Antifungal Substances Produced by Xenorhabdus bovienii and Its Inhibition Mechanism against Fusarium solani.

Fungal colonization can severely damage artifacts. Nematode endosymbiotic bacteria exhibit good prospects in protecting artifacts from fungal damage. We previously found that supernatant from the fermentation of nematode endosymbiotic bacterium, Xenorhabdus bovienii, is effective in inhibiting the growth of Fusarium solani NK-NH1, the major disease fungus in the Nanhai No.1 Shipwreck. Further experiments proved that X. bovienii produces volatile organic compounds (VOCs) that inhibit NK-NH1. Here, using metabolomic analysis, GC-MS, and transcriptomic analysis, we explored the antifungal substances and VOCs produced by X. bovienii and investigated the mechanism underlying its inhibitory effect against NK-NH1. We show that X. bovienii produces several metabolites, mainly lipids and lipid-like molecules, organic acids and derivatives, and organoheterocyclic compounds. The VOCs produced by X. bovienii showed two specific absorption peaks, and based on the library ratio results, these were predicted to be of 2-pentanone, 3-(phenylmethylene) and 1-hexen-3-one, 5-methyl-1-phenyl. The inhibition of F. solani by VOCs resulted in upregulation of genes related to ribosome, ribosome biogenesis, and the oxidative phosphorylation and downregulation of many genes associated with cell cycle, meiosis, DNA replication, and autophagy. These results are significant for understanding the inhibitory mechanisms employed by nematode endosymbiotic bacteria and should serve as reference in the protection of artifacts.

Integration of behavioural tests and transcriptome sequencing of C. elegans reveals how the nematode responds to peanut shell biochar amendment.

Biochars have drawn wide attention as adsorbents, carbon sequesters and soil re-mediators. However, these substances are ambiguous regarding their effects on the motility, phenotypic changes and potential adaptative mechanisms of soil organisms. This study investigated how peanut shell biochar (PBC) affects the C. elegans model via a one-choice selection test and RNA-seq analysis. The results showed that C. elegans were able to select either PBC or a water control, and a clear preference for PBC was observed after 48 h of exposure, with the chemotaxis index (CI) reaching approximately 1.0. The nematode preferences for PBC vs sterile PBC/graphite were not significant, which demonstrated that initial microorganisms and appearances were not the reasons for the worms' selection, but the selection behaviour was instead determined by volatile odours. The treatments also showed that biochar amendment significantly decreased the body length, brood size and superoxide dismutase (SOD) activity of C. elegans to 960.20 ± 15.23 µm, 173.22 ± 4.56, 165.81 ± 3.82 U/mL SOD, respectively. Then, a possible molecular mechanism of PBC-induced developmental and reproductive effects on C. elegans was explored. Differential gene expression analysis was performed, and 1625 genes (1425 up- and 225 downregulated genes) were regulated in response to PBC treatment. The top 20 regulated genes were col genes (col-129; col-81; col-139; col-71), bli-6, perm-4 and his-24, which indicated that cuticle collagen synthesis, eggshell formation and/or heterochromatin in postembryonic growth may be disrupted following exposure to PBC. Therefore, our study suggested that quality standards be used to test nematode preferences and responses to biochar amendment, with the aim of safe application in soils, seedling substrates or fertilizers.

The symbiotic bacteria Alcaligenes faecalis of the entomopathogenic nematodes Oscheius spp. exhibit potential biocontrol of plant- and entomopathogenic fungi.

Soil-dwelling entomopathogenic nematodes (EPNs) kill arthropod hosts by injecting their symbiotic bacteria into the host hemolymph and feed on the bacteria and the tissue of the dying host for several generations cycles until the arthropod cadaver is completely depleted. The EPN-bacteria-arthropod cadaver complex represents a rich energy source for the surrounding opportunistic soil fungal biota and other competitors. We hypothesized that EPNs need to protect their food source until depletion and that the EPN symbiotic bacteria produce volatile and non-volatile exudations that deter different soil fungal groups in the soil. We isolated the symbiotic bacteria species (Alcaligenes faecalis) from the EPN Oscheius spp. and ran infectivity bioassays against entomopathogenic fungi (EPF) as well as against plant pathogenic fungi (PPF). We found that both volatile and non-volatile symbiotic bacterial exudations had negative effects on both EPF and PPF. Such deterrent function on functionally different fungal strains suggests a common mode of action of A. faecalis bacterial exudates, which has the potential to influence the structure of soil microbial communities, and could be integrated into pest management programs for increasing crop protection against fungal pathogens.

ß-Cryptoxanthin suppresses UVB-induced melanogenesis in mouse: involvement of the inhibition of prostaglandin E2 and melanocyte-stimulating hormone pathways.

OBJECTIVE: ß-cryptoxanthin (ß-CPX) is a carotenoid that is widely contained in the fruits of citrus plants. We evaluated the effect of ß-CPX on UVB-induced pigmentation and mRNA expression related to melanogenesis in mouse skin. In addition, changes in melanogenic molecules were evaluated in cultured melanocytes stimulated with prostaglandin (PG) E(2), melanocyte-stimulating hormone (MSH) and endothelin (ET)-1. METHODS: Mice were irradiated with UVB and were given ß-CPX (0.1, 1 and 10 mg/kg) orally for 14 days. Pigmentation was evaluated by skin colour change and microscopic observation. Total RNA was obtained from the skin and the expression of melanogenic mRNA was evaluated by RT-PCR. In cell culture studies, human melanocytes were cultured with ß-CPX and melanogenic stimulants (PGE(2), MSH and ET-1) for 6-10 days. Melanin contents, dendricity, melanogenic mRNA and phosphorylation of cyclic AMP response element-binding protein (CREB) were evaluated. KEY FINDINGS: ß-CPX (10 mg/kg) significantly suppressed skin pigmentation and mRNA expression of cyclooxygenase-2, ET-1 receptors, low-affinity neurotrophin receptor, PGE(2) receptor (EP1), melanocortin 1 receptor (MC1R), tyrosinase (Tyr), tyrosinase-related protein (Tyrp) 1 and microphthalmia transcription factor. ß-CPX (10 µg/ml) suppressed melanogenesis induced by PGE(2), MSH and ET-1. In the PGE(2)-stimulated melanocytes, mRNA expressions of EP-1, Tyr and Tyrp1 and phosphorylation of CREB protein were suppressed. In the ET-1-stimulated cells, only expression of CREB protein was suppressed. In the MSH-induced cells, mRNA expression of MC1R and Tyrp1 and protein expression of CREB were suppressed. CONCLUSION: Oral administration of ß-CPX was found to suppress UVB-induced melanogenesis. Suppression of melanogenic enzymes, receptors of melanogenic stimulators, expression and phosphorylation of CREB are thought to be involved in the mechanism.

Anti-pigmentary activity of fucoxanthin and its influence on skin mRNA expression of melanogenic molecules.

OBJECTIVES: Carotenoids and retinoic acid derivatives are topically applied for sun-protective and whitening purposes. Fucoxanthin is a carotenoid derived from edible sea algae, but its effect on melanogenesis has not been established. Therefore, we examined the effect of fucoxanthin on melanogenesis. METHODS: Inhibitory effects on tyrosinase activity, melanin formation in B16 melanoma and skin pigmentation in UVB-irradiated guinea-pigs were evaluated. To elucidate the action of fucoxanthin on melanogenesis, its effect on skin melanogenic mRNA expression was evaluated in UVB-irradiated mice. Fucoxanthin was given topically or orally to mice once a day and UVB irradiation was applied for 14 days. The effect of fucoxanthin on skin melanogenic mRNA expression was evaluated by real time reverse transcription polymerase chain reaction. KEY FINDINGS: Fucoxanthin inhibited tyrosinase activity, melanogenesis in melanoma and UVB-induced skin pigmentation. Topical application of fucoxanthin (1%) significantly suppressed mRNA expression of cyclooxygenase (COX)-2, endothelin receptor A, p75 neurotrophin receptor (NTR), prostaglandin E receptor 1 (EP1), melanocortin 1 receptor (MC1R) and tyrosinase-related protein 1. The suppression of p75NTR, EP1 and MC1R expressions was observed at 0.01% application. Also, oral application of fucoxanthin (10 mg/kg) significantly suppressed expression of COX-2, p75NTR, EP1 and MC1R. CONCLUSIONS: These results suggest that fucoxanthin exhibits anti-pigmentary activity by topical or oral application in UVB-induced melanogenesis. This effect of fucoxanthin may be due to suppression of prostaglandin (PG) E(2) synthesis and melanogenic stimulant receptors (neurotrophin, PGE(2) and melanocyte stimulating hormone expression).

Anti-inflammatory properties of red ginger (Zingiber officinale var. Rubra) extract and suppression of nitric oxide production by its constituents.

Red ginger (Zingiber officinale var. Rubra) has been prescribed as an analgesic for arthritis pain in Indonesian traditional medicine. The surface color of the rhizome is purple because of the anthocyanidins in its peel. We prepared 40% ethanolic extract from dried red ginger (red ginger extract [RGE]) and evaluated its anti-inflammatory activity using acute and chronic inflammation models. In an acetic acid-induced mouse writhing model, RGE (10-100 mg/kg) suppressed both the frequency of writhing and the increase in permeability of abdominal capillaries. On the other hand, continuous treatment with RGE (10 mg/kg) significantly (P < .05) suppressed footpad edema in a rat adjuvant arthritis model. To clarify the anti-inflammatory mechanism of RGE, we examined the effect on prostaglandin (PG) and nitric oxide (NO) production from mouse leukemic monocytes (RAW264 cells) stimulated by lipopolysaccharide. RGE (3 and 10 microg/mL) significantly (P < .05) suppressed PGE(2) production, while it also suppressed NO production at 100 microg/mL. After bioassay-guided separation of RGE, we found that [6]-shogaol and gingerdiols suppressed NO production. Red dye fractions presumed to be proanthocyanidins also suppressed NO production at 100 microg/mL. Consequently, we found a potent suppressive effect of RGE on acute and chronic inflammation, and inhibition of macrophage activation seems to be involved in this anti-inflammatory effect. [6]-Shogaol, gingerdiols, and proanthocyanidins were identified as constituents that inhibited NO production.

The hypocholesterolemic effects of Cistanche tubulosa extract, a Chinese traditional crude medicine, in mice.

The roots of Cistanche (C.) tubulosa (Orobanchaceae), a parasitic plant that grows in the Taklamakan desert, are traditionally used as medicines and foods in China. We prepared aqueous ethanol extract (CTE) from the roots of C. tubulosa and its hypocholesterolemic effect was evaluated. Using gene chip and RT-PCR analysis of the livers of mice given CTE (400 mg/kg) for 14 days, we found mRNA expression of molecules related to cholesterol transport [apolipoprotein B and very low density lipoprotein (VLDL) receptor] and metabolism [cytochrome P450 side chain cleave (SCC) and steroid 5alpha-reductase 2] were up-regulated. The administration of CTE (400 mg/kg) for 14 days significantly suppressed serum cholesterol elevation in high cholesterol diet-fed mice. The mRNA expressions of VLDL receptor and cytochrome P450 SCC were significantly enhanced. In addition, acteoside, a major constituent of CTE, was found to enhance the mRNA expressions of apolipoprotein B, VLDL receptor, and cytochrome P450 SCC in HepG2 hepatocytes. These results suggest that CTE affects the mRNA expressions of molecules related to cholesterol transport and metabolism and exhibits hypocholesterolemic activity in diet-induced hypercholesterolemia mice. Acteoside was involved in the hypocholesterolemic activity of CTE.

Extract of Yi Zhi Fang improves learning and memory behaviours of mice and its possible mechanisms.

The effects of Yi Zhi Fang extract (YZF) on learning and memory performances were investigated in mice using passive avoidance tasks. Oral administration of YZF improved learning and memory disorders induced by chemicals in both normal and senile mice. The acetylcholine (Ach) concentration, the muscarinic Ach receptors (M-R) and the monoamine oxidase B (MAO-B) activities were analysed by a radioimmunoassay, a radioligand receptor binding assay and UV spectrophotometry, respectively, using senile decapitated mice. The oral administration of YZF to the senile mice increased the M-R concentration, while the concentration of Ach and the activity of MAO-B decreased in senile mice brains. From these results it is evident that YZF promotes the function of the central cholinergic system and inhibits the activity of MAO-B in mice brains resulting in an enhancement of learning and memory.