Cloning and characterization of a sucrase from Leuconostoc mesenteroides

A sucrase gene from Leuconostoc mesenteroides was cloned and expressed in Escherichia coli. The cloned enzyme did not show dextransucrase or sucrose phosphorylase activity. HPLC and GC-MS analyses of the sucrase products indicated the presence of fructose and glucose in equimolar amounts. IPTG induction did not increase sucrase activity in E. coli indicating that the cloned gene may be transcribed from its own promoter. To our knowledge, this is the first sucrase cloned from L. mesenteroides that has invertase activity.     

Inactivation of the postural asymmetry factor at the stage of compensation for the postural disorder induced by unilateral removal of the motor area

The peptide nature of the posture asymmetry factor (PAF) produced in the brain after unilateral removal of the motor region of the neocortex was established. The inactivation of PAF activity in the brain toward the end of the third week after the removal is due to PAF inactivation by the endogenous factor absent from the intact brain. The molecular weight of the inactivation factor exceeds 5.0 kD that makes it possible to separate it from the PAF by gel filtration on Sephadex G-25. The correlation was marked between the increasing activity of the PAF inactivation factor and the recovery of the initial symmetric functioning of the spinal centers during three weeks after unilateral damage to the CNS.     

Role of membranes and energy-producing reactions in cellular processing of insulin in primary cultures of rat hepatocytes

Incubation of primary cultures of rat hepatocytes with the local anesthetics, procaine or lidocaine, had little or no effect on insulin uptake or degradation but caused an inhibition of insulin-stimulated glycogenesis. While exposure of cultures to the amines, monodansylcadaverine or CH3NH2, resulted in significant dose-dependent decreases in glycogenesis, only monodansylcadaverine (an inhibitor of receptor clustering) decreased uptake whereas CH3NH2 (a lysosomotropic agent) caused increases in both insulin uptake and degradation. When cells were treated with agents which inhibit glycolysis (NaF, 2-deoxy-D-glucose) or oxidative metabolism (2,4-dinitrophenol, carbonyl cyanide m-chlorophenyl hydrazone, NaN3, antimycin A), pronounced inhibitions of each of the bioactivities studied (syntheses of glycogen, protein, lipid) were observed, but only the glycolytic inhibitors decreased insulin uptake. These results suggest that insulin is internalized by an endocytotic process involving receptor clustering and requiring metabolic energy derived from glycolysis. The post-receptor biosynthetic processes involved in the expression of the biological activities of insulin (syntheses of glycogen, protein, lipid) require energy produced by oxidative metabolism while the degradation of insulin is carried out by nonlysosomal mechanisms which are not energy-requiring.     

Cyclic nucleotide alterations in mixed leukocyte reaction: a preliminary report

Endogenous cyclic adenosine and guanosine monophosphate (cAMP, cGMP) levels were studied in human peripheral blood lymphocytes during mixed leukocyte reactions (MLR). cAMP level was consistently elevated in one-way MLR, with good correlation to 3H-thymidine uptake in these reactions. In contrast, cGMP level was practically unchanged. Irradiation of reacting cell populations resulted in inhibition of cyclic nucleotide phosphodiesterase (PDE) activity. These results suggest that metabolic alterations in cAMP may be associated with immune reactions of cellular recognition.     

Serum Agglutination and Immunoglobulin Levels of Mice Infected with Naegleria fowleri1

Agglutinins were not detected in sera from mice given one, two, or three intranasal (i.n.) inoculations or a single intravenous (i.v.) inoculation of trophozoites of Naegleria fowleri. However, agglutinins were produced following second and third i.v. inoculations. Serum immunoglobulin levels increased in both i.n.- and i.v.-inoculated mice. IgG and IgM increased substantially more for i.v.-inoculated mice. IgA levels increased more consistently for i.n.-inoculated mice.     

A comparison of the respiratory chain in particles from Paracoccus denitrificans and bovine heart mitochondria by EPR spectroscopy

A study is presented on the EPR characteristics of the paramagnetic groups in the respiratory chain present in membrane particles of Paracoccus denitrificans, the respiratory system of which is very similar to that in submitochondrial particles from beef heart. All paramagnetic prosthetic groups of the mitochondrial system are also found in the bacterial plasma membrane. Their properties suggest that the respiratory groups are embedded in very similar protein environments in the two systems.