Differential expression of multiple protein kinase C subspecies in rat central nervous tissue

Protein kinase C from a number of areas of rat central nervous tissue was resolved into three distinct fractions upon hydroxyapatite column chromatography. One of the enzyme fractions, designated type II, could be further distinguished into two subspecies with polyclonal antisera, which were raised against synthetic peptides specific for the predicted amino acid sequences of two alternative cDNA clones encoding this enzyme type. Using a combination of these biochemical and immunological techniques, the relative activity of the multiple subspecies of protein kinase C was assessed for each brain area. A distinct regional pattern of expression was found, which per se may be an important factor in determining the response of different neuronal cell types to extracellular stimuli.     

Opioid antinociception and positive reinforcement are mediated by different types of opioid receptors

Fentanyl (FEN) and diprenorphine's (DIPR) potentials for analgesia and reinforcement were assayed using rats. Analgesia was measured by the classic tail-flick test. The test germane to opioid reinforcement involved measuring pressing rates for direct electrical stimulation of the lateral hypothalamus and ventral tegmental area. FEN, as does morphine and heroin, produced strong analgesia and enhanced pressing rates for brain stimulation. DIPR produced no analgesia and antagonized FEN's analgesia. DIPR, at doses antagonizing FEN's analgesia, enhanced pressing for brain stimulation. DIPR's enhancement of pressing was antagonized by naloxone (100 micrograms/kg). When FEN and DIPR were given concurrently, pressing for brain stimulation was not reduced and was greater than after FEN alone was given. These data support a conclusion that different types of receptors are associated with opioid analgesia and reinforcement.     

Effect of prostaglandin F2 alpha on the secretion of human prolactin

This study examines the role of PGF2a (prostaglandin F2alpha) in increasing the secretion rate of human prolactin. 11 women (mean gestational period, 18 weeks) seeking pregnancy termination were divided into 4 groups: 1) Group 1 consisted of 6 women who received 30 mg initially of PGF2a injected intramuscularly and an additional 15 mg after 24 hours if abortion had not occured; mean induction to termination period was 38 hours; 2) Group 2 comprised of 3 women who received PGF2a (500-1500 ug) via the transcervical route at 1 to 2 hourly interval; average number of injections was 20; mean induction to termination period, 24 hours; 3) Group 3 had 2 women receiving hypertonic saline by intraamniotic injection; mean induction to termination period was 51 hours; 4) Group 4 had 4 women who served as controls; mean observation period, 20 hours. Venous blood samples were heparinized in tubes at intervals of 2 to 3 hours. A homologous radioimmunoassay using highly purified human prolactin (for iodination and standards) plus rabbit antihuman prolactin measured serum prolactin. Spikes of serum prolactin up to 550 ng/ml were observed at irregular intervals in 5 women in Group 1; the spikes were less frequent and of smaller amplitude in Groups 3 and 4. The increase in serum prolactin was dramatic and more sustained in Group 2 patients and peaked towards the end of the prostaglandin infusion. Serum prolactin of Group 2 patients were significantly higher than those of Groups 3 and 4 (p0.01). 5 of 9 women whose pregnancies were terminated by PGF2a lactated. However, there was no significant difference between the mean serum prolactin levels in women who lactated (136 ng/ml) and those who did not (120 ng/ml). Although PGF2a is not a lactogenic hormone, this study shows that PGF2a stimulates the secretion of human prolactin during second trimester pregnancy. The fact that the transcervical route caused a significant increase in serum prolactin and the intraamniotic route did not is attributed to the increased systemic absorption of PGF2a following transcervical administration. No correlation was seen between the presence or absence of lactation and the serum prolactin level following pregnancy termination with PGF2a.     

The Lactococcus lactis sex-factor aggregation gene cluA

A gene, cluA, was cloned from the chromosomally located sex factor of Lactococcus lactis MG1363. Sequence analysis revealed significant homology with previously described aggregation proteins in Enterococcus and Streptococcus species. The possibility that cluA was an equivalent protein involved in cell aggregation between donor and recipient bacteria during lactococcal conjugation was confirmed by its expression under the control of a heterologous promoter in L. lactis. Analysis of the homology between the CluA protein and the related proteins of Enterococcus and Streptococcus allowed a common structure for these proteins to be postulated. This consisted of five domains. Functionally conserved domains I and V act respectively as a secretary leader and C-terminal membrane anchor. Domains II and IV are conserved at the amino acid level and probably have common structural roles whereas domain III is variable and may control binding specificity.     

Phylogenetic diversification of immunoglobulin genes and the antibody repertoire

Immunoglobulins are encoded by a large multigene system that undergoes somatic rearrangement and additional genetic change during the development of immunoglobulin-producing cells. Inducible antibody and antibody-like responses are found in all vertebrates. However, immunoglobulin possessing disulfide-bonded heavy and light chains and domain-type organization has been described only in representatives of the jawed vertebrates. High degrees of nucleotide and predicted amino acid sequence identity are evident when the segmental elements that constitute the immunoglobulin gene loci in phylogenetically divergent vertebrates are compared. However, the organization of gene loci and the manner in which the independent elements recombine (and diversify) vary markedly among different taxa. One striking pattern of gene organization is the "cluster type" that appears to be restricted to the chondrichthyes (cartilaginous fishes) and limits segmental rearrangement to closely linked elements. This type of gene organization is associated with both heavy- and light-chain gene loci. In some cases, the clusters are "joined" or "partially joined" in the germ line, in effect predetermining or partially predetermining, respectively, the encoded specificities (the assumption being that these are expressed) of the individual loci. By relating the sequences of transcribed gene products to their respective germ-line genes, it is evident that, in some cases, joined-type genes are expressed. This raises a question about the existence and/or nature of allelic exclusion in these species. The extensive variation in gene organization found throughout the vertebrate species may relate directly to the role of intersegmental (V<==>D<==>J) distances in the commitment of the individual antibody-producing cell to a particular genetic specificity. Thus, the evolution of this locus, perhaps more so than that of others, may reflect the interrelationships between genetic organization and function.      

Ferric Leghemoglobin in Plant-Attached Leguminous Nodules

Leghemoglobin (Lb) is essential for nitrogen fixation by intact leguminous nodules. To determine whether ferric Lb (Lb3+) was detectable in nodules under normal or stressed conditions, we monitored the status of Lb in intact nodules attached to sweet clover (Melilotus officinalis) and soybean (Glycine max [L.] Merr.) roots exposed to various conditions. The effects of N2 and O2 streams and elevated nicotinate levels on root-attached nodules were tested to determine whether the spectrophotometric technique was showing the predicted responses of Lb. The soybean and sweet clover nodules' Lb spectra indicated predominantly ferrous Lb and LbO2 in young (34 d) plants. As the nodule aged beyond 45 d, it was possible to induce Lb3+ with a 100% O2 stream (15 min). At 65 d without inducement, the nodule Lb status indicated the presence of some Lb3+ along with ferrous Lb and oxyferrous Lb. Nicotinate and fluoride were used as ligands to identify Lb3+. Computer-calculated difference spectra were used to demonstrate the changes in Lb spectra under different conditions. Some conditions that increased absorbance in the 626 nm region (indicating Lb3+ accumulation) were root-fed ascorbate and dehydroascorbate, plant exposure to darkness, and nodule water immersion.