Transcriptomic and metabolomic profiling reveal the p53-dependent benzeneacetic acid attenuation of silica‐induced epithelial–mesenchymal transition in human bronchial epithelial cells

Background Silica exposure underlies the development of silicosis, one of the most serious occupational hazards worldwide. We aimed to explore the interaction of the silica-induced epithelial–mesenchymal transition (EMT)-related transcripts with the cellular metabolism regulated by p53. Methods We knocked out p53 using CRISPR/Cas9 in the human bronchial epithelial (HBE) cell line. The transcriptomic and metabolomic analyses and integrative omics were conducted using microarrays, GC–MS, and MetaboAnalyst, respectively. Results Fifty-two mRNAs showed significantly altered expression in the HBE p53-KO cells post-silica exposure. A total of 42 metabolites were putatively involved in p53-dependent silica-mediated HBE cell dysfunction. Through integrated data analysis, we obtained five significant p53-dependent metabolic pathways including phenylalanine, glyoxylate, dicarboxylate, and linoleic acid metabolism, and the citrate cycle. Through metabolite screening, we further identified that benzeneacetic acid, a key regulation metabolite in the phenylalanine metabolic pathway, attenuated the silica-induced EMT in HBE cells in a p53-dependent manner. Interestingly, despite the extensive p53-related published literature, the clinical translation of these studies remains unsubstantial. Conclusions Our study offers new insights into the molecular mechanisms by which epithelial cells respond to silica exposure and provide fresh perspective and direction for future clinical biomarker research and potential clinically sustainable and translatable role of p53.

ANALYSIS OF BIO-ACTIVE COMPOUNDS PRESENT IN THE LEAVES AND STEM OF TRICHOSANTHES ROXB. USING GC-MS TECHNIQUE WITH RESPECT TO ITS ANTI-INFLAMMATORY ACTION

Objective: Structural elucidation studies on Trichosanthes lobata ethyl acetate and methanol extracts of leaf and stem parts through Gas Chromatography-Mass Spectrometry (GC-MS) technique with respect to anti-inflammatory potential. Methods: Extracts obtained with shade dried and powdered samples in successive solvent extraction using ethyl acetate and methanol by Soxhlet apparatus and subjected to GC-MS analysis and interpreted for its anti-inflammatory compounds. Results: The study revealed that the extraction solvent used was able to recover compound of classes such as organic acid esters and conjugated alkaloids in larger quantities than other classes of compounds and they varied with leaf and stem and also with the polarity of solvents used. In total compounds identified, GC-MS profile of the Ethyl Acetate leaf extract of T. lobata contained 41 compounds, stem extract contained 45 compounds which have reported bioassays in PubChem. Whereas GC-MS profile of methanol leaf extract of T. lobata contained 66 compounds and stem extract contained 46 compounds having bioassay reports in PubChem. A large number of phytochemical peaks with good area percentage were found in methanolic extract. We were also able to find out potent anti-inflammatory compounds including Octanoic acid, Dodecanoic acid, Octadecane, Enoic acid, Hexanoic acid, Quinazolin-8-one, Ilicic acid, Pentadecanoic acid, Oxaspiro, Benzeneacetic acid, etc. from the extracts. Conclusion: T. lobata contains phytocompounds against inflammation which may serve as a new drug lead of natural products origin in future and make it employable in modern pharmacological practices.

CHROMATOGRAPHY-MASS SPECTROMETRY OF TOLUENE EXTRACT OF YARROW (ACHILLED MILLEFOLIUM L., ASTERACEAE FAMILY) (the message II)

The aim of the study is to study in detail the chemical composition of organic matter of common yarrow in order to establish the main biochemical processes responsible for the formation of the composition of the latter. Materials and methods of research. The chemical composition of toluene extract of yarrow was studied by chromatography-mass spectrometry under the following conditions. GC-2010 gas chromatograph connected to a GCMs-TQ-8030 triple quadrupole mass spectrometer running GCMS Solution 4.11 software. The results of studying the chemical composition of toluene extract-a product of sequential exhaustive extraction of yarrow by chromatography-mass spectrometry, which allowed identifying 129 individual compounds for which the quantitative content was determined, mass spectra and structural formulas were obtained, and the structure-group composition of the extract was calculated. Results and discussion. The basis of the extract is determined by hydrocarbons, esters, sterols and carboxylic acids, which account for: 43.59; 15.47; 15.33 and 7.59 (wt.% of the extract), respectively, the content of ketones, alcohols, aldehydes and organosilicon compounds-3.77; 2.77; 1.68 and 6.49 (wt.% of the extract). The presence of phenols and glycosides has not been established; furan and PYRAN fragments are part of the structures of individual alcohols and ketones. Based on the features of the chemical composition, it can be argued that the pharmacological effect of toluene extract of yarrow is determined precisely by the content of hydrocarbons, with the dominance of alkenes, alkynes, arenes and cycloalkanes; sterols of the following types: Betulin, Lupeol, γ-Sitosterol and Sitostenon, Campesterol, 24-Noroleana-3.12-dien; carboxylic acids containing up to three double and triple bonds in the hydrocarbon chain, as well as esters mainly formed by Oxalic and Benzeneacetic acid. Undoubtedly, a certain contribution to the direction of the pharmacological action of this extract is made by organosilicon compounds, the proportion of which is 6.49 (wt.% of the extract).

Transcriptomic and Metabolomic Profiling Reveal the p53-Dependent Benzeneacetic Acid Attenuation of Silica-Induced Epithelial–Mesenchymal Transition in Human Bronchial Epithelial Cells

Background: Silica exposure underlies the development of silicosis, one of the most serious occupational hazards worldwide. We aimed to explore the interaction of the silica-induced epithelial–mesenchymal transition (EMT)-related transcripts with the cellular metabolism regulated by p53.Methods: We knocked out p53 using CRISPR/Cas9 in the human bronchial epithelial (HBE) cell line. The transcriptomic and metabolomic analyses and integrative omics were conducted using microarrays, GC-MS, and MetaboAnalyst, respectively. Results: Fifty-two mRNAs showed significantly altered expression in the HBE p53-KO cells post-silica exposure. A total of 42 metabolites were putatively involved in p53-dependent silica-mediated HBE cell dysfunction. Through integrated data analysis, we obtained five significant p53-dependent metabolic pathways including phenylalanine, glyoxylate, dicarboxylate, and linoleic acid metabolism, and the citrate cycle. Through metabolite screening, we further identified that benzeneacetic acid, a key regulation metabolite in the phenylalanine metabolic pathway, attenuated the silica-induced EMT in HBE cells ina p53-dependent manner. Interestingly, despite the extensive p53-related published literature, the clinical translation of these studies remains unsubstantial. Conclusions: Our study offers new insights into the molecular mechanisms by which epithelial cells respond to silica exposure and provide fresh perspective and direction for future clinical biomarker research and potential clinically sustainable and translatable role of p53.

Mengungkap Senyawa pada Nata De Coco sebagai Pangan Fungsional

Nata adalah selulosa ekstraseluler yang dihasilkan dari aktiivitas bakteri Acetobacter xylinum dalam proses fermentasi, dan merupakan salah satu makanan kesehatan yang kaya akan serat. Nata yang paling umum ditemukan adalah nata de coco dengan media fermentasi air kelapa. Faktor-faktor dominan dalam pembuatan nata adalah ketersediaan nutrisi (karbon dan nitrogen), derajat keasaman dan media fermentasi yang digunakan. Penentuan kualitas terbaik media air kelapa yang digunakan, penentuan konsentrasi nitrogen dan sukrosa yang ditambahkan serta kondisi pH optimum akan menghasilkan nata yang maksimal.Penelitian ini bertujuan untuk mencari kualitas terbaik pada fermentasi dalam proses pembuatan nata de coco, dan mengetahui senyawa fungsional yang terdapat pada nata de coco sebagai pangan fungsional. Hasil pengujian analisa sampel nata de coco menggunakan Gas chromatography–mass spectrometry (GC-MS) menunjukkan senyawa yang sangat bermaaf bagi kesehatan tubuh manusia. Sepuluh besar senyawa fungsional yag terdapat pada nata de coco yaitu: Benzeneacetic Acid sebagai anti fungal dan scavenger. Hexadecanoid Acid memiliki efek anti-inflamasi, anti bakteri dan anti fungi. 22-Hydroxyhopane, Tetradecanoic Acid yang mempunyai aktivitas antimikroba dan antifungal. 9-Octadecanoid Acid, ρ-Cresol memiliki aktivitas antioksidan. 9-Octadecenamide berfungsi untuk mencegah Alzheimer, menurunkan kolesterol dan menurunkan tekanan darah, Senyawa fungsional yang lain diantaranya (Z), Phenol, 4-(2-aminoethyl), Pentadecanoic Acid, 1-Heptadecanecarboxylic acid. Komposisi kimia nata de coco dihasilkan kadar air 95%. abu 0,35% dan protein 0,45%. Penelitian menunjukkan peningkatkan produksi nata dengan kualitas yang baik sesuai persyaratan standar yang telah ditetapkan dan mempunyai senyawa senyawa yang bermanfaat sebagai pangan fungsional yang bermanfaat bagi kesehatan.

In vitro and in vivo GABAA Receptor Interaction of the Propanidid Metabolite 4-(2-[Diethylamino]-2-Oxoethoxy)-3-Methoxy-Benzeneacetic Acid

Background: Propanidid is a γ-aminobutyric acid type A (GABAA) receptor agonist general anesthetic and its primary metabolite is 4-(2-[diethylamino]-2-oxoethoxy)-3-methoxy-benzeneacetic acid (DOMBA). Despite having a high water solubility at physiologic pH that might predict low-affinity GABAA receptor interactions, DOMBA is reported to have no effect on GABAA receptor currents, possibly because the DOMBA concentrations studied were simply insufficient to modulate GABAA receptors. Our objectives were to measure the propanidid and DOMBA concentration responses on ­GABAA receptors and to measure the behavioral responses of DOMBA in mice at concentrations that affect GABAA receptor currents in vitro. Methods: GABAA receptors were expressed in oocytes using clones for the human GABAA α1, β2 and γ2s subunits. The effects of DOMBA (0.2–10 mmol/L) and propanidid (0.001–1 mmol/L) on oocyte GABAA currents were studied using standard 2-electrode voltage clamp techniques. Based on in vitro results, 6 mice received ­DOMBA 32 mg intraperitoneal and were observed for occurrence of neurologic effects and DOMBA plasma concentration was measured by liquid chromatography tandem mass spectrometry. Results: DOMBA both directly activates GABAA receptors and antagonizes its GABA-mediated opening in a concentration-dependent manner at concentrations between 5–10 and 0.5–10 mmol/L respectively. In vivo, DOMBA produced rapid onset sedation at plasma concentrations that correlate with direct GABAA receptor activation. Conclusion: DOMBA modulation of GABAA receptors is associated with sedation in mice. Metabolites of propanidid analogues currently in development may similarly modulate GABAA, and impaired elimination of these metabolites could produce clinically relevant neurophysiologic effects.