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Biomedical subjects

C Vergara

Publications and source records attributed to C Vergara.

48 records · Page 3Linked to original sources

Phenomenon of formation of giant fat-containing cells in human bone marrow cultures induced by human serum factor: normal and leukemic patterns.

Normal human sera induce the formation of fat-containing cells (FCC) in human bone marrow cultures. A nearly complete monolayer of FCC is formed after 7-14 days of cultivation with 20% human sera in the medium. FCC-inducing activity (FCCIA) is nondialyzable through 14,900-dalton cutoff membrane and is stable at 56 degrees C for 30 min. Abundant FCCIA was found in 83% of normal human sera but in only 20% of sera from untreated patients with different hemopoietic disorders and in 32% of treated leukemic patients. It is suggested that FCCIA may be involved in regulation of the bone marrow microenvironment an that it varies in normal individuals and in patients with different diseases.

Adolescent↗

Kinetics of Ca2+-activated K+ channels from rabbit muscle incorporated into planar bilayers. Evidence for a Ca2+ and Ba2+ blockade.

The interaction of Ca2+ and Ba2+ with a Ca2+-activated K+ channel from rabbit skeletal muscle membranes is studied in planar lipid bilayers. At [Ca2+] greater than or equal to 100 microM in the cis side (the side to which the vesicles are added) and at positive voltages, the channel kinetics consisted of bursts of activity interrupted by long periods of quiescence. We found that the reciprocal of the mean burst time increases linearly with [Ca2+], whereas the mean time for the quiescent (closed) periods is independent of [Ca2+]. The number of quiescent periods is reduced by increasing [K+]. Micromolar amounts of cis Ba2+ do not activate the channel, but induce similar "slow" closings. Also, in this case, the mean burst time is inversely proportional to the [Ba2+] and the mean closed time is independent of [Ba2+]. Raising [K+] either symmetrically or only in the trans side relieved the Ba2+ effect. trans Ba2+ also induces changes in the slow kinetics, but in millimolar amounts. These results suggest that the quiescent periods correspond to a channel blocked by a Ba ion. The voltage dependence of the cis blockade indicates that the Ba2+ binding site is past the middle of the membrane field. The similarities in the slow kinetics induced by Ca2+ and Ba2+ suggest that Ca2+ blocks the channel by binding to the same site. However, binding of Ca2+ to the site is 10(5)-fold weaker.

Animals↗

Uncoupling of Ca2+ transport in sarcoplasmic reticulum as a result of labeling lipid amino groups and inhibition of Ca2+-ATPase activity by modification of lysine residues of the Ca2+-ATPase polypeptide.

Limited labeling of amino groups with fluorescamine in fragmented sarcoplasmic reticulum vesicles inhibits Ca2+-ATPase activity and Ca2+ transport. Under the labeling conditions used, 80% of the label reacts with phosphatidylethanolamine and 20% with the Ca2+-ATPase polypeptide. This degree of labeling does not result in vesicular disruption or in loss of vesicular proteins and does not increase the membrane permeability to Ca2+. Fluorescamine labeling of a purified Ca2+-ATPase devoid of aminophospholipids also inhibits Ca2+-ATPase activity, suggesting that labeling of lysine residues of the enzyme polypeptide is responsible for the inhibition of Ca2+-ATPase activity in sarcoplasmic reticulum. Fluorescamine labeling interferes with phosphoenzyme formation and decomposition in both the native vesicles and the purified enzyme; addition of ATP during labeling, and with less effectiveness ADP or AMP, protects both partial reaction steps. Addition of a nonhydrolyzable ATP analog protects phosphoenzyme formation but not decomposition. The inhibition of Ca2+ transport but not of Ca2+-ATPase occurs in sarcoplasmic reticulum vesicles labeled in the presence of ATP, indicating that the transport reaction is uncoupled from the Ca2+-ATPase reaction. The inhibition of Ca2+ transport but not of Ca2+-ATPase activity is also found in sarcoplasmic reticulum vesicles in which only phosphatidylethanolamine has reacted with fluorescamine. Furthermore, the extent of labeling of phosphatidylethanolamine is correlated with the inhibition of Ca2+ transport rates. The inhibition of Ca2+ transport is a reflection of the inhibition of Ca2+ translocation and is not due to an increase in Ca2+ efflux. We propose that labeling of phosphatidylethanolamine perturbs the lipid environment around the enzyme, producing a specific defect in the Ca2+ translocation reaction.

Animals↗

Reconstitution in planar lipid bilayers of a Ca2+-dependent K+ channel from transverse tubule membranes isolated from rabbit skeletal muscle.

Addition of membrane vesicles prepared from transverse tubule (T-tubule) membranes of rabbit skeletal muscle to the aqueous phase of a planar lipid bilayer induces a stepwise increase in conductance. This conductance is both voltage and Ca2+ dependent. At 1 mM Ca2+, the steady-state conductance is maximal at voltages higher than +20 mV and decreases for more negative voltages. (Voltages refer to the side to which the vesicles are added, cis) Decreasing the Ca2+ concentration reversibly shifts the conductance-voltage curve toward the right along the voltage axis. Furthermore, Ca2+ can activate the conductance only if added to the cis compartment. Neither Mg2+, Ba2+, nor Cd2+ can activate the conductance induced by T-tubule vesicles. Addition of 5 mM tetraethylammonium ion to the trans, but not the cis, side abolishes the T-tubule-induced conductance. The Ca2+-dependent conductance appears as a consequence of ionic channel formation. Single-channel activity appears in bursts followed by periods of time in which the channel remains "silent". The conductance of the open channel averages 226 pS in 0.1 M KC1 and is voltage and Ca2+ independent. However, the fraction of time that the channel remains in the open state is voltage and Ca2+ dependent in a manner that parallels the voltage and Ca2+ dependence of the multichannel membrane. The channel is 6.6 times more permeable to K+ than to Na+ and is impermeable to C1-.

Animals↗

Immunological and biochemical properties of transverse tubule membranes isolated from rabbit skeletal muscle.

A new method for isolating transverse tubule membranes from rabbit skeletal muscle has been developed. This procedure has the advantage of being mild, fast, and producing with good yields a purified membrane fraction. The transverse tubule membranes are purified by a discontinuous sucrose density centrifugation after loading contaminating light sarcoplasmic reticulum vesicles with calcium phosphate in the presence of ATP. Immunofluorescence staining of cryostat sections of rabbit psoas muscle with purified goat antibodies directed against the purified membranes shows that the reacting antigens are distributed at the boundary of the A and I bands of the myofibrils where transverse tubules are localized in mammalian muscle. The purified antibodies showed no cross-reactivity with sarcoplasmic reticulum, nor did they show any fluorescence staining of the muscle plasma membrane, indicating that the isolated membranes indeed originate from the transverse tubules. The transverse tubule fraction has a characteristic protein composition distinguishable from that of sarcoplasmic reticulum, a much higher cholesterol content than that of the crude microsomes, plasma membrane, and sarcoplasmic reticulum, and a phospholipid content about twice as high as that of sarcoplasmic reticulum and plasma membrane. The purified transverse tubule membrane has a distinct phospholipid composition with high contents of sphingomyelin and phosphatidylserine. A Mg2+-activated ATPase characteristic of the transverse tubule fraction undergoes a 20-30-fold increase in specific activity during purification. The levels of Ca2+-ATPase activity present in the purified transverse tubule fraction remain comparable to those of sarcoplasmic reticulum even after extensive removal of the latter.

Adenosine Triphosphatases↗

The extracerebral dopamine antagonist domperidone block the suppressive effect of bromocriptine on prolactin and TSH secretion in man.

The effects of a single oral dose of 2.5 mg bromocriptine on serum level of TSH and prolactin were studied in a group of normal male subjects. Bromocriptine effectively inhibited basal TSH and prolactin concentration as well as the prolactin and TSH response to TRH given 4 hours later. The prior administration of the extracerebral dopamine antagonist domperidone reversed the endocrine effects of bromocriptine. The results suggest that dopamine receptors located at the pituitary may regulate TSH (and prolactin) release in man.

Adult↗

Induction of giant fat cells in human bone marrow culture by human serum factor.

Evidence is presented below that normal human sera contain a potent non-dialyzable factor inducing abundant giant fat cells in human bone marrow culture, normal as well as CML. Media with 20% heated (56 degrees C) human serum induce during 7-14 days almost complete monolayer of fat cells on the bottom of the plastic flasks or dishes. Fetal bovine sera do not exhibit this effect and shift cultures to the proliferation of fibroblasts. We are studying the functions of fat cells in hemopoiesis as well as the biochemical nature of fat cell-inducing factor in human sera.

Adipose Tissue↗

Synthesis and content of polyamines in bloodstream Trypanosma brucei.

The sensitive dansyl procedure was used to detect putrescine and spermidine, but not spermine and cadaverine, in pleomorphic Trypanosoma brucei. The polyamines were synthesized in vitro from [3H]ornithine, [14C]arginine and [14C]methionine. Proline, agmatine, and citrulline, but not glutamine, glutamic or pyroglutamic acids, stimulated spermidine formation from [4C]methionine. Putrescine and sperimidine synthesis occurred rapidly from ornithine: putrescine synthesis peaked in 0.5 h, spermidine in 1 h. Trypanosoma brucei assimilated exogenous 14C-labeled putrescine, spermidine, and spermine; spermidine and spermine were taken up 5 times as rapidly as putrescine. Polyamine syntheses may therefore be a practical target for novel trypanocies.

Amino Acids↗

Extracellular space and diffusion barriers in muscle fibres from Megabalanus psittacus (Darwin).

1. Muscle fibres from Megabalanus psittacus (Darwin) were used to measure the exchange of Na+ and Ca2+ between intracellular and extracellular compartments. 2. The size of the extracellular space was evaluated directly from electron micrographs at 6.1 +/- 0.5% (percentage of the fibre volume). The more conventional estimates using Na+ and 134Cs+ as space markers gave higher values, namely, 8.9 +/- 0.7 and 9.5 +/- 1.5%. 3. An average value of 9.2% was used to correct the total Na+ and K+ in the muscle fibres and to estimate the intracellular concentrations of Na+ and K+ as 39 +/- 4 and 202 +/- 11 mM respectively. 4. Na+ (and similar Ca2+) washout curves could be described using a three compartments diffusion model. The data analysed in terms of this model enabled us to estimate membrane permeabilities as PNa+ = 2.7 x 10(-7) and PCa2+ = 2.7 x 10(-7) cm/sec.

Animals↗

[The context of health care reforms].

In Latin America, health sector reforms have gone hand in hand with social and economic trends during the latter half of the twentieth century and have reflected the particular concept of "development" that has been in vogue at different times. Economic stagnation and increased social spending, both hallmarks of the 1960s, led to the decline of the "import substitution" development model, which had prevailed since the beginning of the century, and slowly gave way in the 1980s to the "globalization" model. From the earlier model, a transition took place toward a restructuring of production and a series of economic adjustment policies that led, ironically, to an increase in poverty in Latin America. Implementation of the new model has occurred in two phases. The first, known as the "social reform" or "first generation" phase, sprang from the notion that poverty is the sum of a number of material shortages that can be corrected through an equitable redistribution of a fixed volume of goods belonging to society. This conceptual framework, which was completely devoid of all historical linkages and separated from economic policy, led to social policies whose entire purpose was to mitigate poverty through subsidies targeting the poorest persons in the society. In the second phase of the globalization model, which arose in the 1990s and became known as the "second generation" or "postadjustment" phase, new economic rules came into play that were based primarily on international competition, efficiency in production, and openness and fairness in the capital markets. And if during the initial stage the conceptual strategy behind all social policy was to fight poverty, in the second stage the strategy became one of achieving equity, which was no longer interpreted as the even distribution of a fixed volume of capital goods, but as the sustained provision of greater and better opportunities for all. Having grown accustomed to the protectionism inherent in the earlier development model, Latin American societies today feel threatened by a new model that offers them no social safety net. The feasibility of economic and social reform policies during the second phase, which reflect the demands of a "globalized" world, thus depends on the ability to overcome people's lack of trust and to garner the support of a political, social, and institutional majority.

Delivery of Health Care↗

Detection of Plasmodium falciparum: a comparison of six cloned DNA probes.

Probes pRepHind, Rep20, p242B1-1, pPF-14, clone 26 and 34 were compared for their applicability to detect P. falciparum in field conditions. Ninety four clinical samples from patients living in the malaria endemic area of Tumaco (Pacific Coast) plus 88 from Villavicencio (Eastern Plains) were tested in "dotblot" hybridization experiments. Probes Rep20, p242B1-1, pRepHind and pPF-14 detected up to 17 pg of purified P. falciparum DNA, while clone 26 and clone 34 detected up to 425 pg DNA. Probes pPF-14, P242B1-1, pRepHind and Rep 20 exhibited comparable detection levels of parasites in infected blood samples. Sensitivity declined from 69-94% in subjects with parasitemias higher than 10.000 par./ul to 15-42% in subjects with parasitemias lower than 100 par./ul. pPF-14 and p24B1-1 showed the highest sensitivity, while clone 26 and 34 presented significantly lower sensitivities. All probes were shown to be highly specific. Detection levels are dependent on specimen treatment. Treatment consisting of serum removal, Triton X-100 lysis, Proteinase K digestion, Phenol and Chloroform extractions followed by Ethanol precipitation yielded 100% sensitivity for specimens with parasite density higher than 1,000 par./l.

Animals↗