD mutants and between FP259 with and without CSP were statistically significant (p < 0.005). Data are from triplicate experiments. (a) CD30L binding activity of vCD30-Fc and (b) recombinant vCD30 expressed from VVCD30. (a) 5 ng vCD30-Fc or mouse CD30âFc were mixed with 150 pM mouse 125I-CD30L in the presence or absence of unlabeled CD30L and added without preincubation to protein Aâcoated FlashPlates. (b) 75 μl supernatants, equivalent to 1.5 à 104 cells, from cultures mock infected or infected with VVCD30 or WR were preincubated with 200 pM mouse 125I-CD30L, mixed with 5 ng vCD30-Fc, and then added to the protein Aâcoated FlashPlates. Bound 125I-CD30L was determined in a Packard Topcount microplate counter. The background radioactivity in the absence of recombinant protein has been subtracted. 125I-CD30L binding of duplicate samples (mean ± SD) is shown. NO exposure results in reduced VASP Ser-239 phosphorylation in diabetic CD34+ cells compared with control cells. The extent of phosphorylation was determined by FACS analysis on CD34+ cells from two diabetic subjects (DA [diabetic donor A] and DB [diabetic donor B]) and cells from a nondiabetic subject (Control [normal donor]). Values represent means ± SD. *P < 0.05. Enhanced food intake by retigabine was reversed by the blocker XE991. The food intake in the baseline group represents the value for normal rats without TMJ inflammation. The effect of retigabine or retigabine and XE991 was tested 24 hours after CFA injection. The food intake in the vehicle group indicates the value for rats with TMJ inflammation induced by CFA. Retigabine at a dose of 7.5 mg/kg, i.p. significantly increased the food intake, as compared with the vehicle group (p < 0.005, one-way ANOVA followed by Dunnett Assay of PNGase activity in an E. coli cell-free extract. The E. coli strain used was BL21(DE3)pLysS. Shown is a paper chromatogram of the reaction product formed after 10 min of incubation of the E. coli extract with labeled substrate. (lane 1) E. coli extract with pET-28b-PNG1; (lane 2) E. coli extract with pET-28b (control vector); and (lane 3) E. coli extract without vector. The extra minor band indicated with an asterisk was confirmed to be [14C]Leu-Asp by analysis by paper electrophoresis, as described earlier (Kitajima, et al. 1995), and may be derived from a contaminating protease activity in the E. coli extract. In fact, this degradation product represents a fraction of the product of PNGase activity because the second amino acid in the substrate was converted into Asp instead of remaining as Asn. P, de-N-glycosylated product ([14C]-Leu-Asp-Asn-Ser-Arg); and S, substrate ([14C]-Leu-Asn(Glc NAc5Man3Gal3)-Asn-Ser-Arg). For details, see Materials and Methods.
Quantitative analysis of cell behavior after injection of the Zyx16-30 peptide. The change in position of the cell edge relative to its original position before injection is shown for representative individual cells injected with the Zyx16-30 peptide (closed squares), the scrambled version of the peptide (open squares), Zyx16-30 peptide preincubated with α-actinin (triangles), and cells injected with the Zyx16-30 peptide in the presence of 10 mM diacetyl monoxime (open circles). The average rate of cell edge retraction upon injection of the Zyx16-30 peptide was determined to be 2.7 microns/min. Adoptively transferred diabetes. 10 à 107 diabetic DQ8+/mIIâ/B7+ splenocytes (depleted red blood cells) were transferred intravenously into irradiated recipients (as indicated). Diabetes was determined as above. Number of mice per group was as follows: n = 9 (4 female, 5 male) DQ8+/mIIâ/B7+; n = 8 (4 female, 4 male) DQâ/mIIâ/B7+; n = 3 (male) DQ8+/mIIâ/B7â; n = 3 (male) DQ8â/mIIâ/B7â; and n = 3 (male) DQ8+/mIIâ/B7+* (these recipients were adoptively transferred with purified CD8+ splenocytes only from diabetic DQ8+/mIIâ/B7+ mice). Mutation of the galU gene increases cytotoxicity of FT. Murine macrophage-like cells (J774) were infected with the WT, galU mutant, the galU-complemented, or wbtA mutant (O-antigen-deficient) FT LVS strains at an MOI of 100. Host cell death was determined by measuring LDH released from infected cells 24-hours post-infection. All data points represent the mean (± SEM) of triplicate samples and the data shown is representative of three experiments of similar design. Statistical analyses were performed via one-way ANOVA with a Bonferroni multiple comparisons post-test (*** indicates a p-value of <0.0001). Peak effect scores for 4 scales of dysphoric effects: A-Muddled/Confused; B-Clumsy; C-Dizzy; D-Nauseous. Participants rated their subjective effects on a 100 point scale from 0 â ânot at allâ to 100 â âextremelyâ.a. Self-reported feelings of âMuddled/Confusedâ.b. Self-reported feelings of âClumsyâ.c. Self-reported feelings of âDizzyâ.d. Self-reported feelings of âNauseousâ.Columns are averages (± sem) by drug condition and progesterone group. *2: Combination drug condition significantly different from nicotine alone and alcohol alone (p<.05). *3: Significant difference between low and high progesterone groups (p<.05). +Statistical trend for combined dose different from nicotine alone condition (p=.062) and for difference between progesterone groups (p=.068). dFOXO-binding sites in adults are distinct from those in larvae or S2 cells. (A) Overlap between the genomic sites bound by dFOXO in larvae and adults. The data for larvae were generated by Teleman et al (2008). The observed overlap was slightly smaller than expected by chance (P=0.02). Expected overlap of nine peaks was determined from simulation of 103 random peak sets, of identical size, length and chromosomal distribution. (B) A schematic of the dInR locus is given with grey boxes representing exons, black marks the P1, P2 and P3 promoters (Casas-Tinto et al, 2007), red boxes the sites bound by dFOXO in adult flies (observed in ChIP-chip data) and green bars the location of amplicons (leftâP1, rightâcoding region) used for ChIP-qPCR shown in (C). (C) dFOXO binding within the dInR locus in adults and S2 cells. The qPCR results show relative enrichment of the P1 promoter and the coding region (CDS) of dInR, or the U6 control, in three biological repeats of adult chromatin, or three IPs from a single chromatin sample from 2 h serum-starved S2 cells. The data are presented as in Figure 1B. ANOVA detected significant differences in enrichment (n=3, P<10â4), with P1 promoter being enriched in S2 cells and the coding region in adults (t-test, P<0.05).
The time of onset of new migraine periods (first headache diary entry of a migraine attack after a headache free period of at least 48 h). The colour code shows the intensities of the first headache diary entry (darker colours indicate strong headaches and light colours indicate milder headache). The start of more than 250 new migraine periods was recorded at 4 a.m., which is strikingly different from all other time points. As expected, the lowest number of new migraine periods (n = 65) began in the evening (8 p.m.) Rewarding effects of methamphetamine (METH) in Wistar and SHR/NCrlCrlj. (A) Place preference data (mean ± SEM) expressed as the difference in the amount of time spent in the METH-paired compartment during the post- and preconditioning days. Data are expressed as mean ± SEM. *p < 0.05, **p < 0.01 when comparing saline (SAL) vs METH-conditioned animals (n = 8). Comparison of triangular vowel space area (tVSA) at first (tVSA_t0) and second (tVSA_t1) examination.PDâ=âParkinson Mean cerebral blood flow velocities (CBFV) expressed in percentages of baseline for patients with immediate and delayed rewarming. Flow velocities remained below the baseline in immediate rewarming group, but for group of delayed rewarming mean flow velocity was comparable with baseline at all postbypass measurement (From 17). Secondary CD8 T-cell response to Listeria is CD28 dependent. Mice were infected with 5000 CFU Lm-OVA, challenged with 105 CFU on day 30, and analyzed for OVA-specific splenic CD8 cells by staining with Kb/SIINFEKL tetramers 5 days later. In A, CD28â/fl C57BL/6 mice were used, and CD28 deletion was induced by tamoxifen in oil feeding (âTMâ, 2.5 mg/dose) on days 24â27; oil only served as control. In B, wt C57BL/6 mice were treated with the blocking CD28-specific mAb E18 (100 μg/dose) on days 29, 31, and 33. ***P < 0.0001
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