A simple and rapid preconcentration technique, based on collecting trace nitrite on a membrane filter and dissolving the membrane filter in an organic solvent, has been applied to its spectrophotometric determination in water. At pH 2.0, nitrous acid diazotizes with p-aminoacetophenone. which is then coupled with N-(1-naphthyl)ethylenediamine, at the same pH. The azo dye formed is collected on a 0.45 urn nitrocellulose filter at pH 4.7 as its ion associate with dodecyl sulfate. The ion associate and filter are dissolved in a small volume of 2-methoxyethanol (methylcellosolve), and acidized with 0.05 ml of 2 M hydrochloric acid and the absorbance of the resulting solution is measured at 555 nm against a reagent blank. Detection limits better than O.1 mug/dm(-3) as NO(2)(-) can be achieved. The ions normally present in water do not interfere when sodium metaphosphate is added as a masking agent. The proposed method has been applied to the analysis of water samples from several sources, the recoveries of the nitrite added to the samples are quantitative, and results found are satisfactory.
Stroke is a leading cause of death, with a continuously increasing incidence. As a metabolic process that catabolizes glucose pyruvate and provides adenosine triphosphate (ATP), glycolysis plays a crucial role in different diseases. Phosphoglycerate kinase 1 (PGK1) facilitates energy production with biosynthesis in many diseases, including stroke. However, the exact role of PGK1/glycolysis in stroke remains to be elucidated. A rat model of middle cerebral artery occlusion (MCAO) was used to mimic ischemia/reperfusion injuries. Oxygen glucose deprivation/re-oxygenation (OGD/R) was used to induce injury to highly aggressively proliferating immortalized (HAPI) rat microglial cells. The extracellular acidification rate (ECAR) was determined using an XFe24 Extracellular Flux Analyzer. ATP, lactate dehydrogenase, tumor necrosis factor-alpha, and interleukin-6 levels were measured using commercial kits. Chromatin immunoprecipitation assay was performed to examine the interaction between H3K27ac or p300 and the PGK1 promoter region. PGK1 was either knocked down or overexpressed by lentivirus. Thus, to examine its role in stroke, real-time polymerase chain reaction and immunoblotting were used to measure gene expression. The expression of PGK1 was increased and associated with M1 polarization and glycolysis in MCAO rat models. OGD/R promoted M1 polarization and HAPI microglial cell inflammation by regulating glycolysis. Silencing PGK1 reduced OGD/R-increased M1 polarization, inflammation, and glycolysis. Conversely, the overexpression of PGK1 promoted HAPI microglial cell inflammation by regulating glycolysis. The mechanism showed that histone acetyltransferase p300 promoted PGK1 expression through H3K27 acetylation. Finally, data indicated that silencing PGK1 inhibited microglia M1 polarization, inflammation, and glycolysis in MCAO rat models. PGK1 could promote ischemia/reperfusion injury-induced microglial M1 polarization and inflammation by regulating glycolysis, which might provide a novel direction in developing new therapeutic medications for preventing or treating stroke.
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