Optimization of the hongqu starter preparation process for the manufacturing of red mold rice with high gamma-aminobutyric acid production by solid-state fermentation.

Red mold rice (RMR) generally contains gamma-aminobutyric acid (GABA), which has several physiological functions. Monascus purpureus M162, with a high GABA production of 15.10 mg/g was generated by atmospheric and room temperature plasma mutation. Furthermore, we conducted a response surface methodology to produce a premium hongqu starter. The results revealed that under optimal conditions, that is, a substrate containing brown rice and bran in a brown rice: bran ratio of 9:1 (wt/wt), an inoculation size of 21.50 mL/100 g, a mixing frequency of one time/9 h, and a cultivation time of 7.20 days, the number of active spores, α-amylase activity, and saccharification power activity was 4.15 × 107 spores/g, 155 U/g, and 3260 U/g in the high-quality starter, respectively. These values were 224.32-fold, 139.64%, and 141.74% higher than those obtained with M. purpureus M162 inoculated into steamed indica rice, respectively, and 153.70-fold, 267.24%, and 151.63% higher than those obtained with the parent strain M. purpureus M1, respectively. The premium hongqu starter of M. purpureus M162 was inoculated into steamed indica rice to produce RMR with 15.93 mg/g of GABA. In conclusion, we proposed a novel strategy for functional RMR production with high GABA concentrations by solid-state fermentation with Monascus spp.

Comprehensive analysis of differentially expressed miRNAs in mice with kidney injury induced by chronic intermittent hypoxia.

Background: miRNAs have been reported to participate in various diseases. Nevertheless, the expression patterns of miRNA in obstructive sleep apnea (OSA)-induced kidney injury remain poorly characterized. In the current study, miRNA sequencing (miRNA-seq) was conducted to investigate miRNA expression profiles in a chronic intermittent hypoxia (CIH)-induced renal injury mouse model. Methods: The mouse model of chronic intermittent hypoxia was established. Differentially expressed miRNAs (DEmiRs) were detected using miRNA-seq technology. The sequencing data were subjected to Gene Ontology (GO) functional enrichment and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses using a bioinformatics approach. RT-qPCR was further used to evaluate the sequencing results. Finally, we created a network for clarifying the relationship between the miRNAs and target genes. Results: In total, nine miRNAs were identified to be upregulated and nine to be downregulated in a mouse model of renal injury induced by chronic intermittent hypoxia. The Kyoto Encyclopedia of Genes and Genomes analyses revealed that the Wnt signaling pathway was involved in the development of chronic intermittent hypoxia-induced renal injury. Subsequently, eight DEmiRs, namely, mmu-miR-486b-3p, mmu-miR-215-5p, mmu-miR-212-3p, mmu-miR-344-3p, mmu-miR-181b-1-3p, mmu-miR-467a-3p, mmu-miR-467 d-3p, and mmu-miR-96-5p, showed a similar trend of expression when verified using RT-qPCR. Finally, five selected DEmiRs were used to construct a miRNA-mRNA network. Conclusion: In conclusion, a total of 18 DEmiRs were identified in the mouse model of chronic intermittent hypoxia-induced renal injury. These findings advance our understanding of the molecular regulatory mechanisms underlying the pathophysiology of obstructive sleep apnea-associated chronic kidney disease.

Effects of post-fermentation on the flavor compounds formation in red sour soup.

Red Sour Soup (RSS) is a traditional fermented food in China. After two rounds of fermentation, sour soup has a mellow flavor. However, the microbial composition and flavor formation processes in post-fermentation in RSS are unclear. This study investigates the bacteria composition of RSS during the post-fermentation stage (0-180 days) using high-throughput sequencing. The results show that lactic acid bacteria (LAB) are dominant during the post-fermentation process, and their abundance gradually increases with fermentation time. Additionally, gas chromatography-mass spectrometry was used to detect volatile flavor compounds in the post-fermentation process. Seventy-seven volatile flavor compounds were identified, including 24 esters, 14 terpenes, 9 aromatic hydrocarbons, 9 alkanes, 6 heterocyclic compounds, 3 alcohols, 3 acids, 3 ketones, 2 phenols, 2 aldehydes, 1 amine, and 1 other. Esters and aromatic hydrocarbons are the main volatile compounds in RSS during the post-fermentation process. Orthogonal partial least squares screening and correlation analysis derived several significant correlations, including 48 pairs of positive correlations and 19 pairs of negative correlations. Among them, Acetobacter spp., Clostridium spp. and Sporolactobacillus spp. have 15, 14, 20 significant correlation pairs, respectively, and are considered the most important bacterial genera post-fermentation. Volatile substances become abundant with increasing fermentation time. LAB are excessive after more than 120 days but cause a drastic reduction in volatile ester levels. Thus, the post-fermentation time should be restricted to 120 days, which retains the highest concentrations of volatile esters in RSS. Overall, these findings provide a theoretical basis to determine an optimal post-fermentation time duration, and identify essential bacteria for manufacturing high-quality starter material to shorten the RSS post-fermentation processing time.

Effect of Ginkgo Biloba Powder on the Physicochemical Properties and Quality Characteristics of Wheat Dough and Fresh Wet Noodles.

Effects of ginkgo biloba powder (GBP) on the chemical, physicochemical properties and quality of dough and fresh wet noodles were investigated. Lower contents of gluten and starch, and higher contents of fibre, amylose and flavonoids in GBP than wheat flour, were detected. Water absorption of dough increased and the development time and stability time of dough were decreased with GBP addition. Meanwhile, the pasting properties results showed that the addition of GBP reduced the aging degree of starch and improved the thermal stability of dough. Scanning electron microscopy results showed that addition of GBP smoothed the surface of raw noodles while increasing the hole size of the cooked noodles. With increased GBP addition (0~40%), the chewiness and extensibility of the fresh wet noodles increased significantly (p < 0.05), and the sensory scores changed, ascending from 0~20% substitution, and then descending from 20~40% substitution. The digestibility and estimated glycemic index (eGI) values of the GBP fresh wet noodles decreased significantly (p < 0.05). In general, 20% GBP addition could improve the chewiness, extensibility, taste and nutrition of fresh wet noodles, and decrease the digestibility and eGI values of noodles. Thus, GBP has potential for application in the noodle industry.

The Correlation Mechanism between Dominant Bacteria and Primary Metabolites during Fermentation of Red Sour Soup.

Chinese red sour soup is a traditional fermented product famous in the southwestern part of China owing to its distinguished sour and spicy flavor. In the present study, the effect of inoculation of lactic acid bacteria (LAB) on the microbial communities and metabolite contents of the Chinese red sour soup was investigated. Traditional red sour soup was made with tomato and red chilli pepper and a live count (108 CFU/mL) of five bacterial strains (including Clostridium intestinalis: Lacticaseibacillus rhamnosus: Lactiplantibacillus plantarum: Lacticaseibacillus casei: Lactobacillus paracei) was added and fermented for 30 days in an incubator at 37 °C. Three replicates were randomly taken at 0 d, 5 d, 10 d, 15 d, 20 d, 25 d and 30 d of fermentation, with a total of 21 sour soup samples. Metabolomic analysis and 16S-rDNA amplicon sequencing of soup samples were performed to determine microbial diversity and metabolite contents. Results revealed that fermentation resulted in the depletion of native bacterial strains as LAB dominated over other microbes, resulting in differences in the relative abundance of bacteria, and types or contents of metabolites. A decrease (p < 0.01) in Shannon and Simpson indices was observed at different fermentation times. The metabolomic analyses revealed a significant increase in the relative content of 10 metabolites (particularly lactic acid, thymine, and ascorbic acid) in fermented samples as compared to the control. The correlation network revealed a positive association of Lacticaseibacillus rhamnosus with differentially enriched metabolites including lactic acid, ascorbic acid, and chlorogenic acid, which can desirably contribute to the flavor and quality of the red sour soup.

Effect of Weekly Long-Acting Growth Hormone Replacement Therapy Compared to Daily Growth Hormone on Children With Short Stature: A Meta-Analysis.

Background: We performed a meta-analysis to evaluate the efficacy and safety of weekly long-acting growth hormone replacement therapy compared to daily growth hormone in children with short stature. Methods: A systematic literature search up to April 2021 was performed and 11 studies included 1,232 children with short stature treated with growth hormone replacement therapy at the start of the study; 737 of them were using weekly long-acting growth hormone replacement therapy and 495 were using daily growth hormone. They were reporting relationships between the efficacy and safety of long-acting growth hormone replacement therapy and daily growth hormone in children with short stature. We calculated the odds ratio (OR), and mean difference (MD) with 95% confidence intervals (CIs) to assess the efficacy and safety of weekly long-acting growth hormone replacement therapy compared to daily growth hormone in children with short stature using the dichotomous or continuous method with a random or fixed-effect model. Results: Long-acting growth hormone replacement therapy had significantly lower height standard deviation scores chronological age (MD, -0.10; 95% CI, -0.13 to -0.08, p <0.001), and insulin-like growth factor binding protein-3 (MD, -0.69; 95% CI, -1.09 to -0.30, p <0.001) compared to daily growth hormone in children with short stature.However, growth hormone replacement therapy had no significantly difference in height velocity (MD, -0.09; 95% CI, -0.69-0.5, p = 0.76), height standard deviation scores bone age (MD, -0.04; 95% CI, -0.10-0.02, p = 0.16), insulin-like growth factor 1 standard deviation scores (MD, 0.26; 95% CI, -0.26-0.79, p = 0.33), and incidence of adverse events (OR, 1.16; 95% CI, 0.90-1.50, p = 0.25) compared to daily growth hormone in children with short stature. Conclusions: Long-acting growth hormone replacement therapy had significantly lower height standard deviation scores chronological age, and insulin-like growth factor binding protein-3 compared to daily growth hormone in children with short stature. However, growth hormone replacement therapy had no significant difference in height velocity, height standard deviation scores bone age, insulin-like growth factor 1 standard deviation scores, and incidence of adverse events compared to daily growth hormone in children with short stature. Further studies are required to validate these findings.

High T3 Induces ß-Cell Insulin Resistance via Endoplasmic Reticulum Stress.

Hyperthyroidism can cause glucose metabolism disorders and insulin resistance. Insulin resistance in muscle and adipose tissues has been extensively studied, whereas investigations on ß-cell insulin resistance are limited. This study preliminarily explored the effects of high T3 levels on ß-cell line (MIN6) insulin resistance, as well as the roles of endoplasmic reticulum stress (ERS). In this study, we treated ß-cell line with T3, with or without an inhibitor of phosphotyrosine phosphatases (PTPs, sodium vanadate) or ERS inhibitor (4-PBA). The results indicated that high levels of T3 significantly inhibited insulin secretion in ß-cell line. In addition, we observed an upregulation of p-IRS-1ser307 and downregulation of Akt. These results can be corrected by sodium vanadate. Moreover, high T3 levels upregulate the ERS-related proteins PERK, IRE1, ATF6, and GRP78, as well as ERS-related apoptosis CHOP and caspase-12. Similarly, this change can be corrected by 4-PBA. These results suggest that high T3 levels can induce insulin resistance in ß-cell line by activating ERS and the apoptotic pathway.

Effects of Concentration and Ionization Degree of Anchoring Cationic Polymers on the Lateral Heterogeneity of Anionic Lipid Monolayers.

We employed coarse-grained Monte Carlo simulations to investigate a system composed of cationic polymers and a phosphatidyl-choline membrane monolayer, doped with univalent anionic phosphatidylserine (PS) and tetravalent anionic phosphatidylinositol 4,5-bisphosphate (PIP2) lipid molecules. For this system, we consider the conditions under which multiple cationic polymers can anchor onto the monolayer and explore how the concentration and ionization degree of the polymers affect the lateral rearrangement and fluidity of the negatively charged lipids. Our work shows that the anchoring cationic polymers predominantly bind the tetravalent anionic PIP2 lipids and drag the PIP2 clusters to migrate on the monolayer. The polymer/PIP2 binding is found to be drastically enhanced by increasing the polymer ionization fraction, which causes the PIP2 lipids to form into larger clusters and reduces the mobility of the polymer/PIP2 complexes. As expected, stronger competition effects between anchoring polymers occur at higher polymer concentrations, for which each anchoring polymer partially dissociates from the monolayer and hence sequesters a smaller PIP2 cluster. The desorbed segments of the anchored polymers exhibit a faster mobility on the membrane, whereas the PIP2 clusters are closely restrained by the limited adhering cationic segments of anchoring polymers. We further demonstrate that the PIP2 molecules display a hierarchical mobility in the PIP2 clusters, which is regulated by the synergistic effect between the cationic segments of the polymers. The PS lipids sequester in the vicinity of the polymer/PIP2 complexes if the tetravalent PIP2 lipids cannot sufficiently neutralize the cationic polymers. Finally, we illustrate that the increase in the ionic concentration of the solution weakens the lateral clustering and the mobility heterogeneity of the charged lipids. Our work thus provides a better understanding of the fundamental biophysical mechanism of the concentration gradients and the hierarchical mobility of the anionic lipids in the membrane caused by the cationic polymer anchoring on length and time scales that are generally inaccessible by atomistic models. It also offers insight into the development and design of novel biological applications on the basis of the modulation of signaling lipids.

Effects of chain rigidity on the adsorption of a polyelectrolyte chain on mixed lipid monolayer: a Monte Carlo study.

We apply Monte Carlo simulation to explore the adsorption of a positively charged polyelectrolyte on a lipid monolayer membrane, composed of electronically neutral, monovalent anionic and mulvitalent anionic phospholipids. We systematically assess the influence of various factors, including the intrinsic rigidity of the polyelectrolyte chain, the bead charge density of the polyelectrolyte, and the ionic strength of the saline solution, on the interfacial structural properties of the polyelectrolyte/monolayer complex. The enhancement of the polyelectrolyte chain intrinsic rigidity reduces the polyelectrolyte conformational entropy loss and the energy gains in electrostatic interaction, but elevates the segregated anionic lipid demixing entropy loss. This energy-entropy competition results in a nonmonotonic dependence of the polyelectrolyte/monolayer association strength on the degree of chain rigidity. The semiflexible polyelectrolyte, i.e., the one with an intermediate degree of chain rigidity, is shown to associate onto the ternary membane below a higher critical ionic concentration. In this ionic concentration regime, the semiflexible polyelectrolyte binds onto the monolayer more firmly than the pancake-like flexible one and exhibits a stretched conformation. When the chain is very rigid, the polyelectrolyte with bead charge density Zb = +1 exhibits a larger tail and tends to dissociate from the membrane, whereas the one with Zb = +2 can still bind onto the membrane in a bridge-like conformation. Our results imply that chain intrinsic rigidity serves as an efficient molecular factor for tailoring the adsorption/desorption transition and interfacial structure of the polyelectrolyte/monolayer complex.

Intracellular iodinated compounds affect sodium iodide symporter expression through TSH-mediated signaling pathways.

The mechanisms by which thyroid stimulating hormone (TSH) regulates the expression and activity of sodium iodide symporter (NIS) through cAMP-PKA have been partially elucidated by many studies. However, the effects of the TSH-mediated PLC-IP3 signaling pathway on the expression of NIS and how intracellular iodinated compounds interfere with these signaling pathways are poorly understood. In this study, we investigated the effects of the TSH-mediated cAMP-PKA and PLC-IP3 pathways on the expression of NIS in the presence of various intracellular iodinated compounds. The intracellular iodinated compounds were formed by treating cells with different concentrations of iodine with or without methimazole (MMI), an inhibitor of iodine organification, in a pig monolayer thyrocyte in vitro. A high concentration of iodine increased NIS expression at the mRNA and protein levels; however, this phenomenon was not observed in the presence of MMI. Both the cAMP-PKA and PLC-IP3 signaling pathways inhibited the expression of NIS at low iodine concentrations; however, in thyrocytes treated with high concentrations of iodine, the effect of cAMP-PKA on the expression of NIS changed from inhibition to promotion, while the PLC-IP3 pathway continued to inhibit NIS expression. These findings indicate that intracellular iodinated compounds affect NIS expression through the TSH-mediated cAMP-PKA and PLC-IP3 pathways.

Inhibition of thyroid-restricted genes by follicular thyroglobulin involves iodinated degree.

Follicular thyroglobulin (TG) reflects the storage of both iodine and thyroid hormone. This is because it is a macromolecular precursor of thyroid hormone and organic iodinated compound in follicular lumen. Thus, it may have an important feedback role in thyroid function. In this study, monolayer cells were cultured and follicles were reconstituted with primary pig thyroid cells in vitro. Reconstituted follicles were treated with iodine and methimazole (MMI), a drug that blocks iodine organification and reduces the degree of TG iodination in follicular lumen. The high degree of iodinated TG in follicular lumen was observed to inhibit thyroid-restricted gene expression. To confirm this finding, monolayer thyroid cells were treated with a different degree of TG iodination at the same concentration. These iodinated TG were extracted from reconstituted follicles of different groups. In this manner, this study provides firsthand evidence suggesting that follicular TG inhibits the expressions of thyroid-restricted genes NIS, TPO, TG, and TSHr.

Upregulation of thyroid transcription factor-1 and human leukocyte antigen class I in Hashimoto's disease providing a clinical evidence for possible triggering autoimmune reaction.

OBJECTIVE: An increase in the expression of autoantigens and their presenting molecules human leukocyte antigen (HLA) class I has been demonstrated to be responsible for autoimmune diseases. Thyroid transcription factor-1 (TTF-1 or NKX2-1) synchronously upregulates both HLA class I and thyroid-specific autoantigen, which may be involved in the pathological process of autoimmune thyroiditis. In this study, the expressions and potential role of TTF-1 and HLA class I in Hashimoto's disease (HT) were examined. PATIENTS: In this study, 22 resection specimens clinically and histopathologically confirmed to have Hashimoto's disease and 30 normal thyroid specimens from adjacent tissues of thyroid adenoma were used. MEASUREMENT: Western blot, real-time PCR, and immunohistochemistry were performed to assay TTF-1 and HLA class I in the thyrocytes of Hashimoto's disease as well as in the normal thyroid from adjacent tissues of thyroid adenoma. RESULTS: The TTF-1 and HLA class I in Hashimoto's disease were significantly higher than those in the controls. CONCLUSION: Upregulation of TTF-1 and HLA class I in Hashimoto's disease provide a clinical evidence for possible triggering of autoimmune reaction.