Effect of thapsigargin on calcium homeostasis in Trypanosoma cruzi trypomastigotes and epimastigotes

By using the fluorescent calcium indicator fura-2, it was found that the concentration of free Ca2+ in the cytoplasm of Trypanosoma cruzi trypomastigotes incubated in the presence or absence of external calcium was maintained at very low levels (10-20 nM). When trypomastigotes were incubated in the presence of succinate and ATP and permeabilized with digitonin, they lowered the medium calcium concentration to a submicromolar level. In the presence of 1 microM FCCP the initial rate of Ca2+ sequestration by these permeabilized cells was very slow. When succinate alone was present, the initial rate of Ca2+ accumulation was slower than with ATP plus succinate, and the calcium set point was about 0.6 microM. The succinate dependence and FCCP sensitivity of the later Ca2+ uptake indicate that it may be exerted by the mitochondria. High concentrations of the tumor promoter thapsigargin slightly increased cytosolic Ca2+ in the presence of extracellular Ca2+ but had no effect on the FCCP- and oligomycin/antimycin A-insensitive Ca2+ pool. In addition, when used at those concentrations (4-20 microM), thapsigargin was shown to release Ca2+ from the mitochondria and to decrease the inner mitochondrial membrane potential of trypomastigotes and epimastigotes as measured using safranine O. Despite the presence of inositol phosphates as determined by [3H]inositol incorporation, no IP3-sensitive Ca2+ release could be detected in trypomastigotes.     

Reversible or irreversible modification of [3H]PAF binding on rabbit platelet membranes differentiates various PAF receptor antagonists

[³H]Platelet-activating factor (PAF) binding to rabbit platelet membranes was examined before and after 20 min preincubation at 25°C in the presence of PAF, lysoPAF, or of five different PAF receptor antagonists (L 652731, BN 52021, WEB 2086, BN 52111 and BN 52115). When platelet membranes were not washed after preincubation with PAF or PAF antagonists, no significant specific binding of [³H]PAF was observed, which suggests full occupancy of the binding sites. When membranes were extensively washed, full recovery of specific [³H]PAF binding was attained with L 652731 and partial recoveries (60%, 55% and 30%) were reached with BN52021, WEB 2086 and PAF, respectively; no recovery was seen with the dioxolanes BN 52111 and BN 52115. Scatchard analysis of the binding data indicated that no significant change in the dissociation constant (K_d) and maximum number of binding sites (B_max) occurred after preincubation of platelet membrane with L 652731, whereas a reduction of B_max was observed when PAF and BN 52021 were measured. When platelet membranes were preincubated with WEB 2086, B_max and K_d significantly increased. The data suggest differing binding properties for PAF and the PAF antagonists. Some of the PAF antagonists may tightly bind to the PAF receptor site(s) and/or irreversibly modify or downregulate PAF recognition sites. Our results also suggest that the interaction of PAF receptor antagonists with PAF receptor can be divided into at least two components, namely a reversible component and an irreversible one. Based on a paper presented at the Third International Conference on Platelet-Activating Factor and Structurally Related Alkyl Ether Lipids, Tokyo, Japan, May 1989.     

Purification of the human anionic polypeptide fraction of the apo-bile lipoprotein complex by zonal ultracentrifugation

The two main proteic constituents of the human Apo-bile lipoprotein complex (BLC), i.e., the anionic polypeptide fraction (APF) and the IgA fragments, were separated by preparative zonal ultracentrifugation using a sucrose gradient containing 1.5 mM glycodesoxycholate. The purification of the APF was verified by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis and immunology, and its amino acid composition then was determined. This procedure was used to obtain a polyclonal antiserum directed solely against the APF.     

Improving Neural Networks for Mechanism Kinematic Chain Isomorphism Identification

Detection of isomorphism among kinematic chains is essential in mechanical design, but difficult and computationally expensive. It has been shown that both traditional methods and previously presented neural networks still have a lot to be desired in aspects such as simplifying procedure of identification and adapting automatic computation. Therefore, a new algorithm based on a competitive Hopfield network is developed for automatic computation in the kinematic chain isomorphism problem. The neural approach provides directly interpretable solutions and does not demand tuning of parameters. We have tested the algorithm by solving problems reported in the recent mechanical literature. Simulation results show the effectiveness of the network that rapidly identifies isomorphic kinematic chains.     

A variable rate multipulse speech coder for CDMA cellular systems

Variable Rate (VR) speech coders are classified into: source-controlled VR coders where the rate is selected depending on the local character of the speech, and network-controlled VR coders where an external control signal selects the coding rate. The first category benefits from the variable rate channels used by Code Division Multiple Access (CDMA) mobile communications. The second category is indispensable for the right behaviour of the CDMA systems under conditions as high traffic levels. The VR speech coder presented in this communication exhibits both types of control. The source control is achieved by means of a Voice Activity Detector (VAD) and a phonetic classifier. The network control acts on the selection procedure of the multipulse excitation sequence to the synthesis filter. This is the main advantage of our VR MultiPulse speech coder because by means of an external signal the bit rate can be changed only every 4 msec, without transitions or distortions. Considering one-way communication, six different operating rates can be externally selected ranging from 4.8 to 9.1 kbps for the active frames; an average bit rate of 380 bps is required for the noise frames.This work has been partly funded by the Spanish Research National Plan under grant no. TIC92-0800-C05-02 and by Northern Telecom.     

Growth inhibitory effect of juvenile hormone analogues on epimastigotes of Trypanosoma cruzi

Several compounds, structurally related to the insect growth regulator Fenoxycarb, exhibited interesting inhibition action to control proliferation of Trypanosoma cruzi, the parasite responsible for Chagas' disease. Some of these drugs were shown to be potent growth inhibitors of this parasite. All of these drugs had previously presented juvenoid activity on several non-related bug species such as Tenebrio molitor, Galleria mellonella, Dysdercus cingulatos, and Pyrrhocoris apterus.     

Behavior of biliary phospholipids in intestinal lumen during fat digestion in rat

The phospholipids present in the intestinal lumen of rats following ingestion of triglycerides are of biliary origin. They consist of lecithins accompanied by a small proportion of lysolecithins. Their behavior in comparison with the other lipid constituents of the intestinal content was studied by subjecting the latter to gel filtration on an agarose column in the presence of a solution of 6 mM sodium taurocholate in 0.1 M NaCl. Part of the phospholipids is present with the triglycerides and diglycerides in the emulsified phase excluded from the gel where pancreatic lipase and colipase also are found. The remainder is found in optically clear fractions containing fatty acids, monoglycerides, and bile salts. These fractions are eluted at 2.0 column volumes, while mixed fatty acids, monoglycerides, bile salts micelles emerge from the column at 2.4 column volumes in the same chromatographic conditions. This difference in behavior may be explained by the presence of biliary lecithins. This presence could have an important bearing upon the mucosal uptake of the lipolysis products of triglycerides.     

Balanced pasture fertilization in the Basque Country: I. Phosphorus and potassium budgets on dairy farms

Dairy farming is the main agricultural activity of the Basque Country. A dairy farm is characterized as a system with soils and crops, forage, cattle and manure as main components, and in such a system, nutrient cycling is very important to maintain soil fertility and optimize forage production. To quantify nutrient transfers in the cycle, a simple system was developed and has been applied to seventeen farms to examine its ability to achieve a balanced P and K fertilization. These farms have provided data on inputs (fertilizer, feeds, concentrates), pasture and manure management, and outputs (milk production), and soil samples have been taken from farm pastures. Phosphorus and K in excreta and uneaten pasture is used with a relatively high efficiency as suggested by the relatively high efficiency of P and K utilization by the pasture that usually ranges from 70 to 90%. Concentrate feeding (3000 kg cow^–1 yr^–1) represents one of the main P and K inputs in Basque Country dairy farms, averaging 26 and 66 kg ha^–1, respectively. Besides, release of K in the soil through slow liberation from non-exchangeable sites was estimated as 30 kg ha^–1. Thus, a high efficiency in excreta recycling would diminish substantially P and K mineral fertilizer needs. Farm nutrient budgets appear to be a convenient tool for determining nutrient shortages and surpluses at farm level, and thus they are considered as a first step to support a better management of maintenance fertilization of permanent pastures.     

Templating of crystallization and shear-induced self-assembly of single-wall carbon nanotubes in a polymer-nanocomposite

We report the appearance of a novel self-assembling of a fraction of single-wall carbon nanotubes (SWCNT) within a SWCNT-polymer nanocomposite subjected to flow fields upon injection molding processing. By combining X-ray diffraction and Raman spectroscopy techniques, both working on a microfocus fashion, we probe that a fraction of the thinnest SWCNT self-assembles into a rectangular lattice in the sample regions where the shear stress induces the highest levels of nanotube aggregation. Additionally, we demonstrate that a modest amount in weight of nanotubes is enough to template the morphology of crystallization during flow providing a method to obtain a highly desirable fiber-like morphology.