Decision Framework for the Optimal Installation of Outriggers of Super-Tall Buildings

This paper was reviewed and accepted by the APCWE-IX Programme Committee for Presentation at the 9th Asia-Pacific Conference on Wind Engineering, University of Auckland, Auckland, New Zealand, held from 3-7 December 2017

Myosin II levels and localization are upregulated in DP-null cells and epidermis.

<p>(A-D) WT (A, C) and DP cKO (B, D) epidermis at E17.5 (A, B) and E18.5 (C, D) were stained for myosin IIA (green). Basement membrane is marked with a dashed line. Scale bar, 10 µm. (E) Myosin II levels in E18.5 WT and DP cKO epidermis were evaluated by Western blot. β-tubulin is the loading control. (F, G) WT and DP-null keratinocytes were stained for myosin IIA (green). (H) Quantitation of cortical/cytoplasmic ratios of myosin IIA in WT and DP null cells. n>60 cells for two independent experiments, p = 0.027.</p

Changes in adherens junctions in DP-null keratinocytes.

<p>(A-B) WT and DP-null keratinocytes stained for E-cadherin (green) and α-catenin (red) after 24 h in Ca²⁺. (C-F) WT (C,D) and DP-null (E,F) keratinocytes stained with the tension-sensitive anti-α-catenin antibody, α18 (red). The α18 epitope is exposed in WT keratinocytes after microtubules are reorganized to the cell cortex with taxol treatment, 1 hour at 10 µM. (D). DP-null keratinocytes have the α18 epitope exposed at steady state (E). The junctional intensity is not significantly changed upon treatment with taxol (F). (G) Quantitation of junctional/cytoplasmic intensity of α18 in indicated samples. n>100 cells from at least two independent experiments. p values as compared to WT DMSO are <0.0001 for both WT with taxol and DP KO DMSO. There was no significant difference between DP KO DMSO verses taxol. (H) α18 (red) staining of DP null keratinocytes treated with the myosin II inhibitor blebbistatin (25 µM for 1 hour). (I,J) E-cadherin (red) and α-catenin (green) localization in DP cells treated either with DMSO (I) or 25 µM blebbistatin (J) for 1 hour. Scale bar, 10 µm.</p

Pathogenic pemphigus antibodies induce increased contractility in WT keratinocytes.

<p>(A-C) WT keratinocytes were stained for myosin IIA (green). Cortical myosin II staining is observed when cells are treated with taxol to increase tension (B), or when treated with pathogenic pemphigus antibodies (AK23, in C). (D-F) WT keratinocytes were stained for α18 (red). The tension-sensitive epitope of α-catenin is exposed after taxol treatment (E) or after pathogenic pemphigus antibody treatment (F). (G,H) Desmoplakin (red) localization in WT cells treated with normal mouse sera (G) or with AK23 antibodies (H). Scale bars, 10 µm. (I) WT and myosin IIA-null keratinocytes were subjected to cell sheet disruption after treatment with control IgG or pathogenic pemphigus antibody. **, p<.005, n = 4.</p

Tight junctions are altered upon loss of desmoplakin.

<p>(A-B′) Calcium was added to WT and DP-null keratinocytes, and cells were fixed at various time points and stained for tight junction proteins occludin (red) and ZO-1 (green). Scale bar, 10 µm. (C,D) ZO-1 staining of WT and DP-null keratinocytes at 3 hour after calcium switch. (E) Co-stain for ZO-1 (red) and E-cadherin (green) in DP null keratinocytes 3 hours after calcium switch. (F) RNA was isolated from WT and DP-null keratinocytes, and RT-PCR for several claudins was performed. ***, p<.0005, **, p<.005. n = 3. (G-I) Western blot analysis of total levels of claudin-1 and β-actin in lysates from cultured keratinocytes (G,H) and from isolated epidermis (I). (J,K) Claudin 1 staining of WT and DP-null keratinocytes 24 hours after calcium shift. (L,M) Immunofluorescence analysis of claudin 1 (green) and β4-integrin (red) in wild type (L) and desmoplakin conditional null epidermis (M). Scale bars are 10 µm.</p

Friedel–Crafts Alkylation of Acylphloroglucinols Catalyzed by a Fungal Indole Prenyltransferase

Naturally occurring prenylated acylphloroglucinol derivatives are plant metabolites with diverse biological and pharmacological activities. Prenylation of acylphloroglucinols plays an important role in the formation of these intriguing natural products and is catalyzed in plants by membrane-bound enzymes. In this study, we demonstrate the prenylation of such compounds by a soluble fungal prenyltransferase AnaPT involved in the biosynthesis of prenylated indole alkaloids. The observed activities of AnaPT toward these substrates are much higher than that of a microsomal fraction containing an overproduced prenyltransferase from the plant hop

Effects of UPLC gradient on chromatography separation of the DXP pathway intermediates.

<p>Different concentrations (0.8, 0.4, 2.6, 2.6, 0.5 and 2 µM respectively) of DXP, MEP, CDP-ME, CDP-MEP, MEC, and HMBPP were prepared in 1 mL acidic extraction solution and purified as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#s4" target="_blank">METHODS AND MATERIALS</a>. Quantification m/z ratio for each compound was extracted from total ion chromatography as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#pone-0047513-t001" target="_blank">Table 1</a> and traces were overlaid. (A) UPLC gradient described was employed as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#pone-0047513-t002" target="_blank">Table 2</a> except 40% aqueous solution was used in step 3 and 4; (B) UPLC gradient described was employed as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#pone-0047513-t002" target="_blank">Table 2</a> except 50% aqueous solution was used in step 3 and 4; (C) UPLC gradient described was employed as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#pone-0047513-t002" target="_blank">Table 2</a>; (D) UPLC gradient described was employed as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#pone-0047513-t002" target="_blank">Table 2</a> except 70% aqueous solution was used in step 3 and 4; (E) UPLC gradient described was employed as <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0047513#pone-0047513-t002" target="_blank">Table 2</a> except 80% aqueous solution was used in step 3 and 4.</p

The cross-lapping in vitro assembly (CLIVA) method.

<p>(A) Illustration of the design at one junction between two modules (blue and red). The cross-lapping primer consists of gene specific sequence (GSS) and tag sequence complementary to adjacent primer’s GSS. The phosphorothioate modifications were indicated as cycles. An “Ox/y” designation was used to define the primers, where O denoted overlap; x was the length of overlap which had one modification at each y base pairs of the sequence. (B) Illustration of assembling of multiple DNA modules into one plasmid. </p

The effects of Fe-S operons on the amorphadiene production.

<p>Different concentrations of IPTG were represented by bars with different colors. The experiment was repeated four times and the standard errors of four replicates were presented as error bars. The two tailed p-values of student’s t-test were carried out to compare certain conditions and presented as P in the figure.</p

Overexpression of ispG to alleviate efflux of MEC for lycopene production.

<p>SIDF: BL21 Gold (DE3) harboring pET-SIDF and pACLYC; SIDFG: BL21 Gold (DE3) harboring pET-SIDFG and pACLYC. (A) Lycopene production as a function of time; (B) Extracellular MEC concentration as a function of time; (C) Intracellular MEC concentration as a function of time; (D) Intracellular HMBPP concentration as a function of time. Presented data were average of triplicates and standard errors were drawn on the plot.</p