Insecta LFM (dry powder type, Nosan Co., Yokohama, Japan) and dry leaf powder were the major solid components of the diet (Table 1 ). Insecta is a commercial diet used for a wide range of phytophagous insects (e.g., common cutworm, legume pod borer, carpenter moth, and yellow-spotted longicorn beetle) without admixture (Kudo et al. 2014 ; Nagamine et al. 2016 (link), 2017 ; Nakanishi et al. 2017 ; Nakano et al. 2018 ), and is also used for oligophagous insects (e.g., fungivorous moth, swallowtail butterfly, and chrysanthemum longicorn beetle) with host powder (Kitajima 2013 , Nishikawa et al. 2013 (link), Shintani 2011 (link)). To prepare leaf powder of C. japonica, foliage was cut from around the campus and the leaves were dried naturally in the laboratory for at least 5 d before being pulverized with an electric blender. Chloramphenicol and propionic acid (Nacalai Tesque, Kyoto, Japan) were added as antibacterial and antifungal agents, respectively.
Propionic acid
Manufactured by Nacalai Tesque
Sourced in Japan
Propionic acid is a chemical compound used in various industrial applications. It is a colorless liquid with a pungent odor. Propionic acid is commonly used as a preservative in food products, animal feed, and pharmaceutical formulations.
Automatically generated - may contain errors
Lab products found in correlation
N butyric acid, by Nacalai Tesque (2 mentions)
Acetic acid, by Nacalai Tesque (2 mentions)
Chloramphenicol, by Nacalai Tesque (1 mentions)
Sodium dihydrogen phosphate, by Fujifilm (1 mentions)
Disodium hydrogen phosphate, by Fujifilm (1 mentions)
Ach chloride, by Kanto Chemical (1 mentions)
2 aminoethyl trimethylammonium chloride hydrochloride, by Merck Group (1 mentions)
Dichloromethane dcm, by Fujifilm (1 mentions)
Deuterium oxide, by Kanto Chemical (1 mentions)
Choline chloride, by Nacalai Tesque (1 mentions)
Arium 611 ultrapure water system, by Sartorius (1 mentions)
Cholesterol, by Fujifilm (1 mentions)
Yeast autolysate, by Merck Group (1 mentions)
Butyl p hydroxybenzoate, by Nacalai Tesque (1 mentions)
Glucose, by Merck Group (1 mentions)
Proteinase inhibitor cocktail, by Nacalai Tesque (1 mentions)
Cellulose, by Merck Group (1 mentions)
Cholesterol, by Nacalai Tesque (1 mentions)
Fructose, by Nacalai Tesque (1 mentions)
Choline bitartrate, by Nacalai Tesque (1 mentions)
6 protocols using propionic acid
1
Insecta LFM Diet for Phytophagous Insects
2
Synthesis and Characterization of Choline Compounds
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Ultrapure water with a specific resistance of 18.2 MΩ/cm was produced in an Arium 611 Ultrapure Water System (Sartorius Co., Goettingen, Germany) and used in our experiments. Methanol (high-performance liquid chromatography [HPLC]-grade), acetonitrile (HPLC-grade), choline chloride, propionic acid, n-butyric acid, ethyl acetate (EtOAc), formic acid and 1-N hydrochloric acid were purchased from Nacalai Tesque, Inc. (Kyoto, Japan). N,N′-dicyclohexylcarbodiimide (DCC) was purchased from Peptide Institute, Inc. (Osaka, Japan); 4-N HCl/dioxane and N,N-dimethylformamide (DMF) were purchased from Watanabe Chemical Industries, Ltd. (Hiroshima, Japan); N,N-dimethyl-4-aminopyridine (DMAP) was purchased from Kokusan Chemical Co., Ltd. (Tokyo, Japan); and ACh chloride, pivalic acid and deuterium oxide were purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). (2-aminoethyl)trimethylammonium chloride hydrochloride was purchased from Sigma-Aldrich, Inc. (Ontario, Canada). All other purchased chemicals—including dichloromethane (DCM), sodium dihydrogen phosphate and disodium hydrogen phosphate—were from Fujifilm Wako Pure Chemical Industries, Ltd. (Osaka, Japan). PCh, BCh, LCh and (2-aminoethyl)trimethylammonium pivaloylamide (EN) were synthesized in our laboratory.
3
Cholesterol-Deficient Low-Sterol Food Protocol
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A recipe for Low-sterol food (LSF) was based on Carvalho et al. (2010) (link) with some modifications. LSF used in this study was a mixture of 10% yeast autolysate (Sigma), 10% glucose, 1% purified agar, 0.3% propionic acid, and 0.06% butyl p-hydroxybenzoate (all Nacalai Tesque). cholesterol-supplemented LSF was prepared by adding cholesterol (Wako, Osaka, Japan) to LSF. The final concentration of cholesterol in this food was 6.2 μg/ml. In the original recipe of Eaton’s group, yeast autolysate and agarose for the lipid-depleted medium were chloroform-extracted (Carvalho et al. 2010 (link)). However, we found that even when we did not treat these materials with chloroform, the control larvae on LSF exhibited a reproducible larval-arrest phenotype (Figure 4A ). More importantly, the larval-arrest phenotype on LSF was restored by adding cholesterol to LSF (Figure 4A ), implying that sterol contents in our LSF were significantly reduced. We should note that the phenotype on our LSF was weaker than the phenotype on Eaton’s lipid-depleted medium (Carvalho et al. 2010 (link)), probably because our LSF recipe omitted the chloroform extraction procedure.
4
Bilberry Anthocyanins Extract Powder Protocol
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Bilberry anthocyanins extract powder (Mirtoselect) was purchased from Indena Japan Co., Ltd. (Tokyo, Japan) and batch number was 32265/M2. Lard and cellulose were purchased from Sigma-Aldrich Co., LLC. (Tokyo, Japan). Soybean oil, cholesterol, choline bitartrate, methionine, fructose, acetic acid, lactic acid, propionic acid, 1,1,3,3-tetraethoxypropane, 2-thiobarbituric acid, and proteinase inhibitor cocktail were purchased from Nacalai Tesque, Inc. (Kyoto, Japan). AIN-93G mineral mix and AIN-93G vitamin mix were purchased from Oriental Yeast Co., Ltd. (Tokyo, Japan). Corn starch was purchased from Sanwa Starch Co., Ltd. (Nara, Japan). Edible Acid Casein 30–60 Mesh was purchased from Meggle GmbH & Co. KG (Wasserburg am Inn., Germany). Sucrose was purchased from Hayashi Pure Chemical Industry., Ltd. (Osaka, Japan). Paraformaldehyde, hexane and 2-ethyl butyric acid were purchased from Wako Pure Chemical Co. (Osaka, Japan). Oil red O reagent was purchased from Muto Pure Chemicals Co., Ltd. (Tokyo, Japan). Antibodies against NF-E2-related factor 2 (Nrf2), Kelch-like ECH-associated protein 1 (Keap1), superoxide dismutase-2 (SOD2), and α-smooth muscle actin (α-SMA) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA).
5
Quantifying Cecal Short-Chain Fatty Acids
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SCFAs in cecal contents were measured by the method of Rémésy and Démigne 15) using a Shimadzu GC-14A equipped with FID and a capillary column (CBP 20, 0.25 mm × 25 m; Shimadzu, Tokyo, Japan), with crotonic acid as the internal standard. Acetic acid, propionic acid, n-butyric acid, and crotonic acid were purchased from Nacalai Tesque (Kyoto, Japan).
6
Comprehensive Analysis of Organic Compounds
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L-Cysteine was purchased from Sigma-Aldrich (St Louis, MO, USA). ATP, AMP and 2-hydroxyBenzoic acid were purchased from Kishida Chemical (Osaka, Japan). Benzoic acid, 4-methylBenzoic acid, 4-cyanoBenzoic acid, phthalic acid, propionic acid, butyric acid, 3-mercaptopropionic acid and L-amino acids (glycine, L-alanine, L-valine, L-leucine, L-isoleucine, L-serine, L-threonine, L-methionine, L-aspartic acid, L-asparagine, L-glutamic acid, L-glutamine, L-arginine, L-lysine, L-histidine, L-phenylalanine, L-tyrosine, L-tryptophan and L-proline) were purchased from Nacalai Tesque (Kyoto, Japan). 3-HydroxyBenzoic acid, 3-cyanoBenzoic acid, 2,6-difluoroBenzoic acid and pentanoic acid were purchased from Tokyo Chemical Industry (Tokyo, Japan). Other reagents were purchased from Wako Pure Chemical Industries (Osaka, Japan).
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