Characterization of toad liver glutathione transferase. (1/78)

The major form of glutathione transferase from the toad liver previously designed as Bufo bufo liver GST-7.6 (A. Aceto, B. Dragani, T. Bucciarelli, P. Sacchetta, F. Martini, S. Angelucci, F. Amicarelli, M. Miranda and C. Di Ilio, Biochem. J. 289 (1993) 417-422) has been characterized. According to its partial amino acid sequence, the toad enzyme may be included in the pi class GST and named bbGST P2-2. However, bbGST P2-2 appears to be immunologically, structurally and kinetically distinct from any other members of pi family, including bbGST P1-1, suggesting that it may constitute a subset of pi class GST. The data support the hypothesis that the transition from aquatic to terrestrial life causes a switch of the GST amphibian pattern promoting the expression of a GST form (bbGST P2-2) able to counteract, with higher efficiency, the toxic effects of reactive metabolites of oxidative metabolism and those of hydrophobic xenobiotics.  (+info)

Chloride dependence of active sodium transport in frog skin: the role of intercellular spaces. (2/78)

1. In agreement with previous observations the replacement of Cl by a nonpenetrating anion in the solution bathing either the outside or both sides of the frog skin causes a fall in the short-circuit current. 2. When Cl is replaced by a non-penetrating anion in the solution bathing the outside of the frog skin the Isc is still a correct measure of the net Na transport. 3. Under the same conditions both active and shunt paths seem to be affected since there is a decrease in Isc, Na influx, amiloride-dependent conductance, and initial Na uptake across the external barrier, together with a decrease in Cl-backfluxes and amiloride-independent conductance. There is also a decrease in water permeability and a reduction in size of the intercellular spaces. 4. The removal of Cl does not appear to affect the entry step of Na but may have an effect on the shunt path. This in turn may change the active Na transport.  (+info)

A visual evoked potential correlate of global figure-ground segmentation. (3/78)

Human observers discriminated the global orientation of a texture-defined figure which segregated from a texture surround. Global figure discriminability was manipulated through within-figure collinearity, figure-surround interaction, and figure connectedness, while the local orientation contrast at edges between figure and surround was kept constant throughout all the experiments. Visual evoked potentials (VEPs) were recorded during onset-offset stimulation in which the figure cyclically appeared and disappeared from a uniform texture background. A difference component was obtained by subtraction of offset-from onset-VEP. Two negative peaks of the difference component are found with latencies around 140-160 and 200-260 ms, respectively. Enhanced discriminability of the global figure reduced (11-25 ms) the latency of the second peak, hence indicating that the 200-260 ms component was produced by global figure-ground segmentation.  (+info)

Seeing better at night: life style, eye design and the optimum strategy of spatial and temporal summation. (4/78)

Animals which need to see well at night generally have eyes with wide pupils. This optical strategy to improve photon capture may be improved neurally by summing the outputs of neighbouring visual channels (spatial summation) or by increasing the length of time a sample of photons is counted by the eye (temporal summation). These summation strategies only come at the cost of spatial and temporal resolution. A simple analytical model is developed to investigate whether the improved photon catch afforded by summation really improves vision in dim light, or whether the losses in resolution actually make vision worse. The model, developed for both vertebrate camera eyes and arthropod compound eyes, calculates the finest spatial detail perceivable by a given eye design at a specified light intensity and image velocity. Visual performance is calculated for the apposition compound eye of the locust, the superposition compound eye of the dung beetle and the camera eye of the nocturnal toad. The results reveal that spatial and temporal summation is extremely beneficial to vision in dim light, especially in small eyes (e.g. compound eyes), which have a restricted ability to collect photons optically. The model predicts that using optimum spatiotemporal summation the locust can extend its vision to light intensities more than 100,000 times dimmer than if it relied on its optics alone. The relative amounts of spatial and temporal summation predicted to be optimal in dim light depend on the image velocity. Animals which are sedentary and rely on seeing small, slow images (such as the toad) are predicted to rely more on temporal summation and less on spatial summation. The opposite strategy is predicted for animals which need to see large, fast images. The predictions of the model agree very well with the known visual behaviours of nocturnal animals.  (+info)

The distribution of sodium, potassium and chloride in the nucleus and cytoplasm of Bufo bufo oocytes measured by electron microprobe analysis. (5/78)

1. Measurements of cytoplasmic and nuclear Na, K and Cl have been made by electron microprobe analysis on freeze-dried sections of oocytes of Bufo bufo, using standards of bovine plasma albumin and gamma-globulin. Concentrations were obtained per kilogram of dry mass, were converted to concentrations per litre of water content using known figures for water and solid concentration of nucleus and cytoplasm, and were then compared with measurements on cells from the same animal obtained by flame photometry. 2. In fresh oocytes concentrations were (mean +/- S.E. of mean in m-mole/l. H2O) in cytoplasm Na 10.9 +/- 1.95, K 70.2 +/- 3.22, Cl 98.8 +/- 11.0, and in nucleus Na 10.4 +/- 1.79, K 266.4 +/- 22.8, Cl 91.3 +/- 9.0. 3. After treatment with Na-free Ringer (Li substituted for Na) for 5 hr, concentrations were in cytoplasm Na 11.1 +/- 2.44, K 64.4 +/- 5.7, Cl 88.7 +/- 8.8, and in nucleus Na 2.4 +/- 0.73, K 141 +/- 13.9, Cl 75.0 +/- 6.7. Na inexchangeable with Li therefore lay in the cytoplasm but not in the nucleus as previously shown by autoradiography. 4. For K electron microscopic analysis measurements agreed well with those obtained by flame photometry but the former measured only 35% of Na measured by flame photometry. This discrepancy may be due either to technical difficulties with the electron microprobe analysis or to localization of Na in the cytoplasm.  (+info)

Pepsin-mediated processing of the cytoplasmic histone H2A to strong antimicrobial peptide buforin I. (6/78)

The intestinal epithelium forms a first line of innate host defense by secretion of proteins with antimicrobial activity against microbial infection. Despite the extensive studies on the antimicrobial host defense in many gastrointestinal tracts, little is known about the antimicrobial defense system of the stomach. The potent antimicrobial peptide buforin I, consisting of 39 aa, was isolated recently from the stomach tissue of an Asian toad, Bufo bufo gargarizans. In this study we examined the mechanism of buforin I production in toad stomach tissue. Buforin I is produced by the action of pepsin isozymes, named pepsin Ca and Cb, cleaving the Tyr39-Ala40 bond of histone H2A. Immunohistochemical analysis revealed that buforin I is present extracellularly on the mucosal surface, and unacetylated histone H2A, a precursor of buforin I, is localized in the cytoplasm of gastric gland cells. Furthermore, Western blot analysis showed that buforin I is also present in the gastric fluids, and immunoelectron microscopy detected localization of the unacetylated histone H2A in the cytoplasmic granules of gastric gland cells. The distinct subcellular distribution of the unacetylated histone H2A and the detection of the unacetylated buforin I both on the mucosal surface and in the lumen suggest that buforin I is produced from the cytoplasmic unacetylated histone H2A secreted into the gastric lumen and subsequently processed by pepsins. Our results indicate that buforin I along with pepsins in the vertebrate stomach may contribute to the innate host defense of the stomach against invading microorganisms.  (+info)

Recombinant human interleukins IL-1alpha, IL-1beta, IL-4, IL-6, and IL-7 show different and specific calcium-independent carbohydrate-binding properties. (7/78)

A method was developed for the determination of putative lectin activities of cytokines. It involved the immunoblotting measurement of the quantity of these cytokines unbound to a series of different immobilized glycoconjugates and displacement of the bound cytokines with oligosaccharides of known structures. This method allows demonstrating that the following interleukins specifically recognize different oligosaccharide structures in a calcium-independent mechanism: interleukin-1alpha binds to the biantennary disialylated N-glycan completed with two Neu5Acalpha2-3 residues; interleukin-1beta to a GM4 sialylated glycolipid Neu5Acalpha2-3Galbeta1-Cer having very long and unusual long-chain bases; interleukin-4 to the 1,7 intramolecular lactone of N-acetyl-neuraminic acid; interleukin-6 to compounds having N-linked and O-linked HNK-1-like epitopes; and interleukin-7 to the sialyl-Tn antigen. Because the glycan ligands are rare structures in human circulating cells, it is suggested that such activities could be essential for providing specific signaling systems to cells having both the receptors and the oligosaccharide ligands of the interleukin at their cell surface.  (+info)

Developmental expression and distribution of amphibian glutathione transferases. (8/78)

This work is aimed at detecting the expression and location of embryonic Bufo bufo GST (bbGSTP1-1) and adult B. bufo GST (bbGSTP2-2) during toad development, in order to assign a putative role to these enzymes also on the basis of their compartmentalization and to verify whether during the premetamorphic liver ontogeny the bbGSTP2-2 form appears. This study was also performed in the adult liver (the primary site of Pi class GST expression) and in the mature ovary, to discern if the embryonic form derives from maternal form. The results show that the embryos and the ovary express only bbGSTP1-1. Moreover, bbGSTP1-1 distribution is the same both in the early embryos and in the ovary: this strongly suggests that bbGSTP1-1 is of maternal origin. As development goes on, a wide distribution of bbGSTP1-1 all over the differentiating organs is observed. The embryonic liver expresses exclusively the bbGSTP1-1 form, while the adult liver is highly positive only towards the bbGSTP2-2 form. This implies that the switch towards the adult bbGSTP2-2 form occurs in metamorphic or postmetamorphic phases and that the detoxication metabolic requirements of the embryo may be completely fulfilled by the bbGSTP1-1 isoenzyme.  (+info)

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