Phenomic profiles are high-dimensional sets of readouts that can comprehensively capture the biological impact of chemical and genetic perturbations in cellular assay systems. Phenomic profiling of ...compound libraries can be used for compound target identification or mechanism of action (MoA) prediction and other applications in drug discovery. To devise an economical set of phenomic profiling assays, we assembled a library of 1,008 approved drugs and well-characterized tool compounds manually annotated to 218 unique MoAs, and we profiled each compound at four concentrations in live-cell, high-content imaging screens against a panel of 15 reporter cell lines, which expressed a diverse set of fluorescent organelle and pathway markers in three distinct cell lineages. For 41 of 83 testable MoAs, phenomic profiles accurately ranked the reference compounds (AUC-ROC ≥ 0.9). MoAs could be better resolved by screening compounds at multiple concentrations than by including replicates at a single concentration. Screening additional cell lineages and fluorescent markers increased the number of distinguishable MoAs but this effect quickly plateaued. There remains a substantial number of MoAs that were hard to distinguish from others under the current study's conditions. We discuss ways to close this gap, which will inform the design of future phenomic profiling efforts.
The significant power dissipation of the on-chip L2 cache is a major concern in modern microprocessors. This Letter proposes a unique low-power technique for a high-associative large L2 cache that ...has fragmented locality by L1 cache. The dynamic per history length adjustment cache (DHL-cache) dynamically selects the qualified way candidates to be accessed and controls the way-prediction window size based on the access history pattern. With a high degree of the way-prediction accuracy, the DHL-cache shows a 55.3% energy-delay product improvement over the location-cache with minimum hardware support. Therefore, the DHL-cache alleviates the power limit issue for L2 cache, even with high associativity and fragmented locality.
Toll-like receptor-4 (TLR4) can be activated by nonbacterial agonists, including saturated fatty acids. However, downstream signaling pathways activated by nonbacterial agonists are not known. Thus, ...we determined the downstream signaling pathways derived from saturated fatty acid-induced TLR4 activation. Saturated fatty acid (lauric acid)-induced NFκB activation was inhibited by a dominant-negative mutant of TLR4, MyD88, IRAK-1, TRAF6, or IκBα in macrophages (RAW264.7) and 293T cells transfected with TLR4 and MD2. Lauric acid induced the transient phosphorylation of AKT. LY294002, dominant-negative (DN) phosphatidylinositol 3-kinase (PI3K), or AKT(DN) inhibited NFκB activation, p65 transactivation, and cyclooxygenase-2 (COX-2) expression induced by lauric acid or constitutively active (CA) TLR4. AKT(DN) blocked MyD88-induced NFκB activation, suggesting that AKT is a MyD88-dependent downstream signaling component of TLR4. AKT(CA) was sufficient to induce NFκB activation and COX-2 expression. These results demonstrate that NFκB activation and COX-2 expression induced by lauric acid are at least partly mediated through the TLR4/PI3K/AKT signaling pathway. In contrast, docosahexaenoic acid (DHA) inhibited the phosphorylation of AKT induced by lipopolysaccharide or lauric acid. DHA also suppressed NFκB activation induced by TLR4(CA), but not MyD88(CA) or AKT(CA), suggesting that the molecular targets of DHA are signaling components upstream of MyD88 and AKT. Together, these results suggest that saturated and polyunsaturated fatty acids reciprocally modulate the activation of TLR4 and its downstream signaling pathways involving MyD88/IRAK/TRAF6 and PI3K/AKT and further suggest the possibility that TLR4-mediated target gene expression and cellular responses are also differentially modulated by saturated and unsaturated fatty acids.
Toll-like receptor 4 (TLR4) and TLR2 agonists from bacterial origin require acylated saturated fatty acids in their molecules. Previously, we reported that TLR4 activation is reciprocally modulated ...by saturated and polyunsaturated fatty acids in macrophages. However, it is not known whether fatty acids can modulate the activation of TLR2 or other TLRs for which respective ligands do not require acylated fatty acids. A saturated fatty acid, lauric acid, induced NFκB activation when TLR2 was co-transfected with TLR1 or TLR6 in 293T cells, but not when TLR1, 2, 3, 5, 6, or 9 was transfected individually. An n-3 polyunsaturated fatty acid (docosahexaenoic acid (DHA)) suppressed NFκB activation and cyclooxygenase-2 expression induced by the agonist for TLR2, 3, 4, 5, or 9 in a macrophage cell line (RAW264.7). Because dimerization is considered one of the potential mechanisms for the activation of TLR2 and TLR4, we determined whether the fatty acids modulate the dimerization. However, neither lauric acid nor DHA affected the heterodimerization of TLR2 with TLR6 as well as the homodimerization of TLR4 as determined by co-immunoprecipitation assays in 293T cells in which these TLRs were transiently overexpressed. Together, these results demonstrate that lauric acid activates TLR2 dimers as well as TLR4 for which respective bacterial agonists require acylated fatty acids, whereas DHA inhibits the activation of all TLRs tested. Thus, responsiveness of different cell types and tissues to saturated fatty acids would depend on the expression of TLR4 or TLR2 with either TLR1 or TLR6. These results also suggest that inflammatory responses induced by the activation of TLRs can be differentially modulated by types of dietary fatty acids.
Background. The prevalence of glomerular diseases differs according to geographic area, race, age and indications for a renal biopsy. This study was conducted to evaluate the distribution and ...changing patterns of renal diseases during the past 20 years in a large patient population in Korea. Methods. Patients aged 16 years or older who underwent a renal biopsy at Severance Hospital in the Yonsei University Health System from 1987 to 2006 were enrolled. All medical records were reviewed retrospectively. Results. In total, 1818 patients (M:F = 1.02:1) were reviewed. Glomerulonephritis (GN) comprised 85.9% of the total biopsied cases. The most common primary GN was IgA nephropathy (IgAN) (28.3%), which was followed by minimal change disease (MCD) (15.5%), membranous nephropathy (MN) (12.3%), focal segmental glomerulosclerosis (FSGS) (5.6%) and membranoproliferative GN (MPGN) (4.0%). The most common secondary GN was lupus nephritis (8.7%). The most common idiopathic nephrotic syndrome was MCD (38.5%), which was followed by MN and IgAN. Among 128 (7.4%) patients who were HBsAg-positive, MN (30.5%) and MPGN (21.1%) were the most common GN. When the incidence rates between 1987–91 and 2002–06 were compared, IgAN increased from 25.6 to 34.5%, while MCD (from 23.2 to 7.0%) and MPGN (from 6.7 to 1.7%) decreased significantly (P < 0.01). Conclusions. IgAN was the most common primary GN, and MCD was the most common cause of nephrotic syndrome. In the 5-year quartile comparison, the relative frequency of IgAN increased, while the relative frequency of MCD and MPGN decreased significantly during the past 20 years.
Solvent hypertolerant Pseudomonas sp. BCNU 106 still has some underlying growth limitation in solvents. Therefore, efficient mass cultivation methods are needed to pursue its applications in ...biotechnology. Pseudomonas sp. BCNU 106 was cultured in a medium supplemented with 0·05 mol l−1 glycerol and cell survival was monitored during its cultivation in the presence of 1% (v/v) toluene. Exogenously supplemented glycerol provided more protection against damage caused by toluene stress and conferred higher solvent tolerance of Pseudomonas sp. BCNU 106 to toluene compared to control Pseudomonas sp. BCNU 106 without the supplementation of glycerol. This low‐cost mass cultivation method can be used to efficiently apply solvent‐tolerant bacteria in biotransformation and biodegradation.
Significance and Impact of the Study
Protection against toluene and improvement in bacterial cell growth by supplementation of glycerol in the presence of toluene are demonstrated in this study. This result can be used to solve growth‐related hindrances of solvent‐tolerant bacteria and establish their low‐cost mass cultivation, thereby broadening their industrial and environmental applications.
Significance and Impact of the Study: Protection against toluene and improvement in bacterial cell growth by supplementation of glycerol in the presence of toluene are demonstrated in this study. This result can be used to solve growth‐related hindrances of solvent‐tolerant bacteria and establish their low‐cost mass cultivation, thereby broadening their industrial and environmental applications.
A
bstract
The results of a search for solar axions from the Korea Invisible Mass Search (KIMS) experiment at the Yangyang Underground Laboratory are presented. Low-energy electron-recoil events would ...be produced by conversion of solar axions into electrons via the axio-electric effect in CsI(Tl) crystals. Using data from an exposure of 34,596 kg · days, we set a 90 % confidence level upper limit on the axion-electron coupling,
g
ae
, of 1
.
39 × 10
−
11
for an axion mass less than 1 keV/c
2
. This limit is lower than the indirect solar neutrino bound, and fully excludes QCD axions heavier than 0.48 eV/c
2
and 140.9 eV/c
2
for the DFSZ and KSVZ models respectively.
We present new constraints on the dark matter-induced annual modulation signal using 1.7 years of COSINE-100 data with a total exposure of 97.7 kg yr. The COSINE-100 experiment, consisting of 106 kg ...of NaI(Tl) target material, is designed to carry out a model-independent test of DAMA/LIBRA's claim of WIMP discovery by searching for the same annual modulation signal using the same NaI(Tl) target. The crystal data show a 2.7 cpd/kg/keV background rate on average in the 2-6 keV energy region of interest. Using a χ-squared minimization method we observe best fit values for modulation amplitude and phase of 0.0092±0.0067 cpd/kg/keV and 127.2±45.9 d, respectively.
As the solvent hyper‐resistant Pseudomonas sp. BCNU 106 experiences limited growth with solvents, a strategy is therefore needed to allow better growth to broaden its performance in biotechnological ...applications. Pseudomonas sp. BCNU 106 was cultivated in a medium supplemented with 0·05 mol l⁻¹ trehalose, and the cell survival was observed during subsequent growth with 1% (v/v) toluene. Exogenously added trehalose was transported into the cells and conferred protection against toluene stress. BCNU 106 grown in the presence of exogenous trehalose showed higher solvent tolerance, it can thus have more potential for biotransformation and biodegradation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that exogenously supplemented trehalose confers protection against toluene stress and enhances the bacterial cell growth in the presence of toluene. This is of importance to the mass cultivation of solvent‐tolerant bacteria, where some of the growth‐related limitations of solvent‐tolerant bacteria can be overcome, and their performance in biotechnological applications for biotransformation and biodegradation broadened.
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