Clinical physiology of late (refractory) gram-negative bacteremic shock

Blair, Emil; Cowley, R. Adams; Wise, Arthur J.; Mackay, Albert G.

doi:10.1016/0002-9610(69)90019-1

Cited by 11 publications

(2 citation statements)

References 22 publications

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“…Gram-negative bacillary sepsis continues to be an important clinical entity with a high mortality rate (1)(2)(3). Based primarily on animal studies (1,(3)(4)(5)(6), it appears that many of the physiological consequences of gram-negative bacteremia are causally related to the presence of bacterial lipopolysaccharides, namely, endotoxin.…”

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confidence: 99%

Determination of lipid A and endotoxin in serum by mass spectroscopy

Maitra

Schotz

Yoshikawa

et al. 1978

Proc. Natl. Acad. Sci. U.S.A.

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ABSTRACrA quantitative technique for determining lipid A content of endotoxin added to serum by combined gas chromatography-mass spectrometry is described. This MATERIALS AND METHODS Serum Samples. New Zealand white rabbits weighing approximately 2 kg, and not fed overnight, were used. Blood was obtained by heart puncture under sterile conditions. Blood was also obtained aseptically from human volunteers by venipuncture. Serum was separated by centrifugation and stored in pyrogen-free tubes at -200.Solvents. All solvents were either freshly distilled or were liquid chromatography grade and purchased from Burdick and Jackson Co. Determination of #(OH) Myristic Acid Content of LipidA. Varying amounts of Salmonella minnesota R595-lipid A or Escherichia coli 0127:B8 endotoxin in 5 ml of pyrogen-free 0.85% wt/vol NaCl or in 3-5 ml of rabbit or human serum were hydrolyzed with 5 ml of 8 M HCI for 4.5 hr at 1100. The free fatty acids of the hydrolysate were extracted three times with 10 ml of diethyl ether. The ether extracts were pooled, washed with 5 ml of pyrogen-free water, and dried under nitrogen. The extracted free fatty acids were methylated by the procedure of Schlenk and Gellerman (27), dried under nitrogen, dissolved in 0.5 ml of pentane, and applied to a dry silica gel column 60-200 mesh (0.5 X 3 cm). The column was eluted with 3% ether in pentane to remove saturated and unsaturated methyl esters of fatty acids (28). The remaining polar fatty acids, including hydroxy fatty acid methyl esters, were eluted quantitatively with 20% ether in pentane and dried under nitrogen. After addition of 0.25 ml of trimethylsilyl reagent (MesSi imidazole, Applied Science), the samples were shaken on a Vortex mixer. After 2 min, 1 ml of water was added. The Me3Si ethers of hydroxy fatty acid methyl esters were extracted three times with 1 ml of pentane. The pentane extracts were pooled and dried carefully under a nitrogen stream at 300. The dried Me3Si ethers were dissolved in 0.1 ml of acetonitrile, mixed on a Vortex, and filtered through pasteur pipettes plugged with silanized glass wool.Two to five microliters of the acetonitrile solution were injected into a gas chromatograph-mass spectrometer (model 5992 A, Hewlett-Packard, Palo Alto) equipped with a membrane separator. The relative abundance at 315.4 atomic mass unit was determined by using a selected ion monitoring program in conjunction with a six-ion data acquisition program available from Hewlett-Packard. The glass column (2 mm inner diameter X 1.8 m) was packed with 3% Silar-lOC on 100/120 Abbreviations: #(OH), fl-hydroxy; Me3Si, trimethylsilyl; Me3Si-Mefl(OH)C14, trimethylsilyl ether of ,B(OH) methyl myristate.

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mentioning

confidence: 99%

Determination of lipid A and endotoxin in serum by mass spectroscopy

Maitra

Schotz

Yoshikawa

et al. 1978

Proc. Natl. Acad. Sci. U.S.A.

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“…This includes urgent focus on ensuring end-organ perfusion, definitive treatment of the underlying cause (from anaphylaxis to sepsis), and providing subsequent physiological restoration. Despite the apparent heterogeneity of "shock" as a clinico-pathological entity, there are some common threads that permeate all forms and manifestations of shock, with apparent increase in observed commonalities in the more advanced (and often irreversible) stages of the systemic syndrome [1][2][3][4][5][6][7].…”

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confidence: 99%