HIF-1α-regulated lncRNA-TUG1 promotes mitochondrial dysfunction and pyroptosis by directly binding to FUS in myocardial infarction.

Myocardial infarction (MI) is a fatal heart disease that affects millions of lives worldwide each year. This study investigated the roles of HIF-1α/lncRNA-TUG1 in mitochondrial dysfunction and pyroptosis in MI. CCK-8, DHE, lactate dehydrogenase (LDH) assays, and JC-1 staining were performed to measure proliferation, reactive oxygen species (ROS), LDH leakage, and mitochondrial damage in hypoxia/reoxygenation (H/R)-treated cardiomyocytes. Enzyme-linked immunoassay (ELISA) and flow cytometry were used to detect LDH, creatine kinase (CK), and its isoenzyme (CK-MB) levels and caspase-1 activity. Chromatin immunoprecipitation (ChIP), luciferase assay, and RNA-immunoprecipitation (RIP) were used to assess the interaction between HIF-1α, TUG1, and FUS. Quantitative real-time polymerase chain reaction (qRT-PCR), Western blotting, and immunohistochemistry were used to measure HIF-1α, TUG1 and pyroptosis-related molecules. Hematoxylin and eosin (HE), 2,3,5-triphenyltetrazolium chloride (TTC), and terminal deoxynucleotidyl transferase dUTP risk end labelling (TUNEL) staining were employed to examine the morphology, infarction area, and myocardial injury in the MI mouse model. Mitochondrial dysfunction and pyroptosis were induced in H/R-treated cardiomyocytes, accompanied by an increase in the expression of HIF-α and TUG1. HIF-1α promoted TUG1 expression by directly binding to the TUG1 promoter. TUG1 silencing inhibited H/R-induced ROS production, mitochondrial injury and the expression of the pyroptosis-related proteins NLRP3, caspase-1 and GSDMD. Additionally, H/R elevated FUS levels in cardiomyocytes, which were directly inhibited by TUG1 silencing. Fused in sarcoma (FUS) overexpression reversed the effect of TUG1 silencing on mitochondrial damage and caspase-1 activation. However, the ROS inhibitor N-acetylcysteine (NAC) promoted the protective effect of TUG1 knockdown on H/R-induced cardiomyocyte damage. The in vivo MI model showed increased infarction, myocardial injury, ROS levels and pyroptosis, which were inhibited by TUG1 silencing. HIF-1α targeting upregulated TUG1 promotes mitochondrial damage and cardiomyocyte pyroptosis by combining with FUS, thereby promoting the occurrence of MI. HIF-1α/TUG1/FUS may serve as a potential treatment target for MI.

In Vitro Evaluation of Enriched Brewers' Spent Grains Using Bacillus subtilis WX-17 as Potential Functional Food Ingredients.

Brewers' spent grains (BSGs) are nutritious food processing by-products generated in the brewing industry. In this study, in vitro digestion-fermentation was employed to examine fermented BSG using Bacillus subtilis WX-17 as functional food ingredients. Insoluble fibers in BSG were converted into soluble fibers after fermentation, giving an increase from 6.13 ± 0.42 to 9.37 ± 0.53 mg/100 g BSG. After in vitro digestion of unfermented and fermented BSG, various nutritional components were found to be higher in fermented BSG. Components such as amino acids and fatty acids gave a concentration of 1.635 ± 0.236 mg/mL and 6.35 ± 0.65 mg/mL, respectively. Additionally, vitamin K2 MK7 was detected in fermented BSG with a concentration of 0.00012 ± 0.000005 mg/mL. Probiotics Bacillus subtilis WX-17 was observed to withstand the in vitro digestion. After in vitro fermentation, various short-chain fatty acids namely acetic acid, propanoic acid, and butyric acid were produced at higher amounts for fermented BSG. The concentrations obtained were 124.11 ± 18.72 mM, 13.18 ± 1.38 mM, and 46.25 ± 7.57 mM respectively. As for gut microbiota profile, differential genera such as Bacteroides and Ruminococcus were detected, showing different effects on the intestinal microbiota. This study demonstrates the potential of using microbial fermentation of underutilized BSG to serve as potential functional food ingredients.

Technology innovations for food security in Singapore: A case study of future food systems for an increasingly natural resource-scarce world.

BACKGROUND: Food security is becoming an increasingly important global issue. Anthropogenic factors such as rapid urbanization and industrialization have strained finite resources like land and water. Therefore, against the impending threat of food security, the world can no longer rely on traditional methods to meet its needs. Instead, more creative and technologically advanced methods must be adopted to maximise diminishing natural resources. Singapore is a good case study of a small city-state that is trying to increase its own self-production of food using technology. SCOPE AND APPROACH: This review highlights the technologies that Singapore have adopted in enhancing food security given its limitation in natural resources. These methodologies serve as a case study that can be used as a reference point in light of the increasingly finite natural resources. The review also presents the advantages of these techniques as well as challenges that need to be overcome for them to be more widely adopted. KEY FINDINGS AND CONCLUSION: To increase self-production of food and enhance its food security, Singapore has employed the use of technologies such as vertical farming and aquaponics in urban farming, nutrient recovery from food waste, biodegradable food packaging from durian rinds, natural preservatives, insect farming, microalgae and cultivated meat as alternative protein sources. These technologies workaround Singapore's land and natural resource constraints, which many countries around the world can adapt. However, many of them are still relatively nascent with numerous challenges, which have to be addressed before they can be widely accepted and implemented.

Effects of submerged liquid fermentation of Bacillus subtilis WX-17 using okara as sole nutrient source on the composition of a potential probiotic beverage.

This work aims to produce a functional probiotic beverage using okara as the sole nutrient source. Hence, okara was fermented with Bacillus subtilis WX-17 in submerged liquid fermentation and the supernatant was tested. Metabolomic analysis showed that the nutritional profile of the beverage was enhanced after fermentation. Essential amino acids as well as short-chain fatty acids were significantly (p < .05) upregulated. Total phenolic content and antioxidant content (in terms of DPPH radical scavenging activity) increased by 6.32 and 1.55 times, respectively. After 6 weeks, probiotic viability remains unchanged when stored at 4°C and the cell count is above the minimum dosage to confer health benefits. Antimicrobial activity was also detected against gram-positive bacteria. The findings of this work showed the potential of submerged liquid fermentation of Bacillus subtilis WX-17 using okara as sole substrate to produce a functional and low-cost probiotic beverage.

Potential novel nutritional beverage using submerged fermentation with Bacillus subtilis WX-17 on brewers' spent grains.

Food processing generates side streams that are not fully utilized and typically treated as waste materials. One of such food by-product, brewers' spent grains (BSG) are disposed in huge quantities from the beer industry annually. Submerged fermentation of BSG using Bacillus subtilis WX-17, without supplementary components, is herein employed. The fermentation products were extracted in the liquid phase, resulting in a potential novel nutritional beverage containing Bacillus subtilis WX-17. Bacillus subtilis WX-17, was still viable after a period of 6 weeks with a final cell count of 9.86 log CFU/mL. Gas chromatography-mass spectrophotometry (GC-MS) was employed for identification of the metabolites produced from the growth of Bacillus subtilis WX-17. Seven essential amino acids and citric acid cycle (TCA) intermediates were found to have increased significantly (p < 0.05) whereas all carbohydrates decreased significantly (p < 0.05) in the beverage after submerged fermentation. Additionally, antioxidant activity quantified using DPPH radical scavenging activity, increased by 2.08-fold while total phenolic content increased from 125.7 ± 0.74 µg/mL to 446.74 ± 1.26 µg/mL. The results proved the potential of employing submerged fermentation of BSG using Bacillus subtilis WX-17 to produce a novel and highly nutritious beverage.

A metabolomic approach to understand the solid-state fermentation of okara using Bacillus subtilis WX-17 for enhanced nutritional profile.

Okara is a major agro-waste produced from the soybean industry. To hydrolyze the okara and enable nutrient release, a strategy to valorize okara using solid-state fermentation with food grade Bacillus subtilis (B. subtilis) WX-17 was carried out. The study showed that fermentation of okara with B. subtilis WX-17 improved its overall nutritional content. The total amino acids content increased from 3.04 ± 0.14 mg/g in unfermented okara to 5.41 ± 1.21 mg/g in okara fermented with B. subtilis WX-17. Total fatty acids content increased from 153.04 ± 5.10 to 166.78 ± 2.41 mg/g okara, after fermentation. Antioxidant content (DPPH) also increased by 6.4 times after fermentation. To gain insight into the mechanism, gas chromatography-mass spectrometry analysis was carried out. In total, 49 metabolites were detected, which could be classified mainly into carbohydrates, TCA cycle metabolites, amino acids and fatty acids. The decrease in carbohydrate metabolites, showed that glycolysis was upregulated. This would have provided the energy and metabolic flux towards the amino acid and fatty acid pathway. This is also in line with the increased amino acids and fatty acid production seen in okara fermented with B. subtilis WX-17. The findings of this work demonstrated the potential of using B. subtilis WX-17 fermentation, to enhance the nutritional profile of okara. This could serve as a potential low-cost animal feed or incorporated into the human diet.

[Effects of nerve growth factor pretreatment on apoptosis of neurons and expression of Bcl-2, Bax protein in brain tissue following cerebral ischemia/reperfusion injury in gerbils].

OBJECTIVE: To study the effect of nerve growth factor (NGF) pretreatment on apoptosis of neurons and the expression of Bcl-2 and Bax protein in cerebral cortex and hippocampus CA1 zone following global cerebral ischemia/reperfusion (I/R) injury in gerbils and to explore the mechanism of protection and the best time window of NGF pretreatment. METHODS: Global cerebral I/R injury model was induced by occlusion of bilateral carotid arteries. NGF was injected into the lateral ventricle. Thirty gerbils were randomly divided into five groups, with six animals in each: sham operation group (A group), I/R injury group (B group), NGF pretreatment 12, 24 and 48 hours groups (C, D and E group). Gerbils in all groups were sacrificed after being subjected to 20 minutes of cerebral ischemia followed by 72 hours reperfusion, except A group. Neural apoptosis was identified by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and immunohistochemistry was used to detect the expression of Bcl-2 and Bax protein in cerebral cortex and hippocampus CA1 zone. RESULTS: Compared with B group, the number of apoptotic neurons and the expression of Bax positive cells in NGF pretreatment groups were decreased significantly (all P<0.05), while the expression of Bcl-2 positive cells was increased significantly (all P<0.05). The apoptotic rate in cerebral cortex and hippocampus CA1 zone and expression rate of Bax protein positive cells were the lowest, but the expression rate of Bcl-2 protein positive cells was the highest at 48 hours. CONCLUSION: NGF pretreatment can significantly decrease the neuronal apoptosis of the cerebral I/R injury in gerbils, and the best time window of NGF pretreatment is 48 hours. The mechanism of protection may be related to induction of Bcl-2 protein expression and inhibition of Bax protein expression by NGF pretreatment, thereby preventing neuronal apoptosis.

[Significance of detecting HBV-DNA by the fluorescence quantitative PCR].

BACKGROUND: To study the correlativity between HBV-DNA and the markers of hepatitis B virus infection and different clinical types of hepatitis B. METHODS: The fluorescence quantitation (FQ) of HBV-DNA of 105 patients with hepatitis B was performed by PCR, and the correlativity between the fluorescence quantitation of HBV-DNA and the markers of hepatitis B virus and different clinical types of hepatitis B was analyzed. RESULTS: Ninety-seven percent of the patients were found HBsAg(+), HBeAg(+), HBcAb(+); 75% were HBsAg(+), HBeAb(+), HBcAb(+); 60% were HBsAg(+), HBcAb(+); 40% were HBsAg(+); in HBsAb(+), HBeAb(+), HBcAb(+) (or both HBsAb and HBcAb were positive) group the HBV DNA was undetectable. The analysis indicated that there was a significant difference among different groups (P less than 0.05).HBV-DNA was detected in 72.2% in acute hepatitis B group, in 75% of chronic hepatitis B group, and in 70% of cases of liver cirrhosis with hepatitis B group. The analysis indicated that there was no significant difference among the different clinical types of hepatitis (P greater than 0.05). CONCLUSION: The levels of viral replication were not correlated with different clinical types of hepatitis B; the concentration of HBV-DNA in serum was related to hepatitis B antigen.