Astrocytic urea cycle detoxifies Aβ-derived ammonia while impairing memory in Alzheimers Disease

Alzheimers disease (AD) is one of the foremost neurodegenerative diseases, characterized by beta-amyloid (Aβ) plaques and significant progressive memory loss. In AD, astrocytes are known to take up and clear Aβ plaques. However, how Aβ induces pathogenesis and memory impairment in AD remains elusive. We report that normal astrocytes show non-cyclic urea metabolism, whereas Aβ-treated astrocytes show switched-on urea cycle with upregulated enzymes and accumulated entering-metabolite aspartate, starting-substrate ammonia, end-product urea, and side-product putrescine. Gene-silencing of astrocytic ornithine decarboxylase-1 (ODC1), facilitating ornithine-to-putrescine conversion, boosts urea cycle and eliminates aberrant putrescine and its toxic by-products ammonia, H2O2, and GABA to recover from reactive astrogliosis and memory impairment in AD model. Our findings implicate that astrocytic urea cycle exerts opposing roles of beneficial Aβ detoxification and detrimental memory impairment in AD. We propose ODC1-inhibition as a promising therapeutic strategy for AD to facilitate removal of toxic molecules and prevent memory loss.

The Effect of Dietary Protein Imbalance during Pregnancy on the Growth, Metabolism and Circulatory Metabolome of Neonatal and Weaned Juvenile Porcine Offspring

Protein imbalance during pregnancy affects women in underdeveloped and developing countries and is associated with compromised offspring growth and an increased risk of metabolic diseases in later life. We studied in a porcine model the glucose and urea metabolism, and circulatory hormone and metabolite profile of offspring exposed during gestation, to maternal isoenergetic low–high (LP-HC), high–low (HP-LC) or adequate (AP) protein–carbohydrate ratio diets. At birth, LP-HC were lighter and the plasma acetylcarnitine to free carnitine ratios at 1 day of life was lower compared to AP offspring. Plasma urea concentrations were lower in 1 day old LP-HC offspring than HP-LC. In the juvenile period, increased insulin concentrations were observed in LP-HC and HP-LC offspring compared to AP, as was body weight from HP-LC compared to LP-HC. Plasma triglyceride concentrations were lower in 80 than 1 day old HP-LC offspring, and glucagon concentrations lower in 80 than 1 day old AP and HP-LC offspring. Plasma urea and the ratio of glucagon to insulin were lower in all 80 than 1 day old offspring. Aminoacyl-tRNA, arginine and phenylalanine, tyrosine and tryptophan metabolism, histidine and beta-alanine metabolism differed between 1 and 80 day old AP and HP-LC offspring. Maternal protein imbalance throughout pregnancy did not result in significant consequences in offspring metabolism compared to AP, indicating enormous plasticity by the placenta and developing offspring.

iTRAQ-Based Quantitative Proteomics Analysis of Sprague-Dawley Rats Liver Reveals Perfluorooctanoic Acid-Induced Urea Metabolism Dysfunction

Perfluorooctanoic acid (PFOA) is a typical C8 representative compound of perfluoroalkyl and polyfluoroalkyl substances (PFAS) widely used in industrial and domestic products. It is a persistent organic pollutant found in the environment as well as in the tissues of humans and wildlife. Despite emerging scientific and public interest, the precise mechanisms of PFOA toxicity remain unclear. In this study, male rats were exposed to 1.25, 5, and 20 mg PFOA/kg body weight/day for 14 days. Urine samples were also collected and monitored by raising rats in metabolic cages. In vivo results demonstrate that PFOA exposure induces significant hepatocellular hypertrophy and reduced urea metabolism. iTRAQ-based quantitative proteomics analysis of Sprague-Dawley (SD) rats livers identified 3,327 non-redundant proteins of which 112 proteins were significantly upregulated and 80 proteins were downregulated. Gene ontology analysis revealed proteins are primarily involved in cellular, metabolic and single − organism processes. Among them, eight proteins (ACOX1, ACOX2, ACOX3, ACSL1, EHHADH, GOT2, MTOR and ACAA1) were related to oxidation of fatty acids and two proteins (ASS1 and CPS1) were found to be associated with urea cycle disorder. The downregulation of urea synthesis proteins ASS1 and CPS1 after exposure to PFOA was then confirmed through qPCR and western blot analysis. Together, these data demonstrate that PFOA exposure directly influences urea metabolism and identify CPS1 as a potential regulatory target.

RUMINAL AMMONIA AND BLOOD UREA METABOLISM IN SHEEP FED HAY WITH OR WITHOUT CONCENTRATE SUPPLEMENT

Metabolism of ruminal ammonia and blood urea was investigated in West African Dwarf ewes and wethers, fed a low quality hay with or without concentrate supplementation, using single injection of [15N] ammonium chloride (HN4Cl) or [15N] urea into the rumen and blood respectively. The percentages of 15N administered intraruminal as HN4Cl recovered in the urine, faeces and milk of the ewes were 4.3, 9.0 and 3.1, Also 32.3 and 28.7% of [15Nl urea administered into the blood were recovered in the urine of the wethers. Ruminal ammonia contributed 50.6% or protozoal-N in sheep fed hay and 14.2 78.7 and 35.0% respectively in sheep fed hay and concentrate. Also, 59.0 and 7.9% of ruminal ammonia-N was derived from blood urea of sheep fed hay and hay plus concentrate respectively. The inclusion of concentrate in the diet increased the extent of ruminal bacteria protein synthesis but not that of the protozoa, However, the contributions of ruminal ammonia to blood urea synthesis and of blood urea to ruminal ammonia were Sharply decreased in the presence of the concentrate.

Chinese Yellow Rice Wine Processing with Reduced Ethyl Carbamate Formation by Deleting Transcriptional Regulator Dal80p in Saccharomyces cerevisiae

Ethyl carbamate (EC) is a potential carcinogen that forms spontaneously during Chinese rice wine fermentation. The primary precursor for EC formation is urea, which originates from both external sources and arginine degradation. Urea degradation is suppressed by nitrogen catabolite repression (NCR) in Saccharomyces cerevisiae. The regulation of NCR is mediated by two positive regulators (Gln3p, Gat1p/Nil1p) and two negative regulators (Dal80p/Uga43p, Deh1p/Nil2p/GZF3p). DAL80 revealed higher transcriptional level when yeast cells were cultivated under nitrogen-limited conditions. In this study, when DAL80-deleted yeast cells were compared to wild-type BY4741 cells, less urea was accumulated, and genes involved in urea utilization were up-regulated. Furthermore, Chinese rice wine fermentation was conducted using dal80Δ cells; the concentrations of urea and EC were both reduced when compared to the BY4741 and traditional fermentation starter. The findings of this work indicated Dal80p is involved in EC formation possibly through regulating urea metabolism and may be used as the potential target for EC reduction.

Codium fragile Ameliorates High-Fat Diet-Induced Metabolism by Modulating the Gut Microbiota in Mice

Codium fragile (CF) is a functional seaweed food that has been used for its health effects, including immunostimulatory, anti-inflammatory, anti-obesity and anti-cancer activities, but the effect of CF extracts on obesity via regulation of intestinal microflora is still unknown. This study investigated anti-obesity effects of CF extracts on gut microbiota of diet-induced obese mice. C57BL/6 mice fed a high-fat (HF) diet were given CF extracts intragastrically for 12 weeks. CF extracts significantly decreased animal body weight and the size of adipocytes, while reducing serum levels of cholesterol and glucose. In addition, CF extracts significantly shifted the gut microbiota of mice by increasing the abundance of Bacteroidetes and decreasing the abundance of Verrucomicrobia species, in which the portion of beneficial bacteria (i.e., Ruminococcaceae, Lachnospiraceae and Acetatifactor) were increased. This resulted in shifting predicted intestinal metabolic pathways involved in regulating adipocytes (i.e., mevalonate metabolism), energy harvest (i.e., pyruvate fermentation and glycolysis), appetite (i.e., chorismate biosynthesis) and metabolic disorders (i.e., isoprene biosynthesis, urea metabolism, and peptidoglycan biosynthesis). In conclusion, our study showed that CF extracts ameliorate intestinal metabolism in HF-induced obese mice by modulating the gut microbiota.

Urea Addition Promotes the Metabolism and Utilization of Nitrogen in Cucumber

Nitrogen (N) forms include ammonium [NH4+-N], nitrate [NO3−-N], and urea [CO(NH2)2]. Urea is the most common nitrogen fertilizer in agriculture due to its inexpensive price and high N content. Although the reciprocal influence between NO3−-N and NH4+-N is well known, CO(NH2)2 interactions with these inorganic N forms have been poorly studied. We studied the effects of different nitrogen forms with equal nitrogen on dry matter, yield, enzyme activity, and gene expression levels in cucumber. NO3−-N treatment with equal CO(NH2)2 promoted nitrate reduction, urea utilization, and the GS/GOGAT cycle but reduced the nitrate content. UR-2, NR-2, NR-3, NiR, GOGAT-1-1, and GS-4 were upregulated in response to these changes. NH4+-N treatment with equal CO(NH2)2 promoted nitrogen metabolism and relieved the ammonia toxicity of pure NH4+-N treatment. UR-2, GOGAT-2-2, and GS-4 were upregulated, and GDH-3 was downregulated in response to these changes. Treatment with both NO3−-N with added equal CO(NH2)2 and NH4+-N with added equal CO(NH2)2 enhanced the activities of GOGAT, GS, and UR and the amino acid pathway of urea metabolism; manifested higher glutamate, protein, chlorophyll, and nitrogen contents; and improved dry matter weight. A greater proportion of dry matter was distributed to the fruit, generating significantly higher yields. Therefore, the addition of urea to ammonium or nitrate promoted N metabolism and N utilization in cucumber plants, especially treatments with 50% NO3−-N + 50% CO(NH2)2, as the recommended nitrogen form in this study.