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It is commonly considered that the corrosion of steel in concrete is controlled by the oxygen content of the pore solution and there are service life models that relate the corrosion rate to the amount of oxygen. It is also commonly believed that in water saturated conditions the oxygen content in the pores is negligible and that underwater there is no risk of depassivation and the corrosion rate is very low. However, the available data on corrosion rates in immersed conditions do not indicate such performance; on the contrary corrosion develops when sufficient chloride reaches the reinforcement. In the present paper, results are presented for tests performed in alkaline chloride solutions that were purged with nitrogen to reduce the oxygen content. The results indicate that at very low oxygen concentrations, corrosion may develop in the presence of chlorides. The presence or absence of corrosion is influenced by the amount of chloride, the corrosion potential and the steel surface condition.

Large conductance voltage and calcium-activated potassium (MaxiK) channels are activated by membrane depolarization and elevated cytosolic Ca(2+). In the brain, they localize to neurons and astrocytes, where they play roles such as resetting the membrane potential during an action potential, neurotransmitter release, and neurovascular coupling. MaxiK channels are known to associate with several modulatory proteins and accessory subunits, and each of these interactions can have distinct physiological consequences. To uncover new players in MaxiK channel brain physiology, we applied a directed proteomic approach and obtained MaxiK channel pore-forming α subunit brain interactome using specific antibodies. Controls included immunoprecipitations with rabbit immunoglobulin G (IgG) and with anti-MaxiK antibodies in wild type and MaxiK channel knockout mice (Kcnma1(-/-)), respectively. We have found known and unreported interactive partners that localize to the plasma membrane, extracellular space, cytosol and intracellular organelles including mitochondria, nucleus, endoplasmic reticulum and Golgi apparatus. Localization of MaxiK channel to mitochondria was further confirmed using purified brain mitochondria colabeled with MitoTracker. Independent proof of MaxiK channel interaction with previously unidentified partners is given for GABA transporter 3 (GAT3) and heat shock protein 60 (HSP60). In human embryonic kidney 293 cells containing SV40 T-antigen (HEK293T) cells, both GAT3 and HSP60 coimmunoprecipitated and colocalized with MaxiK channel; colabeling was observed mainly at the cell periphery with GAT3 and intracellularly with HSP60 with protein proximity indices of ∼ 0.6 and ∼ 0.4, respectively. In rat primary hippocampal neurons, colocalization index was identical for GAT3 (∼ 0.6) and slightly higher for HSP60 (∼ 0.5) association with MaxiK channel. The results of this study provide a complete interactome of MaxiK channel the mouse brain, further establish the localization of MaxiK channel in the mouse brain mitochondria and demonstrate the interaction of MaxiK channel with GAT3 and HSP60 in neurons. The interaction of MaxiK channel with GAT3 opens the possibility of a role of MaxiK channel in GABA homeostasis and signaling.

Fast inactivating Shaker H4 potassium channels and nonconducting pore mutant Shaker H4 W434F channels have been used to correlate the installation and recovery of the fast inactivation of ionic current with changes in the kinetics of gating current known as "charge immobilization" (Armstrong, C.M., and F. Bezanilla. 1977. J. Gen. Physiol. 70:567-590.). Shaker H4 W434F gating currents are very similar to those of the conducting clone recorded in potassium-free solutions. This mutant channel allows the recording of the total gating charge return, even when returning from potentials that would largely inactivate conducting channels. As the depolarizing potential increased, the OFF gating currents decay phase at -90 mV return potential changed from a single fast component to at least two components, the slower requiring approximately 200 ms for a full charge return. The charge immobilization onset and the ionic current decay have an identical time course. The recoveries of gating current (Shaker H4 W434F) and ionic current (Shaker H4) in 2 mM external potassium have at least two components. Both recoveries are similar at -120 and -90 mV. In contrast, at higher potentials (-70 and -50 mV), the gating charge recovers significantly more slowly than the ionic current. A model with a single inactivated state cannot account for all our data, which strongly support the existence of "parallel" inactivated states. In this model, a fraction of the charge can be recovered upon repolarization while the channel pore is occupied by the NH2-terminus region.

Prolonged depolarization induces a slow inactivation process in some K+ channels. We have studied ionic and gating currents during long depolarizations in the mutant Shaker H4-Delta(6-46) K+ channel and in the nonconducting mutant (Shaker H4-Delta(6-46)-W434F). These channels lack the amino terminus that confers the fast (N-type) inactivation (Hoshi, T., W.N. Zagotta, and R.W. Aldrich. 1991. Neuron. 7:547-556). Channels were expressed in oocytes and currents were measured with the cut-open-oocyte and patch-clamp techniques. In both clones, the curves describing the voltage dependence of the charge movement were shifted toward more negative potentials when the holding potential was maintained at depolarized potentials. The evidences that this new voltage dependence of the charge movement in the depolarized condition is associated with the process of slow inactivation are the following: (a) the installation of both the slow inactivation of the ionic current and the inactivation of the charge in response to a sustained 1-min depolarization to 0 mV followed the same time course; and (b) the recovery from inactivation of both ionic and gating currents (induced by repolarizations to -90 mV after a 1-min inactivating pulse at 0 mV) also followed a similar time course. Although prolonged depolarizations induce inactivation of the majority of the channels, a small fraction remains non-slow inactivated. The voltage dependence of this fraction of channels remained unaltered, suggesting that their activation pathway was unmodified by prolonged depolarization. The data could be fitted to a sequential model for Shaker K+ channels (Bezanilla, F., E. Perozo, and E. Stefani. 1994. Biophys. J. 66:1011-1021), with the addition of a series of parallel nonconducting (inactivated) states that become populated during prolonged depolarization. The data suggest that prolonged depolarization modifies the conformation of the voltage sensor and that this change can be associated with the process of slow inactivation.

Abstract: Objective: To evaluate the sensitivity and specificity of three laboratory techniques for the diagnosis of Giardia intestinalis. Materials and Methods: 31 fecal samples from preschoolers were submitted to microscopic examination with SSF-Lugol, the Ritchie concentration method and the “Giardia-Strip” immunological method. The Ritchie technique was considered the “gold standard. “Results: G. intestinalis cysts were identified in 6 samples (19.35%) using the Ritchie technique. The fresh test identified 5 cases of protozoan cysts (16.1%), showing a sensitivity of 83 % and specificity of 100 %. The Giardia-Strip identified cysts in 4 samples (12.9%) with 66.66 % sensitivity and 100% specificity. The positive predictive value for the “Giardia Strip” method was 14% and the negative predictive value was 93%. The fresh test showed a positive predictive value of 100% and a negative predictive value of 96 %. Conclusions: Ritchie concentrate and the fresh test showed higher sensitivity and specificity in the diagnosis of Giardia intestinalis, when samples contain only cysts. However, the Giardia-Strip method proved faster in obtaining results. Resumen: Evaluar la sensibilidad y especificidad de tres técnicas de laboratorio para el diagnóstico de Giardia intestinalis. Materiales y Métodos: 31 muestras de heces provenientes de niños en edad preescolar, se procesaron a través del examen microscópico con SSF-lugol, método de concentración de Ritchie y método inmunológico “Giardia-Strip”. La Técnica de Ritchie fue considerada como “Gold Standard”. Resultados: Se identificaron quistes de Giardia intestinalis en 6 muestras (19,35%) a través de la técnica de Ritchie. La técnica del examen al fresco identificó en 5 de ellas quistes del protozoario (16,1%), mostrando una sensibilidad de 83% y especificidad del 100%. La técnica Giardia-Strip identificó quistes en 4 muestras (12,9%) con sensibilidad de 66,66% y especificidad del 100%. El valor predictivo positivo para el método de “Giardia-Strip” fue 14% y valor predictivo negativo de 93%. El examen al fresco mostró valor predictivo positivo del 100% y valor predictivo negativo de 96%. Conclusiones: El concentrado de Ritchie y el examen al fresco mostraron mayor sensibilidad y especificidad en el diagnóstico de Giardia intestinalis, cuando las muestras contienen solo quistes. Sin embargo, el método Giardia-Strip, demostró mayor rapidez en la obtención de los resultados.