(Z)-ethyl cinnamate

New technical approaches for 3D morphological imaging and quantification of measurements.[Pubmed:32533657]

Anat Rec (Hoboken). 2020 Jun 12.

3D imaging is becoming more and more popular, as it allows us to identify interactions between structures in organs. Furthermore, it gives the possibility to quantify and size these structures. To allow 3D imaging, the tissue sample has to be transparent. This is usually achieved by using optical tissue clearing protocols. Although using optical tissue clearing often results in perfect 3D images, these protocols have some pitfalls, like long duration of sample preparation (up to several weeks), use of toxic substances, damage to antibody staining, fluorescent proteins or dyes, high refractive indices, and high costs of sample processing.Recently we described [Huang et al., Scientific Reports 9(1): 521 (2019)] a fast, safe, and inexpensive ethyl cinnamate (ECi) based optical tissue clearing protocol. Here, we present extensions of our protocol with respect to the deparaffinization of old paraffin-embedded samples allowing 3D imaging of the blocks. In addition, we learned to remove ECi from the samples allowing the use of routine immunolabeling protocols. Furthermore, we demonstrate new pictures of lungs after expansion microscopy and adaptation of already existing protocols. The aim of our work is, in summary, to describe the advances in these methodologies, focusing on the morphological imaging of kidneys and lungs.

Characterization of key aroma compounds in Huangjiu from northern China by sensory-directed flavor analysis.[Pubmed:32517941]

Food Res Int. 2020 Aug;134:109238.

Huangjiu (yellow rice wine) from Shanxi province, with a dominant Zao-aroma (fermented grain aroma), is particularly popular in northern China. The key aroma compounds in Huangjiu from Shanxi province were characterized by sensory-directed flavor analysis. A total of 106 compounds separated with solvent-assisted flavor evaporation (SAFE) distillation were identified by gas chromatography-olfactometry (GC-O) coupled with aroma extract dilution analysis (AEDA) and odor specific magnitude estimation (Osme). Forty-seven of them were further quantitated and 21 odorants had odor activity values (OAVs) >/= 1. The aroma profiles of three recombination models had >95.84% similarities to their corresponding samples. And then, the omission/addition tests further confirmed that beta-phenylethanol, 3-methylbutanoic acid, ethyl lactate, sotolon, 2-acetyl-1-pyrroline, vanillin, ethyl cinnamate, ethyl 3-phenylpropionate, 3-methylbutanal, and methional were the key odorants for Huangjiu from Shanxi province. Meanwhile, it was firstly confirmed that ethyl cinnamate and ethyl 3-phenylpropionate played the key roles in the overall aroma of Huangjiu.

3D Imaging of the Transparent Mycobacterium tuberculosis-Infected Lung Verifies the Localization of Innate Immune Cells With Granuloma.[Pubmed:32500041]

Front Cell Infect Microbiol. 2020 May 15;10:226.

Using a novel tissue-clearing method, we aimed to visualize the three-dimensional (3D) distribution of immune cells within Mycobacterium tuberculosis (Mtb)-infected mice lungs. Ethyl cinnamate-based tissue clearing of Mtb-infected mice lungs was performed to obtain transparent lung samples, which were then imaged using a light sheet fluorescence microscope. Using the 3D images, we performed quantitative analysis of the immune cell population within multiple granulomas. In addition, to compare the data from the tissue clearing method, we performed histopathological and immunofluorescence analyses, and flow cytometry. We then created 3D images of the Mtb-infected lung that successfully demonstrated the distribution of blood vessels, immune cells, and granulomas. Since the immune cells within a granuloma could be separately selected and counted, the immune cell population within a specific lesion could be quantified. In addition, macroscopic analysis, e.g., the size or shape of a granuloma, as well as microscopic analysis could be performed as intact lung samples were used. The use of the tissue clearing method in infected lungs could be a novel modality for understanding the role of the immune system in the pathogenesis of tuberculosis.

Physicochemical and aromatic characterization of carob macerates produced by different maceration conditions.[Pubmed:32148803]

Food Sci Nutr. 2020 Jan 9;8(2):942-954.

Carob liqueur is an alcoholic drink (minimum 15% v/v of ethanol and 100 g/L of sugar) typical for the Mediterranean countries. In the current work, carob macerate produced by maceration of carob pods in hydroalcoholic base at different maceration conditions was characterized for the first time based on its aroma compounds/profile, physicochemical parameters, and chromatic characteristics. The results confirm the migration process of bioactive compounds, aroma compounds, and sugars flowing from the carob pod to the hydroalcoholic base. Changes in ethanol concentration modify the physical properties of the solvent and influence the phenolic and aroma compounds extraction, color, and acidity of the obtained samples. The higher content of phenolic compounds was determinate in the samples obtained in the darkness. The amounts of phenols were in the range of some red fruit liqueurs or walnut liqueurs, and sugars (mostly sucrose) ranging between 96 and 107 g/L. Twenty-six (out of total 94) aroma compounds were detected in all samples, of which 17 esters, 3 alcohols, 4 ketones, and 2 acids. Low molecular weight ethyl esters, ethyl hexanoate, ethyl 2-methyl propanoate, ethyl octanoate, ethyl benzoate, ethyl butanoate, and ethyl cinnamate, were the most abundant. Carob pod maceration in 50% v/v hydroalcoholic base (1:5 solid to liquid ratio) in darkness at room temperature during 8 weeks can be recommended as optimal maceration conditions for production of the aromatic carob macerate with functional properties.

Phytochemistry, pharmacological activities and uses of traditional medicinal plant Kaempferia galanga L. - An overview.[Pubmed:32061673]

J Ethnopharmacol. 2020 May 10;253:112667.

ETHNOPHARMACOLOGICAL RELEVANCE: Kaempferia galanga L. is a stemless, rhizomatous, aromatic, perennial and indigenous herb. It is native to India and distributed in China, Bangladesh, Myanmar, Sri Lanka, Japan, Thailand, Indonesia, Malaysia, Vietnam, Laos, Sudan, Nigeria and South Africa. It is an important Indian medicinal herb that has a long history of use in the treatment of several kinds of human ailments including vata ailments like cough and cold, fever, headache, pains disorders, skin diseases, rheumatic diseases, arthritis, joint fractures, vertigo, wounds, gastritis, antidote for snake venoms, inflammation, blood vomiting, mouth sores and tongue blisters in infants. Moreover, the rhizomes of this plant are highly aromatic and have been used widely as spices, in food flavoring, pickles, cosmetics and in perfumery products. AIM OF THE REVIEW: This paper aimed to provide a critical review of current scenario on K. galanga. This review provides a current data on diversity, phytochemistry, pharmacological activities and traditional uses of K. galanga. MATERIALS AND METHODS: The information and data on K. galanga were collated from various resources like ethnobotanical textbooks and literature databases such as PubMed, Science Direct, Wiley, Springer, Tailor and Francis, Scopus, Inflibnet, Google and Google Scholar. RESULTS: The forty-nine phytochemicals including esters, terpenoids, flavonoids, thiourea derivatives, polysaccharides, diarylheptanoids, phenolic acids, phenolic glycoside and cyclic lipodepsipeptide have been hitherto isolated and characterized. The major bioactive compounds extracted from the rhizome of K. galanga were ethyl p-methoxycinnamate, ethyl cinnamate, kaempferol, kaempferide, kaempsulfonic acids, kaemgalangol A, xylose, cystargamide B and 3-caren-5-one. Various studies demonstrated that the K. galanga and its constituents possess several pharmacological activities like antimicrobial, antioxidant, amebicidal, analgesic, anti-inflammatory, anti-tuberculosis, anti-dengue, anti-nociceptive, anti-angiogenic, anticancer, hyperlipidemic, hypopigmentary, osteolysis, larvicidal, insecticidal and mosquito repellent, nematocidal, sedative, sniffing, vasorelaxant and wound healing. CONCLUSION: Kaempferia galanga L. is a valuable medicinal plant which is used traditionally in India to treat a wide variety of ailments. A number of bioactive phytochemicals like esters, terpenoids, flavonoids, polysaccharides, diarylheptanoids, cyclic lipodepsipeptide, phenolic acids and glucoside have been isolated from the rhizomes of K. galanga by several researchers. These phytochemicals are highly bioactive and exhibit various pharmacological activities.

Treatments with native Coleus forskohlii endophytes improve fitness and secondary metabolite production of some medicinal and aromatic plants.[Pubmed:31823202]

Int Microbiol. 2020 May;23(2):345-354.

Endophytes have been shown to play a crucial role in determining the fitness of host plant during their association, yet the cross-functional effect of endophytes of one plant on another plant remains largely uncharacterized. In this study, we attempt to analyze the effect of native endophytes of Coleus forskohlii (Phialemoniopsis cornearis (SF1), Macrophomina pseudophaseolina (SF2), and Fusarium redolens (RF1), isolated from stem and root parts) on plant growth and secondary metabolite enhancement in medicinal plant Andrographis paniculata, and aromatic plants Pelargonium graveolens and Artemisia pallens. Here, we report, endophytic treatments with SF2 (21%) and RF1 (9%) in A. paniculata resulted in significant enhancement of andrographolide along with plant primary productivity. Correspondingly, application of fungal endophytes RF1, SF1, and SF2 significantly improved the plant growth (11 to 40%), shoot weight (28 to 34%), oil content (44 to 58%), and oil yield (72 to 122%) in P. graveolens. Interestingly, treatment of A. pallens with three fungal endophytes resulted in significant enhancement of plant productivity and oil content (12 to 80%) and oil yield (32 to 139%). Subsequently, the endophyte treatments RF1 and SF1 enhanced davanone (13 to 22%) and ethyl cinnamate (11 to 22%) content. However, SF2 endophyte-treated plants did not show any improvement in ethyl cinnamate content but enhanced the content of davanone (10%), a signature component of davana essential oil. Overall, results depict cross-functional role of native endophytes of C. forskohlii and repurposing of functional endophytes for sustainable cultivation of economically important medicinal and aromatic crops.

Phytochemical and spectral analysis of the methanolic extracts of leaves of Murraya koenigii of Jazan, Saudi Arabia.[Pubmed:31631708]

Nat Prod Res. 2019 Oct 21:1-5.

The present study aimed to explore the biocomponents of Murraya koenigii leaves through hot continuous percolation method using methanol as solvent. The spectral analysis by GC-MS and FTIR study demonstrated the presence of alkaloids, tannins, fatty acids, and steroids. The extract was characterized by the presence of unique compounds such as epiyangambin, stigmasterol, alpha-terpineol, eucalyptol, ethyl cinnamate, fatty acids, and other steroids.

Optical Clearing and Imaging of Immunolabeled Kidney Tissue.[Pubmed:31380853]

J Vis Exp. 2019 Jul 22;(149).

Optical clearing techniques render tissue transparent by equilibrating the refractive index throughout a sample for subsequent three-dimensional (3-D) imaging. They have received great attention in all research areas for the potential to analyze microscopic multicellular structures that extend over macroscopic distances. Given that kidney tubules, vasculature, nerves, and glomeruli extend in many directions, which have been only partially captured by traditional two-dimensional techniques so far, tissue clearing also opened up many new areas of kidney research. The list of optical clearing methods is rapidly growing, but it remains difficult for beginners in this field to choose the best method for a given research question. Provided here is a simple method that combines antibody labeling of thick mouse kidney slices; optical clearing with cheap, non-toxic and ready-to-use chemical ethyl cinnamate; and confocal imaging. This protocol describes how to perfuse kidneys and use an antigen-retrieval step to increase antibody- binding without requiring specialized equipment. Its application is presented in imaging different multicellular structures within the kidney, and how to troubleshoot poor antibody penetration into tissue is addressed. We also discuss the potential difficulties of imaging endogenous fluorophores and acquiring very large samples and how to overcome them. This simple protocol provides an easy-to-setup and comprehensive tool to study tissue in three dimensions.

Contemporaneous 3D characterization of acute and chronic myocardial I/R injury and response.[Pubmed:31127113]

Nat Commun. 2019 May 24;10(1):2312.

Cardioprotection by salvage of the infarct-affected myocardium is an unmet yet highly desired therapeutic goal. To develop new dedicated therapies, experimental myocardial ischemia/reperfusion (I/R) injury would require methods to simultaneously characterize extent and localization of the damage and the ensuing inflammatory responses in whole hearts over time. Here we present a three-dimensional (3D), simultaneous quantitative investigation of key I/R injury-components by combining bleaching-augmented solvent-based non-toxic clearing (BALANCE) using ethyl cinnamate (ECi) with light sheet fluorescence microscopy. This allows structural analyses of fluorescence-labeled I/R hearts with exceptional detail. We discover and 3D-quantify distinguishable acute and late vascular I/R damage zones. These contain highly localized and spatially structured neutrophil infiltrates that are modulated upon cardiac healing. Our model demonstrates that these characteristic I/R injury patterns can detect the extent of damage even days after the ischemic index event hence allowing the investigation of long-term recovery and remodeling processes.

Effect of Simultaneous Snail Slime-aided Degradation and Yeast Fermentation on Terpenoid Composition of Plantain Pseudostem Waste.[Pubmed:30961480]

Curr Pharm Biotechnol. 2019;20(6):459-464.

BACKGROUND: In this study, local sustainable enzyme sources involving excised digestive juice of African land snail and yeast were utilized to achieve the simultaneous saccharification (SSF) and fermentation (SSF) of plantain pseudostem (PPS) waste, and afterwards their effects on terpenoids using gas chromatography coupled to a flame ionization detector (GC-FID), were examined. METHODS: The most abundant terpenoids were found in the order alpha-pinene > borneol > camphor > humulene > beta-caryophellene, while the least in abundance were cis ocimene (8.78x10-6 mg/100g), and cyperene (1.81x10-5 mg/100g). The application of exclusive fermentation and SSF respectively elevated azuluene by 95.46 and 99.6%, while pinene-2-ol was elevated by 83.02 and 98.57%, respectively. RESULTS: Both exclusive fermentation and SSF had no effect on myrcene, cyperene, ethyl cinnamate, germacrene b, valencene, beta selinene, aromadendrene, and taraxerol, while the degree of degradation of some of the terpenoids by both processes was respectively as follows; gama muurolene (100%), beta-caryophyllene (97.60 and 93.14%), alpha-terpinenyl acetate (91.95 and 83.16%), geranyl acetate (94.81 and 43.87%), and terpinen-4-ol (94.40 and 57.00%). CONCLUSION: The findings of this study encourage the imminent application of simultaneous saccharification and fermentation for the enhancement of bioactivities of terpenoids.

Ester flavorants detection in foods with a bienzymatic biosensor based on a stable Prussian blue-copper electrodeposited on carbon paper electrode.[Pubmed:30952296]

Talanta. 2019 Jul 1;199:541-546.

A stable and reproducible layer of Prussian blue (PB) modified with copper was electrodeposited on carbon paper electrodes for the multiple detection of ester flavorants with a bienzymatic biosensor. Carbon fiber composite paper was investigated as high-surface, low-cost substrate for biosensor development. The pre-activation of the electrode surface by cyclic voltammetry was necessary to improve the electrochemical properties before the electrochemical deposition of Prussian blue-copper film (PB-Cu). The stability and the reproducibility of the obtained PB-Cu carbon paper electrode was demonstrated at pH 7.4, optimum for biosensor development. The developed biosensor is based on the immobilization of two enzymes (carboxyl esterase and alcohol oxidase) by cross-linking with glutaraldehyde onto PB-Cu carbon paper electrode. A mixture of key aroma ester compounds (methyl butyrate, ethyl butyrate, methyl cinnamate and ethyl cinnamate) was detected in several food samples with low interferences.

Broad applicability of a streamlined ethyl cinnamate-based clearing procedure.[Pubmed:30665888]

Development. 2019 Feb 1;146(3). pii: dev.166884.

Turbidity and opaqueness are inherent properties of tissues that limit the capacity to acquire microscopic images through large tissues. Creating a uniform refractive index, known as tissue clearing, overcomes most of these issues. These methods have enabled researchers to image large and complex 3D structures with unprecedented depth and resolution. However, tissue clearing has been adopted to a limited extent due to a combination of cost, time, complexity of existing methods and potential negative impact on fluorescence signal. Here, we describe 2Eci (2nd generation ethyl cinnamate-based clearing), which can be used to clear a wide range of tissues in several species, including human organoids, Drosophila melanogaster, zebrafish, axolotl and Xenopus laevis, in as little as 1-5 days, while preserving a broad range of fluorescent proteins, including GFP, mCherry, Brainbow and Alexa-conjugated fluorophores. Ethyl cinnamate is non-toxic and can easily be used in multi-user microscope facilities. This method opens up tissue clearing to a much broader group of researchers due to its ease of use, the non-toxic nature of ethyl cinnamate and broad applicability.