Effect of Chitosan and Hyperbranched Poly-L-Lysine Treatment on Quality of Cucumber (Cucumis sativus L.) during Storage.

To enhance the storage time of cucumbers, this research investigated the impact of chitosan (CS) and hyperbranched poly-L-lysine (HBPL) on the quality and nutritional attributes of cucumbers when stored at a temperature of 25 °C. The results demonstrated that sensory evaluation scores for cucumbers treated with a CS-HBPL combination were significantly higher than the control (CK), CS, and HBPL groups. On the 18th day of storage, cucumbers in the CK group exhibited significant decay and softening; however, there was a decrease in hardness observed in the CS-HBPL group and no decay or noticeable sour taste was detected. Furthermore, compared to the CK group, treatment with CS-HBPL effectively delayed cucumber decay and weight loss rate while significantly inhibiting decreases in cucumber hardness and growth of surface microorganisms. Additionally, it substantially reduced losses of soluble protein content as well as vitamin C (Vc), reducing sugars, and total phenolic compounds within cucumbers, which were 4.7 mg/g, 4.7 mg/g, 0.94 mg/g, and 0.52 mg/kg, respectively. Moreover, compared to the CK group, combined treatment with CS-HBPL significantly inhibited malondialdehyde (MDA) accumulation and reducing relative electrolyte permeability within cucumbers, which were 1.45 µmol·g-1FW and 29.82%. Furthermore, it notably enhanced activities of superoxide dismutase (SOD) and catalase (CAT), while exerting a significant inhibitory effect on polyphenol oxidase (PPO). In summary, the combined CS-HBPL treatment successfully prolonged cucumber shelf life at room temperature, enabling new possibilities for extending cucumber shelf life.

UV-ARTP-DES compound mutagenesis breeding improves natamycin production of Streptomyces natalensis HW-2 and reveals transcriptional changes by RNA-seq.

Natamycin is widely used in food, medical and health, agriculture, and animal husbandry. In this study, Streptomyces natalensis HW-2 was used as the research object, and a mutant DES-26 with stable genetic characters was selected by UV-ARTP-DES compound mutation. The natamycin yield was 1.64 g/L, 86.36% higher than original strain. Differential expression genes were analyzed by transcriptomics, and results showed that 295 and 860 genes were significantly differentially expressed at fermentation for 48 h and 72 h. GO and KEGG analysis showed that compound mutagenesis had a significant impact on glycolysis, pentose phosphate, TCA cycle, fatty acid metabolism pathways, and several key enzyme genes in the pathways were up-regulated, and genes related to natamycin biosynthesis (pimB-pimI) and transcriptional regulator (pimR) were also up-regulated. qRT-PCR results confirmed that expression levels of these genes were consistent with transcriptional changes of RNA-Seq. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01191-z.

Effect of Combined Chitosan and Hyperbranched Poly-L-Lysine Based Coating on Prolonging the Shelf Life of Oyster Mushroom (Pleurotus ostreatus).

To extend the shelf life of oyster mushroom (Pleurotus ostreatus), the effects of chitosan (CS) and hyperbranched poly-L-lysine (HBPL) combined treatment on quality characteristics, nutritional quality, storage characteristics, and enzyme activity of oyster mushroom during postharvest storage at 4 °C were investigated. The results showed that CS-HBPL combined treatment could significantly reduce rot degree and weight loss and significantly inhibit the browning of oyster mushroom. At the same time, the loss of reducing sugar, vitamin C, soluble protein, and total phenolic was significantly reduced. Compared with the control, CS-HBPL combined treatment could also significantly inhibit an increase in malondialdehyde (MDA) and significantly decrease the relative electrolyte leakage of oyster mushroom. In addition, the activities of catalase (CAT), superoxide dismutase (SOD), phenylalnine ammonialyase (PAL), and peroxidase (POD) were significantly improved, and the activity of polyphenol oxidase (PPO) was significantly inhibited in oyster mushroom. In conclusion, CS-HBPL combined treatment had a good protective effect on the membrane permeability damage of oyster mushroom and could effectively delay the oxidation of phenolic substances and browning of oyster mushroom. Therefore, CS-HBPL combined treatment can be used as a potential strategy to extend the storage time of oyster mushroom.

Association of Vegetable and Fruit Consumption with Urinary Oxidative Biomarkers in Teenaged Girls: A School-Based Pilot Study in Japan.

Hexanoyl-lysine (HEL), 8-hydroxy-2'deoxyguanosine (8-OHdG), and dityrosine (DT) have served as potential biomarkers for detecting oxidative modified lipids, DNA, and proteins in biological samples, respectively. Whether regular higher levels of consumption of vegetables/fruit (V/F) would decrease oxidative modification of these biomolecules in the body remain unelucidated. To examine the association of regular V/F consumption with the generation of these reactive oxygen species-induced biomarkers, this study evaluated V/F consumption in a school-based sample of teenaged girls (mean age 15.6 ± 1.7 years, n = 103), and quantified the formation of oxidative stress biomarkers in their urine. Only 19.4% and 23.3% of participants reported that they consumed the recommended daily amount of vegetables and fruits, respectively. Individuals who consumed lower levels of fruit (<100g/day) or vegetables (<250g/day) had significantly higher HEL excretion in their urine than those who consumed higher levels of fruit (≥100g/day) (p < 0.05) or vegetables (≥250g/day) (p = 0.057). The results of a multiple regression analysis showed that vegetable consumption was an important inhibiting factor of early lipid peroxidation measured as HEL in urine, independent of various confounders (ß = - 0.332, p < 0.05). The findings suggest that relatively higher consumption of vegetables would help in the prevention of early lipid peroxidation in adolescents.

Microwave Irradiation: Effects on the Change of Colour Characteristics and Main Phenolic Compounds of Cabernet Gernischt Dry Red Wine during Storage.

Wine colour is an essential organoleptic property considered by consumers. In this paper, the potential effects on colour characteristics and the content of main phenolic compounds in red wine under microwave irradiation were investigated during wine storage. The results showed that the changing trend of colour characteristics of microwave-treated and untreated wines was very similar. Moreover, total phenolic compounds, total monomeric anthocyanins, main anthocyanins, main flavonoids, and main phenolic acids (gallic acid; caffeic acid; syringic acid; (+)-catechin; Cy-3-glu; Mv-3-glu) also showed similar change trends during storage. In other words, microwave irradiation had a long-term effect on the colour properties and main phenolic compounds of red wine, changes that require long-time aging in traditional processing. In terms of the studied parameters, the changes in microwave-treated wine were faster than those in untreated wine. These results showed that microwave technology, as a promising artificial aging technology, could in a short time produce red wine of similar quality to traditional aging.

Biotechnological production and application of epsilon-poly-L-lysine (ε-PL): biosynthesis and its metabolic regulation.

Epsilon-poly-L-lysine (ε-PL) is an unusual biopolymer composed of L-lysine produced by several microorganisms, especially by the genus Streptomyces. Due to its excellent antimicrobial activity, good water solubility, high safety, and biodegradable nature, ε-PL with a GRAS status has been widely used in food and pharmaceutical industries. In the past years, studies have focused on the biotechnological production of ɛ-PL, the biosynthetic mechanism of microbial ɛ-PL, and its application. To provide new perspectives from recent advances, the review introduced the methods for the isolation of ɛ-PL producing strains and the biosynthetic mechanism of microbial ɛ-PL. We summarized the strategies for the improvement of ɛ-PL producing strains, including physical and chemical mutagenesis, ribosome engineering and gene engineering, and compared the different metabolic regulation strategies for improving ɛ-PL production, including medium optimization, nutrient supply, pH control, and dissolved oxygen control. Then, the downstream purification methods of ɛ-PL and its recent applications in food and medicine industries were introduced. Finally, we also proposed the potential challenges and the perspectives for the production of ε-PL.

Effects of ethanol stress on epsilon-poly-l-lysine (ε-PL) biosynthesis in Streptomyces albulus X-18.

The aim of the study was to reveal the effects of ethanol stress on the production of epsilon-poly-l-lysine (ε-PL) in Streptomyces albulus X-18. The results showed that biomass and the utilization of glucose were respectively increased by ethanol stress. The ε-PL yield was increased by 41.42 % in the shake flask and 37.02 % in 10 L fermenter with 1% (v/v) ethanol. The morphology of colonies and mycelia showed significant differences. The intracellular reactive oxygen species level was increased by about 100 %. The ratio of unsaturated fatty acids to saturated fatty acids in the cell membrane was increased by ethanol stress. Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) proteomic profile showed that 265 identified proteins were differentially expressed. The differentially expressed proteins (DEPs) were mainly involved in biological processes. The up-regulated DEPs were mainly involved in the redox reaction and stress response. The metabolic flux of l-Asp was shifted to l-Lys biosynthesis, and the DAP pathway was strengthened. Protein-protein interaction analysis showed that 30 DEPs interacted with l-Lys biosynthesis. The changes of ten proteins by Parallel Reaction Monitoring (PRM) were consistent with those by iTRAQ. The study provided valuable clues to better understand the mechanism of ε-PL biosynthesis improvement by ethanol stress.

Blue Light from Cell Phones Can Cause Chronic Retinal Light Injury: The Evidence from a Clinical Observational Study and a SD Rat Model.

BACKGROUND: To investigate the chronic photodamage induced by the low-intensity blue light of phones, we carried out a clinical pilot study and established an animal model by irradiating SD rats with a homemade illuminator. METHODS: Clinical investigation: A total of 25 clinical medical workers in our hospital were selected and divided into a control group and an observation group according to the daily video terminal use time. Multifocal electrophysiological system (Mf-ERG) was used for retinal functional examination. Animal experiment: A total of sixty SD rats were randomly divided into a control group (n = 6) and an experimental group (n = 54). The experimental rats were divided into nine groups, which were exposed to the blue light illuminator of the simulated cell phone array for different time. The visual electrophysiology of the rats was tested, and changes in structure were observed by H&E staining and transmission electron microscopy. RESULTS: In clinical investigation, macular centers near the concave area retinal photoreceptor cells have reduced amplitude. In animal experiments, the amplitude of photoreceptor cells decreased, the peak time was delayed, and the amplitudes were lower in the experimental groups. H&E staining and transmission electron microscope showed retinal tissue structure and functional damage in experimental groups. CONCLUSIONS: Long-term exposure to low-illuminance blue light can cause retinal tissue structure and functional damage, and the chronic damage due to low-illuminance light warrants attention. The clinical registration number is 2018-KY-KS-LHL.

[Advances in the biosynthesis of natamycin and its regulatory mechanisms].

Natamycin is a polyene macrolide antibiotics with strong and broad spectrum antifungal activity. It not only effectively inhibits the growth and reproduction of fungi, but also prevents the formation of some mycotoxins. Consequently, it has been approved for use as an antifungal food preservative in most countries, and is also widely used in agriculture and healthcare. Streptomyces natalensis and Streptomyces chatanoogensis are the main producers of natamycin. This review summarizes the biosynthesis and regulatory mechanism of natamycin, as well as the strategies for improving natamycin production. Moreover, the future perspectives on natamycin research are discussed.

Analysis of chemical compounds and toxicological evaluation of Forsythia suspensa leaves tea.

To determine the compositions of Forsythia suspensa leaves tea (FSLT) and its safety, the chemical compounds were analysed with some methods, and the toxicity was evaluated in Kunming mice and Wistar rats. The results showed that FSLT contained rich flavonoid, lignans, triperpene acids, amino acids, and mineral elements. In the acute toxicity study, none of the mice died, and no obvious poisoning symptoms were observed after 14 days in mice at the dose of 15 mg/g·body weight (bw) FSLT; in the sub-chronic toxicity, no abnormal or dead rat was found at the dose of 1, 3, and 10 mg/g·bw during 90 days feeding administration; there was no significant difference in bw and food consumption; no significant differences were found in each hematology and serum biochemistry parameter and organ/body weight ratio comparing with the control experimental group. The results revealed that the FSLT has low or no toxicity via oral administration. Therefore, FSLT is very suitable and safe to be used as a new resource food.

Psychosocial Determinants of Fruit and Vegetable Intake in Japanese Adolescents: A School-Based Study in Japan.

A few studies in Japan have demonstrated positive attitudes, self-efficacy, social support, and perceived barrier were associated with fruit and vegetable (F&V) intake in adults; however, limited evidence addresses the association of psychosocial factors with F&V intake in adolescents. A cross-sectional study through a questionnaire survey was conducted at junior and senior high schools, and 933 students completed the questionnaire. Data were analyzed by X2 test and Student t-test. The findings demonstrated 2.7% of participants were aware of the current recommendations for vegetable and 2.0% for fruit. Only 4% and 8.1% of participants reported they consumed recommended amount of vegetables and fruits. In comparison with males, females showed higher scores of attitude (p < 0.01), responsibility (p < 0.01), and social support (p < 0.01). The barriers to vegetable intake were "I'm eating enough now", "not always available when eating away from home"; the barriers to fruit intake were "don't have a habit of having 100% juice or fruit in the morning", and "cost too much". The findings suggest the change of adolescents' knowledge about what they should eat is needed in boosting F&V consumption. The development of an intervention program for adolescents needs to target socio-environmental factors such as family support, and nutritional education for early healthy habit formation.

Effects of a Forest Walk on Urinary Dityrosine and Hexanoyl-Lysine in Young People: A Pilot Study.

A few studies indicate exposure to forests may alleviate oxidative stress in the body. However, more evidence is needed to support this potentiality. The purpose of the current study aimed at examining whether there is any difference in urinary levels of oxidatively modified proteins or lipids-dityrosine (DT) and hexanoyl-lysine (HEL), respectively, after a forest or urban walk. The study was performed on 29 university students who took part in forest walks (Shinjo Village) in Okayama Prefecture of Japan and on 42 university students who took part in urban walks in the downtown area of Okayama City. Urine samples before and after the walks were analyzed for DT and HEL excretion. Air phytoncides during the walks were also measured. We found a decreased tendency in urinary DT and HEL (p < 0.05) in most participants after the forest walks, but not after the urban walks. We further found the total levels of air phytoncides in the forest field were 1.50 times higher compared with those in the urban field. This study suggests the possibility that regular immersion in a forest environment might contribute toward weakening of the oxidative modifications of proteins or lipids in the body.

Mutated SASH1 promotes Mitf expression in a heterozygous mutated SASH1 knock­in mouse model.

The SAM and SH3 domain­containing 1 (SASH1) genes have been identified as the causal genes of dyschromatosis universalis hereditaria (DUH); these genes cause the pathological phenotypes of DUH, and SASH1 variants have been shown to regulate the abnormal pigmentation phenotype in human skin in various genodermatoses. However, investigations into the mutated SASH1 gene have been limited to in vitro studies. In the present study, to recapitulate the molecular pathological phenotypes of individuals with DUH induced by SASH1 mutations, a heterozygous BALB/c mouse model, in which the human SASH1 c.1654 T>G (p. Tyr 551Asp, Y551D) mutation was knocked in was first generated. The in vivo functional experiments on Y551D SASH1 indicated that the increased expression of microphthalmia­associated transcription factor (Mitf) was uniformly induced in the tails of heterozygous BALB/c mice, and an increased quantity of Mitf­positive epithelial cells was also detected. An increased expression of Mitf­ and Mitf­positive cells was also demonstrated in the epithelial tissues of Y551D­SASH1 affected individuals. In the present study, Mitf expression was also found to be increased by Y551D SASH1 in vitro. Taken together, these findings indicate that the upregulation of Mitf is the bona fide effector of the Y551D SASH1­mediated melanogenesis signaling pathway in vivo. SASH1 may function as a scaffold molecule for the assembly of a SASH1­Mitf molecular complex to regulate Mitf expression in the cell nucleus and thus to promote the hyperpigmented phenotype in the pathogenesis of DUH and other genodermatoses related to pigment abnormalities.

Screening of cardioprotective diseases bioactive components from Danshen extracts and LC-MS analysis.

A rapid and efficient analysis and screening method is adopted for cell affinity capture coupled with HPLC-MS (CAC-HPLC-MS) analysis of bioactive components that have possible efficiency against cardiovascular diseases. This method involves affinity capture, concentration, and separation of bioactive components from Danshen library using oxidatively damaged endothelial cells induced by H2 O2 , as well as analysis and identification of targeted compounds with HPLC and MS. It combines the specific interaction between cell membrane receptors and bioactive components with the powerful analysis and identification function of HPLC-MS. The CAC-HPLC-MS method was also used for analysis and screening of bioactive components from crude extracts of Danshen. A total of 19 components were found to be bound to oxidatively damaged endothelial cells with seven of these identified. Existing literature confirms that these seven components have many activities related to cardioprotective diseases. Therefore, the combination of biological affinity capture with HPLC-MS should be regarded as an attractive method with great potential for rapid and efficient screening of bioactive components related to anti-cardiovascular diseases from natural product libraries.

Fungal elicitor-induced transcriptional changes of genes related to branched-chain amino acid metabolism in Streptomyces natalensis HW-2.

Natamycin is a polyene macrolide antibiotic and widely used as a natural food preservative. Fungal elicitor had positive effects on the natamycin biosynthesis in Streptomyces natalensis HW-2. However, the global gene expression in response to fungal elicitor is not still reported. In the study, RNA-Seq was used to check the change of transcriptome by fungal elicitor in S. natalensis HW-2. The results showed that there were 1265 differential expression genes (DEGs) at 40 h and 2196 DEGs at 80 h. Most of the genes involved in natamycin biosynthesis were upregulated. KEGG pathway analysis showed that fungal elicitor had strong effects on the transcriptional levels of genes related to branch-chained amino acid (BCAA) metabolism. There were 23 upregulated or downregulated DEGs involved in BCAA biosynthesis and degradation at 40 h and 80 h. To confirm whether the improvement of BCAA biosynthesis could produce more natamycin, metabolic engineering was used to homologously overexpress the gene ilvH which encoded the regulatory subunit of acetolactate synthase (ALS) in S. natalensis. The results showed that overexpression of ilvH in S. natalensis HW-2 increased natamycin production to 1.25 g/L in the flask, which was a 32% increase compared with that of the parent strain. Real-time quantitative PCR analysis showed that the transcriptional level of ilvH in mutant strain S. natalensis ZS101 was significantly increased. Acetyl-CoA content was also raised. The results suggested that the fungal elicitor enhanced natamycin biosynthesis by improving precursor supply via BCAA metabolism. This study will open a new avenue for enhancing natamycin production by metabolic engineering and adding fungal elicitor. KEY POINTS: • The fungal elicitor had strong effects on the transcriptional levels of genes related to branch-chained amino acid metabolism by RNA-Seq. • The homologous overexpression of gene ilvH increased natamycin production by 32% and acetyl-CoA content was raised in mutant strain S. natalensis ZS101.

Effect of hydrogen peroxide on normal and acatalasemic mouse erythrocytes.

OBJECTIVES: Normal and acatalasemic mouse erythrocytes were used to clarify the relationship between oxidative damage in H2O2-treated erythrocytes and catalase activity. DESIGN & METHODS: Generation of hydrolysis-resistant erythrocytes and hemolysis were examined. The osmotic fragility test, the negative charges and the number of membrane-flickering erythrocytes among the H2O2-treated erythrocytes were investigated. RESULTS: Small amounts of hydrolysis-resistant mouse erythrocytes were generated by treatment with 0.1 mM H2O2, and the amount of acatalasemic erythrocytes was larger than untreated controls. Hemolysis in the acatalasemic erythrocytes was observed 30 min after the addition of the H2O2. A drastic increase in hydrolysis-resistant erythrocytes and a loss of membrane proteins in the acatalasemic erythrocytes were found as a result of the addition of 1 mM H2O2. Hemolysis in normal erythrocytes was observed at 3 mM H2O2. CONCLUSIONS: Catalase is a potent H2O2-scavenger even in acatalasemic mouse erythrocytes. It is concluded that the drastic increase of hydrolysis-resistant erythrocytes is induced by a loss of membrane function and is associated with the low catalase activity in these cells.

Lsi1 modulates the antioxidant capacity of rice and protects against ultraviolet-B radiation.

Silicon (Si) enhances the resistance of rice to biotic and abiotic stress. In rice, the accumulation of Si is controlled by the low silicon rice 1 (Lsi1) gene; overexpression of Lsi1 (Lsi1-OX) increases Si uptake and accumulation, while the reverse is observed in Lsi1-RNA interference (Lsi1-RNAi) transgenic rice. When the two transgenic rice lines and wild-type (WT) rice were exposed to ultraviolet (UV)-B radiation, the Lsi1-OX or Lsi1-RNAi rice showed differential microRNA (miRNA) expression, compared to WT rice. These miRNAs were predicted to target genes involved in light signal transduction and cell detoxification. The greatest capacities of ascorbate peroxidase, superoxide dismutase, peroxidase, and phenylalanine ammonia lyase (PAL) and highest contents of phenolics, flavonoids, and proline were found in Lsi1-OX rice, followed by WT rice and Lsi1-RNAi transgenic rice. A further comparison of the transcript levels of individual PAL genes revealed that the expression of PAL2-2 (Os02g0626400) was positively regulated by Lsi1. Our results demonstrate that Lsi1 overexpression or interference causes changes in both miRNA expression and antioxidant capacity in rice, and therefore modulates rice tolerance to UV-B radiation. Furthermore, we demonstrated that PAL2-2 was positively regulated by Lsi1 during this process.

Changes in Urinary Hydrogen Peroxide and 8-Hydroxy-2'-Deoxyguanosine Levels after a Forest Walk: A Pilot Study.

Some studies have shown that exposure to forests has positive effects on human health, although the mechanisms underlying the health benefits of a forest environment have not been elucidated yet. The current study was aimed at examining how the levels of urinary hydrogen peroxide (H2O2) and 8-hydroxy-2'deoxyguanosine (8-OHdG) change after a forest or urban walk in healthy subjects. Twenty-eight volunteers (19 men and 9 women) participated in the study. The forest walks were carried out in a forest in Okayama Prefecture, Japan, and the urban walks (15 men and 7 women) were carried out in the downtown area of Okayama city, each for two hours. Spot urine samples were collected before the walk, the next day and one week after the forest or urban walk. Compared with pre-forest walk levels, urinary H2O2 (p < 0.1) and 8-OHdG (p < 0.1) concentrations significantly decreased in the participants the day after the forest walk; furthermore, urinary 8-OHdG remained at a low level even at one week after the forest walk (p < 0.05). However, there were no significant changes in the concentrations of these oxidative biomarkers after the urban walk. These findings suggest the possibility that exposure to forests may alleviate oxidative stress in the body.

Physicochemical and microbial responses of Streptomyces natalensis HW-2 to fungal elicitor.

The effects of fungal elicitor on the physicochemical and microbial responses of Streptomyces natalensis HW-2 were investigated. The results showed that the elicitor could decrease dry cell weight (DCW) by 17.7% and increase the utilization of glucose, while the curve of pH was not obviously altered. The elicitor enhanced the yield of natamycin from 1.33 to 2.49 g/L. The morphology of the colony and the mycelium treated with elicitor showed significant differences from that of control. The level of intracellular reactive oxygen species (ROS) increased to 333.8 ng/L, which was a twofold increase comparing with the control. The concentration of Ca2+ reached 421.1 nmol/L, which increased by 32.8% after the addition of the elicitor. The activities of pyruvic carboxylase and phosphoenol pyruvate carboxylase were enhanced by 27.8 and 11.9%, respectively, while citrate synthase activity decreased by 23.1% in comparison with the control.

Psychosocial Determinants of Fruit and Vegetable Consumption in a Japanese Population.

There is limited evidence in Japan regarding the psychosocial determinants of fruit/vegetable intake. We performed a cross-sectional study of people aged 18 years or older in four regions of Japan; 2308 (men: 1012, women: 1296) individuals who completed the questionnaires were included. We found that 24.8% of people were aware of the current recommendations for vegetables and 13.2% for fruit and that "ability to design meals" and "availability when eating outside of the home" were the most important factors related to self-efficacy and barriers to fruit and vegetable intake, respectively. People with high self-efficacy (OR: 3.16; 95% CI: 2.17, 4.60 for fruit; OR: 4.52; 95% CI: 3.08, 6.64 for vegetables) were more likely to consume more fruit and vegetables. People with high scores on attitude (OR: 1.54; 95% CI: 1.06, 2.24) and social support (OR: 1.59; 95% CI: 1.11, 2.27) were more likely to consume more fruit. People with high perceived barriers (OR: 0.69; 95% CI: 0.48, 0.98) were less likely to consume fruit. This study suggests a need to increase the general population's awareness of the fruit and vegetable intake recommendations; facilitating positive attitudes, self-efficacy, and social support for individuals and strengthening the ability of individuals to design meals with more vegetables and fruit might be useful intervention programs.