Plasma 5-Hydroxyindoleacetic Acid as an Indicator of Monoamine Oxidase-A Inhibition in Rat Brain and Peripheral Tissues

We have examined the changes induced by the monoamine oxidase (MAO; EC 1.4.3.4) inhibitors tranylcypromine, clorgyline, and deprenyl on MAO activity and 5-hydroxytryptamine (serotonin, 5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) content in rat brain and blood (plasma and whole blood). The decreases of MAO-A activity observed in the liver and lungs after different doses of clorgyline or tranylcypromine correlated significantly (r > 0.80 in all cases) with the decline of plasma 5-HIAA. This was unaffected by 0.25 and 5 mg kg^?1 of deprenyl, indicating that 5-HT was deaminated exclusively in the periphery by MAO-A. It is interesting that very potent and significant correlations (r > 0.75) were found between plasma 5-HIAA and MAO-A activity, 5-HIAA and 5-HT content in brain tissue. These results suggest that plasma 5-HIAA can be used confidently as a peripheral indicator of the inhibition of MAO-A in brain. This may represent a favorable alternative to the analysis of 5-HIAA in CSF in psychiatric patients undergoing antidepressant treatment with nonspecific MAO inhibitors or with the new selective MAO-A inhibitors.     

In Vivo Brain Dialysis Study of the Somatodendritic Release of Serotonin in the Raphe Nuclei of the Rat: Effects of 8-Hydroxy-2-(Di-n-Propylamino)tetralin

Abstract: The characteristics of the serotonin (5-HT) output in the dorsal and median raphe nuclei of the rat were studied using in vivo microdialysis. The basal output of 5-HT increased after KC1 was added to the perfusion fluid. In contrast, neither the omission of calcium ions nor the addition of 0.5 nM tetrodotoxin affected dialysate 5-HT or 5-hy-droxyindoleacetic acid (5-H1AA). Reserpine did not decrease the output of 5-HT and 5-HIAA 24 h later and p-chloroamphetamine increased 5-HT in both vehicle- and reserpine-treated rats severalfold. 8-Hydroxy-2-(di-n-pro-pylamino)tetralin (8-OH-DPAT), at 1 or 10 μM, perfused into the raphe did not change the outputs of 5-HT or 5-HIAA. Higher doses (0.1, Land 10 mM) increased extracellular 5-HT in the raphe, probably via an inhibition of uptake. In animals bearing two probes (raphe nuclei and ventral hippocampus), only the 10 vaM dose of 8-OH-DPAT perfused into the raphe decreased the hippocampal output of 5-HT and 5-HIAA. The systemic injection of 0.1 mg/kg 8-OH-DPAT decreased dialysate 5-HT and 5-HIAA in the raphe and hippocampus. These results suggest that extracellular 5-HT in raphe nuclei originates from a cytoplasmic pool and is not dependent on either nerve impulse of 5-HT neurons or local activation of 5-HT_1A receptors.     

Serum selenium concentration of a healthy northwest Spanish population

Selenium (Se) is an essential element, cofactor for glutathione peroxidase (GSHPx) activity, whose deficiency may induce modifications in the cellular antioxidative status and induce the appearance of different diseases. Current views suggest that a serum Se concentration inferior to 45 μg/L may correlate with an increased risk of coronary hearth diseases, coronary atherosclerosis and cancer. Since the Se concentration in human blood varies between geographical areas, we initiated a study to evaluate the Se status in the general healthy population of Barcelona. Serum Se concentration was investigated in a random sample of 150 subjects (age range 18–70 yr) by graphite furnace atomic spectrometry (FLAAS). L'vov platform, Zeeman background correction, and other specifications of stabilized temperature platform furnace (STPF) concept were followed. The results show that in the general population of Barcelona, Se serum concentration ranges between 60 and 106 μg/L (X=80.7±10 μg/L). These values can be considered within the safe limits, since no subject was found with a concentration lower than the threshold of 45 μg/L.     

Injection of anticentromere antibodies in interphase disrupts events required for chromosome movement at mitosis

We have used autoantibodies to probe the function of three human centromere proteins in mitosis. These antibodies recognize three human polypeptides in immunoblots: CENP-A (17 kD), CENP-B (80 kD), and CENP-C (140 kD). Purified anticentromere antibodies (ACA-IgG) disrupt mitosis when introduced into tissue culture cells during interphase. We have identified two execution points for antibody inhibition. Antibodies injected into the nucleus greater than or equal to 3 h before mitosis prevent the chromosomes from undergoing normal prometaphase movements in the subsequent mitosis. Antibodies injected in the nucleus during late G2 cause cells to arrest in metaphase. Surprisingly, antibodies introduced subsequent to the beginning of prophase do not block mitosis. These results suggest that the CENP antigens are involved in two essential interphase events that are required for centromere action in mitosis. These may include centromere assembly coordinate with the replication of alpha-satellite DNA at the end of S phase and the structural maturation of the kinetochore that begins at prophase.     

Chromosomal passengers: Toward an integrated view of mitosis

Conclusions The immunocytochemical taxonomy experiments cited above have identified a class of chromosomal antigens whose properties were not predicted by earlier models of mitosis. Our theory describing one possible explanation for the transfer of these antigens from the chromosomes to the spindle midzone at the metaphase: anaphase transition must now be subjected to further experimental tests. The phenotypes of cells microinjected with antibodies to passenger proteins should enable us to identify mitotic processes dependent on these proteins, as in the example of CHO1 antibody blocking mitotic progression (Nislow et al. 1990). In addition, the availability of cDNA clones and high titer antibodies may enable homologues of these components to be identified in organisms in which they can be subjected to genetic analysis.For the time being, we suggest that current views of the relative roles of chromosomes and cytoskeletal components in mitosis may require revision. Our hypothesis takes the current model for the role of the kinetochores in organizing the bipolar mitotic spindle (Kirschner and Mitchison 1986) a step further. The process of assembling a functional spindle and positioning the cleavage furrow may entail a degree of functional cooperation between chromosomes and cytoskeletal components far beyond that envisioned before now.     

Design of Ca-alginate immobilized yeast cell beads with controlled low density to enhance their fluidization behaviour in bioreactors

Summary The incorporation of small glass hollow spheres into Ca-alginate beads allows the density of immobilized cell preparations to be decreased. For immobilized yeast, glass hollow spheres at 14% (v/v) decrease the density from 1088 to 996 kg m^-3, thus greatly changing their hydrodynamic properties. As a consequence, the operation of a three-phase fluidized bed reactor is clearly improved.     

Synthesis of novel proteins

De novo and rational protein design are progressing towards the chemical synthesis of proteins with pre-selected structure and function. The data illustrate diverse experimental and computational approaches which test our comprehension of protein structure, hydrophobic core packing and global stability, especially of coiled-coil proteins. The incorporation of biologial cofactors, including hemes, as well as active sites, such as tht of iron superoxide dismutase, into designed proteins provides an exciting next step towards the synthesis of proteins with enzymatic function.