Alterations in high-affinity binding characteristics and levels of opioids in invertebrate ganglia during aging: Evidence for an opioid compensatory mechanism

In Mytilus and Leucophaea the high-affinity binding site density is significantly lower in old animals than in young animals, whereas the low-affinity site density remains unchanged. In Mytilus the estimated met-enkephalin and met-enkephalin-Arg6-Phe7 levels are significantly higher in old than in young animals. In Leucophaea only the met-enkephalin level can be determined, and it is also higher in old animals. The decrease in the high-affinity binding site density and the corresponding increase in endogenous enkephalin levels suggest the existence of an opioid compensatory mechanism associated with the aging process. In Mytilus there is a demonstrated decrease with age in intraganglionic dopamine levels in response to applied opiates. In addition, the inhibition of dopamine-stimulated adenylate cyclase activity by opiates also decreases in older animals. In Leucophaea the sex difference in opioid binding densities diminishes with age.     

Effects of chronic alcoholism on the amygdaloid complex. A study in human and rats

The effect of chronic alcoholism on the amygdaloid complex was studied in 16 humans and 10 rats. Eighteen patients whose death was due to extraneural causes were selected as controls with 3 rats. The alcoholic cases, in addition to the data collected in their clinical history, showed, microscopically confirmed, liver cirrhosis or steatosis. The alcoholics and controls were divided into 4 groups: 35-44 years old (4 cases), 45-54 (5 cases), 55-64 (5 cases) over 65 (2 alcoholics and 4 controls). The alcoholic ingestion in the rats (Wistar, 10 weeks old) was 3 ml at a concentration of 30% in water solution administered by esophagic intubation, for 48 (5 rats) or 58 weeks (5 rats). To judge the state of the amygdaloid nuclei, a neuronal count and caryometry were carried out. The numerical data obtained in this study were analyzed statistically. The results in humans have paralleled those obtained in rats and the behaviour of the different nuclei of the amygdala was uniform and can be summarized as follows: 1) ethanol provoked a prominent and early loss of neurons, and 2) the remainder of non-affected neurons did not react in order to compensate for this neuronal loss.     

The prophylactic effect of diphenylhidantoin in the kindling effect on reptiles (Gallotia galloti)

1. Lizards Gallotia galloti received daily 3 mg/kg body wt of diphenylhidantoin (DPH) over a period of 15 days and at the same time the animals were kindled. 2. The progression of the kindling effect was evaluated by counting the number of spontaneous epileptiform potentials, the duration of afterdischarges and the duration of electroencephalographic spontaneous seizures. 3. The diphenylhidantoin treated group, relative to controls presented: (a) significant reduction in the duration of afterdischarges and spontaneous electroencephalographic seizures; and (b) increased frequency of the spontaneous epileptiform potentials.     

Phospholipid-Sensitive Ca2+-Dependent Protein Kinase Preferentially Phosphorylates Serine-115 of Bovine Myelin Basic Protein

Phospholipid-sensitive Ca2+ -dependent protein kinase (PL-Ca-PK) and cyclic AMP-dependent protein kinase (A-PK) both preferentially phosphorylated serine residues of bovine myelin basic protein (MBP). Tryptic peptide maps of MBP phosphorylated by PL-Ca-PK or A-PK, however, revealed different phosphopeptides, suggesting a difference in the intramolecular substrate specificity for the two enzymes. Serine-115 of MBP, in the sequence (-Arg-Phe-Ser(115)-Trp-), was found to be a preferred and probably major phosphorylation site for PL-Ca-PK. Because serine-115 of bovine MBP corresponds to serine-113 of rabbit MBP, an in vivo phosphorylation site reported by Martenson et al. (1983), and PL-Ca-PK is present at a very high level in brain and myelin, it is suggested that the enzyme may be responsible for the in vivo phosphorylation of this and other sites in MBP.     

Substrate-dependent degradation patterns in the decay of wheat straw and beech wood by ligninolytic fungi

Summary The ability of 45 fungal strains to degrade wheat straw and beech wood was studied. Degradation patterns were defined in terms of chemical evolution of substrates and changes in lignin and polysaccharides. Trametes versicolor produced an important degradation of lignin and increased substrate digestibility, but it caused high weight losses and gave rise to similar decay patterns on both substrates. A preferential degradation of lignin was produced during straw transformation by Pleurotus eryngii. The increase of soluble lignin and decreases of lignin content and H/C ratio defined the degradation tendency after principal component analysis. The cation exchange capacity and water and alkali solubility presented the highest loading factors for the characterization of fungal transformation of beech wood. Offprint requests to: A. T. Martínez     

Artificial hybridization within Saxifraga pentadactylis (Saxifragaceae)

Saxifraga pentadactylis subsp. almanzorii , an endemic to the subalpine nucleus of Sierra de Gredos (central Spain), differs from its closest relative, subsp. willkommiana , by its less showy petals. An artificial crossing program was carried out in order to assess the degree of reproductive isolation between the subspecies. To facilitate interpretation of the results, the program was extended to 10 other interspecific hybrid combinations within sect. Saxifraga . All the data gathered are congruent with the occurrence of two evolutionary scenarios. Interspecific crossings, rendering moderate to high seed-set (in obtaining the F₁), and vigorous but relatively sterile F₁ offspring, reveal reproductive barriers at the level of the F₁ fertility, probably originated as a byproduct of divergent evolution. In contrast, intraspecific crossings within S. pentadactylis resulted in seed-set values lower than expected (in obtaining the F₁), in a majority of weak non-viable F₁ offspring but also in a few fertile F₁ hybrid specimens which were able to originate F₂ offspring. This second pattern reveals reproductive barriers at the level of the F₁ vitality, probably arisen in a quite abrupt fashion. The lower P/O for subsp. almanzorii as compared to subsp. willkommiana , together with the rest of the evidence suggest that the reproductive barriers between them might be the product of active selection against hybridization achieved by incrementing the levels of autogamy in the former.     

Inhibition of pea ferredoxin-NADP(H) reductase by Zn-ferrocyanide.

Ferredoxin-NADP(H) reductases (FNRs) represent a prototype of enzymes involved in numerous metabolic pathways. We found that pea FNR ferricyanide diaphorase activity was inhibited by Zn2+ (Ki 1.57 microM). Dichlorophenolindophenol diaphorase activity was also inhibited by Zn2+ (Ki 1.80 microM), but the addition of ferrocyanide was required, indicating that the inhibitor is an arrangement of both ions. Escherichia coli FNR was also inhibited by Zn-ferrocyanide, suggesting that inhibition is a consequence of common structural features of these flavoenzymes. The inhibitor behaves in a noncompetitive manner for NADPH and for artificial electron acceptors. Analysis of the oxidation state of the flavin during catalysis in the presence of the inhibitor suggests that the electron-transfer process between NADPH and the flavin is not significantly altered, and that the transfer between the flavin and the second substrate is mainly affected. Zn-ferrocyanide interacts with the reductase, probably increasing the accessibility of the prosthetic group to the solvent. Ferredoxin reduction was also inhibited by Zn-ferrocyanide in a noncompetitive manner, but the observed Ki was about nine times higher than those for the diaphorase reactions. The electron transfer to Anabaena flavodoxin was not affected by Zn-ferrocyanide. Binding of the apoflavodoxin to the reductase was sufficient to overcome the inhibition by Zn-ferrocyanide, suggesting that the interaction of FNRs with their proteinaceous electron partners may induce a conformational change in the reductase that alters or completely prevents the inhibitory effect.     

Lethality and mutagenicity inHalobacterium mediterranei caused by N-methyl-N′-nitro-N-nitrosoguanidine

Compared withEscherichia coli, Halobacterium mediterranei was highly resistant to the lethal effect of N-methyl-N-nitro-N-nitrosoguanidine (nitrosoguanidine), but it was sensitive to the mutagenic action of this chemical agent. Nitrosoguanidine at 500 g ml^–1 gave a cell survival level between 1% and 10%, and this allowed us to obtain more Josamycin-resistant mutants compared with lower concentrations, which gave higher survival rates but fewer mutants. The efficiency of the mutagenicity obtained with the nitrosoguanidine treatment was examined under a variety of conditions. The optimal conditions for obtaining Josamycinresistant mutants were achieved by exposing, in darkness and without shaking, a suspension of about 10⁸ log-phase cells to 500 g nitrosoguanidine in 1 ml of 50 mM modified saline Tris-maleate buffer at pH 7.5, or in 1 ml of 5 mM modified saline Tris-citrate-maleate for 30 min at 37°C.