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Evaluation of spice and herb as phyto-derived selective modulators of human retinaldehyde dehydrogenases using a simple in vitro method.

ABSTRACT: Selective modulation of retinaldehyde dehydrogenases (RALDHs)-the main aldehyde dehydrogenase (ALDH) enzymes converting retinal into retinoic acid (RA), is very important not only in the RA signaling pathway but also for the potential regulatory effects on RALDH isozyme-specific processes and RALDH-related cancers. However, very few selective modulators for RALDHs have been identified, partly due to variable overexpression protocols of RALDHs and insensitive activity assay that needs to be addressed. In the present study, deletion of the N-terminal disordered regions is found to enable simple preparation of all RALDHs and their closest paralog ALDH2 using a single protocol. Fluorescence-based activity assay was employed for enzymatic activity investigation and screening for RALDH-specific modulators from extracts of various spices and herbs that are well-known for containing many phyto-derived anti-cancer constituents. Under the established conditions, spice and herb extracts exhibited differential regulatory effects on RALDHs/ALDH2 with several extracts showing potential selective inhibition of the activity of RALDHs. In addition, the presence of magnesium ions was shown to significantly increase the activity for the natural substrate retinal of RALDH3 but not the others, while His-tag cleavage considerably increased the activity of ALDH2 for the non-specific substrate retinal. Altogether we propose a readily reproducible workflow to find selective modulators for RALDHs and suggest potential sources of selective modulators from spices and herbs.

SUBMITTER: Bui TBC

PROVIDER: S-EPMC8493444 | biostudies-literature |

REPOSITORIES: biostudies-literature

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A commentary on the paper: 'Evaluation of spice and herb as phytoderived selective modulators of human retinaldehyde dehydrogenases using a simple in vitro method'.

Project description:It is commonly known that aldehyde dehydrogenases (ALDHs) are a promising therapeutic target in many diseases. Bui et al.-the authors of the paper I am discussing here (Biosci Rep (2021) 41(5): BSR20210491 https://doi.org/10.1042/BSR20210491)-point that there is a lack of research on the use of spices and herbs as the sources of naturally occurring modulators of ALDH activity. In order to carry out this type of research, the authors prepared ethanolic extracts of 22 spices and herbs. The main objective of the study was to investigate retinaldehyde dehydrogenases (RALDHs), of which retinal is the main substrate and ALDH2, the mitochondrial isoform, having acetaldehyde as the main substrate. The obtained results indicated that the tested extracts exhibited differential regulatory effects on RALDHs/ALDH2 and some of them showed a potential selective inhibition of the activity of RALDHs.

| S-EPMC8753342 | biostudies-literature

'BhAVI-23'-A spice-herb based dietary infusion possessing in-vitro anti-viral potential.

Project description:BackgroundViruses cause many life threatening human diseases. Recently, COVID-19 pandemic has challenged the health care systems worldwide. As a disease preventive approach and to bring relief to the severity of the symptoms, a infusion termed as Bhabha Anti-Viral Infusion-23 ('BhAVI-23') was conceptualized and formulated which comprised of 23 selected spices and herbals.ObjectiveThe present study was conducted to assess the in vitro antiviral potential of the formulation, BhaAVI-23.Material and methodsThe in-vitro anti-viral potential of BhAVI-23 was assessed through inhibition of HIV1 reverse transcriptase (RT) as well as through a novel P1 (virulent) bacteriphage based screening assay system. Anti-diabetic potential was assessed by non-enzymatic glycosylation of haemoglobin and the bioactive volatile components were detected through headspace gas chromatography followed by molecular docking analysis.ResultsThe infusion displayed prominent anti-viral activity as evident from significant (57%) inhibition of the HIV1-RT as well as through reduction in the infectivity of P1 (virulent) bacteriophage. The infusion also exerted profound protection (∼64%) to non-enzymatic glycosylation of haemoglobin. Headspace gas chromatography and mass spectrometric analysis confirmed the presence of at least 47 major compounds. Docking analysis indicated possible interaction of α-pinene and eugenol with SARS-CoV spike protein.ConclusionThis 'BhAVI-23' infusion displayed prominent in-vitro anti-viral and anti-diabetic potential in different model systems. These attributes have relevance as diabetic patients are more prone to COVID-19 morbidity. 'BhAVI-23' opens the avenue for its potential inclusion as a supportive health care system upon due regulatory approval during the current pandemic.

| S-EPMC7783457 | biostudies-literature

Profiling of retinaldehyde dehydrogenases in cancer cells

Project description:Retinaldehyde dehydrogenases convert retinal into all-trans retinoic acid (ATRA), thereby regulating cell differentiation and cancer stem cell proliferation. Chemical tools and methods are valuable commodities to study aldehyde dehydrogenase (ALDH) activity in human cells. Here, we describe the design, synthesis and application of LEI-945, a first-in-class activity-based probe modeled on retinal that reports on individual ALDHs in cancer cells. Comparative chemical proteomics with LEI-945 explained the ability of various breast cancer cell lines to produce ATRA via retinaldehyde dehydrogenases isozymes, whereas the ALDEFLUORTM assay could not. Competitive chemical proteomics using LEI-945 revealed that the ALDH inhibitor NCT-505 inhibited both ALDH1A1 and ALDH1A3 in MDA-MB-468 breast cancer cells inducing a cell cycle arrest in the G1 phase as well as cell death through necrosis. Our results show that LEI-945 holds promise to guide the discovery of ALDH-based therapeutics

2019-12-18 | PXD015495 | Pride

Development of a Retinal-Based Probe for the Profiling of Retinaldehyde Dehydrogenases in Cancer Cells.

Project description:Retinaldehyde dehydrogenases belong to a superfamily of enzymes that regulate cell differentiation and are responsible for detoxification of anticancer drugs. Chemical tools and methods are of great utility to visualize and quantify aldehyde dehydrogenase (ALDH) activity in health and disease. Here, we present the discovery of a first-in-class chemical probe based on retinal, the endogenous substrate of retinal ALDHs. We unveil the utility of this probe in quantitating ALDH isozyme activity in a panel of cancer cells via both fluorescence and chemical proteomic approaches. We demonstrate that our probe is superior to the widely used ALDEFLUOR assay to explain the ability of breast cancer (stem) cells to produce all-trans retinoic acid. Furthermore, our probe revealed the cellular selectivity profile of an advanced ALDH1A1 inhibitor, thereby prompting us to investigate the nature of its cytotoxicity. Our results showcase the application of substrate-based probes in interrogating pathologically relevant enzyme activities. They also highlight the general power of chemical proteomics in driving the discovery of new biological insights and its utility to guide drug discovery efforts.

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Spice and Herb Frauds: Types, Incidence, and Detection: The State of the Art.

Project description:There is a necessity to protect the quality and authenticity of herbs and spices because of the increase in the fraud and adulteration incidence during the last 30 years. There are several aspects that make herbs and spices quite vulnerable to fraud and adulteration, including their positive and desirable sensorial and health-related properties, the form in which they are sold, which is mostly powdered, and their economic relevance around the world, even in developing countries. For these reasons, sensitive, rapid, and reliable techniques are needed to verify the authenticity of these agri-food products and implement effective adulteration prevention measures. This review highlights why spices and herbs are highly valued ingredients, their economic importance, and the official quality schemes to protect their quality and authenticity. In addition to this, the type of frauds that can take place with spices and herbs have been disclosed, and the fraud incidence and an overview of scientific articles related to fraud and adulteration based on the Rapid Alert System Feed and Food (RASFF) and the Web of Science databases, respectively, during the last 30 years, is carried out here. Next, the methods used to detect adulterants in spices and herbs are reviewed, with DNA-based techniques and mainly spectroscopy and image analysis methods being the most recommended. Finally, the available adulteration prevention measurements for spices and herbs are presented, and future perspectives are also discussed.

| S-EPMC10528162 | biostudies-literature

Processes to Preserve Spice and Herb Quality and Sensory Integrity During Pathogen Inactivation.

Project description:Selected processing methods, demonstrated to be effective at reducing Salmonella, were assessed to determine if spice and herb quality was affected. Black peppercorn, cumin seed, oregano, and onion powder were irradiated to a target dose of 8 kGy. Two additional processes were examined for whole black peppercorns and cumin seeds: ethylene oxide (EtO) fumigation and vacuum assisted-steam (82.22 °C, 7.5 psia). Treated and untreated spices/herbs were compared (visual, odor) using sensory similarity testing protocols (α = 0.20; β = 0.05; proportion of discriminators: 20%) to determine if processing altered sensory quality. Analytical assessment of quality (color, water activity, and volatile chemistry) was completed. Irradiation did not alter visual or odor sensory quality of black peppercorn, cumin seed, or oregano but created differences in onion powder, which was lighter (higher L* ) and more red (higher a* ) in color, and resulted in nearly complete loss of measured volatile compounds. EtO processing did not create detectable odor or appearance differences in black peppercorn; however visual and odor sensory quality differences, supported by changes in color (higher b* ; lower L* ) and increased concentrations of most volatiles, were detected for cumin seeds. Steam processing of black peppercorn resulted in perceptible odor differences, supported by increased concentration of monoterpene volatiles and loss of all sesquiterpenes; only visual differences were noted for cumin seed. An important step in process validation is the verification that no effect is detectable from a sensory perspective.

| S-EPMC5435955 | biostudies-literature

Evaluation of a DNA-based method for spice/herb authentication, so you do not have to worry about what is in your curry, buon appetito!

Project description:It is long believed that some spices may help protect against certain chronic conditions. Spices are usually parts of plants that have been powdered into small pieces. Have you ever wondered what the curry powder in your dish is made of? The aim of this work was to develop an appropriate DNA-based method for assessment of spice identity. Selecting the best marker for species recognition in the Zingiberaceae family. Six DNA regions were investigated in silico, including ITS, matK, rbcL, rpoC, trnH-psbA and trnL. Then, only four regions (ITS, matK, rbcL and trnH-psbA) were included in the simulated HRM (High-resolution Melting) analysis as the results from previous analysis showed that rpoC and trnL may not be suitable to be used to identify Zingiberaceae species in HRM analysis based on both the percentage of nucleotide variation and GC content. Simulated HRM analysis was performed to test the feasibility of Bar-HRM. We found that ITS2 is the most effective region to be used for identification of the studied species and thus was used in laboratory HRM analysis. All seven tested Zingiberaceae plants were then able to be distinguished using the ITS2 primers in laboratory HRM. Most importantly the melting curves gained from fresh and dried tissue overlapped, which is a crucial outcome for the applicability of the analysis. The method could be used in an authentication test for dried products. In the authentication test, only one of seven store-sold Zingiberaceae products that were tested contained the species listed on their labels, while we found substitution/contamination of the tested purchased products in the rest.

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| S-EPMC4145728 | biostudies-literature

Mutation detection by real-time PCR: a simple, robust and highly selective method.

Project description:BackgroundMolecular tests for diagnosis of disease, particularly cancer, are gaining increased acceptance by physicians and their patients for disease prognosis and selection of treatment options. Gene expression profiles and genetic mutations are key parameters used for the molecular characterization of tumors. A variety of methods exist for mutation analysis but the development of assays with high selectivity tends to require a process of trial and error, and few are compatible with real-time PCR. We sought to develop a real-time PCR-based mutation assay methodology that successfully addresses these issues.Methodology/principal findingsThe method we describe is based on the widely used TaqMan real-time PCR technology, and combines Allele-Specific PCR with a Blocking reagent (ASB-PCR) to suppress amplification of the wildype allele. ASB-PCR can be used for detection of germ line or somatic mutations in either DNA or RNA extracted from any type of tissue, including formalin-fixed paraffin-embedded tumor specimens. A set of reagent design rules was developed enabling sensitive and selective detection of single point substitutions, insertions, or deletions against a background of wild-type allele in thousand-fold or greater excess.Conclusions/significanceASB-PCR is a simple and robust method for assaying single nucleotide mutations and polymorphisms within the widely used TaqMan protocol for real time RT-PCR. The ASB-PCR design rules consistently produce highly selective mutation assays while obviating the need for redesign and optimization of the assay reagents. The method is compatible with formalin-fixed tissue and simultaneous analysis of gene expression by RT-PCR on the same plate. No proprietary reagents other than those for TaqMan chemistry are required, so the method can be performed in any research laboratory with real-time PCR capability.

| S-EPMC2642996 | biostudies-literature

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