Heat Stress and Microbial Stress Induced Defensive Phenol Accumulation in Medicinal Plant Sparganium stoloniferum.

An approach based on the heat stress and microbial stress model of the medicinal plant Sparganium stoloniferum was proposed to elucidate the regulation and mechanism of bioactive phenol accumulation. This method integrates LC-MS/MS analysis, 16S rRNA sequencing, RT-qPCR, and molecular assays to investigate the regulation of phenolic metabolite biosynthesis in S. stoloniferum rhizome (SL) under stress. Previous research has shown that the metabolites and genes involved in phenol biosynthesis correlate to the upregulation of genes involved in plant-pathogen interactions. High-temperature and the presence of Pseudomonas bacteria were observed alongside SL growth. Under conditions of heat stress or Pseudomonas bacteria stress, both the metabolites and genes involved in phenol biosynthesis were upregulated. The regulation of phenol content and phenol biosynthesis gene expression suggests that phenol-based chemical defense of SL is stimulated under stress. Furthermore, the rapid accumulation of phenolic substances relied on the consumption of amino acids. Three defensive proteins, namely Ss4CL, SsC4H, and SsF3'5'H, were identified and verified to elucidate phenol biosynthesis in SL. Overall, this study enhances our understanding of the phenol-based chemical defense of SL, indicating that bioactive phenol substances result from SL's responses to the environment and providing new insights for growing the high-phenol-content medicinal herb SL.

De novo genome assembly of Bradysia cellarum (Diptera: Sciaridae), a notorious pest in traditional special vegetables in China.

Bradysia cellarum (Diptera: Sciaridae) is a destructive vegetable insect pest infesting more than 30 species of host plants from seven families in Asia and Europe. B. cellarum causes grave problems in Chinese chive, which originated in China and is cultivated widely in East Asia. The B. cellarum infestation results in economic losses and subsequent severe food safety problems in farm productions, insecticide resistance and environmental pollution. The genomic and molecular information of B. cellarum to delineate the biological features, insecticide resistance, evolution remains poorly understood. Herein, we decode the whole genome of B. cellarum to delineate the underlying molecular mechanisms causing insecticide resistance. We constructed a highly reliable genome for B. cellarum using PacBio, Illumina and 10X Genomics sequencing platforms. The genome size of B. cellarum was 375.91 Mb with a contig N50 of 1.57 Mb. A total of 16,231 genes were identified, among which 93.8% were functionally annotated, and 42.06% were repeat sequences. According to phylogenetic analysis, B. cellarum diverged from the common ancestor of Drosophila melanogaster and Musca domestica ~139.3-191.0 million years ago. Moreover, some important genes responsible for significant insecticide resistance, such as cytochrome P450s, ABC transporters and those involved in glutathione metabolism, were expanded in B. cellarum. We assembled a high-quality B. cellarum genome to provide valuable insights into their life history strategies, insecticide resistance and biological behaviours. It also lays the foundation for exploring gene structure and functional evolution, as well as comparative genomics of B. cellarum and other model insect species.

Rare Diseases Related with Lipoprotein Metabolism.

Rare diseases are gathering increasing attention in last few years, not only for its effects on innovation scientific research, but also for its propounding influence on common diseases. One of the most famous milestones made by Michael Brown and Joseph Goldstein in metabolism field is the discovery of the defective gene in familial hypercholesterolemia, a rare human genetic disease manifested with extreme high level of serum cholesterol (Goldstein JL, Brown MS, Proc Natl Acad Sci USA 70:2804-2808, 1973; Brown MS, Dana SE, Goldstein JL, J Biol Chem 249:789-796, 1974). Follow-up work including decoding the gene function, mapping-related pathways, and screening therapeutic targets are all based on the primary finding (Goldstein JL, Brown MS Arterioscler Thromb Vasc Biol 29:431-438, 2009). A series of succession win the two brilliant scientists the 1985 Nobel Prize, and bring about statins widely used for lipid management and decreasing cardiovascular disease risks. Translating the clinical extreme phenotypes into laboratory bench work has turned out to be the first important step in the paradigm conducting translational and precise medical research. Here we review the main categories of rare disorders related with lipoprotein metabolism, aiming to strengthen the notion that human rare inheritable genetic diseases would be the window to know ourselves better, to treat someone more efficiently, and to lead a healthy life longer. Few rare diseases related with lipoprotein metabolism were clustered into six sections based on changes in lipid profile, namely, hyper- or hypocholesterolemia, hypo- or hyperalphalipoproteinemia, abetalipoproteinemia, hypobetalipoproteinemia, and sphingolipid metabolism diseases. Each section consists of a brief introduction, followed by a summary of well-known disease-causing genes in one table, and supplemented with one or two diseases as example for detailed description. Here we aimed to raise more attention on rare lipoprotein metabolism diseases, calling for more work from basic research and clinical trials.

Tat-CIRP Peptide Facilitates Frozen Wound Healing by Ameliorating Inflammation and Promoting Angiogenesis.

Background: Frostbite is a chemia resulting from cold-induced skin damage. The process of frostbite is often accompanied by inflammation, and the therapeutic strategies focusing on anti-inflammation are the main direction to data. Tat-CIRP is a 15 amino acid peptide containing HIV protein and cold-inducible RNA-binding protein (CIRP), which is believed to compete with endogenous CIRP for myeloid differentiation 2 (MD2) binding. This study aims to investigate the efficacy of Tat-CIRP in the treatment of frostbite. Methods: A mouse model of frostbite was established, and on the first day after frostbite occurrence, Tat-CIRP peptide was administered intravenously via the tail with a dosage interval of one day for a total of three doses. Frozen mouse skin sections were subjected to histological analysis, including hematoxylin-eosin (HE) staining, Masson staining, and immunohistochemical examination. Western blotting was performed to detect the expression level of Ki-67 in mouse skin tissue. Results: One day after frostbite, mice exhibited skin swelling and a solid appearance. From day 1 to 5 after frostbite, MD2 expression was significantly upregulated, while CIRP expression was downregulated. Compared to the frostbite group, mice treated with Tat-CIRP showed accelerated frostbite recovery, reduced levels of inflammatory factors and MD2. Furthermore, the expression of cell proliferation-associated protein Ki-67 and angiogenesis-related protein CD31 was upregulated. Conclusion: Tat-CIRP promotes frozen wound healing via inhibiting inflammation and promoting angiogenesis in frostbitten mice.

Effects of the addition of leucine on flavor and quality of sausage fermented by Lactobacillus fermentum YZU-06 and Staphylococcus saprophyticus CGMCC 3475.

Methyl-branched aldehydes, especially 3-methylbutanal, have been reported to be perceived either as a malty or as a nutty/chocolate-like aroma and were considered an important flavor contributor in fermented meat products. Decomposition of leucine (Leu) by branched-chain amino acid transaminase (BACT) is a crucial step in the metabolism of Leu to 3-methylbutanal. This study was conducted to explore the effects of mixed-starter culture (Lactobacillus fermentum YZU-06 and Staphylococcus saprophyticus CGMCC 3475) and addition of Leu (0, 1, and 3 mM) on the flavor and quality of fermented sausages. The pH, water activity, texture profile analysis, color, counts of lactic acid bacteria (LAB) and staphylococci, peptide, and flavor compounds were detected during fermentation. The results showed that the starter culture group increased hardness, elasticity, the counts of LAB and staphylococci, peptide content, volatile flavor compounds, as well as the sensorial scores of sausage, while decreasing pH, a w , and L* and b* values compared with the non-inoculation group. The mixed starter of adding with 3 mM Leu enhanced the content of 3-methylbutanal and overall flavor of fermented sausages. It is applicable to directionally produce methyl-branched aldehydes and improve the overall quality of fermented sausage by the addition of Leu and using starter of L. fermentum YZU-06 and S. saprophyticus CGMCC 3475.

Characterization of a lactic acid bacteria using branched-chain amino acid transaminase and protease from Jinhua Ham and application in myofibrillar protein model.

This study was to screen a strain with both branched-chain amino acid transaminase (BCAT) and protease activities for producing methyl-branched flavor compounds in a myofibrillar protein extract model. Lactic acid bacteria (LAB) was isolated from Jinhua ham and screened with BCAT activity by an electrochemical sensor and protease activity by the agar plate method. In the culture medium, strain L6 was selected with high utilization rate and characteristic metabolites content of branched-chain amino acids (BCAAs), and identified as Lactobacillus fermentum YZU-06 (L. fermentum). Compared with the previously reported L. plantarum, L. fermentum exhibited an excellent capacity of hydrolyzing myofibrillar protein with the higher contents of free amino acids, peptides and small molecular weight proteins. Moreover, L. fermentum group presented more BCAAs metabolites of 2-methylbutanal and 3-methylbutanal than that of L. plantarum group. In conclusion, L. fermentum YZU-06 is a promising starter culture to improve the flavor of fermented meat products.

Impaired Insulin Clearance as the Initial Regulator of Obesity-Associated Hyperinsulinemia: Novel Insight Into the Underlying Mechanism Based on Serum Bile Acid Profiles.

OBJECTIVE: To investigate the roles of insulin clearance and insulin secretion in the development of hyperinsulinemia in obese subjects and to reveal the association between insulin clearance and bile acids (BAs). RESEARCH DESIGN AND METHODS: In cohort 1, insulin secretion, sensitivity, and endogenous insulin clearance were evaluated with an oral glucose tolerance test in 460 recruited participants. In cohort 2, 81 participants underwent an intravenous glucose tolerance test and a hyperinsulinemic-euglycemic clamp to assess insulin secretion, endogenous and exogenous insulin clearance, and insulin sensitivity. Based on insulin resistance levels ranging from mild to severe, obese participants without diabetes were further divided into 10 quantiles in cohort 1 and into tertiles in cohort 2. Forty serum BAs were measured in cohort 2 to examine the association between BAs and insulin clearance. RESULTS: All obese participants had impaired insulin clearance, and it worsened with additional insulin resistance in obese subjects without diabetes. However, insulin secretion was unchanged from quantile 1 to 3 in cohort 1, and no difference was found in cohort 2. After adjustments for all confounding factors, serum-conjugated BAs, especially glycodeoxycholic acid (GDCA; ß = -0.335, P = 0.004) and taurodeoxycholic acid (TDCA; ß = -0.333, P = 0.003), were negatively correlated with insulin clearance. The ratio of unconjugated to conjugated BAs (ß = 0.335, P = 0.002) was positively correlated with insulin clearance. CONCLUSIONS: Hyperinsulinemia in obese subjects might be primarily induced by decreased insulin clearance rather than increased insulin secretion. Changes in circulating conjugated BAs, especially GDCA and TDCA, might play an important role in regulating insulin clearance.

Targeted lipidomics reveals associations between serum sphingolipids and insulin sensitivity measured by the hyperinsulinemic-euglycemic clamp.

AIMS: Sphingolipids(SPs) and their substrates and constituents, fatty acids (FAs), are implicated in the pathogenesis of various metabolic diseases associated. This study aimed to systematically investigate the associations between serum sphingolipids and insulin sensitivity as well as insulin secretion. METHODS: We conducted a lipidomics evaluation of molecularly distinct SPs in the serum of 86 consecutive Chinese adults using LC/MS. The glucose infusion rate over 30 min (GIR30) was measured under steady conditions to assess insulin sensitivity by the gold standard hyperinsulinemic-euglycemic clamp. We created the ROC curves to detect the serum SMs diagnostic value. RESULTS: Total and subspecies of serum SMs and globotriaosyl ceramides (Gb3s) were positively related to GIR30, free FAs (FFA 16:1, FFA20:4), some long chain GM3 and complex ceramide GluCers showed strong negative correlations with GIR30. Notably, ROC curves showed that SM/Cer and SM d18:0/26:0 may be good serum lipid predictors of diagnostic indicators of insulin sensitivity close to conventional clinical indexes such as 1/HOMA-IR (areas under the curve > 0.80) based on GIR30 as standard diagnostic criteria, and (SM/Cer)/(BMI*LDLc) areas under the curve = 0.93) is the best. CONCLUSIONS: These results provide novel associations between serum sphingolipid between insulin sensitivity measured by the hyperinsulinemic-euglycemic clamp and identify two specific SPs that may represent prognostic biomarkers for insulin sensitivity.

[Effect of quercetin on bone formation in the mid-palatal suture of rats during rapid maxillary expansion].

PURPOSE: This study was aimed to investigate the effect of quercetin on rats' palatal suture during rapid maxillary expansion(RME). METHODS: Eighteen male 6-week-old（specific pathogen free，SPF） rats were randomly divided into 3 groups. Group A was control group, group B underwent expansion alone, and group C underwent expansion plus quercetin administration. Each group had 6 rats. Neither expansion nor quercetin was given to group A. Each rat in group B and C was set an orthodontic appliance with an initial expansive force (0.98 N). The rats in group C were administered with quercetin (100 mg/kg) at the same time every day after RME, while equal normal saline was given to rats in group A and B. Then the rats were sacrificed on day 14. Sections were cut perpendicular to the midpalatal suture and stained with hematoxylin and eosin (H-E) for observation of celluar response， and immunostained for evaluation of bone formation．The changes of collagen were observed through Masson's staining. Image-pro plus adhesive was used to analyze the slice absorbance, and SPSS19.0 software package was used to analyze the data. RESULTS: The results showed that small amount of fibrous tissue was found in the palate of group A, and there were chondrocytes, mesenchymal cells and osteoblasts. There were more fibrous tissues in the palate of group C, fibroblasts and cartilage cells increased, and osteoblasts were seen near the bone region of the palate. The number of osteoblasts in group C was significantly higher than that in group B, and there were new bone calcification deposits. The expression of BMP-2 in the midpalatal suture was significantly greater in group B than in group A at day 14 (P<0.01); the expression of BMP-2 in the midpalatal suture in group C was significantly greater than in group B (P<0.05). CONCLUSIONS: Rapid maxillary expansion can enlarge the middle palatal suture of rats during growth and development, quercetin can promote the expression of BMP-2 in the middle palatal suture of rats during rapid maxillary expansion, make bone deposition and calcification, and accelerate new bone formation.