Measurement and Correlation of Solubility of Loratadine in Different Pure Solvents and Binary Mixtures

The solubility of loratadine in pure solvents and binary mixed solvents is measured at the temperature range from 283.15 to 323.15 K using a laser technique. The results indicate that the solubility of loratadine increases with the increasing temperature in all these selected solvents. The solubility of loratadine reaches the maximum value when the mole fraction of methanol is 0.7 in the binary solvent mixtures of methanol + acetonitrile. Also, in the system of <i>n</i>-pentanol + acetonitrile mixed solvent the solubility reaches the maximum value when the mole fraction of <i>n</i>-pentanol is 0.6. The modified Apelblat equation and three parameters van’t Hoff equation are applied to correlate the experimental solubility in pure solvents. In binary mixed solvents, the modified Apelblat equation and Jouyban-Acree model are used to correlate the solubility data

Determination and Correlation of Solubility of Phenylbutazone in Monosolvents and Binary Solvent Mixtures

In this paper, we focused on the solubility of phenylbutazone, which was characterized by FT-IR spectra and X-ray diffraction. By the method of a laser technique, the solubility of phenylbutazone was measured in 10 monosolvents (methanol, ethanol, 2-propanol, <i>n</i>-butanol, 1-pentanol, <i>n</i>-hexanol, acetonitrile, ethyl acetate, acetone, and DMF) and 2 binary solvent mixtures of methanol + acetone and ethanol + acetonitrile from 283 to 323 K at about 5 K intervals under atmospheric pressure. The results show that the solubility of phenylbutazone in monosolvents and mixed solvents increases with the increase of temperature. However, it decreases with the increasing initial mole fraction of alcohols in the selected mixed solvents at constant temperature. The experimental data were correlated by the modified three-parameter van’t Hoff equation and the modified Jouyban–Acree equation

Measurement and Correlation of Solubility on Reactive Crystallization of Methyl D‑(−)-4-Hydroxy-phenylglycinate

In this paper, the solubility of methyl D-(−)-4-hydroxy-phenylglycinate was measured under the different pH levels, pure solvents, mixed solvents, ionic strength, and impurities in the temperature range from 283 to 323 about 5 K intervals by using laser method at atmospheric pressure. The results reveal that the solubility of methyl D-(−)-4-hydroxy-phenylglycinate increases with increasing temperature in all selected solvents, which decreases with increasing mole fraction composition of water in the mixed solvents. The solubility curve of methyl D-(−)-4-hydroxy-phenylglycinate in the aqueous solution at different pH is “U” shape and the solubility of methyl D-(−)-4-hydroxy-phenylglycinate increases with concentrations of ammonium chloride and has no obvious changes when its concentration increases up to 1.25 mol/kg. It is beneficial to maximize the reaction conversion rate of D-4-hydroxyphenylglycine methyl ester hydrochloride and reduce the residual D-4-hydroxyphenylglycine methyl ester hydrochloride on reactive crystallization of methyl D-(−)-4-hydroxy-phenylglycinate. Furthermore, the modified Apelblat equation and <i>C</i>_T = <i>C</i>₀(α_H⁺/<i>k</i>₁ + <i>k</i>₂/α_H⁺ + 1) type correlation regression model have made good correlation of the experimental solubility in pure solvents, mixed solvents, and the aqueous solution at different pH, respectively

Influence of Human p53 on Plant Development

<div><p>Mammalian p53 is a super tumor suppressor and plays a key role in guarding genome from DNA damage. However, p53 has not been found in plants which do not bear cancer although they constantly expose to ionizing radiation of ultraviolet light. Here we introduced <i>p53</i> into the model plant Arabidopsis and examined p53-conferred phenotype in plant. Most strikingly, p53 caused early senescence and fasciation. In plants, fasciation has been shown as a result of the elevated homologous DNA recombination. Consistently, a reporter with overlapping segments of the <i>GUS</i> gene (1445) showed that the frequency of homologous recombination was highly induced in <i>p53</i>-transgenic plants. In contrast to p53, SUPPRESSOR OF NPR1-1 INDUCIBLE 1 (SNI1), as a negative regulator of homologous recombination in plants, is not present in mammals. Comet assay and clonogenic survival assay demonstrated that SNI1 inhibited DNA damage repair caused by either ionizing radiation or hydroxyurea in human osteosarcoma U2OS cancer cells. RAD51D is a recombinase in homologous recombination and functions downstream of SNI1 in plants. Interestingly, p53 rendered the <i>sni1</i> mutants madly branching of inflorescence, a phenotype of fasciation, whereas <i>rad51d</i> mutant fully suppressed the p53-induced phenotype, indicating that human p53 action in plant is mediated by the SNI1-RAD51D signaling pathway. The reciprocal species-swap tests of p53 and SNI1 in human and Arabidopsis manifest that these species-specific proteins play a common role in homologous recombination across kingdoms of animals and plants.</p></div

Human p53 acts through the SNI1-RAD51D signaling pathway in plant.

<p>(A) Fascinated inflorescence of <i>sni1</i> mutant. Inset (box in red): part of the fascinated inflorescence is enlarged. (B) A single plant of five-week-old <i>sni1</i> mutant and four-month-old <i>p53</i>-transgenic <i>sni1</i> mutant (<i>sni1</i>/<i>p53</i>). (C) Three-week-old WT, <i>p53</i>-transgenic (<i>p53</i>), <i>rad51d</i> and <i>p53</i>-transgenic <i>rad51d</i> (<i>rad51d</i>/<i>p53</i>) plants. Arrows indicate cotyledons. (D) Number (#) of secondary inflorescences of WT, <i>p53</i>, <i>rad51d</i> and <i>rad51d</i>/<i>p53</i> plants was plotted. The letter above the bar indicates a statistically significant difference between groups at p value < 0.01. Experiments were conducted in triplicate (n > 30) with similar results.</p

The reciprocal species-swap test of p53 and SNI1 between Arabidopsis and human.

<p>(A) Somatic recombination in wild type (WT) and <i>p53</i>-transgenic (<i>p53</i>) plants is shown in blue sectors by a reporter with overlapping segments of the <i>GUS</i> gene (1445). (B) Quantitative result of panel A. Experiments were performed in three <i>p53</i>-transgenic lines (n = 50 ~ 100) (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0162840#pone.0162840.s005" target="_blank">S1 Table</a>). The result of line 1 is shown. Error bars represent SEs. ***, p value < 0.001 compared to WT by binomial test. (C) Human osteosarcoma U2OS cancer cells transfected with empty vector (EV) or hemagglutinin (HA)-tagged SNI1 (SNI1). Proteins extracted from the transfected U2OS cancer cells were blotted with anti-HA antibody (abcam, ab1265). Anti-α-tubulin was used as an internal loading control. (D) The comet assay was carried out on the transfected U2OS cancer cells which were treated with 10 Gy of ionizing radiation (IR) and recovered with indicated time. The level of DNA break repair was visualized with the length of comet tail. (E) Images in panel B were analyzed using CometScore software (Tritek) to quantify the comet tail moment of at least 75 cells for each sample. Error bars represent SEs. ***, p value < 0.001, compared to EV by binomial test. Experiments were performed three times with similar results. (F) The transfected U2OS cancer cells were pulse-treated with hydroxyurea (HU) for 24 hours to introduce DNA damage and recovered in drug-free medium. (G) Quantitative results of panel D. After 14 days of culture, colonies were counted and normalized to untreated control. Error bars represent SEs. Experiments were carried out in triplicate.</p

Influence of p53 on plant transcriptome.

<p>(A) Gene Ontology (GO) analysis of microarray data. Ten-day-old wild type (WT) and <i>p53</i>-transgenic (<i>p53</i>) seedlings were used for microarray analysis (GEO accession number: GSE79678). The differential expressed genes (t test, p value < 0.05 and fold change > 2) were analyzed for enriched biological processes by Gene Ontology (GO: <a href="https://www.arabidopsis.org/tools/bulk/go/index.jsp" target="_blank">https://www.arabidopsis.org/tools/bulk/go/index.jsp</a>). Experiments were performed in triplicate. (B) The expressions of <i>SNI1</i>, 7 <i>SSN</i>s (<i>SUPPRESSORS OF SNI1</i>s) and 3 fascination-associated genes in <i>p53</i>-transgenic plants were compared to those in WT plants. The red line indicates the expression with no change. (C) Ten-day-old wild type (WT) and <i>p53</i>-transgenic (<i>p53</i>) seedlings were used for RNA extraction. <i>RAD51D</i> transcripts were quantified by qPCR. <i>UBQ5</i> was used as an internal control. Error bars represent SEs. Experiments were conducted in triplicate.</p

Human p53-conferred phenotype in plant.

<p>(A) Leaves of 3-week-old wild type (WT) and <i>p53</i>-transgenic plants (<i>p53</i>). Arrows indicate cotyledons. (B) Four-week-old WT and <i>p53</i> plants. Arrows indicate the first pair of true leaves. (C) Bolting WT and <i>p53</i> plants. (D) Left panel: inflorescences of WT and <i>p53</i>. Insets show enlarged stems (in yellow box). Right panel: number (#) of secondary inflorescences. Error bars represent standard errors (SEs). ***, p value < 0.001, compared to WT by binomial test. Experiments were carried out in triplicate (n > 30) with similar results. (E) Siliques of WT and <i>p53</i>. Arrow indicates clustered (fascinated) siliques.</p

Phenformin induces epithelial features in breast cancer cells.

<p>(A) MCF7, ZR-75-1, MDA-MB-231 and SUM1315 cells were treated with or without phenformin for 24 hours. Cell extracts were analyzed by western blotting to detect the expression of E-cadherin, vimentin and GAPDH. (B) Expression ratios of E-cadherin to GAPDH, (C) vimentin to GAPDH. The data are presented as the mean±SEM of three replicates per group. Asterisks indicate significant differences at p<0.05 by Student’s t test. Phenformin-treated cells were labeled as P and control cells were labeled as C.</p

Phenformin inhibits MDA-MB-231 cells migration.

<p>(A) After incubation with phenformin for 24 hours, MDA-MB-231 cells (25,000 cells per chamber) were seeded in the upper chamber in serum free medium. The lower chamber contained medium with 10% FBS. After incubation for 16 hours, the cells were removed from the upper surface of the chamber membrane, and the cells on the lower surface of the chamber were stained with crystal violet and counted using a microscope(100X). (B) The number of cells/five fields was plotted. The data are presented as the mean±SEM of three replicates per group. Asterisks indicate significant differences at p<0.05 by Student’s t test.</p