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Thus, while we conclusively identify the existence of an oxygen-poor, carbon-rich watermass in the Pacific Ocean, future work to quantitatively determine the oxygen content (as previously done for the Atlantic56,57,58) is essential for determining the precise timing and spatio-temporal extent of respiratory carbon storage.
After 14 days of differentiation, cells were fixed with 4% formaldehyde and stained overnight with Oil Red O (Sigma-Aldrich) to visualize neutral lipid content as previously described.35 Cellular triacylglycerides (TGs) were quantified using a TG Assay Kit (Zenbio).
Sugar and moisture content, as previously reported for Lazio honeys, are strictly correlated [38].
Sera from mice immunized with irradiated sporozoites were tested by immunofluorescence using air-dried methanol-fixed or unfixed ("wet") sporozoites from the different Plasmodium lines, in order to detect total or surface antigen content as previously described [24].
Liver and spleen tissue specimens were analyzed for non-heme iron content as previously reported.
Aliquots of supernatants were used for measurement of total protein content as previously described [ 18].
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The amounts of glucose and fructose before and after acid hydrolysis were measured, using the hexokinase phosphoglucose isomerase glucose-6-phosphate dehydrogenase system, by calculating the sucrose and fructan contents, as previously described [ 13].
In our study, P-deficient leaves had decreased sucrose (Fig. 5C and 5G) and starch (Fig. 5D and 5H) contents, as previously found for trifoliate orange (Poncirus trifoliata (L).
Human serum HDL were isolated, and their protein content quantified, as previously described [ 23].
Edema in the hypothalamus was determined by measuring brain water content (BWC) as previously described [ 30].
Images obtained by light microscopy were digitized and analyzed using ImageJ v1.46 software to quantify the percentage of collagen content (fibrosis), as previously described [ 20].
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