Serum Metabolic Profiling Reveals Potential Anti-Inflammatory Effects of the Intake of Black Ginseng Extracts in Beagle Dogs.

Black ginseng (BG) has better health benefits than white ginseng. The intake of BG changes the levels of metabolites, such as amino acids, fatty acids, and other metabolites. However, there is no research on the effect of BG extract intake on the metabolic profile of dog serum. In this study, serum metabolic profiling was conducted to investigate metabolic differences following the intake of BG extracts in beagle dogs. The beagle dogs were separated into three groups and fed either a regular diet (RD, control), RD with a medium concentration of BG extract (BG-M), or RD with a high concentration of BG extract (BG-H). Differences were observed among the three groups after the dogs ingested the experimental diet for eight weeks. The concentrations of alanine, leucine, isoleucine, and valine changed with the intake of BG extracts. Furthermore, levels of glycine and ß-alanine increased in the BG-H group compared to the control and BG-M groups, indicating that BG extracts are associated with anti-inflammatory processes. Our study is the first to demonstrate the potential anti-inflammatory effect of BG extract in beagle dogs. Glycine and ß-alanine are proposed as candidate serum biomarkers in dogs that can discriminate between the effects of ingesting BG-H.

In Vitro Antioxidant, Anti-Diabetes, Anti-Dementia, and Inflammation Inhibitory Effect of Trametes pubescens Fruiting Body Extracts.

Trametes pubescens, white rot fungus, has been used for folk medicine in Asian countries to treat ailments such as cancer and gastrointestinal diseases. This study was initiated to evaluate the in vitro antioxidant, anti-diabetes, anti-dementia, and anti-inflammatory activities of T. pubescens fruiting bodies. The 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging activities of T. pubescens methanol (ME) and hot water (HWE) extracts (2.0 mg/mL) were comparable to butylated hydroxytoluene (BHT), the positive control. However, the chelating effects of ME and HWE were significantly higher than that of BHT. The HWE (6 mg/mL) also showed comparable reducing power to BHT. Eleven phenol compounds were detected by high performance liquid chromatography (HPLC) analysis. The α-amylase and α-glucosidase inhibitory activities of the ME and HWE of the mushroom were lower than Acarbose, the standard reference; however, the inhibitory effects of the mushroom extracts at 2.0 mg/mL were moderate. The acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory effects of ME and HWE were moderate and comparable with galanthamine, the standard drug to treat early stages of Alzheimer's disease (AD). The ME had a neuroprotective effect against glutamate-induced PC-12 cell cytotoxicity at the concentration range of 2-40 µg/mL. The mushroom extracts also showed inflammation inhibitory activities such as production of nitric oxide (NO) and expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-induced murine macrophage-like cell lines (RAW 264.7) and significantly suppressed the carrageenan-induced rat paw-edema. Therefore, fruiting body extracts of T. pubescens demonstrated antioxidant related anti-diabetes, anti-dementia and anti-inflammatory activities.

Appraisal of antioxidant and anti-inflammatory activities of various extracts from the fruiting bodies of Pleurotus florida.

Pleurotus florida has been widely used for nutritional and medicinal purposes. The present study was conducted to evaluate the antioxidant and anti-inflammatory effects of the fruiting bodies of P. florida extracted with acetone, methanol, and hot water. The antioxidant activities of the acetone and methanol extracts of P. florida showed stronger inhibition of ß-carotene-linoleic acid compared to that of the hot water extract. The acetone extract (8 mg/mL) showed a high reducing power of 1.86. The acetone and methanol extracts showed more effective DPPH radical scavenging activities than the hot water extract. The chelating effect of the extracts at lower concentrations was significantly effective compared to that of the positive control. Thirteen phenolic compounds were detected from acetonitrile and hydrochloric acid solvent extracts. Nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipolysaccahride (LPS) stimulated RAW 264.7 cells, a murine macrophage cell line, were inhibited significantly by the mushroom extracts in a concentration dependent manner. The anti-inflammatory activity on carrageenan-induced edema in the rat hind-paw reduced significantly by the mushroom extracts. Therefore, we have demonstrated that P. florida fruiting bodies possess antioxidant and anti-inflammatory activites related to their inhibitory activities on NO production, iNOS protein expression, and carrageenan-induced paw edema in rats. The results suggest that the fruiting bodies of P. florida are a good source of natural antioxidant and anti-inflammatory agents.

Four new and three newly recorded species of Diduga Moore, [1887] (Lepidoptera, Erebidae, Arctiinae) from China.

Four new species, Diduga luteogibbosa n. sp., D. allodubatolovi n. sp., D. scalprata n. sp. and D. hainanensis n. sp. are described from China along with three newly recorded species, D. albicosta Hampson, 1891, D. nigridentata Bayarsaikhan Bae, 2019 and D. hanoiensis Bayarsaikhan Bae, 2019. A key to the Chinese species of the genus Diduga, with illustrations of adults and genitalia of examined species are presented.

In Vitro and In Vivo Medicinal Value of Culinary-Medicinal Lung Oyster Mushroom Pleurotus pulmonarius var. stechangii (Agaricomycetes).

Pleurotus pulmonarius var. stechangii is a culinary-medicinal mushroom commonly cultivated in subtropical countries in Asia. In this study, the in vitro antixanthine oxidase, antihyperglycemic, and in vivo anti-inflammatory activities of a methanol extract (ME) of P. pulmonarius var. stechangii fruiting bodies were evaluated. The xanthine oxidase inhibitory activity of the ME of P. pulmonarius var. stechangii was lower than that of allopurinol, a xanthine oxidase inhibitor used as a positive control. Eleven phenolic compounds were identified from the fruiting bodies of P. pulmonarius var. stechangii by HPLC analysis. The inhibitory effects of ME on α-amylase and α-glucosidase were moderate and lower than that of acarbose, a positive control. The ME inhibited the production of nitric oxide (NO) and nitric oxide synthases (iNOS) protein expression in lipopolysaccharide-induced RAW 264.7 macrophages. It also exhibited an inhibitory effect on carrageenan-induced hind paw edema in a rat model. Taken together, our experimental results demonstrated that the fruiting bodies of P. pulmonarius var. stechangii might be a good natural source to promote human health through its antixanthine oxidase, antihyperglycemia, and anti-inflammatory activities.

Investigations on Metabolic Changes in Beagle Dogs Fed Probiotic Queso Blanco Cheese and Identification of Candidate Probiotic Fecal Biomarkers Using Metabolomics Approaches.

Intake of probiotic cheese improves the intestinal health of humans and animals. However, metabolic changes in the intestines of dogs in response to the ingestion of probiotic cheese have not been evaluated. Thus, we aimed to determine the metabolic changes in healthy beagle dogs fed queso blanco cheese with added Lactobacillus reuteri KACC 92293 and Bifidobacterium longum KACC 91563 (QCLB) and to identify potential fecal biomarkers to distinguish the metabolic changes based on intake of probiotic cheese through metabolomics approaches. The dogs were randomly divided into three groups and fed a regular diet without any cheese (control), a diet with queso blanco cheese (QC), or one with QCLB for eight weeks. The concentrations of acetic, propionic, and 4-aminobutyric acids were increased in the QCLB group compared to those in the control group. Additionally, higher levels of propionic acid and lower levels of xylose were found in the QCLB group compared to those in the QC group. This is the first report on the identification of metabolic changes in beagle dogs fed queso blanco cheese with added L. reuteri KACC 92293 and B. longum KACC 91563. We also found that metabolomics approaches can be useful for identifying potential fecal markers in dogs fed probiotic cheese.

Two new species and a newly recorded species of the genus Stictane Hampson (Lepidoptera, Erebidae, Arctiinae) from Cambodia.

Two new species of the genus Stictane, S. heppneri n. sp. and S. transversana n. sp. are described from Cambodia along with one newly recorded species, S. obscura (Inoue, 1976). A key to the Cambodian species of the genus Stictane with figures of adults and genitalia are presented. Moreover, adults of the little known three Vietnamese species of Diduga are illustrated herein.

Barsine insolita, a new species from Indochina and India (Lepidoptera, Erebidae, Arctiinae).

Barsine Walker, 1854 is one of the largest quadrifid Erebidae genera within subtribe Nudariina (Erebidae, Arctiinae, Lithosiini). It was established for its type species Barsine defecta Walker, 1854 (by subsequent designation, Kirby (1892)) from Nepal. The genus has in the past been treated as a synonym or subgenus of Miltochrista Hübner, [1819] (Hampson 1900; Strand 1917; Reich 1937; Daniel 1951; 1952; 1955; Inoue 1980; Holloway 1982; Fang 1991; 2000; Cerný 1995). In 2001, J.D. Holloway revived Barsine as a distinct genus. The genus is widely distributed in eastern and southeastern Palaearctic and Oriental tropics and more than a hundred of valid species and subspecies were worked on by Fang (2000), Holloway (2001), Kaleka (2003), Cerný & Pinratana (2009), Cerný (1995, 2016), Bucsek (2012, 2014), Dubatolov et al. (2012), Dubatolov & Bucsek (2013), Wu et al. (2013), Kirti & Singh (2015, 2016), Volynkin & Cerný (2016a, 2016b, 2016c, 2017a, 2017b, 2017c, 2017d, 2018a, 2018b; 2019), Bayarsaikhan et al. (2018), Joshi et al. (2018), Spitsyn et al. (2018), Volynkin (2018), Volynkin et al. (2018; 2019a; 2019b; 2019c) and Huang et al. (2018).

Antioxidant, Anti-Melanogenic and Anti-Wrinkle Effects of Phellinus vaninii.

In this study, the antioxidant, anti-xanthine oxidase, anti-melanogenic and anti-wrinkle effects of methanol (ME) and hot water (HE) extracts from the fruiting bodies of Phellinus vaninii were investigated. The 1,1-diphenyl-2-picryl-hydrazyl free radical scavenging activity of 2.0 mg/mL HE (95.38%) was comparable to that of butylated hydroxytoluene (96.97%), the reference standard. The hydroxyl radical scavenging activities of ME (98.19%) and HE (97.55%) were higher than that of butylated hydroxytoluene (92.66%) at 2.0 mg/mL. Neither ME nor HE was cytotoxic to murine melanoma B16-F10 cells at 25-750 µg/mL. Although the xanthine oxidase (XO) inhibitory effects of ME and HE were significantly lower than that of allopurinol, the values were higher than 84 percent. The in vitro tyrosinase inhibitory activities of ME and HE were comparable to kojic acid at 2.0 mg/mL. The cellular tyrosinase and melanin synthetic activities of ME and HE on B16-F10 melanoma cells at 500 µg/mL were higher than arbutin, indicating that the inhibitory effects of arbutin on the tyrosinase and melanin synthesis were higher than those of ME and HE. The collagenase inhibitory activity of HE was comparable to EGCG at 2.0 mg/mL, however, the elastase inhibitory activity of ME and HE was lower than EGCG at the concentration tested. The study results demonstrated that the fruiting bodies of Ph. vaninii possessed good antioxidant, anti-xanthine oxidase, cell-free anti-tyrosinase, cellular anti-tyrosinase, anti-collagenase, and moderate anti-elastase activities, which might be used for the development of novel anti-gout, skin-whitening, and skin anti-wrinkle agents.

Miltochrista nigrococcinea, a new species from Vietnam (Lepidoptera, Erebidae, Arctiinae).

Miltochrista Hübner is one of the largest genera of lichen-moths (family Erebidae, subfamily Arctiinae, tribe Lithosiini) including more than 160 valid species and widely distributed in Oriental and Palaearctic Regions. The genera Miltochrista and the related Asura Walker for a long time were treated in a wide sense. Hampson (1900) considered the generic names Barsine Walker, Ammatho Walker, Sesapa Walker, Cabarda Walker, Mahavira Moore, Korawa Moore and Gurna Swinhoe as synonyms of Miltochrista, and genera such as Lyclene Moore, Nepita Moore, Cyme Felder and Adites Moore under Asura. Holloway (2001) revived the genera Barsine (=Ammatho, Mahavira and Korawa), Cabarda, Lyclene, Nepita, Cyme and Adiles and transferred most of species of Miltochrista to Barsine and Cabarda, and most of species of Asura to Lyclene, Adiles, Cyme and Nepita. Later, the genera Gurna and Sesapa were also revalidated by Volynkin (2016; 2017a). According to Holloway (2001) the main differences between Miltochrista and Lyclene are wing venation and number of cornuti. However, these features may vary within the genus, while the ground plan of clasping apparatuses of the two taxa is the same. For this reason, Kirti Singh (2016) synonymized Lyclene with Miltochrista. Volynkin et al. (2018) confirmed this synonymy and considered Miltochrista sensu Holloway (2001) as the M. miniata (Forster) species-group only.

Dynamics of Short-Term Metabolic Profiling in Radish Sprouts (Raphanus sativus L.) in Response to Nitrogen Deficiency.

Nitrogen (N) is a macronutrient important for the survival of plants. To investigate the effects of N deficiency, a time-course metabolic profiling of radish sprouts was performed. A total of 81 metabolites-including organic acids, inorganic acid, amino acids, sugars, sugar alcohols, amines, amide, sugar phosphates, policosanols, tocopherols, phytosterols, carotenoids, chlorophylls, and glucosinolates-were characterized. Principal component analysis and heat map showed distinction between samples grown under different N conditions, as well as with time. Using PathVisio, metabolic shift in biosynthetic pathways was visualized using the metabolite data obtained for 7 days. The amino acids associated with glucosinolates accumulated as an immediate response against -N condition. The synthesis of pigments and glucosinolates was decreased, but monosaccharides and γ-tocopherol were increased as antioxidants in radish sprouts grown in -N condition. These results indicate that in radish sprouts, response to N deficiency occurred quickly and dynamically. Thus, this metabolic phenotype reveals that radish responds quickly to N deficiency by increasing the content of soluble sugars and γ-tocopherol, which acts as a defense mechanism after the germination of radish seeds.

Hyperlipidemic Inhibitory Effects of Phellinus pini in Rats Fed with a High Fat and Cholesterol Diet.

This study evaluated the in vitro and in vivo hypolipidemic effects of the medicinal mushroom Phellinus pini. The methanol extract (ME) of the fruiting body of Ph. pini was active against pancreatic lipase and cholesterol esterase with 99.14% and 67.23% inhibited activity at 1.0 mg/mL, respectively. It also inhibited 81.81% and 55.33% of α-glucosidase and α-amylase activities, respectively, at 2.0 mg/mL. Hyperlipidemia as induced by feeding rats with a high fat and cholesterol diet (HFC). HFC supplemented with a 5% fruiting body powder of Ph. pini (HFC + PhP) significantly reduced plasma total cholesterol, low-density lipoprotein cholesterol, and triglycerides in rats compared with HFC. The reduced levels were comparable to rats fed the normal control diet (NC). The atherogenic index of HFC + PhP rats was significantly lower than that of the HFC rats. The excretion of fecal total lipid and cholesterol in the HFC + PhP rats was significantly higher than those in the NC and HFC rats. Histopathological examinations demonstrated scant deposition of lipids in the liver of rats fed HFC + PhP. The dietary supplementation with the fruiting body powder provided natural plasma lipid and glucose lowering effects in experimental rats without adverse effects on the plasma biochemical parameters and liver function related enzyme activities. Therefore, the hypolipidemic effects of Ph. pini may be due to the inhibitory effects on pancreatic lipase, cholesterol esterase, α-glucosidase, and α-amylase, and excretion of excess lipids and cholesterol in the feces.

Ethanol Production from Various Sugars and Cellulosic Biomass by White Rot Fungus Lenzites betulinus.

Lenzites betulinus, known as gilled polypore belongs to Basidiomycota was isolated from fruiting body on broadleaf dead trees. It was found that the mycelia of white rot fungus Lenzites betulinus IUM 5468 produced ethanol from various sugars, including glucose, mannose, galactose, and cellobiose with a yield of 0.38, 0.26, 0.07, and 0.26 g of ethanol per gram of sugar consumed, respectively. This fungus relatively exhibited a good ethanol production from xylose at 0.26 g of ethanol per gram of sugar consumed. However, the ethanol conversion rate of arabinose was relatively low (at 0.07 g of ethanol per gram sugar). L. betulinus was capable of producing ethanol directly from rice straw and corn stalks at 0.22 g and 0.16 g of ethanol per gram of substrates, respectively, when this fungus was cultured in a basal medium containing 20 g/L rice straw or corn stalks. These results indicate that L. betulinus can produce ethanol efficiently from glucose, mannose, and cellobiose and produce ethanol very poorly from galactose and arabinose. Therefore, it is suggested that this fungus can ferment ethanol from various sugars and hydrolyze cellulosic materials to sugars and convert them to ethanol simultaneously.

Evaluation of Antioxidant, Anti-cholinesterase, and Anti-inflammatory Effects of Culinary Mushroom Pleurotus pulmonarius.

Culinary mushroom Pleurotus pulmonarius has been popular in Asian countries. In this study, the anti-oxidant, cholinesterase, and inflammation inhibitory activities of methanol extract (ME) of fruiting bodies of P. pulmonarius were evaluted. The 1,1-diphenyl-2-picryl-hydrazy free radical scavenging activity of ME at 2.0 mg/mL was comparable to that of butylated hydroxytoluene, the standard reference. The ME exhibited significantly higher hydroxyl radical scavenging activity than butylated hydroxytoluene. ME showed slightly lower but moderate inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase than galantamine, a standard AChE inhibitor. It also exhibited protective effect against cytotoxicity to PC-12 cells induced by glutamate (10~100 µg/mL), inhibitory effect on nitric oxide (NO) production and inducible nitric oxide synthase protein expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, and carrageenan-induced paw edema in a rat model. High-performance liquid chromatography analysis revealed the ME of P. pulmonarius contained at least 10 phenolic compounds and some of them were identified by the comparison with known standard phenolics. Taken together, our results demonstrate that fruiting bodies of P. pulmonarius possess antioxidant, anti-cholinesterase, and inflammation inhibitory activities.

Evaluation of Anticholinesterase and Inflammation Inhibitory Activity of Medicinal Mushroom Phellinus pini (Basidiomycetes) Fruiting Bodies.

Phellinus pini, a medicinal mushroom, has been used as folk medicine in Asian countries for treating ailments such as cancer and gastrointestinal diseases. In this study we evaluated in vitro the antidementia and anti-inflammatory activities of Ph. Pini fruiting bodies. Eleven phenol compounds were detected by high-performance liquid chromatography analysis. Acetylcholinesterase and butyrylcholinesterase inhibitory effects of a methanol extract and a hot water extract were moderate and comparable with those of galanthamine, the standard drug used to treat the early stages of Alzheimer's disease. The methanol extract had a neuroprotective effect against glutamate-induced cytotoxicity on PC-12 cells at concentration ranging from 20 to 40 µg/mL. The mushroom extracts also inhibited the production of nitric oxide and the expression of inducible nitric oxide synthase in lipopolysaccharide-induced RAW 264.7 macrophages, and they significantly inhibited in vivo carrageenan-induced hind-paw edema in rats. Therefore, it is suggested that Ph. Pini fruiting bodies possess anticholinesterase and anti-inflammatory effects.

Effects of abiotic stresses on cell cycle progression in tobacco BY-2 cells.

Mild stresses such as high temperature (30 degrees C) or a low H2O2 concentration induced transient cell cycle arrest at G1/S or G2/M depending on the cell cycle stage at which the stress was applied. When stresses were introduced during G0 or G1, the G1/S checkpoint was mainly used; when stresses were introduced after S phase, G2/M was the primary checkpoint. The slowing of cell cycle progression was associated with transient delays in expression of A-, B-, and D-type cyclins. The delay in expression of NtcycA13, one of the A-type cyclins, was most pronounced. The levels of expression of Ntcyc29 (a cyclin B gene) and of CycD3-1 differed most depending on the applied stress, suggesting that different cellular adjustments to mild heat and a low concentration of H2O2 are reflected in the expression of these two cyclins.

Expression of sucrose-phosphate synthase (SPS) in non-photosynthetic tissues of maize.

Sucrose phosphate synthase (SPS) is responsible for sucrose synthesis in photosynthetic tissues. We have detected SPS expression in non-photosynthetic tissues. It was predominantly expressed in the basal region of developing endosperm, suggesting that sucrose is re-synthesized in this region, where sucrose is unloaded from the phloem and hydrolyzed into glucose and fructose. The SPS transcript in endosperm was approximately 300 nt smaller than in leaf. However, the size of the SPS protein was similar to that of leaf but had higher activity. SPS expression was also detected in developing and germinating embryos, indicating that sucrose resynthesis also occurs in embryos. Although the level of SPS mRNA and protein was lower in embryos than in leaf, enzymatic activity was higher. Similarly, the level of SPS transcript was 10-fold lower in endosperm than in leaf but the level of SPS protein was comparable, and activity was 2 fold higher. Thus, SPS expression was evident in maize kernels, and its expression and regulation were different from the SPS in leaf.

Isolation of defense-related genes differentially expressed in the resistance interaction between pepper fruits and the anthracnose fungus Colletotrichum gloeosporioides.

Unripe mature green fruits of pepper (Capsicum annuum) are susceptible to Colletotrichum gloeosporioides, whereas ripe red fruits are not. We established this pepper-C. gloeosporioides interaction as a model system to study the fungal resistance that develops during ripening of nonclimacteric fruit. Histochemical examination of transverse sections suggested that fungal invasion 24 h after inoculation (HAI) and colonization 48 HAI are critical events that differentiate between resistant and susceptible interactions. Based on this observation, we used messenger RNA differential display to isolate defense-related genes differentially expressed at 24 and 48 HAI. RNA gel blot analysis showed that six out of eighty cloned cDNAs were differentially expressed after infection of ripe fruit. The proteins encoded by these six clones, ddP1, ddP3, ddP4, ddP6, ddP13, and ddP47, showed significant homology to aldehyde dehydrogenase, P23 protein, NP24 protein, cytochrome P450 protein, esterase, and MADS-box protein, respectively, and may be involved in the resistance of ripe fruit to C. gloeosporioides infection.

Cloning of genes differentially expressed during the initial stage of fruit development in melon (Cucumis melo cv. Reticulatus).

We have cloned genes involved in the initial stage of fruit development in the melon by suppression subtractive hybridization. A cDNA library of unfertilized ovules was subtracted from that of fruit 9 days after pollination (DAP); 10 of the 40 selected cDNA clones were identified by reverse Northern analysis as genes differentially expressed in fruit at 9 DAP. Seven of the ten genes were homologous to genes of known function; two were related to genes with unknown functions, and one was novel. With the exception of cucumisin, none of the cDNAs had been previously identified in melon. According to Northern analyses, six of the genes were expressed at high levels early in fruit development. Expression of cucumisin, Cmf-25, Cmf-30, and Cmf-124 was highest at 9 DAP, implying that these genes are involved in the initial stage of fruit development. Cmf-30, a seed nucellus-specific gene, was also expressed early in seed development. The other genes were expressed at a moderate level throughout fruit development, with the highest expression occurring in fruit at 9 and 18 DAP. In conclusion, nine new genes involved in early fruit development in melon were cloned, and their temporal and spatial expression patterns indicate that they are preferentially expressed during the active growing stage of fruit.

Overexpression of the downward leaf curling (DLC) gene from melon changes leaf morphology by controlling cell size and shape in Arabidopsis leaves.

A plant-specific gene was cloned from melon fruit. This gene was named downward leaf curling (CmDLC) based on the phenotype of transgenic Arabidopsis plants overexpressing the gene. This expression level of this gene was especially upregulated during melon fruit enlargement. Overexpression of CmDLC in Arabidopsis resulted in dwarfism and narrow, epinastically curled leaves. These phenotypes were found to be caused by a reduction in cell number and cell size on the adaxial and abaxial sides of the epidermis, with a greater reduction on the abaxial side of the leaves. These phenotypic characteristics, combined with the more wavy morphology of epidermal cells in overexpression lines, indicate that CmDLC overexpression affects cell elongation and cell morphology. To investigate intracellular protein localization, a CmDLC-GFP fusion protein was made and expressed in onion epidermal cells. This protein was observed to be preferentially localized close to the cell membrane. Thus, we report here a new plant-specific gene that is localized to the cell membrane and that controls leaf cell number, size and morphology.