Hepatic malonyl-CoA synthesis restrains gluconeogenesis by suppressing fat oxidation, pyruvate carboxylation, and amino acid availability.

Acetyl-CoA carboxylase (ACC) promotes prandial liver metabolism by producing malonyl-CoA, a substrate for de novo lipogenesis and an inhibitor of CPT-1-mediated fat oxidation. We report that inhibition of ACC also produces unexpected secondary effects on metabolism. Liver-specific double ACC1/2 knockout (LDKO) or pharmacologic inhibition of ACC increased anaplerosis, tricarboxylic acid (TCA) cycle intermediates, and gluconeogenesis by activating hepatic CPT-1 and pyruvate carboxylase flux in the fed state. Fasting should have marginalized the role of ACC, but LDKO mice maintained elevated TCA cycle intermediates and preserved glycemia during fasting. These effects were accompanied by a compensatory induction of proteolysis and increased amino acid supply for gluconeogenesis, which was offset by increased protein synthesis during feeding. Such adaptations may be related to Nrf2 activity, which was induced by ACC inhibition and correlated with fasting amino acids. The findings reveal unexpected roles for malonyl-CoA synthesis in liver and provide insight into the broader effects of pharmacologic ACC inhibition.

Glycogen phosphorylase inhibition improves cognitive function of aged mice.

Inhibition of glycogen breakdown blocks memory formation in young animals, but it stimulates the maintenance of the long-term potentiation, a cellular mechanism of memory formation, in hippocampal slices of old animals. Here, we report that a 2-week treatment with glycogen phosphorylase inhibitor BAY U6751 alleviated memory deficits and stimulated neuroplasticity in old mice. Using the 2-Novel Object Recognition and Novel Object Location tests, we discovered that the prolonged intraperitoneal administration of BAY U6751 improved memory formation in old mice. This was accompanied by changes in morphology of dendritic spines in hippocampal neurons, and by "rejuvenation" of hippocampal proteome. In contrast, in young animals, inhibition of glycogen degradation impaired memory formation; however, as in old mice, it did not alter significantly the morphology and density of cortical dendritic spines. Our findings provide evidence that prolonged inhibition of glycogen phosphorolysis improves memory formation of old animals. This could lead to the development of new strategies for treatment of age-related memory deficits.

Response regulator PorX coordinates oligonucleotide signalling and gene expression to control the secretion of virulence factors.

The PglZ family of proteins belongs to the alkaline phosphatase superfamily, which consists of metallohydrolases with limited sequence identity but similar metal-coordination architectures in otherwise divergent active sites. Proteins with a well-defined PglZ domain are ubiquitous among prokaryotes as essential components of BREX phage defence systems and two-component systems (TCSs). Whereas other members of the alkaline phosphatase superfamily are well characterized, the activity, structure and biological function of PglZ family proteins remain unclear. We therefore investigated the structure and function of PorX, an orphan response regulator of the Porphyromonas gingivalis TCS containing a putative PglZ effector domain. The crystal structure of PorX revealed a canonical receiver domain, a helical bundle, and an unprecedented PglZ domain, similar to the general organization of the phylogenetically related BREX-PglZ proteins. The PglZ domain of PorX features an active site cleft suitable for large substrates. An extensive search for substrates revealed that PorX is a phosphodiesterase that acts on cyclic and linear oligonucleotides, including signalling molecules such as cyclic oligoadenylates. These results, combined with mutagenesis, biophysical and enzymatic analysis, suggest that PorX coordinates oligonucleotide signalling pathways and indirectly regulates gene expression to control the secretion of virulence factors.

Validated liquid chromatography-mass spectrometry method for the quantification of glycogenolysis phosphorylase inhibitor in mouse tissues - 5-isopropyl-4-(2-chlorophenyl)-1-ethyl-1,4-dihydro-6-methyl-2,3,5-pyridinetricarboxylic acid ester disodium salt hydrate.

5-Isopropyl-4-(2-chlorophenyl)-1-ethyl-1,4-dihydro-6-methyl-2,3,5-pyridinetricarboxylic acid ester disodium salt hydrate, is a noncompetitive inhibitor of glycogen phosphorylase - a critical enzyme in the process of glycogenolysis. This chemical compound is most widely used in studies focused on the inhibition of liver and muscle glycogenolysis. However, there are also reports linking phosphorylase inhibitor action with cognitive function and glycogen metabolism in the brain. The aim of this study was to develop and validate the liquid chromatography-mass spectrometry method for quantitative analysis of present chemical compound in mouse tissues including different brain regions. Obtained linearity was in the range of 10-550 ng/mL with a correlation coefficient of 0.9996. In tissue matrix samples the limit of detection was 7.76 ng/mL, while the limit of quantification was 23.29 ng/mL. The coefficient of variation values did not exceed ±15% for either within a run or between run precision quality control samples. The extraction recovery was between 89.44% and 98.70% for various validation analyte concentrations. The present method was successful in the quantitative determination of the presented analyte in mouse tissues and provided evidence that the compound is not only present in the liver, heart, and skeletal muscle but also in different regions of brain tissue such as the hippocampus, cerebellum, and cortex.

Hydrogel Alginate Seed Coating as an Innovative Method for Delivering Nutrients at the Early Stages of Plant Growth.

Seed coating containing fertilizer nutrients and plant growth biostimulants is an innovative technique for precision agriculture. Nutrient delivery can also be conducted through multilayer seed coating. For this purpose, sodium alginate with NPK, which was selected in a preliminary selection study, crosslinked with divalent ions (Cu(II), Mn(II), Zn(II)) as a source of fertilizer micronutrients, was used to produce seed coating. The seeds were additionally coated with a solution containing amino acids derived from high-protein material. Amino acids can be obtained by alkaline hydrolysis of mealworm larvae (Gly 71.2 ± 0.6 mM, Glu 55.8 ± 1.3 mM, Pro 48.8 ± 1.5 mM, Ser 31.4 ± 1.5 mM). The formulations were applied in different doses per 100 g of seeds: 35 mL, 70 mL, 105 mL, and 140 mL. SEM-EDX surface analysis showed that 70 mL of formulation/100 g of seeds formed a continuity of coatings but did not result in a uniform distribution of components on the surface. Extraction tests proved simultaneous low leaching of nutrients into water (max. 10%), showing a slow release pattern. There occurred high bioavailability of fertilizer nutrients (even up to 100%). Pot tests on cucumbers (Cornichon de Paris) confirmed the new method's effectiveness, yielding a 50% higher fresh sprout weight and four times greater root length than uncoated seeds. Seed coating with hydrogel has a high potential for commercial application, stimulating the early growth of plants and thus leading to higher crop yields.

Gender-Specific Metabolomics Approach to Kidney Cancer.

Renal cell carcinoma (RCC) is the most common form of kidney malignancy. RCC is more common among men with a 2/1 male/female incidence ratio worldwide. Given the underlying epidemiological differences in the RCC incidence between males and females, we explored the gender specific 1H NMR serum metabolic profiles of RCC patients and their matched controls. A number of differential metabolites were shared by male and female RCC patients. These RCC specific changes included lower lactate, threonine, histidine, and choline levels together with increased levels of pyruvate, N-acetylated glycoproteins, beta-hydroxybutyrate, acetoacetate, and lysine. Additionally, serum lactate/pyruvate ratio was a strong predictor of RCC status regardless of gender. Although only moderate changes in metabolic profiles were observed between control males and females there were substantial gender related differences among RCC patients. Gender specific metabolic features associated with RCC status were identified suggesting that different metabolic panels could be leveraged for a more precise diagnostic.