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Biomedical subjects

J Bustamante

Publications and source records attributed to J Bustamante.

At least 55 records · Page 3Linked to original sources

Purification and analysis of plasmatic components of middle molecular weight in patients with uremic syndrome.

Many hormonal, metabolic and enzymatic alterations have been described in patients with chronic renal failure (CRF) after prolonged hemodialysis treatment. This toxicity has been associated with the accumulation of middle molecular weight components, called 'middle molecules', in their blood. To investigate the chemical nature of these molecules, 20 CRF patients were studied and compared with 10 control subjects. Sera from these individuals were treated with molecular exclusion chromatography, the components of high molecular weight were eliminated from the elution peaks, and the remainder was then concentrated by lyophilization. The zones of greatest concentration were fractioned by ionic exchange chromatography and by HPLC in reverse phase. Infrared spectroscopy was also performed on the most relevant zones, as well as SDS/PAGE. Finally, the amino acid sequence of a peak presenting the best conditions was analyzed. The results indicate that the majority of the compounds isolated are peptides, amino acids or amino alcohols, and that many of these 'middle molecules' are joined to trace metals.

Adult↗

Role of melanin as a scavenger of active oxygen species.

The protective role of melanin, either synthetic or derived from a metastatic lung melanoma nodule, was studied in terms of its ability to interact with active oxygen species (O2., H2O2, RO., ROO., etc.). Both melanins showed the ability to react with O2.. The superoxide dismutase-like activity corresponds to 21 and 10 U/mg for synthetic and tumor melanin, respectively. The latter value accounts for about 8% of the superoxide dismutase activity of cultured melanoma cells. Neither type of melanin showed catalase-like or glutathione peroxidase-like activity. Both types of melanin reacted with RO. and ROO. radicals as determined by inhibition of the lipid peroxidation reaction of rat liver homogenates. The spontaneous lipid peroxidation of rat liver homogenate was inhibited up to 90% and 80% by synthetic and tumor melanin with half-maximal effects at 2.5 and 5.5 micrograms melanin/ml, respectively. The 2,2-azo-bis-(2 amidino propane) (AAPH)-initiated lipid peroxidation of rat liver homogenate was inhibited up to 30% and 20% by synthetic and tumor melanin, with half maximal effect at 120 and 500 micrograms melanin/ml, respectively. Both types of melanin were able to protect the in vitro inactivation of glucose oxidase, which occurs in the presence of AAPH-generated radicals.

Amidines↗

Signal transduction and nonlinearities revealed by white noise inputs in the fast adapting crayfish stretch receptor.

Input-output relations were investigated in the fast adapting stretch receptor organ (RM2) of the crayfish by matching gaussian white noise (GWN) length inputs, with the resulting spike output. The analysis revealed the expected sensitivity to lengthening velocity, a behavior termed phasic. It also disclosed a sensitivity to sustained elongation, a performance termed tonic and previously not recognized in the RM2. Spectral analysis indicated the properties of a low-pass filter, confirming the tonic sensitivity. A variety of individual length trajectories could lead to a spike. The average trajectory consisted in a biphasic shortening-lengthening wave. The range of possible trajectories and their averages changed with stimulus prestretch and GWN amplitude, indicating that system properties depended on the input characteristics; i.e., a nonlinear operation. Length waveforms in the GWN were isolated by computing methods and the corresponding responses were calculated. Symmetric stimuli led to responses that reflected magnitudes and velocities asymmetrically. Nonlinear interactions between responses in the past and present were negligible. In conclusion, depending on the input, the RM2 modifies its operation to enhance the detectability of the functionally relevant signal in each natural situation.

Animals↗

Melanin content and hydroperoxide metabolism in human melanoma cells.

Human melanoma cells were grown to exponential and stationary phases showing melanin contents of 4.2 +/- 0.3 and 11.3 +/- 0.6 micrograms/10(6) cells, respectively. The cells were separated in four subpopulations by a Percoll gradient; the subpopulation of density 1.07 (g/ml) was the most enriched in pigmented cells and produced 28 and 58% of the cells in exponential and stationary phases, respectively. Melanoma cells had similar superoxide dismutase and glutathione peroxidase activities in exponential and stationary phases. Moreover melanoma cells exhibited a higher catalase activity in the stationary phase: whole homogenate and cytosol activities were 7.0 +/- 0.3 and 10.8 +/- 0.6 U/mg protein, whereas in exponential phase the activities were 4.9 +/- 0.1 and 7.6 +/- 0.3 U/mg protein for whole homogenate and cytosol, respectively. The intracellular H2O2 steady-state concentration was 3.3 +/- 0.2 and 2.1 +/- 0.2 microM H2O2 for exponential and stationary phases, respectively. The spontaneous chemiluminescence of the two culture phases was 169 +/- 27 cps/10(6) cells (exponential) and 78 +/- 24 cps/10(6) cells (stationary). The cytotoxicity of H2O2 generated extracellularly by glucose oxidase was determined after 60 min of exposure. IC50 values for exponential and stationary cell cultures were 0.9 and 2.4 mU/ml of glucose oxidase, respectively. The increased catalase activities in the stationary phase as compared with the exponential phase are consistent with the decreased intracellular H2O2, with the decreased spontaneous chemiluminescence, and with the increased resistance to exogenous H2O2.

Catalase↗

Adriamycin effects on hydroperoxide metabolism and growth of human breast tumor cells.

Human breast tumor cells MCF-7 were grown during 5 days in the presence of Adriamycin and the IC50 was 50 nM with the highest sublethal concentration 0.1 microM. At this latter concentration Adriamycin produced a complete inhibition of cell division and a partial reversion to a normal breast epithelial appearance. Similar effects of Adriamycin were observed in cells cultured in the presence of 10% FBS and in a chemically defined medium, with Se-glutathione peroxidase activities of 3.8 and 1.3 U/mg of protein, respectively. Cell size and cell oxygen uptake were increased by 41% and by 50%, respectively, in Adriamycin-treated cells. The spontaneous chemiluminescence of monolayers of intact MCF-7 cells (81 +/- 9 cps/mg protein) was increased by 48% in the Adriamycin-treated cultures (120 +/- 11 cps/mg of protein) in agreement with a 91% higher concentration of malondialdehyde in the same cultures. Adriamycin treatment produced a 71% increase in the steady state concentration of H2O2, which was estimated assuming diffusion equilibrium with the external medium, from 1.38 microM in the control cells to 2.38 microM in the treated cells. Cyanide-insensitive respiration was also higher in the cells exposed to the drug than in the control cells. Adriamycin did not affect the activity of the antioxidant enzymes, Cu-Zn and Mn-superoxide dismutase, Se and non-Se-glutathione peroxidase, and catalase. These results contribute to the current hypothesis that oxygen free radicals produced by Adriamycin redox cycling are responsible for at least part of the cytotoxic effects due to this drug.

Analysis of Variance↗

White noise analysis of pace-maker-response interactions and non-linearities in slowly adapting crayfish stretch receptor.

Input-output relations were investigated in the slowly adapting stretch receptor organ of crayfish using a Gaussian white noise length input with a 0.03-12.5 Hz band width and the resulting action potential output. The noise input was presented to the de-efferented receptor in situ, at three mean elongations and at four different amplitudes. The three mean elongations were set within the normal range in vivo, two at the extremes close to the minimum and maximum physiological lengths and the other in the mid-range. With white noise inputs there is a finite probability that the system will be tested in all possible conditions within the chosen band width because white noise has the advantage that it contains, with a finite probability, all possible stimulus wave forms at random. The analysis indicated similarities between the effects of the input variables, namely white noise amplitude and mean elongation. With low input variables the activity was periodic. With larger inputs, impulse rates were higher and irregular. The average length trajectories leading to a spike (i.e. the average stimulus) were either biphasic with high inputs or multiphasic and periodic with lower input variables. The frequency of periodicity increased with mean elongation. Although for a given length and noise amplitude a variety of individual length trajectories preceded spikes, the final biphasic shortening-lengthening average stimulus sequence before a spike was similar in all cases irrespective of the input variables. The number of possible trajectories decreased with increments in the input variables. The standard deviation of length values for each average stimulus was computed and displayed as a function of time relative to the spike. It was first constant, and decreased gradually to a minimum value at the spike reference. Standard deviation values were lower for higher white noise amplitudes and mean elongation. Simple, short-lasting stimulus wave forms in the white noise were isolated and the corresponding responses were computed. Responses were periodic at low input variables and aperiodic with larger inputs and also asymmetric, being larger and briefer for lengthenings than for shortenings. Asymmetries augmented with input increments. Investigation of the interactions between the pace-maker and the white noise effects showed that lengthenings were more effective when they occurred just after a spike while the effectiveness of shortenings was greater just before a spike.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Modifications of dynamic and static behavior by small-length perturbations in crayfish stretch receptor organs.

To investigate whether static and dynamic sensitivities of slowly and rapidly adapting stretch receptor organs (SAO and RAO, respectively) or crayfish are different when perturbed compared with those in conventional laboratory experiments, receptors were submitted to ramplike length changes of different velocities separated by long-duration, constant lengths of different values. They were perturbed at random by fast, small-amplitude length variations called "jitter." First-order afferent discharges were recorded extracellularly. Quantifications involved the separate estimation of static and dynamic response components. In the SAO, jitter (1) augmented the static sensitivity, (2) decreased the dynamic sensitivity, (3) simplified response profiles by decreasing nonlinearities and increasing transduction fidelity in terms of coding length. In the RAO, jitter (1) changed the behavior from phasic to tonic with length sensitivity, (2) decreased the dynamic sensitivity to values close to those of the unperturbed SAO, (3) increased transduction fidelity for stimulus length and decreased nonlinearities. Perturbation effects suggest that differences between SAO and RAO are more quantitative than qualitative. Moreover, they have general implications and are relevant to other mechanoreceptors at other levels in the CNS.

Animals↗

Effects of feedback and reinforcement in tachistoscopic training on a fault-detection task.

This study looked at possible differential effects of feedback and reinforcement on a stimulated fault-detection task. Volunteer college students, 11 females and 13 males, participated in the study. Subjects were assigned to three different groups and tachistoscopically presented with series of slides of intact and faulty cups randomly arranged. Subjects in Group 1 were shown a highly preferred slide following each correct detection (positive reinforcement). In Group 2, subjects received a second, longer presentation of every item responded to and every critical one omitted (knowledge of results). Subjects in Group 3 faced a blank screen during inter-stimulus intervals (practice control). Detection scores along five decreasing stimulus presentation times were recorded. Data indicate the treatment conditions as equally efficient and significantly better than the control condition. However, performance of the control group did improve significantly throughout training which suggests the use of tachistoscope as a training device per se. Commission and omission errors did not differ significantly within treatments, though both types of error were significantly elevated for the control group. A possible dissociation between the effects of reinforcers and feedback is discussed, and experiments which would tax higher levels of information processing are suggested to probe this issue further.

Adult↗

Rate-dependent action potential changes in rat atrium.

Experiments in isolated left atria from rat hearts were performed in order to study the effects of stimulation rate on the transmembrane action potential. 1. Two components (fast and slow) of the action potential upstroke could be differentiated by adding MnCl2 to the perfusion solution. 2. With the increase in rate of stimulation over the control cycle length (500 msec), amplitude, Vmax and action potential duration at 80% of repolarization (D80) diminished in normal Krebs. In Mn-containing Krebs, only a slight reduction in amplitude was recorded. Resting potential and action potential duration at 20% (D20) and 50% (D50) of repolarization were only slightly affected in normal Krebs and not at all in Mn-Krebs. 3. Low rates of stimulation in normal Krebs increased D50 only slightly; however, D80 increased significantly while other parameters remained constant. No effects were seen in Mn-Krebs. 4. The results with Mn-Krebs indicate the importance of slow inward current on changes induced by stimulation rates. A possible mechanism relating intracellular calcium concentration and the outward K current, depending on the rate of stimulation, is discussed.

Animals↗

Changes in copper and ceruloplasmin in chronic renal insufficiency treated by hemodialysis and peritoneal dialysis.

Serum copper and ceruloplasmin as well as other parameters in 68 patients with chronic renal insufficiency were studied. It was found that chronic renal insufficiency increases neither the serum copper nor the serum ceruloplasmin. In the group on hemodialysis, there is a progressive increase of copper and ceruloplasmin in serum due to a liberation of 44.44% of the copper content of the membranes. In the group on periodic peritoneal dialysis, there is also an increase of serum copper during the dialysis. We do not have an explanation for this finding.

Adolescent↗