Ca-bound pectin was plentiful in the boundary cell layers between the skin and mesocarp.

<p>A, Ca content in tomato fruit tissues. B, Ca content in the tomato fruit cell wall. The alcohol-insoluble residue fraction was extracted from fruit tissues for analysis. Ca content in each fraction was determined by inductively coupled plasma atomic emission spectroscopy (ICP-AES). C, Ca determination between the skin and mesocarp. Quantitative imaging by secondary ion-microprobe mass spectrometry (SIMS). Ruthenium: showing ruthenium, which binds with pectin. Calcium: showing calcium content. Merge: showing overlap of ruthenium (pectin) and calcium.</p

Preparation for tissue-specific analysis.

<p>A, The fruit ripening stages of cv. Micro Tom. The six stages included immature green (I), mature green (M), breaker (B), turning (T), red ripe (R) and overripe (O). B, The fruit tissues of cv. Micro Tom. The eight tissues included skin, mesocarp, endocarp, septum, locular tissue, seed, placenta and core. These were separated by hand-sectioning.</p

Immunolocalization of Homogalacturonan (HG) epitopes in tomato fruit longitudinal section of fruit.

<p>A, Light microscopy images with Toluidine Blue as a control. B, Negative control. C, Immunolabeled with LM19, which labelled de-methyl-esterified HG. D, Immunolabeled with LM20, which labelled methyl-esterified HG. Ripening stage: I, immature green; M, mature green; B, breaker; T, turning; R, red ripe. E, Immunolabeled with LM19, which labelled de-methyl-esterified HG. Top row micrographs indicate Sepal side of pericarp (Se), middle row micrographs indicate Middle of pericarp (Mi), and bottom row micrographs indicate Stigma side of pericarp (St).</p

Pectin biosynthesis/degradation-related gene expression patterns differed among tissues.

<p>Gene expression was analysed by RT-PCR. A, PE2, pectin methyl-esterase 2 (25 cycles); B, PG2, polygalacturonase 2 (25 cycles); C, GAUT1 family <i>Arabidopsis</i> pectin homogalacturonan galacturonosyltransferase-like gene family (25 cycles); D, rRNA, as a control (20 cycles). Expression levels were compared to <i>rRNA</i> in the same assay. The eight tissues analyzed in these assays included skin, mesocarp, endocarp, septum, locular tissue, seed, placenta, and core. Ripening stages were the following: I, immature green; M, mature green; B, breaker; T, turning; R, red ripe; O, overripe.</p

PE activity and a decreasing degree of pectin methyl-esterification were specific to pericarp tissues.

<p>A, PE activity. Total protein in the cell wall was extracted from each fruit tissue and assayed for PE activity. One unit means a decrease in OD₆₂₀ per second. B, Degree of pectin methyl-esterification. The pectin fraction was extracted from fruit tissues for analysis. The five tissues analysed in these assays included skin, mesocarp/endocarp, septum, locular tissue and seed. Ripening stage: I, immature green; M, mature green; B, breaker; T, turning; R, red ripe; O, overripe. ±SD of three independent replicates. C, Pectin localisation between the skin and the mesocarp. Light microscopy images with Toluidine Blue (a-e) as a control, Ruthenium Red after NaOH treatment for 5 min (f-j) staining of pectin and Ruthenium Red (k-o) staining of de-methyl-esterified pectin. Ripening stage: I, immature green; M, mature green; B, breaker; T, turning; R, red ripe.</p

Changes in the cell wall and pectin content differed in fruit tissues during ripening.

<p>A, Dry weight of the alcohol-insoluble residue (AIR) per 1 g fresh weight from each fruit tissue. B, Pectin content per 1 g fresh weight from each fruit tissue. C, Relative pectin content in the cell wall. The five tissues analysed in this assay included skin, mesocarp/endocarp, septum, locular tissue and seed. D, Suger content in pectin fraction extracted from skin cell wall. Ripening stage: I, immature green; M, mature green; B, breaker; T, turning; R, red ripe; O, overripe. ±SD of three independent replicates.</p

Image_1_Palmitoylethanolamide Ameliorates Carbon Tetrachloride-Induced Liver Fibrosis in Rats.PDF

<p>Background: Liver fibrosis is a complex inflammatory and fibrogenic process, and the progression of fibrosis leads to cirrhosis. The only therapeutic approaches are the removal of injurious stimuli and liver transplantation. Therefore, the development of anti-fibrotic therapies is desired. Palmitoylethanolamide (PEA) is an endogenous fatty acid amide belonging to the N-acylethanolamines family and contained in foods such as egg yolks and peanuts. PEA has therapeutic anti-inflammatory, analgesic, and neuroprotective effects. However, the effects and roles of PEA in liver fibrosis remain unknown. Here we investigated the therapeutic effects of PEA in rats with liver fibrosis.</p><p>Methods: We conducted in vitro experiments to investigate the effects of PEA on the activation of hepatic stellate cells (HSCs, LX-2). Liver fibrosis was induced by an intraperitoneal injection of 1.5 mL/kg of 50% carbon tetrachloride twice a week for 4 weeks. Beginning at 3 weeks, PEA (20 mg/kg) was intraperitoneally injected thrice a week for 2 weeks. Then rats were sacrificed and we performed histological and quantitative reverse-transcription polymerase chain reaction analyses.</p><p>Results: The expression of α-smooth muscle actin (SMA) induced by transforming growth factor (TGF)-β1 in HSCs was significantly downregulated by PEA. PEA treatment inhibited the TGF-β1-induced phosphorylation of SMAD2 in a dose-dependent manner, and upregulated the expression of SMAD7. The reporter gene assay demonstrated that PEA downregulated the transcriptional activity of the SMAD complex upregulated by TGF-β1. Administration of PEA significantly reduced the fibrotic area, deposition of type I collagen, and activation of HSCs and Kupffer cells in rats with liver fibrosis.</p><p>Conclusion: Activation of HSCs was significantly decreased by PEA through suppression of the TGF-β1/SMAD signaling pathway. Administration of PEA produced significant improvement in a rat model of liver fibrosis, possibly by inhibiting the activation of HSCs and Kupffer cells. PEA may be a potential new treatment for liver fibrosis.</p

Induction of Reg IV expression by CDX2.

<p>A: Western blot analysis of CDX2, Reg IV, and β-actin and qRT-PCR analysis of <i>REG4</i> in 9 GC cell lines. B: Western blot analysis of CDX2, Reg IV, and β-actin and qRT-PCR analysis of <i>REG4</i> in HT-29/PGS-CDX2, HT-29/PGS-neo, SW480/PGS-CDX2, and SW480/PGS-neo. C: Western blot analysis of Reg IV and β-actin and qRT-PCR analysis of <i>REG4</i> in HT-29/CDX2-ER. Time course of <i>REG4</i> gene induction in response to activation of a CDX2-ER fusion protein by 4-OHT was analyzed. D: Western blot analysis of CDX2, Reg IV, and β-actin and qRT-PCR analysis of <i>REG4</i> in HSC-39 cells transfected with CDX2 siRNA (siRNA1 and siRNA2) and the negative control siRNA. The units of <i>REG4</i> mRNA expression level are arbitrary. <i>P</i> values were calculated using Student’s t-test. * N.S. = not significant.</p

Interaction of arc magmas with subvolcanic hydrothermal systems: insights from compositions and metasomatic textures of olivine crystals in fresh basalts of Daisen and Mengameyama, Western Honshu, Japan

<p>Pleistocene basalts from Daisen and Mengameyama in the SW Japan volcanic arc of western Honshu are characterized by an abundance of olivine crystals with Fe-rich rims. At Daisen, these have previously been interpreted to have formed from their host melt by equilibrium crystal fractionation and by disequilibrium fractionation during supercooling. Here we use combined electron probe microanalysis, isotopography, transmission electron microscopy and selected area electron diffraction to show that crystal rims are significantly enriched in aluminium (up to <em>c.</em> 1 wt%) and hydrogen (up to <em>c.</em> 10 000 ppm) hosted in oriented low-density amorphous domains. These domains are interpreted to have formed by melting of deuteric and/or post-deuteric metasomatic alteration minerals upon uptake of older olivine crystals into fresh, initially aphyric host melts up to a few hours prior to eruption. It is argued that uptake of variably altered crystals into initially aphyric or sparsely phyric melts may be a common process at subduction zones, and can account for typical disequilibrium textures displayed by arc magmas erupted in SW Japan and elsewhere. Analyses of the altered crystal cargo in arc volcanic rocks therefore provides an important tool for understanding subvolcanic hydrothermal systems and the interaction of ascending melts with such systems. </p

<i>REG4</i> promoter analysis.

<p>Localization of regulatory elements and CDX2 binding sites in the 5′-flanking region of the <i>REG4</i> gene. A: Schematic representation of the 5′-flanking region of the <i>REG4</i> gene. The location and sequence of 4 consensus CDX2-binding sites in the 5′-flanking region of <i>REG4</i> (i.e., sites A, B, C, and D) is indicated. B: Schematic representation of <i>REG4</i> reporter gene constructs. The <i>REG4</i> genomic DNA sequences present in the reporter gene vectors are indicated. Key sequences for <i>REG4</i> transcription reside between base pairs −634 and −116. Reporter assays with the series of <i>REG4</i> deletion constructs were performed in the CDX2-expressing GC cell line, HSC-39. The luciferase activity of the empty pGL4.10 basic vector was assigned a value of 1. The reporter assays were performed in triplicate, and mean and SD values of luciferase activity are shown. C: Localized mutations in the candidate CDX2-binding sites (i.e., sites A, B, C, and D) were introduced into the −2019/+58 construct, and the series of constructs generated is shown. The CDX2 candidate binding site designated as “C” plays critical roles in <i>REG4</i> transcription. Reporter assays were performed in CDX2-expressing GC cell line, HSC-39. The activity of the pGL4.10 basic vector was assigned a value of 1. Assays were performed in triplicate. Mean and SD luciferase activity values are shown.</p