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The myocardial sialic acid content, ATPase activities, and Ca2+ level were investigated in rats with full thickness burn of 30% TBSA. The results showed that the burned rats had a decreased level of myocardial sialic acid which was only 58.8% of controls at 3 h postburn. Myocardial Na+, K(+)-ATPase, Ca2+, Mg(2+)-ATPase and Ca(2+)-ATPase activities were inhibited in burned rats. As compared to controls, the burned rats showed a higher level of Ca2+ in heart tissue. This study indicated that there were abnormal energy metabolism, dysfunction of ion pump, and paradox Ca2+ overload in burned rats, which may be associated with the decrease in sialic acid content in myocardium.

The inducible nuclear transcription factor complex, AP-1, typically consists of heterodimers between Jun and Fos proteins. Although components are drawn from families of related molecules, little is known about the physiologic regulation of jun- and fos-related gene products. In particular, it is not known whether expression of individual family members is stimulus-specific or whether the same signaling pathways are responsible for induction of all subunits. To clarify these issues, AP-1 components were examined following activation of primary B lymphocytes through two separate receptors, the surface Ig Ag receptor, and the CD40 receptor for T cell influences. Both forms of stimulation led to expression of JunB and JunD mRNA and protein; however, induction of JunB mediated by anti-Ig Ab was protein kinase C (PKC)-dependent, whereas induction mediated by CD40 ligand was resistant to PKC depletion. The two forms of stimulation diverged even further with respect to Fos expression. Although both stimuli induced c-Fos, expression of FosB was stimulus specific at both the mRNA and protein levels, in that this component was induced by anti-Ig but not by CD40 ligand. Stimulated expression of c-Fos and FosB was in all cases PKC-independent. These results provide evidence for receptor-specific differences in the expression of AP-1 components, primarily with respect to FosB. They also indicate that separate intracellular pathways may be used for induction of jun and fos gene products and that the same transcription factor (junB) may be triggered by two surface receptors through separate pathways that differ in PKC dependence.

We demonstrate herein that resting primary B lymphocytes do not contain detectable levels of AP-1 (TRE)-binding activity. Upon cross-linking of surface immunoglobulin (sIg) receptors, TRE-binding activity is induced within 2 hr and its appearance requires de novo protein synthesis. Antisera to Jun-B inhibits the vast majority of TRE-binding activity, indicating that Jun-B is a primary component of B cell TRE-binding complexes. In keeping with this, Jun-B protein is not detectable in cytosol or nuclear extracts from resting B lymphocytes, as determined by immunoblotting with Jun-B antisera. However, the nuclear expression of Jun-B is induced within 2 hr following sIg cross-linking and is completely blocked by cycloheximide. 35S-labeling studies suggest that the increase in Jun-B expression results from de novo protein synthesis. Moreover, Jun-B migrates in SDS-polyacrylamide gels as two distinct electrophoretic proteins that correspond to a 41-kDa species and a phosphorylated 47-kDa form. These results suggest that the induction of AP-1-binding activity in primary B lymphocytes following sIg cross-linking does not result from post-translational phosphorylation of a preexisting cellular pool of Jun-B protein, but rather is coupled to the stimulation of de novo Jun-B synthesis. Thus Jun-B synthesis represents an integral event in the production of receptor-mediated AP-1 in B cells. The significance of these results with respect to the role of Jun-B in controlling gene expression during the activation of primary B cells is discussed.

We report a DNA sequence encoding the nuclear factor NF-kappa B p50 precursor from primary murine B-lymphocytes that differs from that previously published for the murine 22D6 B-cell line. The variation is located in one of the ankyrin-like repeats at the C terminus of the molecule.

DNA looping between the araO2 and araI sites holds the uninduced or basal level of expression of the araBAD genes from the pBAD promoter at a low level. Despite the presence of another and closer site potentially capable of mediating looping to araI, no repression from this site, called araO1, is observed. Here we show, using both in vivo and in vitro experiments, that the araO1 site is not normally occupied by AraC protein under repressing conditions, but that if AraC protein is overproduced and the araO2 site is absent, araO1 is then occupied and repression of pBAD can be observed.

The araCBAD regulatory region of Escherichia coli contains two divergently oriented promoters and three sites to which AraC, the regulatory protein of the operon, can bind. This paper presents the results of in vivo dimethyl sulfate "footprinting" experiments to monitor occupancy of the three AraC sites and measurements of activity of the two promoters. These measurements were made both in the absence of the inducer arabinose and at various times after arabinose addition to growing cells containing the wild-type ara regulatory region or the regulatory region containing various deletions and point mutations. The data lead to the conclusion that two different DNA loops can form in the ara regulatory region. These loops are generated by AraC protein molecules binding to two different DNA sites and binding to each other. One of these loops predominates in the absence of arabinose and plays a major role in repressing activity of one of the promoters. Upon the addition of arabinose the amount of the first loop type, the repression loop, decreases and the amount of a second loop increases. Formation of this second loop precludes the counterproductive formation of the repression loop.

Two sets of experiments have been performed to test the DNA loop model of repression of the araBAD operon of Escherichia coli. First, dimethyl sulfate methylation protection measurements on normally growing cells show that the AraC regulatory protein occupies the araI site in the presence and absence of the inducer arabinose. Similarly, the araO2 site is shown to be occupied by AraC protein in the presence and absence of arabinose; however, its occupancy by AraC is greatly reduced when araI and adjacent sequences are deleted. Thus, AraC protein binds to araO2 cooperatively with some other component of the ara system located at least 60 base pairs away. Second, the mutational analysis presented here shows that the DNA components required for repression of araBAD are araI, araO2, and perhaps the araBAD operon RNA polymerase binding site.