Cyclo(His-Pro) up-regulates heme oxygenase 1 via activation of Nrf2-ARE signalling

Paraquat (1,1'-dimethyl-4,4'-bipyridinium), a widely used non-selective herbicide, is a redox cycling agent with adverse effects on dopamine systems. Epidemiological data have shown that exposure to paraquat is one of the several risk factors for Parkinson's disease. We have already shown that cyclo(His-Pro), an endogenous cyclic dipeptide produced by the cleavage of the thyrotropin releasing hormone, has a cytoprotective effect through a mechanism involving Nrf2 activation that decreases production of reactive oxygen species and increases glutathione synthesis. Using primary neuronal cultures and PC12 cells as targets of paraquat neurotoxicity, we addressed whether and how cyclo(His-Pro) causes cellular protective response against paraquat-mediated cell death. We found that cyclo(His-Pro) attenuated reactive oxygen species production, and prevented glutathione depletion by up-regulating Nrf2 gene expression, triggering its nuclear accumulation and activating the expression of heme oxygenase1. These protective effects were abolished by RNA interference-mediated Nrf2 knock down whereas were unaffected by RNA interference-mediated Keap1 knock down. Inhibition of heme oxygenase activity decreased cyclo(His-Pro)-induced neuroprotection. These results suggest that cyclo(His-Pro), acting as a selective activator of the brain modulable Nrf2 pathway, may be a promising candidate as neuroprotective agent that act through induction of phase II genes.     

Oligomeric State and Thermal Stability of Apo- and Holo- Human Ornithine δ-Aminotransferase

Human ornithine δ-aminotransferase (hOAT) (EC 2.6.1.13) is a mitochondrial pyridoxal 5′-phosphate (PLP)-dependent aminotransferase whose deficit is associated with gyrate atrophy, a rare autosomal recessive disorder causing progressive blindness and chorioretinal degeneration. Here, both the apo- and holo-form of recombinant hOAT were characterized by means of spectroscopic, kinetic, chromatographic and computational techniques. The results indicate that apo and holo-hOAT (a) show a similar tertiary structure, even if apo displays a more pronounced exposure of hydrophobic patches, (b) exhibit a tetrameric structure with a tetramer-dimer equilibrium dissociation constant about fivefold higher for the apoform with respect to the holoform, and (c) have apparent T_m values of 46 and 67?°C, respectively. Moreover, unlike holo-hOAT, apo-hOAT is prone to unfolding and aggregation under physiological conditions. We also identified Arg217 as an important hot-spot at the dimer–dimer interface of hOAT and demonstrated that the artificial dimeric variant R217A exhibits spectroscopic properties, T_m values and catalytic features similar to those of the tetrameric species. This finding indicates that the catalytic unit of hOAT is the dimer. However, under physiological conditions the apo-tetramer is slightly less prone to unfolding and aggregation than the apo-dimer. The possible implications of the data for the intracellular stability and regulation of hOAT are discussed.     

Long‐term population dynamics of a coral reef gastropod and responses to disturbance

The age‐specific density of the red‐lipped stromb Strombus luhuanus (Mollusca: Gastropoda) was monitored over 13 years (1981–1993) at four locations on the intertidal reef flat at Heron Island, Great Barrier Reef. Densities were highly variable, but there were persistent, location‐specific differences in population density, age structure and adult body size, the latter indicating that the populations were not extensively linked by adult movement. There was relatively high recruitment at most locations in 1984, 1989 and 1993, each occurring approximately 2 years after El Niño/Southern Oscillation events, although recruit density during these years varied in both space and time. The studied strombs experienced three disturbance events: (i) experimental harvesting at two locations (1984–1985); (ii) siltation from a harbour dredging operation (1987–1988); and (iii) a severe cyclone (1992). Resilience to harvesting at a local scale (0.5–2 ha) was high: density had recovered within a year, due to immigration of adults and older juveniles. Strombus luhuanus responded much more strongly to broad‐scale changes to its environment than to localized harvesting. After dredging, there was a progressive density decline coupled with low recruitment at two locations, and a later decline at a third location, followed by a recruitment‐driven rebound after the cyclone. Generalized environmental effects of siltation and the cyclone were also reflected in substantial changes in algal cover. Long‐term variations in environmental conditions probably cause high temporal variation over large spatial scales through effects on the survival of larvae or recruits. Localized short‐term field monitoring of such species would give a misleading picture of key factors affecting population dynamics.     

Renal oncocytoma: origin from the collecting duct

The histo- and cytogenesis of two cases of renal oncocytoma have been studied by cytomorphological and cytochemical methods. Transitions from collecting ducts into oncocytic tubules were observed at the light and electron microscopic levels. The fine structure of the oncocytes in tubules and tumors is described in detail. Cytochemically, the oncocytic tubules and oncocytomas share many characteristics with the distal nephron, especially the collecting ducts. A striking difference is the enhanced activity of succinic dehydrogenase which corresponds to the increase in the number of mitochondria in oncocytes. All the results suggest that renal oncocytoma originates from the collecting duct.     

A transient voltage-dependent outward current in cultured cerebellar granules

Granule cells were dissociated from rat cerebella with a procedure that yields a 98% pure cell population. Potassium currents in these cells were studied using the patch-clamp technique. Depolarizing pulses of 10 mV step and 100 ms duration from a holding potential of –80 mV elicited two different potassium outward currents: a transient, low-voltage activated component and a long lasting, high-voltage activated component. At +30 mV, the total current reached an amplitude of 2 nA (mean value of 15 experiments). The reversal potential of the transient current, estimated by measuring tail currents, was –77 mV, close to that predicted by the Nernst equation. The transient current was half inactivated with a holding potential of –78 mV and completely inactivated with –50 mV or more positive holding potentials. Finally, the current decay could be fitted by the sum of two exponentials with time constants of about 20 and 250 ms.     

The cytotaxonomy ofEmilia spp. (Asteraceae: Senecioneae) occurring in Brazil

Analysis of several populations in a large part of the distribution area of the genusEmilia in Brazil has revealed only two species: the diploidE. sonchifolia and the tetraploidE. fosbergii. The more widely reportedE. coccinea was not found. They show a karyotype constancy in morphology and chromosome number (2n = 10 and 2n = 20, respectively), C-banding pattern and number of secondary constrictions. Some indications were found thatE. fosbergii may be an allopolyploid and that its ancestors had different genome sizes.     

Histopathology of experimental Aspergillus fumigatus keratitis

Histopathological studies in rabbit's eyes, 7 and 14 days after intracorneal inoculation with 1×10⁵ Aspergillus fumigatus conidia have been performed.Similar lesions were found in both periods with fungal hyphae in the anterior third of corneal stroma, round cell infiltration from the sclero-corneal edge and in the anterior chamber and, neovascularization.No lesions were found in the Descemet's membrane.Gomori silver-methenamine stain with hematoxiline-eosine counter-stain was found to be the most reliable stain to detect fungal presence in corneal stroma, and Masson's trichromic stain in the study of pathological changes in ocular elements.     

Gating properties of channels formed by colicin Ia in planar lipid bilayer membranes

Summary Colicin Ia forms voltage-dependent channels when incorporated into planar lipid bilayers. A membrane containing many Colicin Ia channels shows a conductance which is turned on when high positive voltages (>+10 mV) are applied to thecis side (side to which the protein is added). The ionic current flowing through the membrane in response to a voltage step shows at first an exponential and then a linear rise with time. The relationship between the steady-state conductance, achieved immediately after the exponential portion, and voltage is S-shaped and is adequately fit by a Boltzmann distribution. The time constant () of the exponential is also dependent on voltage, and the relation between these two parameters is asymmetric aroundV _o (voltage at which half of the channels are open). In both cases the steepness of the voltage dependence, a consequence of the number of effective gating particles (n) present in the channel, is greatly influenced by the pH of the bathing solutions. Thus, increasing the pH leads to a reduction inn, while acidic pH's have the opposite effects. This result is obtained either by changing the pH on both sides of the membrane or on only one side, be itcis orrans. On the other hand, changing pH on only one side by addition of an impermeant buffer fails to induce any change inn. At the single-channel level, pH had an effect both on the unitary conductance, doubling it in going from pH 4.5 to 8.2, as well as on the fraction of time the channels stay open,F _(v). For a given voltage,F _(v) is clearly diminished by increasing the pH. This titration of the voltage sensitivity leads to the conclusion that gating in the Colicin Ia molecule is accomplished by charged amino-acid residues present in the protein molecule. Our results also support the notion that these charged groups are inside the aqueous portion of the channel.