Role of GPx4 in human vascular endothelial cells, and the compensatory activity of brown rice on GPx4 ablation condition.

Oxidative stress is implicated in the pathologies of vascular endothelial cells. However, the importance of specific antioxidant enzymes in vascular endothelial cells is not fully understood. The purpose of this study was to elucidate the importance of Glutathione peroxidase 4 (GPx4), and the involvement of ferroptosis on cell death induced by GPx4 loss in human vascular endothelial cells. In addition, we examined the compensatory activity of brown rice on GPx4 ablation condition. Human umbilical vein endothelial cells were transfected with GPx4 or scramble control siRNA. GPx4 knockdown caused the increase in the levels of lipid oxidation, and induced cytotoxicity. On the other hand, α-tocopherol (vitamin E) and extract of brown rice, ameliorated lipid peroxidation, cytotoxicity, and delay of proliferation induced by GPx4 knockdown. Furthermore, ferrostatin-1, inhibitor of ferroptosis, also prevented cytotoxicity and delay of proliferation. In conclusion, our data demonstrated that GPx4 is an essential antioxidant enzyme for protecting lipid peroxidation, and is a regulator of ferroptosis in vascular endothelial cells. Furthermore, vitamin E rich food, such as brown rice, can compensate for GPx4 loss by protecting cells against lipid peroxidation.

Effects of Astaxanthin, Lutein, and Zeaxanthin on Eye-Hand Coordination and Smooth-Pursuit Eye Movement after Visual Display Terminal Operation in Healthy Subjects: A Randomized, Double-Blind Placebo-Controlled Intergroup Trial.

(1) Background: The impairment of eye-hand coordination and smooth-pursuit eye movement caused by visual display terminal (VDT) operation is thought to impair daily living activities, for which no effective methods are currently known. On the other hand, various food ingredients, including astaxanthin, lutein, and zeaxanthin, are known to help improve the eye health of VDT operators. This study aimed to test the hypothesis that the combination of astaxanthin, lutein, and zeaxanthin can prevent the impairment of eye-hand coordination and smooth-pursuit eye movement caused by VDT operation. (2) Methods: We conducted a randomized, placebo-controlled, parallel-group clinical trial. Healthy subjects who regularly worked with VDTs were randomly assigned to the active and placebo groups. All of the subjects took soft capsules containing 6 mg of astaxanthin, 10 mg of lutein, and 2 mg of zeaxanthin or placebo soft capsules once daily for eight weeks. We evaluated the eye-hand coordination, smooth-pursuit eye movements, and macular pigment optical density (MPOD) at 0, two, four, and eight weeks after soft-capsule intake. (3) Results: The active group showed significantly improved eye-hand coordination after VDT operation at eight weeks. However, there was no clear improvement in the effect of the supplementation on smooth-pursuit eye movements. The active group also showed a significant increase in MPOD levels. (4) Conclusions: Consumption of a supplement containing astaxanthin, lutein, and zeaxanthin mitigates the decline of eye-hand coordination after VDT operation.

Phylogenetic analysis of carotenoid-producing marine microorganisms around the coral reefs of the Kerama Islands of Okinawa.

Seawater sample from the coral reefs of the Kerama Islands of Okinawa were assessed for the presence of carotenoid-producing bacteria. Results of 16S rDNA analysis of the bacteria obtained from the isolated bacteria showed unique patterns that were different from those of the bacteria obtained from the ordinary marine area. Phylogenetic analysis revealed a slight correlation with the statistical analysis of the PDA chart patterns. The results suggest that useful materials for human health such as carotenoids can be extracted from many carotenoid-producing bacteria such as those found the coral reefs the Kerama Islands.

Antibacterial activity of stilbene oligomers against vancomycin-resistant Enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) and their synergism with antibiotics.

Two resveratrol trimers, gnemonol B isolated from Gnetum gnemon and gnetin E obtained from the Gnetum species, were found to exhibit strong antibacterial activities against vancomycin-resistant Enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA). The MIC values of gnemonol B against five strains of VRE and nine strains of MRSA were 12.5 and 6.25 microg/ml, respectively. The MIC values of gnetin E against five strains of VRE and nine strains of MRSA ranged from 12.5 to 25microg/ml. These compounds also showed synergistic effects when used in combination with commercially available antibiotics according to the evaluation method using FIC indices. These findings suggested that the application of the test compounds alone or in combination with antibiotics might be useful in controlling and treating VRE and MRSA infections

Inhibitory actions of several natural products on proliferation of rat vascular smooth muscle cells induced by Hsp60 from Chlamydia pneumoniae J138.

Atherosclerosis is a vascular disorder involving inflammation, a narrowed vascular lumen in the entire tunica intima, and reduced elasticity of the arterial wall. It has been found that Hsp60 from Chlamydia pneumoniae, an obligate bacterial pathogen associated with atheroma lesions, mimics human Hsp60, thereby causing attacks by immune cells on stressed endothelial cells expressing endogenous Hsp60 on their surface. Furthermore, Hsp60 from C. pneumoniae has been shown to promote the growth of vascular smooth muscle cells (VSMCs). To explore probes that can be used for studying signal transduction elicited by the chlamydial Hsp60, we have tested several natural products for their inhibitory actions on the Hsp60-induced proliferation of rat arterial smooth muscle cells. Sesamol, vanillyl alcohol, and trans-ferulic acid exhibited moderate inhibitory actions on the Hsp60-induced cell proliferation; zerumbone, humulene, and caryophylene effectively inhibited it at low concentrations with IC(50) values of 529, 122, and 110 nM, respectively. The results indicated that the 11-membered alicyclic ring is favorable for interactions with receptors involved in the Hsp60-induced VSMC proliferation.

The feasibility of using mosquitofish (Gambusia affinis) for detecting endocrine-disrupting chemicals in the freshwater environment.

We evaluated the utility of gene-transcriptional responses in the liver of mosquitofish (Gambusia affinis), a species introduced to many countries and therefore widely available, for detecting endocrine-disrupting activity in water. Exposure to ß-naphthoflavone, an aryl hydrocarbon receptor (AhR) agonist, significantly increased the transcript of the cytochrome P4501A gene (cyp1a), peaking at 24 h, in both sexes at concentrations of 10 µg/L or more. 17ß-Estradiol (E(2) ) at 500 ng/L increased the number of males showing gene transcription of precursors of yolk protein, vitellogenin (Vtga, Vtgb, and Vtgc), at 24, 48, and 72 h. Exposure for 48 h to bisphenol A (BPA), an estrogen mimic, also increased vtg-positive males at 1 mg/L or more. Leachate from a Japanese stable-type landfill significantly increased vtg-positive males after 48 h exposure, and the in vitro activity of the leachate against the estrogen receptor (ER) was estimated as an E(2) equivalent of 240 ng/L by yeast transfected with the ER. Chemical analysis showed that major contributors to the ER activation were BPA and 4-tert-octylphenol. This leachate and drainage water from a control-type landfill had AhR activities, estimated by yeast with the AhR, but had no significant effect on cyp1a transcription. These results showed that mosquitofish are suitable for detecting in vivo AhR and ER effects, but are insensitive to E(2).

Culture characteristics of carotenoid-producing filamentous fungus T-1, and carotenoid production.

The culture characteristics, carotenoid production, and associated biosynthetic pathway of strain T-1 were examined. As a result of examining the culture temperature and light irradiation, an increase of neurosporaxanthin and neurosporaxanthin beta-D-glucopyranoside was observed at a low temperature and 0 lx. It was suggested that highly polar carotenoids, such as neurosporaxanthin, and carotenoid glycosides were involved in the stabilization of membrane during nutrition storage other than the defense function of fungus bodies. Strain T-1 produced lycopene, beta-carotene, gamma-carotene, torulene, neurosporaxanthin, and neurosporaxanthin beta-D-glucopyranoside, as assessed by HPLC, LC-MS, and NMR analysis. Carotenoid biosynthesis begins with neurosporene, passing to lycopene and gamma-carotene through cyclization, and produces beta-carotene. In addition, it is saturated, gamma-carotene is converted to torulene, and neurosporaxanthin is produced. Thus, the carotenoid biosynthetic pathway in strain T-1 was estimated.

Elucidation of a carotenoid biosynthesis gene cluster encoding a novel enzyme, 2,2'-beta-hydroxylase, from Brevundimonas sp. strain SD212 and combinatorial biosynthesis of new or rare xanthophylls.

A carotenoid biosynthesis gene cluster mediating the production of 2-hydroxyastaxanthin was isolated from the marine bacterium Brevundimonas sp. strain SD212 by using a common crtI sequence as the probe DNA. A sequence analysis revealed this cluster to contain 12 open reading frames (ORFs), including the 7 known genes, crtW, crtY, crtI, crtB, crtE, idi, and crtZ. The individual ORFs were functionally analyzed by complementation studies using Escherichia coli that accumulated various carotenoid precursors due to the presence of other bacterial crt genes. In addition to functionally identifying the known crt genes, we found that one (ORF11, named crtG) coded for a novel enzyme, carotenoid 2,2'-beta-hydroxylase, which showed intriguingly partial homology with animal sterol-C5-desaturase. When this crtG gene was introduced into E. coli accumulating zeaxanthin and canthaxanthin, the resulting transformants produced their 2-hydroxylated and 2,2'-dihydroxylated products which were structurally novel or rare xanthophylls, as determined by their nuclear magnetic resonance and high-performance liquid chromatography/photodiode array detector/atmospheric pressure chemical ionization mass spectrometry spectral data. The new carotenoid produced was suggested to have a strong inhibitory effect on lipid peroxidation.

A new carotenoid glycosyl ester isolated from a marine microorganism, Fusarium strain T-1.

A new carotenoid glycosyl ester, neurosporaxanthin beta-D-glucopyranoside (2), together with neurosporaxanthin (1), beta-carotene, gamma-carotene, and torulene were isolated from cultured cells of a marine microorganism, strain T-1, which was identified as Fusarium sp. Their structures were determined by chemical and spectral data.