A single high-cholesterol, high-fat meal preferentially increases low molecular weight apolipoprotein B concentration in rat plasma.

Rats were fed either rat chow (control), chow + 20% olive oil (olive oil), or chow + 20% olive oil + 2% cholesterol (olive oil/cholesterol) as a single meal to study the short-term effects of fat and the above combination of fat/cholesterol-containing diets on plasma apoB concentration and its influence on the distribution of apoB subspecies. Rats were given their meals and allowed to consume them ad libitum until they were killed, 3 hr or 9 hr afterwards. Three hours after feeding, serum triglyceride concentrations were increased to the same extent in both the olive oil and olive oil/cholesterol-fed rats as compared with concentrations in control rats, but serum apoB concentrations did not differ among the groups. Nine hours after feeding, serum triglyceride concentrations were still equally elevated in both experimental groups; however, in the olive oil/cholesterol-fed rats, total serum apoB as well as total serum cholesterol were increased above both the control and olive oil groups. In addition, the d less than 1.21 g/ml lipoprotein apoBl/apoBh ratio of the olive oil/cholesterol-fed rats was greatly increased at 9 hr, whereas apoBl/apoBh ratio in the d less than 1.21 g/ml fraction of the olive oil group was unchanged, despite the increase in plasma triglyceride concentration. In the olive oil/cholesterol-fed rats at 9 hr, cholesterol, total apoB, apoBl, and apoBh of both VLDL and IDL were greater than in the control or olive oil rats. In d less than 1.21 g/ml lipoproteins, VLDL, and IDL, the increases in apoBl concentrations were of a greater magnitude than the increases in apoBh.(ABSTRACT TRUNCATED AT 250 WORDS)

Distribution of apolipoprotein A-IV between the lipoprotein and the lipoprotein-free fractions of rat plasma: possible role of lecithin:cholesterol acyltransferase.

Plasma samples were incubated under various conditions to study the effect of in vitro incubation on apolipoprotein A-IV distribution between the lipoprotein and lipoprotein-free fractions. When plasma was fractionated immediately after bleeding, apolipoprotein A-IV was present in equal concentrations in the lipoprotein and lipoprotein-free fractions. After a 4-hr, 37 degrees C incubation, greater than 90% of total plasma apolipoprotein A-IV was present in the lipoprotein fraction and the percentage of plasma cholesterol present as cholesteryl ester increased from 58% to 74%. When plasma was incubated for 4 hr at 37 degrees C in the presence of 1.5 mM 5,5-dithiobis(2-nitrobenzoic acid) (DTNB), greater than 90% of total plasma apoA-IV was present in the lipoprotein-free fraction, whereas plasma cholesteryl ester concentration did not change. Incubating heat-inactivated plasma for 4 hr also resulted in the redistribution of apolipoprotein A-IV from the lipoprotein fraction to the lipoprotein-free fraction, concurrent with no change in cholesterol esterification. When heat-inactivated plasma was incubated in the presence of a purified lecithin:cholesterol acyltransferase preparation, cholesterol esterification was restored and apolipoprotein A-IV was redistributed from the lipoprotein-free fraction to the lipoprotein fraction in such a manner that greater than 90% was present in the lipoprotein fraction. No changes in apolipoprotein A-I and apolipoprotein E distributions were found under any of the above conditions. Thus, the in vitro plasma incubations show that apolipoprotein A-IV can move bidirectionally between lipoprotein and lipoprotein-free fractions; the direction of this movement depends on the condition of the incubation.(ABSTRACT TRUNCATED AT 250 WORDS)

Maintenance and genetic stability of vector plasmids pBR322 and pBR325 in Escherichia coli K12 strains grown in a chemostat.

The maintenance and genetic stability of the vector plasmids pBR322 and pBR325 in two genetically different Escherichia coli hosts were studied during chemostat cultivation with glucose and ammonium chloride limitation and at two different dilution rates. The plasmid pBR322 was stably maintained under all growth conditions tested. However pBR325 segregated from both hosts preferentially during glucose limitation and at low dilution rate. In addition to this general segregation process a separate loss of tetracycline resistance was observed. The remaining plasmid conferred resistance to ampicillin and chloramphenicol only, without any remarkable alteration of its molecular weight. Cultivation conditions in the chemostat were found that allowed the stable genetic inheritance of both plasmids in the hosts studied.