Mutations which affect the inhibition of protein phosphatase 2A by simian virus 40 small-t antigen in vitro decrease viral transformation.

Three independent point mutations within residues 97 to 103 of the simian virus 40-small-t antigen (small-t) greatly reduced the ability of purified small-t to inhibit protein phosphatase 2A in vitro. These mutations affected the interaction of small-t antigen with the protein phosphatase 2A A subunit translated in vitro, and a peptide from the region identified by these mutations released the A subunit from immune complexes. When introduced into virus, the mutations eliminated the ability of small-t to enhance viral transformation of growth-arrested rat F111 cells.

Effects of apomorphine and L-methionine sulphoximine on the release of excitatory amino acid neurotransmitters and glutamine in the striatum of the conscious rat.

The effects of apomorphine, a D1-, D2-dopamine receptor agonist, on the extracellular concentrations of excitatory amino acids, glutamic and aspartic acids, and on that of their precursor, glutamine, were investigated using an intracerebral perfusion system. Apomorphine produced a concentration-related rise in glutamic acid concentration in cerebral perfusates (P < 0.01) whereas only the highest concentration of apomorphine (3 x 10(-3) micrograms/microliters) increased the concentration of aspartic acid (P < 0.

Establishment of permanent brown adipocyte cell lines achieved by transfection with SV40 large T antigen and ras genes.

We established multiple clonal cell lines by transfection of primary rat fetal brown adipocytes (BAT) with constitutive or inducible gene constructs of SV40 large T antigen (SVLTag) alone or in combination with transforming or normal ras genes. While primary BAT cell cultures stop growing and expressing uncoupling protein (UCP) after several passages in culture, our transfected cell lines could be propagated indefinitely and kept properties specific of BAT cells. Interestingly, cells transfected with ras genes alone did not survive as permanent cell lines.

Simian virus 40 small-t antigen binds two zinc ions.

Six cysteine residues of the simian virus 40 small-t antigen (small-t) are important for stability of the protein. Stability has been shown to be related to the ability of small-t to bind zinc ions in vitro. Purified small-t expressed either in bacteria or from baculovirus vectors binds two molecules of zinc per molecule of protein.

The ras signaling pathway mimics insulin action on glucose transporter translocation.

Recent observations suggest that insulin increases cellular levels of activated, GTP-bound Ras protein. We tested whether the acute actions of insulin on hexose uptake and glucose-transporter redistribution to the cell surface are mimicked by activated Ras. 3T3-L1 fibroblasts expressing an activated mutant (Lys-61) N-Ras protein exhibited a 3-fold increase in 2-deoxyglucose uptake rates compared with non-transfected cells.

Effects of amphetamine on the release of excitatory amino acid neurotransmitters in the basal ganglia of the conscious rat.

The effects of systemic injections of amphetamine sulfate on the release of aspartic acid, glutamic acid, and glutamine were studied using a push-pull perfusion system in the conscious rat. Amphetamine produced a dose-related increase of the extracellular levels of aspartic acid and glutamic acid. The mean time effect of amphetamine was 40 min, followed by a recovery to baseline levels.

Amphetamine-induced release of diadenosine polyphosphates--Ap4A and Ap5A--from caudate putamen of conscious rat.

The release of diadenosine polyphosphates--diadenosine tetraphosphate (Ap4A) and diadenosine pentaphosphate (Ap5A)--was measured by intracerebral push-pull perfusion in conscious rats after systemic amphetamine injection. Samples were collected from the caudate putamen, and nucleotide compounds were analyzed by HPLC. The presence of Ap4A and Ap5A was demonstrated by their retention times and phosphodiesterase digestion.

p21ras-induced meiotic maturation of Xenopus oocytes in the absence of protein synthesis: MPF activation is preceded by activation of MAP and S6 kinases.

Microinjection of transforming p21ras into Xenopus oocytes caused a time-dependent increase in the level of total cell protein phosphorylation that culminated with germinal vesicle breakdown (GVBD). The same pattern of phosphorylation was observed in oocytes matured by either progesterone or insulin. Treatment with cycloheximide (CHX) completely blocked both GVBD and the associated de novo phosphorylations induced by the hormones, but did not abolish p21ras-induced maturation nor the occurrence of associated maturation promoting factor (MPF)-dependent and -independent phosphorylations.

Dopamine receptor antagonist blocks the release of glycine, GABA, and taurine produced by amphetamine.

The effects of systemic injections of amphetamine sulfate on the extracellular levels of glycine, GABA, and taurine in the neostriatum of awake rats were studied using a push-pull perfusion system. Amphetamine produced a dose-related increase in glycine levels. Amphetamine also produced an enhancement on GABA and taurine levels, although these increases did not follow a dose-related curve.

Clinical pharmacology of alendronate sodium.

Clinical studies have been performed to investigate the pharmacokinetics and pharmacodynamics of alendronate, an inhibitor of bone resorption for the treatment of osteoporosis. Alendronate is one of the most potent bisphosphonates currently undergoing clinical investigation (> 100-fold more potent than etidronate in vivo). The pharmacokinetics of alendronate are similar to those of other bisphosphonates.

Release of amino acid neurotransmitters in different cortical areas of conscious adult and aged rats.

The aim of the present study was to investigate whether or not the levels of amino acid neurotransmitters change during the normal process of aging in the cerebral cortex. In vivo push-pull perfusions were performed in four different areas of the cortex of young (3-4 months) and aged (24-26 months) rats: medial prefrontal cortex, sulcal prefrontal cortex, parieto-temporal cortex and occipital cortex. Extracellular levels of Asp, Glu, Ser, Gln, and Gly were analyzed by HPLC-fluorimetric detection.

Activation of Ras by insulin in 3T3 L1 cells does not involve GTPase-activating protein phosphorylation.

Insulin-induced differentiation of 3T3 L1 cells to adipocytes can be mimicked by the expression of transfected ras oncogenes but not of the tyrosine-kinase oncogenes src and trk. Expression of two different transfected, dominant inhibitory ras mutants resulted in significant inhibition of insulin-induced differentiation, suggesting that endogenous Ras proteins are mediators of insulin signaling in these cells. Exposure of untransfected 3T3 L1 cells to insulin resulted in significant formation of the active Ras.

Pharmacokinetics and dose proportionality of D2-agonist MK-458 (HPMC) in parkinsonism.

To investigate the pharmacokinetic profile, bioavailability, and dose proportionality of the D2-agonist MK-458 (hydroxypropylmethylcellulose tablet, a sustained release formulation), a 4-period crossover study was conducted in 10 patients with mild to moderate Parkinson's disease (mean age = 63 y; 1 woman, 9 men). Following a titration phase to induce tolerance, each patient was given single oral doses of 6, 12 and 18 mg and a single intravenous 40 micrograms dose (5 micrograms/h over 8h). The maximum concentrations of MK-458 observed in plasma after oral administration were 139, 240 and 344 ng/L for the 6, 12 and 18 mg doses, respectively, and occurred after 8.

Effect of the collection method on semen characteristics of Zebu and European type cattle in the tropics.

The effect of the collection method on the characteristics of fresh semen and the recovery of spermatozoa after thawing was studied in 30 Zebu bulls (Bos indicus) and in 30 Brown Swiss (Bos taurus) bulls. Semen was collected by using an artificial vagina and by electroejaculation; the ejaculates were individually evaluated. Semen was diluted for freezing in skimmed milk and stored in 0.

Differentiation of 3T3-L1 fibroblasts to adipocytes induced by transfection of ras oncogenes.

Mammalian 3T3-L1 cells differentiate into adipocytes after continuous exposure to pharmacological doses of insulin or physiological doses of insulin-like growth factor I (IGF-1). Expression of transfected ras oncogenes led to differentiation of these cells into adipocytes in the absence of externally added insulin or IGF-I. Cells transfected with normal ras genes or the tyrosine kinase trk oncogene did not differentiate.

Effect of aspirin and sulindac on methotrexate clearance.

The pharmacokinetics of low dose methotrexate (MTX) were evaluated in 12 rheumatoid arthritis patients in the presence and absence of steady-state levels of salicylic acid (ASA) and sulindac (SU). Using a Latin square design, patients were given MTX plus ASA (mean 3.4 g/day), MTX plus SU (mean 400 mg/day), or MTX alone.

Development of the uncoupling protein in the rat brown-adipose tissue during the perinatal period. Its relationship with the mitochondrial GDP-binding and GDP-sensitive ion permeabilities and respiration.

In this paper we report the development of the immunologically detected uncoupling protein (UCP) in brown adipose tissue during the perinatal period in the rat and its relationship with its functional activity expressed in terms of GDP-binding capacity, GDP-sensitive permeabilities and GDP-sensitive respiration. Immunologically detected UCP increased during the last 2 days of foetal life (under euthermic conditions) and after birth (after postnatal hypothermia) during the early suckling period, reaching its maximum value on day 10 after birth. This increase in UCP is accompanied by parallel increases in the GDP-binding capacity, GDP-sensitive permeabilities to protons and chloride ions and GDP-inhibitable respiration.

Adrenergic regulation of the uncoupling protein expression in foetal rat brown adipocytes in primary culture.

The adrenergic and T3 modulation of UCP expression in non-proliferative foetal brown adipocyte primary cultures were studied. The UCP in the cultured cells was determined by immunological detection of the protein and by quantification of the mitochondrial GDP-binding. Our results showed a relative increase of 65-75% in UCP levels and 60-80% in the mitochondrial GDP-binding capacity under beta-adrenergic stimulatory conditions, while neither alpha 1-adrenergic agonists nor T3 showed an effect.

Simulation of the effect of population size on the evolution of the recombination fraction.

A study, by means of computer simulation, has been performed on the evolution of recombination rate modifier genes in a system with three diallelic loci (A, B and C). The locus C, selectively neutral, is responsible for the modification of the recombination fraction between the major loci (A and B) which are subjected to selection. Two models have been analysed, the modifier allele being recessive in one of them, and codominant in the other, with infinite and finite populations.

[Electrocardiographic analysis of auricular electric systole in the sheep].

The different components of the electric auricular systole, as well as the possible relationship between them have been studied in 302 ECG from Merino Precoz sheep breed. A gradual increase in the P wave duration, RR interval, PQ interval and PQ segment accompanying the growth of the animal, has been obtained, whereas the P wave voltage decreases both gradually and significantly. An analysis of the P wave morphology shows a higher incidence of double waves over simple waves in 1 year old animals than in younger ones.

Fungal endocarditis caused by Arnium leporinum following cardiac surgery.

A case of fungal endocarditis caused by the coprophilous ascomycete Arnium leporinum is reported in a 19-year-old male living in Colombia. The fungus was seen in tissue and recovered in pure culture from peripheral emboli, a mitral valve, pus from an osteolytic lesion, and kidney. The patient died 21 months after cardiac surgery.

Ligand binding to heme proteins. An evaluation of distal effects.

The O2, CO, and alkyl isocyanide-binding properties of a variety of vertebrate and invertebrate heme proteins have been compared in detail to those of protoheme mono-3-(1-imidazoyl)-propylamide monomethyl ester in aqueous suspensions of soap micelles. The proteins examined include: cytochrome P-450cam from Pseudomonas putida, beef heart cytochrome c oxidase, yeast cytochrome c peroxidase, alpha and beta subunits of human hemoglobin, sheep hemoglobin, carp hemoglobin, sperm whale myoglobin, horse heart myoglobin, a monomeric hemoglobin from Glycera dibranchiata, erythrocruorin from Chironomusthummii, soybean leghemoglobin, and several hemoglobins that lack distal histidines. The smallest bimolecular rates were observed for cytochrome P-450 containing bound camphor, cytochrome c oxidase, and cytochrome c peroxidase.

The room temperature potentiometry of xanthine oxidase. pH-dependent redox behavior of the flavin, molybdenum, and iron-sulfur centers.

A series of potentiometric titrations of xanthine oxidase have been performed at room temperature in the pH range 6.1-9.9.