Control of anthocyanin biosynthesis pathway gene expression by eutypine, a toxin from Eutypa lata, in grape cell tissue cultures

Eutypine, 4-hydroxy-3-(3-methyl-3-butene-1-ynyl) benzaldehyde, is a toxin produced by Eutypa lata, the causal agent of Eutypa dieback in grapevine. The effect of the toxin on anthocyanin synthesis has been investigated in Vitis vinifera cv. Gamay cell cultures. At concentrations higher than 200 μmol/L, eutypine reduced anthocyanin accumulation in cells. The reduction in anthocyanin accumulation was proportional to the eutypine concentrations and HPLC analysis showed that eutypine affected the levels of all anthocyanins. The effect of eutypine application on the expression of five genes of the anthocyanin biosynthesis pathway, including chalcone synthase (CHS), flavonone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX), and UDP glucose- flavonoid 3-O-glucosyl transferase (UFGT) was determined. Expression of CHS, F3H, DFR and LDOX was not affected by the addition of eutypine to grapevine cell cultures. In contrast, expression of the UFGT gene was dramatically inhibited by the toxin. These results suggest that in grapevine cell cultures, eutypine strongly affects anthocyanin accumulation by inhibiting UFGT gene expression. The mechanism of action of eutypine is discussed

Vineyard light manipulation and silicon enhance ethylene-induced anthocyanin accumulation in red table grapes

Red color resulted from anthocyanin pigment, is an essential trait for premium table grape production. Anthocyanin biosynthesis occurs through the flavonoid pathway which includes several enzymatic reactions coded by different genes. The expression of these genes is regulated by different cultural practices, cultivars, environmental conditions, and plant hormones. Recently, we reported that the anthocyanin pathway is regulated by several factors such as light and antioxidant activity. Despite the advances in cultural practices, it is still challenging to produce table grapes with high coloration, especially under the current and expected global climate change in warmer areas such as California. In the current study, we deployed two approaches to improve the accumulation of red pigment in table grapes. The first approach involves improving the expression of critical genes involved in the anthocyanin pathway through hormonal treatments and light manipulation using a reflective ground cover (RGC). The second approach was to reduce the negative effect of heat stress through stimulation of the antioxidant pathway to help remove free radicals. Treatments included ethephon (ET) at 600 mg/L, silicon (Si) at 175 mg/L, and a commercial light-reflective white ground cover (RGC) alone and in various combinations. Treatments were conducted either with or without a combination of cluster-zone leaf removal at veraison (LR) on Flame seedless (Vitis vinifera L.). Data collected in 2019 and 2020 showed that the best treatment to improve berry coloration was using ET in combination with Si and RGC, applied at veraison. Adding the LR to this combination did not improve berry color any further, but rather caused a reduction in color development. RGC without conducting LR at veraison significantly increased the quantity of reflected blue and red lights as well as the red (R) to far-red (FR) ratio (R: FR) around clusters. Results were in accordance with the increase in gene expression of flavonoid-3-O-glucosyltransferase (UFGT), a key gene in the anthocyanin biosynthesis pathway, as well as Peroxidase dismutase (POD). Manipulating the light spectrum and application of silicon in combination with the ethephon treatment could be used in table grape vineyards to improve the ethylene-induced anthocyanin accumulation and coloration

Metabolisation of eutypine by plant tissues: an HPLC determination

Eutypine, 4-hydroxy-3-(3-methyl-3-butene-1-ynyl) benzaldehyde, is a toxin produced by Eutypa lata, the causal agent of eutypa dieback of grapevine. The tolerance of some grapevine cultivars to the disease has been ascribed to the potential reduction of eutypine into its corresponding non-toxic alcohol, eutypinol. In the present study, eutypine biotransformation in different tissues of grapevine was investigated by HPLC and LC–MS. Grape callus tissues were able to biotransform eutypine into eutypinol within the first 3 h of culture. The grape plantlets cultured in vitro can also transform eutypine into eutypinol. Grape plantlet leaves do not have any effect on the uptake of eutypine, which goes through the tissues following a concentration gradient. Results revealed that the toxicity of eutypine in grape tissues is an active process showing that eutypinol is rapidly metabolised into other compounds. The use of micro-cuttings and in vitro plants showed that eutypine strongly accumulates in the bottom part of the diseased plant stems

Nanosuspension: An Emerging Trend for Bioavailability Enhancement of Etodolac

Etodolac (ET) (poorly soluble drug) nanosuspensions were prepared by both pH shift method and antisolvent techniques in order to increase its dissolution rate. Various stabilizers were used, namely, Tween 20 and 80, HPMC, PVP K44, PVA, PEG 400, NaCMC, and β-cyclodextrin. The prepared nanosuspensions were characterized by Fourier transform infrared spectroscopy (FTIR) and scanning electron microscope (SEM) and evaluated for their particle size, particle size distribution, and in vitro dissolution rate. In general, it was found that the antisolvent method for the preparation of ET nanosuspensions reduced the drug particle size to a higher extent compared to the pH shift method. The dissolution rate of ET in distilled water was markedly enhanced in the nanosized system, as more than 65% of drug dissolved in 10 min from all the nanosuspension formulations except F5 (stabilized with PVP K44) and F8 (stabilized with Tween 20), as compared to less than 20% of crude drug. Nanoparticles prepared by antisolvent method using Tween 80 as a stabilizer were selected for further in vivo study. The in vivo test demonstrated that nanoparticles of ET were well absorbed with a percentage drug absorption value 2.7 times more than that of micrometric size of crude ET

Adolescent male with anorexia nervosa: a case report from Iraq

This is the first reported case of an adolescent male with anorexia nervosa in Iraq. This disorder is believed to be rare in males across cultures and uncommon for both genders in Arab countries. The patient met the DSM-IV diagnostic criteria for anorexia nervosa. He was hospitalized and received medical and psychiatric treatment at local facilities as discussed below and responded well to treatment

Métabolisation de l'eutypine chez la vigne (Effet sur la biosynthèse des anthocyanes)

aL'eutypiose est une grave maladie de dépérissement de la vigne provoquée par le champignon Eutypa lata. Ce parasite agit à distance sur les organes herbacés par l'intermédiaire d'une toxine, la 4-hydroxy-3-(3-méthyl-3-butène-1-ynyl) benzaldéhyde, appelée eutypine. Les cellules de vigne sont capables de transformer l'eutypine en un composé non toxique pour la vigne, l'alcool 4-hydroxy-3-(3-méthyl-3-butène-1-ynyl) benzylique, appelé eutypinol. Le premier objectif de ce travail a été de mieux caractériser ce mécanisme de résistance à la toxine et d'étudier l'influence de différents facteurs sur la métabolisation de l'eutypine en eutypinol. Les résultats présentés indiquent que l'eutypine est une molécule photostable. Nous avons montré que la concentration d'agar dans le milieu de culture influence l'absorption de l'eutypine dans les tissus de Gamay. Dans les cellules de vigne l'eutypine pénètre rapidement puis est métabolisée très rapidement en eutypinol. Cet alcool est excrété dans le milieu de culture et également métabolisé en d'autres composés. L'utilisation de microboutures et de vitroplants a permis de montrer que l'eutypine circule dans la plante et s'accumule plus fortement dans les tissus du bas de la tige. De plus, les feuilles interviendraient comme les tiges dans l'accumulation et la métabolisation de l'eutypine. Les analyses par spectrométrie de masse ont permis de détecter d'autres composés susceptibles de dériver de l'eutypinol.Dans la deuxième partie de cette étude les résultats obtenus montrent très clairement que l'eutypine inhibe fortement l'accumulation des anthocyanes dans les tissus de Gamay cultivés in vitro. Nous avons décrit l'inhibition de l'expression du gène UFGT par l'eutypine pour la première fois dans la vigne. Nos résultats suggèrent que dans les ceps de vigne contaminés par Eutypa lata, l'eutypine peut certainement inhiber également l'expression du gène UFGT et induire une diminution de l'accumulation des anthocyanes dans les cellules de la pellicule des baies de raisin.TOULOUSE-ENSAT-Documentation (315552324) / SudocSudocFranceF

Comparative Evaluation for the Effect of Green tea, Aloe vera and Chlorhexidine on Dentin Erosion

ABSTRACT: Purpose: This study aimed to compare the anti-collagenolytic effect of Green tea, Aloe vera and chlorhexidine on artificial erosion in human dentin using microhardness test and environmental scanning electron microscope (ESEM). Materials and Methods: For this study, 60 extracted sound human maxillary premolars were subjected to artificial erosion and randomly divided into three equal groups (20 premolars each) according to immersion in the tested anti-collagenolytic agents; group (A1) Green tea, (A2) Aloe vera and group (A3) Chlorhexidine (CHX). The microhardness analysis and (ESEM) photomicrographs were taken for the surface of dentin specimens three times, at base line, after the artificial erosion and after immersion in each treatment solution. Bonferroni’s post-hoc test was used for pair-wise comparisons. Statistical analysis was performed with IBM SPSS Statistics for Windows. Results: Results showed no statistically significant difference in microhardness mean values between Aloe vera and (CHX), However, both showed statistically significant increase in microhardness values than Green tea. These results were also supported by (ESEM) photomicrographs. Conclusion: Chemical gold standard CHX could be substituted by natural safe herbal sources and hence Aloe vera opens a new perspective in management of dental erosion

An educational intervention to promote appropriate antibiotic use for acute respiratory infections in a district in Egypt- pilot study

Abstract Background Antibiotic overuse is the most important modifiable factor contributing to antibiotic resistance. We conducted an educational campaign in Minya, Egypt targeting prescribers and the public through communications focused on appropriate antibiotic use for acute respiratory infections (ARIs). Methods The entire population of Minya was targeted by the campaign. Physicians and pharmacists were invited to participate in the pre-intervention assessments. Acute care hospitals and a sample of primary healthcare centers in Minya were randomly selected for a pre-intervention survey and all patients exiting outpatient clinics on the day of the survey were invited to participate. The same survey methodology was conducted for the post-intervention assessments. Descriptive comparisons were made through three assessments conducted pre- and post-intervention. We quantitated antibiotic prescribing through a survey administered to patients with an ARI exiting outpatient clinics. Additionally, physicians, pharmacists, and patients were interviewed regarding their attitudes and beliefs towards antibiotic prescribing. Finally, physicians were tested on three clinical scenarios (cold, bronchitis, and sinusitis) to measure their knowledge on antibiotic use. Results Post-intervention patient exit surveys revealed a 23.1% decrease in antibiotic prescribing for ARIs in this population (83.7 to 64.4%) and physicians and pharmacists self-reported less frequently prescribing antibiotics for ARIs on their follow-up surveys. We also found an increase in correct responses to the clinical scenarios and in attitude and belief scores for physicians, pharmacists, and patients regarding antibiotic use in the post-intervention sample. Conclusions Overall, the samples surveyed after the community-based educational campaign reported a lower frequency of antibiotic prescribing and improved knowledge and attitudes regarding antibiotic misuse compared to the samples surveyed before the campaign. Ongoing interventions educating providers and patients are needed to decrease antibiotic misuse and reduce the spread of antibiotic resistance in Egypt