Human Brain Phenol Sulfotransferase: Biochemical Properties and Regional Localization

Phenol sulfotransferase (PST) catalyzes the sulfate conjugation of catecholamines and phenol and catechol drugs. The human blood platelet contains a thermolabile (TL) form of PST that catalyzes the sulfate conjugation of dopamine and other monoamines and a thermostable (TS) form that catalyzes the sulfate conjugation of micromolar concentrations of phenol and p-nitrophenol. Experiments were performed to determine whether the brain contains forms of PST analogous to the TL and TS forms found in the human platelet, and to determine whether there are regional variations in human brain PST activity. We found that the human brain contains at least two forms of PST, forms that are similar to the platelet TS and TL forms of the enzyme with respect to substrate specificity, apparent Km constants, thermal stability, and sensitivity to inhibitors. Optimal conditions were determined for the measurement of these two activities in brain homogenates. The stability of PST activities in the human brain after death was determined in five samples of cerebral cortex that were obtained during clinically indicated neurosurgical procedures. An average of 76 +/- 8% and 80 +/- 9% (mean +/- SEM) of the basal TL and TS PST activities, respectively, remained in these five samples of cerebral cortex after 8 h of storage under simulated post-mortem conditions. Six human brains were then obtained less that 8 h after death from patients who had no neurological disease prior to death. The mean activities of the TL and TS forms of PST were measured in 17 different regions of the six brains. If the pituitary was excluded from consideration, TL and TS PST activities both varied approximately fivefold among these regions, and both activities were highest in cerebral cortex. However, the average TS activity in the anterior pituitary, a tissue of non-neural origin embryologically, was 6.5-fold greater than the highest average TS PST activity found in cerebral cortex.     

Temperature Dependence of Catecholamine Secretion from Cultured Bovine Chromaffin Cells

Abstract: Secretion of both epinephrine and norepinephrine by cultured chromaffin cells was studied at temperatures ranging from 0°C to 37°C. The percentage of epinephrine secreted was always lower than that of norepinephrine when the cells were stimulated with either acetylcholine or high K⁺ at any temperature. When the cells were stimulated with acetylcholine or carbachol the percentage of catecholamine secreted at 10 min increased with temperature from 4°C to 24°C and then decreased from 24°C to 37°C. Potassium-stimulated cells secreted increasing amounts of catecholamine as the temperature was increased to 37°C. We found, however, that the initial rates of secretion increased continuously as temperature increased throughout the range for both carbachol-and K⁺-stimulated cells. The temperature maximum of acetylcholine-stimulated secretion is caused by a faster shut-off of secretion at higher temperature. The Arrhenius plots of initial rates show an inflection point at approximately 17°C for carbachol-stimulated cells. The plot for K⁺-stimulated cells is a straight line over the entire temperature range. The transition could be caused by a conformational change in the cholinergic receptor/ion channel molecule.     

Characterization of Cholecystokinin from the Human Brain

Human forms of cholecystokinin have not previously been characterized chemically. In this study, we have extracted and purified the predominant molecular form of cholecystokinin present in human cerebral cortex. The peptide was characterized by amino acid analysis, automated peptide sequencing, and fast atom bombardment mass spectrometry. It appears to be identical to porcine cholecystokinin-octapeptide, with the sequence of Asp-Tyr(SO3)-Met-Gly-Trp-Met-Asp-Phe(NH2). This structural identity is consistent with the observations that the peptide in human brain and porcine cholecystokinin-octapeptide are recognized similarly by a battery of antisera to porcine cholecystokinin; that they coelute from several chromatographic systems, including gel filtration, ion exchange, and reversed-phase; and that they possess similar biological activities.     

The effect of nematode parasitism on the osmolality and major cation concentration in the haemolymph of three larval mosquito species

Parasitism by a mermithid nematode, Romanomermis culicivorax, causes severe depletion of haemolymph carbohydrates and proteins in mosquito larvae. We undertook a study to determine if haemolymph osmolality and cation concentrations were affected also by mermithid parasitism. The haemolymph osmolality of R. culicivorax-infected and control Aedes taeniorhynchus and Culex pipiens fourth-instar larvae was not significantly different. However, the haemolymph osmolality decreased significantly in infected Anopheles quadrimaculatus. Each mosquito species demonstrated significant alterations in the haemolymph concentration of at least one cation when infected although the cation concentrations affected differed for each species. The changes observed were statistically significant but the magnitude of change was not great. Overall, despite the severe nutritional burden of the mermithid nematode, these species of mosquito larvae can continue to maintain osmoregulation.     

Some aspects of social problems facing conservation in Brazil

There has been growing concern in Brazil for environmental issues in the last two decades. The conservation policies for Amazonia, which still represents the largest portion of forests of the country, are still based on isolated decisions made in the late 1970's. Among these policies there is, for instance, the plan for the establishment of a net of National Parks, proposed by Wetterberg et al. (1), based on the 'Pleistocene refugia' model. These refugia are areas of high species endemism, representing forest islands formed during the dry periods of the Pleistocene age, constituting the center of evolution and dispersal of Amazonian species (2). A number of parks and biological reserves have since been established and the decreese of laws protecting some elements of the fauna have been implemented. In 1979, studies for a more comprehensive plan for the conservation and development of Brazilian Amazonia were carried out in several institutions committed to research in Amazonia. As a result, several documents were handed to the government, but nothing has yet been implemented. Indeed, no environmental policy for Amazonia will succeed without an effective and comprehensive social plan, and the latter has yet to be formulated.     

AMP deaminase in Dictycostelium discoideum: Increase in activity following nutrient deprivation induced by starvation or hadacidin

Summary AMP deaminase, the activity that catalyzes the deamination of AMP to form IMP and NH₃ has been measured in Dictyostelium discoideum. A new procedure to assay the activity of this enzyme was developed using formycin 5-monophosphate, a fluorescent analog of AMP as the substrate, and ionpaired reverse phase HPLC to separate the reactants and products. Quantitation of the formycin containing compounds was accomplished at 290 nm. At this wavelength adenosine containing compounds were not detected and activity could be monitored in the presence of its activator ATP. The AMP deaminase activity in vegetative cells was 7.4 nmols/min/mg proteins while the activity in cells measured at 2 and 6 hrs after starvation-induced growth-arrest was 376 nmols/min/mg protein... a 51-fold increase. When vegetative cells were treated with hadacidin, a drug that restricts de novo AMP synthesis and pinocytosis, the activity of the AMP deaminase was 511 nmols/min/mg protein... a 70-fold increase compared to that in untreated vegetative cells. Smaller increases were noted following the inhibition of growth with the drugs cerulenin and vinblastine, as well as after the inhibition of de novo GMP synthesis with the drug mycophenolic acid or the partial inhibition of de novo AMP synthesis with analogs of hadacidin, N-hydroxyglycine and N-formylglycine. In addition, when the activity of two other enzymes involved in purine metabolism, namely adenosine kinase and hypoxanthine-guanine phosphoribosyl transferase, was measured in vegetative cells, and the activity of both compared to that measured in starvation and hadacidin induced growth-arrested cells, showed no significant changes. These data suggest that the changes in the activity of the AMP deaminase are in response to nutrient deprivation and further, that as a consequence of the increase in AMP deaminase activity, ammonia will be produced and an increase in pH should follow. The production of ammonia and its effect on development implicates the AMP deaminase in the early differentiation of this organism.     

Proteolysis in quaking mouse brain and spinal cord

Six proteolytic enzymes were assayed for activity in quaking CNS in examining the hypothesis that increased proteolytic activity contributes to the hypomyelination characteristic of this mutant. Cathepsin B-like enzyme, cathepsin D, neutral proteinase, calcium-activated neutral proteinase, prolyl endopeptidase, and diaminopeptidase II were assayed in whole homogenate of brain or spinal cord and each was found to have activity similar to that in normal mice. These results do not support a relationship between proteolysis and the genetic defect and suggest that other factors should be investigated to delineate the pathogenesis of this mutant.     

Resistance of wild wheat to stripe rust: Predictive method by ecology and allozyme genotypes

From 114 accessions of wild emmer wheat from 11 sites in Israel, known for their allozymic variation (Nevo & al. 1982), individual genotypes were tested for resistance to one isolate of stripe rust both in the seedling stage in a growth chamber and in the adult plant stage in the field. The results indicate that resistance to stripe rust in seedlings and adults are significantly correlated (r_s = 0.40, p < 0.001). Genetic polymorphisms of resistance to stripe rust vary geographically and are predictable by climatic, as well as allozymic markers. Three variable combinations of rainfall, evaporation, and temperature explain significantly 0.40–0.53 of the spatial variance in disease resistance to stripe rust, suggesting the operation of natural selection. Several allozyme genotypes are significantly associated with disease resistance. We conclude that natural populations of wild emmer wheat in Israel contain large amounts of disease resistance genes. These populations could be effectively screened and then utilized by the phytopathologist for identifying resistant genotypes and producing new resistant cultivars.Patterns of Resistance of Wild Wheat to Pathogens in Israel II.     

Larvae of Euphausia superba in the Scotia Sea and Bransfield Strait in March 1984 — Development and abundance compared with 1981 larvae

Summary Larvae of Euphausia superba in the Atlantic sector of the Antarctic in March 1984 averaged 580 per 1000 m³ of water. This is 100 times less than we observed in March 1981, but more than the average of 250 larvae per 1000 m³ found in January–February 1981. There was one 1984 high-abundance sample, accounting for 85% of all larvae caught, from the eastern area of confluence of Drake Passage and Weddell Sea waters. Abundances in 1984 near the South Shetland Islands were commonly 5 to 10 larvae per 1000 m³, and younger by 2 to 4 developmental stages than in March 1981. Body lengths of given stages were generally less in 1984 than in 1981. Advanced furcilia stages, particularly, in the 1984 samples tended to be smaller than the same stages in March 1981, indicating relatively poor growth during February 1984. However, the 1984 younger larvae (calyptopes and the developmental forms of furcilia stages 1 and 2) indicated that, in 1984, recent (March) growth had been good — probably better than in February. Dierct observation of the development of calyptopis stage 3 to furcilia stage yielded a development time of 7.7 days, which compares favorably to the 8-day period estimated from field samples. Reduced food availability did not affect the development rate nor give rise to a clearly higher incidence of indirect pathways of development. It is postulated that recruitment was about of month later in 1984 than in 1981 in most of the area studied and was probably going to be less successful.