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Biomedical subjects

E Diaz

Publications and source records attributed to E Diaz.

At least 55 records · Page 3Linked to original sources
PubMed1995

Lipolytic effects of beta1, beta2 and beta3-adrenergic agonists in isolated human fat cells from omental and retroperitoneal adipose tissues.

The presence of beta1- and beta2-adrenoceptors has been clearly established in human fat cells. There is some controversy about the presence and function of beta3-adrenoceptors. It is well established that there are marked regional variations in catecholamine-induced lipolysis. In this work the possibility that a beta3-adrenoceptor plays a significant role in the control of lipid mobilization is studied and also its importance in comparison to beta1- and beta2-adrenoceptors in isolated human fat cells, is evaluated, by measuring the in vitro lipolysis induced by dobutamine, salbutamol, metaproterenol, BRL 37344 and CGP 12177A. Human adipocytes from omental and retroperitoneal fat deposits exhibited an "atypical" beta-adrenergic response but, given the small lipolytic effect initiated by BRL 37344 and CGP 12177A, they are probably poorly equipped in functional beta3-adrenoceptors.

Adipose Tissue↗
PubMed1994

Potentiation of insulin stimulation of phosphatidylinositol 3-kinase by thiazolidinedione-derived antidiabetic agents in Chinese hamster ovary cells expressing human insulin receptors and L6 myotubes.

Thiazolidinedione derivatives are insulin-sensitizing agents with proven antidiabetic activities in vivo. To explore the mechanism of action of this class of compounds, the effects of pioglitazone, CP-86,325, and AD-5075 on elements of the insulin signal transduction pathways were studied in Chinese hamster ovary cells overexpressing human insulin receptor (CHO.T) and L6 myotubes. In CHO.T cells, the binding of insulin to its receptor and the insulin-stimulated tyrosine kinase activity of the receptor were not altered by pioglitazone or CP-86,325. In contrast, treatment of CHO.T cells with the compounds resulted in significant increases in insulin-stimulated phosphatidylinositol (PI) 3-kinase activity. This insulin-enhancing effect was also observed in L6 myotubes treated with CP-86,325. The augmentations in kinase activity observed in CHO.T cells correlated with increases in the amount of PI 3-kinase (p85 subunit) in anti-phosphotyrosine immunoprecipitates of cell lysates. No gross changes in the tyrosine phosphorylation state of the insulin receptor substrate-1 were detected in insulin-stimulated CHO.T cells following treatment with the compounds. Furthermore, the compounds did not enhance insulin stimulation of mitogen-activated protein kinase or DNA synthesis in CHO.T cells. Thus, thiazolidinedione-derived antidiabetic agents may act as insulin sensitizers by augmenting insulin stimulation of PI 3-kinase activity in a rather specific manner.

Animals↗
PubMed1994

Carboxy-terminal deletion analysis of the major pneumococcal autolysin.

Autolysins are endogenous enzymes that specifically degrade the covalent bonds of the cell walls and eventually can induce bacterial lysis. One of the best-characterized autolysins, the major pneumococcal LytA amidase, has evolved by the fusion of two domains, the N-terminal catalytic domain and the C-terminal domain responsible for the binding to cell walls. The precise biochemical role played by the six repeat units that form the C-terminal domain of the LytA amidase has been investigated by producing serial deletions. Biochemical analyses of the truncated mutants revealed that the LytA amidase must contain at least four units to efficiently recognize the choline residues of pneumococcal cell walls. The loss of an additional unit dramatically reduces its hydrolytic activity as well as the binding affinity, suggesting that the catalytic efficiency of this enzyme can be considerably improved by keeping the protein attached to the cell wall substrate. Truncated proteins lacking one or two repeat units were more sensitive to the inhibition by free choline than the wild-type enzyme, whereas the N-terminal catalytic domain was insensitive to this inhibition. In addition, the truncated proteins were inhibited by deoxycholate (DOC), and the expression of a LytA amidase lacking the last 11 amino acids in Streptococcus pneumoniae M31, a strain having a deletion in the lytA gene, conferred to the cells an atypical phenotype (Lyt+ DOC-) (cells autolysed at the end of the stationary phase but were not sensitive to lysis induced by DOC), which has been previously observed in some clinical isolates of pneumococci. Our results are in agreement with the existence of several choline-binding sites and suggest that the stepwise acquisition of the repeat units and the tail could be considered an evolutionary advantage for the enzyme, since the presence of these motifs increases its hydrolytic activity.

Base Sequence↗
PubMed1994

Achilles tendon ossification.

The authors describe the case of a patient who had an ossified Achilles tendon without fracture. Such ossification is uncommon, occurring most often after trauma or surgery.

Achilles Tendon↗
PubMed1992

Instruments that measure beliefs about cancer from a cultural perspective.

Understanding beliefs about cancer from a cultural perspective is of great importance as the population mix of this country continues to shift and change. In order to assess the cancer beliefs held by diverse populations, culturally specific tools are needed. The purpose of this article is to identify and describe instruments that have been used to assess beliefs about cancer. Eight published and unpublished instruments were identified from a review of the literature. These tools address attitudes and beliefs toward cancer, cancer prevention, and cancer treatment within a variety of age groups and ethnic populations. One tool was related to adolescents, whereas the remainder focused on adults; three were translated into other languages; four tools involved questionnaire format and four interview format. Although it is encouraging to report that culturally specific tools are being developed, there is a demonstrated need to increase the expertise in developing such instruments.

Attitude to Health↗
PubMed1991

Alkylation of an active-site cysteinyl residue during substrate-dependent inactivation of Escherichia coli S-adenosylmethionine decarboxylase.

S-Adenosylmethionine decarboxylase from Escherichia coli is a member of a small class of enzymes that uses a pyruvoyl prosthetic group. The pyruvoyl group is proposed to form a Schiff base with the substrate and then act as an electron sink facilitating decarboxylation. We have previously shown that once every 6000-7000 turnovers the enzyme undergoes an inactivation that results in a transaminated pyruvoyl group and the formation of an acrolein-like species from the methionine moiety. The acrolein then covalently alkylates the enzyme [Anton, D. L., & Kutny, R. (1987) Biochemistry 26, 6444]. After reduction of the alkylated enzyme with NaBH4, a tryptic peptide with the sequence Ala-Asp-Ile-Glu-Val-Ser-Thr-[S-(3-hydroxypropyl)Cys]-Gly-Val-Ile-Ser-Pro - Leu-Lys was isolated. This corresponds to acrolein alkylation of a cysteine residue in the second tryptic peptide from the NH2 terminal of the alpha-subunit [Anton, D. L., & Kutny, R. (1987) J. Biol. Chem. 262, 2817-2822]. The modified residue derived is from Cys-140 of the proenzyme [Tabor, C. W., & Tabor, H. (1987) J. Biol. Chem. 262, 16037-16040] and lies in the only sequence conserved between rat liver and E. coli S-adenosylmethionine decarboxylase [Pajunen et al. (1988) J. Biol. Chem. 263, 17040-17049]. We suggest that the alkylated Cys residue could have a role in the catalytic mechanism.

Adenosylmethionine Decarboxylase↗
PubMed1991

Chimeric pneumococcal cell wall lytic enzymes reveal important physiological and evolutionary traits.

Two novel chimeric pneumococcal cell wall lytic enzymes, named LC7 and CL7, have been constructed by in vitro recombination of the lytA gene encoding the major autolysin (LYTA amidase) of Streptococcus pneumoniae, a choline-dependent enzyme, and the cpl7 gene encoding the CPL7 lysozyme of phage Cp-7, a choline-independent enzyme. In remarkable contrast with previous chimeric constructions, we fused here two genes that lack nucleotide homology. The CL7 enzyme, which contains the N-terminal domain of CPL7 and C-terminal domain of LYTA, exhibited a choline-dependent lysozyme activity. This experimental rearrangement of domains might mimic the process that have generated the choline-dependent CPL1 lysozyme of phage Cp-1 during evolution, providing additional support to the modular theory of protein evolution. The LC7 enzyme, built up by fusion of the N-terminal domain of LYTA and the C-terminal domain of CPL7, exhibited an amidase activity capable of degrading ethanolamine-containing cell walls. The chimeric amidase behaved as an autolytic enzyme when it was cloned and expressed in S. pneumoniae. The chimeric enzymes provided new insights on the mechanisms involved in regulation of the host pneumococcal autolysins and on the participation of these enzymes in the process of cell separation. Furthermore, our experimental approach confirmed the basic role of the C-terminal domains in substrate recognition and revealed the influence of these domains on the optimal pH for catalytic activity.

Base Sequence↗
PubMed1991

Effects of dietary supplementation on work performance in Gambian laborers.

The effect of food supplementation on productivity of Gambian subsistence farmers was studied during a natural food shortage induced by the annual hungry season. Sixteen men in two groups matched for weight, height, hemoglobin, and physical-work capacity followed a crossover protocol with 6-wk supplementation and control periods. Productivity was assessed over the final 3 wk of each period by quantifying piece-rate-paid road building. Supplementation comprised an ad libitum supply of highly palatable energy-dense food provided three times daily. Outcome variables were total loads transported, loads per working hour, time per load, total energy expenditure assessed by doubly labeled water and heart-rate monitoring, postwork activity, and anthropometry. Body weight decreased during control periods, indicating a real energy deficit, and increased during supplementation in both groups. Supplementation had no significant impact on productivity variables. Energy-deficient men can maintain maximal productivity over short periods if sufficiently motivated but at the expense of body weight.

Adult↗
PubMed1991

Free-living energy expenditure measured by two independent techniques in pregnant and nonpregnant Gambian women.

Free-living energy expenditure (EE) was assessed in 37 young pregnant Gambian women at the 12th (n = 11, 53.5 +/- 1.7 kg), 24th (n = 14, 54.7 +/- 2.1 kg), and 36th (n = 12, 65.0 +/- 2.6 kg) wk of pregnancy and was compared with nonpregnant nonlactating (NPNL) control women (n = 12, 50.3 +/- 1.6 kg). The following two methods were used to assess EE: 1) the heart rate (HR) method using individual regression lines (HR vs EE) established at different activity levels in a respiration chamber and 2) the doubly labeled water (2H2(18)O) method in a subgroup of 25 pregnant and 7 control women. With the HR method the EE during the agricultural rainy season was found to be 2,408 +/- 87, 2,293 +/- 122, and 2,782 +/- 130 kcal/day at 12, 24, and 36 wk of gestation and were not significantly different from the control group (2,502 +/- 133 kcal/day). These findings were confirmed by the 2H2(18)O measurements, which failed to show any effect of pregnancy on EE. Expressed per unit body weight, the free-living EE was found to be lower (P less than 0.01 with 2H2(18)O method) at 36 wk of gestation than in the NPNL group. It is concluded that, in these Gambian women, energy-sparing mechanisms that contribute to meet the additional energy stress of gestation are operating during pregnancy (e.g., diminished spontaneous physical activity).

Adolescent↗
PubMed1990

Fast processing of black and white films used in the EM lab.

Kodak black and white film emulsions 4489, 4162, 2415, TMAX-100, TMAX-400, MPD-4 and RPC-651 can be developed in less than 10 minutes on a film processor, quickly printed on resin coated paper and dried with auto-processor and dryer. Adjustments to the ASA (ISO) to compensate for increased contrast with auto-processing are described.

Microscopy, Electron↗
PubMed1990

Evaluation of near infra-red interactance as a method for predicting body composition.

The ability of near infra-red interactance (NIRI) to predict body composition, as measured by whole-body densitometry, was assessed in a group of 29 healthy volunteers; 15 m, 14 f, aged 18-40 years, body mass index (BMI) 18.3-28.5 kg/m2. The results were compared with those obtained by five other predictive methods: whole-body impedance/resistance, skinfold thicknesses, and methods based on weight and height or BMI. The correlation between densitometry and the alternative methods, including NIRI, were found to be remarkably similar, both with respect to fat (r = 0.90-0.92, s.e.e. = 2.12-2.47 kg), and fat-free mass (r = 0.96-0.97, s.e.e. = 2.13-2.71 kg). Percentage body fat (densitometry) correlated better with the near infrared measurements made in the biceps area than with those made in the triceps or thigh areas, or with a combination of measurements made at two or three sites. The 95 per cent limits of agreement between NIRI and densitometry for the estimation of body constituents were slightly greater than those between densitometry and the other methods. NIRI was also found to underestimate body fat increasingly as the degree of adiposity increased. This under-estimation was found to be particularly marked (16 per cent body weight) in a small and separate group of grossly obese women, BMI greater than 50 kg/m2, whose body composition was assessed by total body potassium as well as by densitometry. This study suggests that, in the healthy group of subjects examined, NIRI has little or no advantage over other simple methods in predicting body composition measured by classical whole-body densitometry.

Adolescent↗
PubMed1989

Energy expenditure of Gambian women during peak agricultural activity measured by the doubly-labelled water method.

The doubly-labelled water (2H218O) method was used to measure total energy expenditure (TEE) in ten non-pregnant, non-lactating (NPNL), six pregnant (P) and fourteen lactating (L) women in a rural Gambian community. Measurements were made on free-living subjects at a period of peak energetic stress when high agricultural work loads coincided with a hungary season to induce moderately severe negative energy balance. TEE averaged 10.42 (SD 2.08) MJ/d, equivalent to 1.95 (SD 0.38) times resting metabolic rate (RMR). The energy cost of physical activity plus thermogenesis, derived as TEE-RMR, averaged 4.94 (SD 1.96) MJ/d. Expressed per kg body-weight (103 kJ/kg per d) this component of expenditure was 2.5 times greater than comparative values from inactive, affluent women studied previously (39 kJ/kg per d). Estimated energy intake (EI) in a subset of the women (n 13) was only 4.80 (SD 1.58) MJ/d, yielding an apparent deficit of 6.08 MJ/d between EI and TEE. Weight changes suggested that endogenous fat oxidation accounted for only about 0.85 MJ/d, leaving an unexplained difference of over 5 MJ/d. Critical analysis of possible errors suggests that the new doubly-labelled water method has provided the most reliable estimates and that the estimates of EI were substantially in error. This finding has important consequences for other food intake studies.

Adult↗
PubMed1988

Sorting of chromosomes by magnetic separation.

Chromosomes were isolated from Chinese hamster x human hybrid cell lines containing four and nine human chromosomes. Human genomic DNA was biotinylated by nick translation and used to label the human chromosomes by in situ hybridization in suspension. Streptavidin was covalently coupled to the surface of magnetic beads and these were incubated with the hybridized chromosomes. The human chromosomes were bound to the magnetic beads through the strong biotin-streptavidin complex and then rapidly separated from nonlabeled Chinese hamster chromosomes by a simple permanent magnet. The hybridization was visualized by additional binding of avidin-FITC (fluorescein) to the unoccupied biotinylated human DNA bound to the human chromosomes. After magnetic separation, up to 98% of the individual chromosomes attached to magnetic beads were classified as human chromosomes by fluorescence microscopy.

Animals↗
PubMed1983

Effect of histrionicotoxin on ion channels in synaptic and conducting membranes of electroplax of Electrophorus electricus.

Histrionicotoxin (HTX) at low concentrations of 5-10 microM blocks the postsynaptic potential of the electroplax of Electrophorus electricus. At 100-fold higher concentrations, HTX blocks the directly evoked action potentials of the conducting membrane. The pH dependence of the blockade by HTX at synaptic channels is different from that at the conducting membrane. At the synapse HTX is more potent at acid pH, while at the conducting membrane it is more potent at basic pH. HTX at high concentrations antagonizes the effects of batrachotoxin, indicative of an effect on the batrachotoxin-sensitive sodium channels involved in action potential generation. While the effects of HTX on the synaptic channels are concentration, time, and pH dependent, the effects on the channels of the conducting membrane are, in addition, use dependent, suggesting interactions of HTX with the activated forms of these channels.

Amphibian Venoms↗
PubMed1982

Phosphoenol-3-bromopyruvate. A mechanism-based inhibitor of phosphoenolpyruvate carboxylase from maize.

Phosphoenol-3-bromopyruvate is an excellent competitive inhibitor (versus phosphoenolpyruvate) of maize phosphoenolpyruvate carboxylase (Ki = 30 microM), provided that preincubation of enzyme with inhibitor is avoided. If the enzyme is preincubated with inhibitor in the presence of Mn2+ and HCO3(-), complete inactivation occurs over the course of about 1 h. The inactivation is first order in enzyme and is saturable with respect to inhibitor, with an apparent Michaelis constant of 67.5 microM and a half-life at high inhibitor concentration of 13.4 min. The inactivation is inhibited by phospholactate and is much slower at low HCO3(-) concentration. Incubation of the enzyme with phosphoenol-3-bromopyruvate in the presence of H14CO3(-) and Mn2+ followed by treatment with NaBH4 leads to incorporation of 14C into the inactive enzyme. If treatment with NaBH4 is omitted, the enzyme is inactivated, but no 14C is incorporated into the inactive product. It appears that phosphoenol-3-bromopyruvate is a mechanism-based inactivator of phosphoenolpyruvate carboxylase, probably because of enzyme-catalyzed conversion to 3-bromooxalacetate, which alkylates the enzyme.

Bicarbonates↗