Optimized extraction technology of glutathione from 'Haidao 86' germ rice by response surface methodology.

Glutathione is an important functional component of 'Haidao 86', which has many important physiological functions in organisms and is widely used in medicine and other industries. In this study, the effects of four extraction methods (hot water extraction, formic acid extraction, ethanol extraction, and sulfuric acid extraction) on the yield of glutathione in 'Haidao 86' germ powders were studied by high-performance liquid chromatography, and the yield of glutathione in hot water extraction was the highest. The effects of material-liquid ratio, temperature, pH, and time on the extraction rate of glutathione from 'Haidao 86' were investigated by single-factor experiment and Box-Behnken combined experiment. The results showed that the order of influence on GSH yield was pH, temperature, material-liquid ratio, and time, and the interaction of extraction time and pH had a significant influence on glutathione yield of 'Haidao 86' germ powders. The optimum parameters for hot water extraction of glutathione from 'Haidao 86' germ powders were determined as follows: material-liquid ratio of 1:12, pH value of 2.8, temperature of 84.9°C, time of 14 min, and the extraction rate of glutathione was 139.68 mg/100 g. It provided the scientific proof for the development and industrial production of functional products of 'Haidao 86'.

Protection efficacy of the H1 and H3 bivalent virus-like particle vaccine against swine influenza virus infection.

Swine influenza (SI) is widely prevalent in pig herds worldwide, causing huge economic losses to the pig industry and public health risks. The traditional inactivated swine influenza virus (SIV) vaccines are produced in chicken embryos, and egg-adaptive substitutions that occur during production process can impact vaccine effectiveness. Thus, developing an SI vaccine that can decrease the dependence on chicken embryos with a high immunogenicity is urgently needed. In this study, the utility of insect cell-derived SIV H1 and H3 bivalent virus-like particle (VLP) vaccines containing HA and M1 proteins of Eurasian avian-like (EA) H1N1 SIV and recent human-like H3N2 SIV were assessed in piglets. Antibody levels were monitored, and the protection efficacy of the vaccine after viral challenge was evaluated and compared with the inactivated vaccine. Results show that piglets produced high hemagglutination inhibition (HI) titers of antibodies against H1 and H3 SIV after immunization with SIV VLP vaccine. The neutralizing antibody level was significantly higher in SIV VLP vaccine than in the inactivated vaccine at 6 weeks post vaccination (p < 0.05). Furthermore, piglets immunized with the SIV VLP vaccine were protected against the challenge of H1 and H3 SIV, displaying inhibition of viral replication in piglets, and reduced lung damage. These results show that SIV VLP vaccine has good application prospects, thus laying the foundation for further research and commercialization of SIV VLP vaccine.

Vitamin D supplementation and lower respiratory tract infection in infants: a nested case-control study.

PURPOSE: To assess the association between vitamin D (VD) supplementation and the risk of lower respiratory tract infection (LRTI) among infants. METHODS: This is a nested case-control study from an ongoing prospective birth cohort in Wuhan from 2013. Cases were subjects free of neonatal pneumonia but later developed LRTI during infancy, who were matched with five randomly selected controls by infant sex, birth year, and birth season. We included 190 cases and 950 controls in the final analysis. The primary outcome was the first LRTI incident and the exposure was VD supplementation from birth to the index endpoint. The association between VD supplementation and LRTI risk was assessed using the Cox proportional-hazards regression model. RESULTS: Infants taking supplements had a 59% relative reduction in the hazard ratio of LRTI (HR = 0.41; 95% CI 0.26, 0.64) compared to those not supplemented. There was a linear relationship between LRTI risk and VD supplementation within range of 0-603 IU/day: for each 100 IU per day increment in VD supplementation, infants experienced a 21% lower risk of developing LRTI (adjusted HR: 0.79; 95% CI 0.71, 0.89). The linear relationship was stably observed in the sensitivity analyses as well. CONCLUSIONS: VD supplementation was associated with the reduced risk of LRTI throughout infancy, and the optimal supplementation dose for infants may be beyond the current recommendation.

Protective effect of bivalent H1N1 and H3N2 VLP vaccines against Eurasian avian-like H1N1 and recent human-like H3N2 influenza viruses in a mouse model.

Eurasian avian-like (EA) H1N1 swine influenza viruses (SIVs) are currently the most prevalent SIVs in Chinese swine populations, but recent human-like H3N2 SIV subtypes have also been frequently isolated. Hence, there is an urgent need to develop an effective vaccine against both EA H1N1 and recent human-like H3N2 infections. In this study, we utilized the baculovirus expression system to produce virus-like particles (VLPs) containing hemagglutinin protein (HA) and matrix protein (M1) based on A/Swine/Guangdong/YJ4/2014 (H1N1) and A/swine/Guangdong/L22/2010 (H3N2). An immunological experiment showed that in a mouse model, bivalent VLP vaccines against H1N1 and H3N2 can induce stronger humoral and cellular immune responses than whole influenza virus vaccines. Compared with monovalent inactivated vaccines that cannot offer protection against different SIV subtypes, monovalent H1N1 or H3N2 VLP vaccines can provide partial protection against lethal challenge by viruses of different subtypes. Meanwhile, bivalent VLP vaccines against H1N1 and H3N2 can provide full protection against lethal doses of homologous and heterologous viruses belonging to the EA H1N1 or recent human-like H3N2 lineage. These results suggest a promising approach to the development of vaccines against SIVs.

Association of vitamin D supplementation with respiratory tract infection in infants.

Vitamin D deficiency has been reported to be associated with respiratory tract infection (RTI). However, evidence regarding the effects of vitamin D supplementation on susceptibility of infants to RTI is limited. In this prospective birth cohort study, we examined whether vitamin D supplementation reduced RTI risk in 2,244 infants completing the follow-up from birth to 6 months of age. The outcome endpoint was the first episode of paediatrician-diagnosed RTI or 6 months of age when no RTI event occurred. Infants receiving vitamin D supplements at a daily dose of 400-600 IU from birth to the outcome endpoint were defined as vitamin D supplementation and divided into four groups according to the average frequency of supplementation: 0, 1-2, 3-4, and 5-7 days/week. We evaluated the relationship between vitamin D supplementation and time to the first episode of RTI with Kaplan-Meier plots. The associations of vitamin D supplementation with infant RTI, lower RTI (LRTI), and RTI-related hospitalization were assessed using modified Poisson regression. The median time to first RTI episode was 60 days after birth (95% CI [60, 90]) for infants without supplementation and longer than 6 months of age for infants with supplementation (p < .001). We observed inverse trends between supplementation frequency and risk of RTI, LRTI, and RTI-related hospitalization (p for trend < .001), with the risk ratios in the 5-7 days/week supplementation group of 0.46 (95% CI [0.41, 0.50]), 0.17 (95% CI [0.13, 0.24]), and 0.18 (95% CI [0.12, 0.27]), respectively. These associations were significant and consistent in a subgroup analysis stratified by infant feeding.

The association between caesarean delivery and the initiation and duration of breastfeeding: a prospective cohort study in China.

BACKGROUND/OBJECTIVES: Breastfeeding is of great benefit to infants and their mothers. Caesarean delivery (CD) have increased worldwide in recent years and emerging evidence has implied that CD may influence the initiation and duration of breastfeeding. However, the findings are inconsistent and intricate. The aim of this study is to illuminate the association between the CD and the initiation and duration of breastfeeding in the first 6 months postpartum. SUBJECTS/METHODS: A total of 2058 mother-infant pairs were studied in this prospective study. Delivery information was obtained from birth records. Feeding information in the first 6 months postpartum were collected from face-to-face interviews. Logistic regression was used to explore the association between CD and the initiation and duration of breastfeeding. RESULTS: After adjustment for potential confounders, CD was significantly associated with the unsuccessful initiation of breastfeeding [odds ratio (OR): 1.943, 95% confidence interval (CI): 1.050-3.597] and delayed initiation of breastfeeding [OR: 1.450, (95% CI: 1.041-2.019)], when compared with vaginal delivery (VD). More importantly, for mothers who had initiated breastfeeding, CD was associated with significantly higher risks of an inability to sustain full breastfeeding (OR: 1.369, 95% CI: 1.128-1.662), any breastfeeding at 3 months postpartum (OR: 1.715, 95% CI: 1.265-2.325) and any breastfeeding at 6-month postpartum (OR: 1.462, 95% CI: 1.174-1.820). CONCLUSIONS: CD is an independent risk factor for the inability to initiate and sustain breastfeeding. It is desirable to reduce the CD rate and provide specific breastfeeding support during early postpartum period to CD mothers.

Early feeding of larger volumes of formula milk is associated with greater body weight or overweight in later infancy.

BACKGROUND: The relation between infant feeding and growth has been extensively evaluated, but studies examining the volume of formula milk consumption on infant growth are limited. This study aimed to examine the effects of early feeding of larger volumes of formula on growth and risk of overweight in later infancy. METHODS: In total, 1093 infants were studied prospectively. Milk records collected at 3 mo of age were used to define the following 3 feeding groups: breast milk feeding (BM, no formula), lower-volume formula milk feeding (LFM, <840 ml formula/d), and higher-volume formula milk feeding (HFM, ≥840 ml formula/d). Body weight and length were measured at 3 time points of 3, 6 and 12 mo of age. RESULTS: The results showed that the difference in weight and length between the HFM and BM infants was significant at 3 mo of age (P < 0.05) and continued until 12 mo of age (P < 0.001). The adjusted mean changes in weight-for-length z-scores (WLZ) and BMI-for-age z-scores (BAZ) from 3 to 6 mo of age were significantly higher in HFM and LFM group than in BM group. Two-way interactions between feeding practice and age intervals were significant for WLZ changes (P = 0.002) and BAZ changes (P = 0.017). Compared with BM-fed infants, infants fed with HFM had 1.60-fold (95% CI 1.05-2.44) higher odds of greater body weight (1SD < WLZ ≤2 SD) at the age of 6 mo and 1.55-fold (95% CI 1.01-2.37) higher odds of greater body weight and 2.13-fold (95% CI 1.03-4.38) higher odds of overweight (WLZ > 2 SD) at the age of 12 mo. CONCLUSION: Feeding higher volumes of formula in early infancy is associated with greater body weight and overweight in later infancy.

Leucine Supplementation Differently Modulates Branched-Chain Amino Acid Catabolism, Mitochondrial Function and Metabolic Profiles at the Different Stage of Insulin Resistance in Rats on High-Fat Diet.

The available findings concerning the association between branched-chain amino acids (BCAAs)-particularly leucine-and insulin resistance are conflicting. BCAAs have been proposed to elicit different or even opposite effects, depending on the prevalence of catabolic and anabolic states. We tested the hypothesis that leucine supplementation may exert different effects at different stages of insulin resistance, to provide mechanistic insights into the role of leucine in the progression of insulin resistance. Male Sprague-Dawley rats were fed a normal chow diet, high-fat diet (HFD), HFD supplemented with 1.5% leucine, or HFD with a 20% calorie restriction for 24 or 32 weeks. Leucine supplementation led to abnormal catabolism of BCAA and the incompletely oxidized lipid species that contributed to mitochondrial dysfunction in skeletal muscle in HFD-fed rats in the early stage of insulin resistance (24 weeks). However, leucine supplementation induced no remarkable alternations in BCAA catabolism, but did enhance mitochondrial biogenesis with a concomitant improvement in lipid oxidation and mitochondrial function during the hyperglycaemia stage (32 weeks). These findings suggest that leucine trigger different effects on metabolic signatures at different stages of insulin resistance, and the overall metabolic status of the organisms should be carefully considered to potentiate the benefits of leucine.

Dietary apigenin potentiates the inhibitory effect of interferon-α on cancer cell viability through inhibition of 26S proteasome-mediated interferon receptor degradation.

BACKGROUND: Type I interferons (IFN-α/ß) have broad and potent immunoregulatory and antiproliferative activities. However, it is still known whether the dietary flavonoids exhibit their antiviral and anticancer properties by modulating the function of type I IFNs. OBJECTIVE: This study aimed at determining the role of apigenin, a dietary plant flavonoid abundant in common fruits and vegetables, on the type I IFN-mediated inhibition of cancer cell viability. DESIGN: Inhibitory effect of apigenin on human 26S proteasome, a known negative regulator of type I IFN signaling, was evaluated in vitro. Molecular docking was conducted to know the interaction between apigenin and subunits of 26S proteasome. Effects of apigenin on JAK/STAT pathway, 26S proteasome-mediated interferon receptor stability, and cancer cells viability were also investigated. RESULTS: Apigenin was identified to be a potent inhibitor of human 26S proteasome in a cell-based assay. Apigenin inhibited the chymotrypsin-like, caspase-like, and trypsin-like activities of the human 26S proteasome and increased the ubiquitination of endogenous proteins in cells. Results from computational modeling of the potential interactions of apigenin with the chymotrypsin site (ß5 subunit), caspase site (ß1 subunit), and trypsin site (ß2 subunit) of the proteasome were consistent with the observed proteasome inhibitory activity. Apigenin enhanced the phosphorylation of signal transducer and activator of transcription proteins (STAT1 and STAT2) and promoted the endogenous IFN-α-regulated gene expression. Apigenin inhibited the IFN-α-stimulated ubiquitination and degradation of type I interferon receptor 1 (IFNAR1). Apigenin also sensitized the inhibitory effect of IFN-α on viability of cervical carcinoma HeLa cells. CONCLUSION: These results suggest that apigenin potentiates the inhibitory effect of IFN-α on cancer cell viability by activating JAK/STAT signaling pathway through inhibition of 26S proteasome-mediated IFNAR1 degradation. This may provide a novel mechanism for increasing the efficacy of IFN-α/ß.

Emodin potentiates the antiproliferative effect of interferon α/ß by activation of JAK/STAT pathway signaling through inhibition of the 26S proteasome.

The 26S proteasome is a negative regulator of type I interferon (IFN-α/ß) signaling. Inhibition of the 26S proteasome by small molecules may be a new strategy to enhance the efficacy of type I IFNs and reduce their side effects. Using cell-based screening assay for new 26S proteasome inhibitors, we found that emodin, a natural anthraquinone, was a potent inhibitor of the human 26S proteasome. Emodin preferably inhibited the caspase-like and chymotrypsin-like activities of the human 26S proteasome and increased the ubiquitination of endogenous proteins in cells. Computational modeling showed that emodin exhibited an orientation/conformation favorable to nucleophilic attack in the active pocket of the ß1, ß2, and ß5 subunits of the 26S proteasome. Emodin increased phosphorylation of STAT1, decreased phosphorylation of STAT3 and increased endogenous gene expression stimulated by IFN-α. Emodin inhibited IFN-α-stimulated ubiquitination and degradation of type I interferon receptor 1 (IFNAR1). Emodin also sensitized the antiproliferative effect of IFN-α in HeLa cervical carcinoma cells and reduced tumor growth in Huh7 hepatocellular carcinoma-bearing mice. These results suggest that emodin potentiates the antiproliferative effect of IFN-α by activation of JAK/STAT pathway signaling through inhibition of 26S proteasome-stimulated IFNAR1 degradation. Therefore, emodin warrants further investigation as a new means to enhance the efficacy of IFN-α/ß.

Genome-wide transcriptome and expression profile analysis of Phalaenopsis during explant browning.

BACKGROUND: Explant browning presents a major problem for in vitro culture, and can lead to the death of the explant and failure of regeneration. Considerable work has examined the physiological mechanisms underlying Phalaenopsis leaf explant browning, but the molecular mechanisms of browning remain elusive. In this study, we used whole genome RNA sequencing to examine Phalaenopsis leaf explant browning at genome-wide level. METHODOLOGY/PRINCIPAL FINDINGS: We first used Illumina high-throughput technology to sequence the transcriptome of Phalaenopsis and then performed de novo transcriptome assembly. We assembled 79,434,350 clean reads into 31,708 isogenes and generated 26,565 annotated unigenes. We assigned Gene Ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations, and potential Pfam domains to each transcript. Using the transcriptome data as a reference, we next analyzed the differential gene expression of explants cultured for 0, 3, and 6 d, respectively. We then identified differentially expressed genes (DEGs) before and after Phalaenopsis explant browning. We also performed GO, KEGG functional enrichment and Pfam analysis of all DEGs. Finally, we selected 11 genes for quantitative real-time PCR (qPCR) analysis to confirm the expression profile analysis. CONCLUSIONS/SIGNIFICANCE: Here, we report the first comprehensive analysis of transcriptome and expression profiles during Phalaenopsis explant browning. Our results suggest that Phalaenopsis explant browning may be due in part to gene expression changes that affect the secondary metabolism, such as: phenylpropanoid pathway and flavonoid biosynthesis. Genes involved in photosynthesis and ATPase activity have been found to be changed at transcription level; these changes may perturb energy metabolism and thus lead to the decay of plant cells and tissues. This study provides comprehensive gene expression data for Phalaenopsis browning. Our data constitute an important resource for further functional studies to prevent explant browning.

Luteolin sensitizes the antiproliferative effect of interferon α/ß by activation of Janus kinase/signal transducer and activator of transcription pathway signaling through protein kinase A-mediated inhibition of protein tyrosine phosphatase SHP-2 in cancer cells.

New negative regulators of interferon (IFN) signaling, preferably with tissue specificity, are needed to develop therapeutic means to enhance the efficacy of type I IFNs (IFN-α/ß) and reduce their side effects. We conducted cell-based screening for IFN signaling enhancer and discovered that luteolin, a natural flavonoid, sensitized the antiproliferative effect of IFN-α in hepatoma HepG2 cells and cervical carcinoma HeLa cells. Luteolin promoted IFN-ß-induced Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway activation by enhancing the phosphorylation of Jak1, Tyk2, and STAT1/2, thereby promoting STAT1 accumulation in the nucleus and endogenous IFN-α-regulated gene expression. Of interest, inhibition of phosphodiesterase (PDE) abolished the effect of IFN-ß and luteolin on STAT1 phosphorylation. Luteolin also increased the cAMP-degrading activity of PDE bound with type I interferon receptor 2 (IFNAR2) and decreased the intracellular cAMP level, indicating that luteolin may act on the JAK/STAT pathway via PDE. Protein kinase A (PKA) was found to negatively regulate IFN-ß-induced JAK/STAT signaling, and its inhibitory effect was counteracted by luteolin. Pull-down and immunoprecipitation assays revealed that type II PKA interacted with IFNAR2 via the receptor for activated C-kinase 1 (RACK-1), and such interaction was inhibited by luteolin. Src homology domain 2 containing tyrosine phosphatase-2 (SHP-2) was further found to mediate the inhibitory effect of PKA on the JAK/STAT pathway. These data suggest that PKA/PDE-mediated cAMP signaling, integrated by RACK-1 to IFNAR2, may negatively regulate IFN signaling through SHP-2. Inhibition of this signaling may provide a new way to sensitize the efficacy of IFN-α/ß.