Microscale investigations of Ni uptake by cement using a combination of scanning electron microscopy and synchrotron-based techniques.

Cement is used to condition waste materials and for the construction and backfilling of repositories for low-and intermediate-level radioactive waste. In this study, Ni uptake by hardened cement paste has been investigated with the aim of improving our understanding of the immobilization process of Ni(ll) in cement on the microscale. Information on the cement microstructure, Ni distribution, Ni concentration, and speciation of the Ni phases formed in the cement system and their association with specific cement minerals has been gained by using scanning electron microscopy (SEM) and synchrotron-based micro-X-ray fluorescence (micro-XRF) and micro-X-ray absorption spectroscopy (micro-XAS). The Ni-doped samples were prepared at a water/cement ratio of 0.4 using a sulfate-resisting Portland cement and were hydrated for 30 days. The metal loadings of the system were varied from 50 up to 5000 mg/kg. SEM investigations show that for all metal loadings the Ni phases form rims around inner-calcium silicate hydrates, suggesting a direct association with this cement phase. The micro-XAS measurements further reveal that a mixture of Ni phases form at Ni-enriched regions. Data analysis indicates that Ni(ll) is predominantly immobilized in a layered double hydroxide-type phase (Ni-Al LDH) and only to a minor extent precipitates as Ni-hydroxides (alpha-Ni(OH)2 and beta-Ni(OH)2). At 50 mg/kg Ni loading, however, the p-XAS measurements suggest the presence of an additional Ni species. In the latter system Ni-Al LDH is found in Ni-rich regions, whereas at Ni-poor regions an unknown species is formed.

Channel formation by salmon and human calcitonin in black lipid membranes.

In this study we investigated the interaction of salmon and human calcitonin (Ct) with artificial lipid bilayer membranes. Both peptides were able to form either transient or permanent channels in the model membranes. The channels formed by salmon Ct at concentration (125 nM) had, on average, a single-channel conductance of 0.58 +/- 0.04 nS in 1M KCl (+10 mV), which is voltage-dependent at lower voltages. Human Ct forms at the same concentration channels with a much lower probability, and high voltages of up to +150 mV were needed to initiate channel formation. The estimated single-channel conductance formed under these conditions was approximately 0.0119 +/- 0.0003 nS in 1 M KCl. Both salmon and human Ct channels were found to be permeable to calcium ions. The possibility is discussed that the superior therapeutic effect of salmon Ct as a tool to treat bone disorders, including Paget disease, osteoporosis, and hypercalcemia of malignancy, rather than human Ct is related to the lack of the fibrillating property of salmon Ct. Preliminary data indicate that also eel and porcine Ct and carbocalcitonin form channels in model membranes.

Studies of mitochondrial porin incorporation parameters and voltage-gated mechanism with different black lipid membranes.

Our work in general aims to clarify the mechanism of what can be considered as a process of the kinetics of porin incorporation into bilayer planar membranes and to identify the parameters involved. In this paper, we report the results of systematically investigating the kinetics of porin incorporation into bilayer membranes made up of phosphatidylinositol or oxidized cholesterol using a simple and low-cost ac method. By means of a mathematical model, we provide evidence that two concurrent processes are present during the kinetics which can be interpreted as positive/negative cooperativity, and we investigate the parameters' dependence on external applied voltages. We observed a phase transition (or similar phenomenon) which seems to take place during the insertion process. The conductance measurement obtained by using data at the steady state conditions, provided indirect indications of two possible gating mechanisms.

Pore formation in lipid bilayer membranes made of phosphatidylinositol and oxidized cholesterol followed by means of alternating current.

The kinetics of porin incorporation into black lipid membranes (BLM) made of phosphatidylinositol (PI) or oxidized cholesterol (Ox Ch) were studied by means of alternating current; the set-up was able to acquire resistance and capacitance simultaneously by means of a mixed double-frequency approach at 1 Hz and 1 KHz, respectively. Conductance was dependent on the interaction between protein-forming pores and lipids. For PI membranes below a porin concentration of 12.54 ng/ml, there was no membrane conductivity, whereas at 200 ng/ml a steady-state value was reached. Different behavior was displayed by Ox Ch membranes, in which a concentration of 12.54 ng/ml was sufficient to reach a steady state. The incorporation kinetics when porin was added after membrane formation were sigmoidal. When porin was present in the medium before membrane formation, the kinetics were sigmoidal for PI membranes but became exponential for Ox Ch membranes. Furthermore, for BLM made of PI, the conductance-versus-porin concentration relationship is sigmoidal, with a Hill coefficient of 5.6 +/- 0.07, which is functional evidence corroborating the six-channel repeating units seen previously. For BLM made of Ox Ch, this relationship followed a binding isotherm curve with a Hill coefficient of 0.934 +/- 0.129.

[Hepatic artery aneurysm]. / Aneurisma de la arteria hepática.

We report the case of a 45-year old man who presented a common hepatic artery aneurysm. He had alcoholic habits and after an episode of upper gastrointestinal tract bleeding underwent an abdominal ultrasound study. This showed a large fluid-filled upper abdominal mass related to the pancreatic head. Both T scan and an angiogram showed an hepatic artery aneurysm. It was resected and the artery repaired interposing a saphenous vein graft. The patient was discharged a week after surgery. Most of the hepatic artery aneurysms are free of symptoms and difficult to palpate and only a few patients can be operated before the occurrence of aneurysmal fissure. Broad use of ultrasound and selective angiography has increased the number of cases diagnosed before complications, reducing the mortality rate, which, in the past, has been reported up to 59%.

Potentiation by m-cresol on transepithelial sodium transport across frog skin induced by insulin.

1. In this study we found that insulin mixed with m-cresol, normally used as pharmaceutical preparation, shows an earlier and larger stimulating effect on transepithelial sodium transport than insulin alone. 2. The action displayed by the m-cresol seems to be specific for insulin because m-cresol mixed with ADH, a hormone known to stimulate sodium transport, failed to show the potentiation seen for insulin. 3. It is proposed that m-cresol could facilitate the interaction with its receptor by modifying the insulin molecule. 4. This finding could be of biological and pharmacological importance.

The effect of ADH and insulin on active sodium transport across frog skin in the presence of alternariol mycotoxin.

1. The effect of vasopressin and insulin on active sodium transport across frog skin in the presence of internal alternariol mycotoxin was studied, using the short-circuit technique. 2. Vasopressin stimulates sodium transport across frog skin by decreasing the resistance to sodium entry into the epithelial cells, thus partially removing the inhibition on the short-circuit current due to the action of Alternariol mycotoxin. 3. Even insulin which is known to increase the short-circuit current by a different mechanism, determines a rapid reversal effect on the inhibition due to Alternariol. 4. These data confirm the different action of the two hormones on active sodium transport across frog skin, and furthermore are indicative of an inhibition of transepithelial sodium transport by Alternariol mycotoxin probably via the sodium pump.

The effect of T-2 toxin on active sodium transport across frog skin in the presence of ADH and amphotericin B.

1. The effect of T-2 toxin on active sodium transport across frog skin both in the presence and in the absence of stimulants of sodium transport, such as Amphotericin B and ADH, was studied using the short circuit current technique with the following results. 2. T-2 toxin produces inhibition of active sodium transport in a dose-response correlation. 3. This effect is irreversible since the washing out of the tissue does not restore its functionality. This indicates that the micotoxin may cross the cellular membrane and act on the internal site. 4. ADH partially removes the inhibitory effect of T-2 toxin. 5. The increase of the sodium pool in the cell as determined by Amphotericin B does not reverse the inhibitory effect of T-2 toxin. 6. The biological significance of these data is discussed in regard to the possible effect of T-2 toxin on Na+, K+-ATPase activity either directly or by a reduction in the metabolic supply of substrates, or by a modified stoichiometry of the pump reaction.

[Effects of dihydrostreptomycin and gentamycin on platelet membrane Na+-K+ATPase]. / Azione della diidrostreptomicina e della gentamicina sulla Na+-K+ATPasi di membrana piastrinica.

The use of aminoglycosides is characterised by a bound linkage between therapeutic and toxical levels. The aim of our study was to define the influence of diidrostreptomicin and gentamicin over Na+-K+ATPase of membranes of platelets. In this study, Bounting's and Caravaggio's method has been used, while ATPase activity was expressed in n mol of hydrolized ATP/min/mg of proteins. The data so obtained showed that the antibiotics were able to block up ATPase activity.

Effect of aminoglycoside antibiotics on sodium transport across the Rana esculenta urinary bladder.

The aminoglycoside antibiotics inhibit the short circuit current (s.c.c.) in the urinary bladder of Rana esculenta. This effect is due to a direct inhibition on the sodium pump; ADH or db-AMPc are not able to reverse the inhibition of the sodium current, but are effective in stimulating the urea fluxes in the presence of antibiotics. In correlation with the number of the NH+2 groups in the antibiotic molecule there is a different pattern of inhibitory effect, i.e. neomycin greater than gentamycin greater than dihydrostreptomycin greater than kanamycin greater than spectinomycin.

Interaction of the aminoglycoside antibiotic dihydrostreptomycin with the H+-ATPase of mitochondria.

In this paper a study is presented of the effect of dihydrostreptomycin on the H+-ATPase of the inner mitochondrial membrane. The antibiotic caused at concentrations of 1-5 X 10(-3)M a marked enhancement of the hydrolytic activity of the H+-ATPase complex in intact mitochondria and submitochondrial particles which was accompanied, in the latter, by enhancement of passive transmembrane proton conduction by the complex. The stimulation by dihydrostreptomycin of ATP hydrolysis resulted in a suppression of the sensitivity of this activity to inhibition by oligomycin. On the other hand the dihydrostreptomycin-promoted proton conduction in submitochondrial particles was suppressed by oligomycin. At concentrations above 10(-2)M dihydrostreptomycin caused inhibition of the activity of both membrane bound and isolated H+-ATPase. In submitochondrial particles devoid of the catalytic moiety (F1) of the H+-ATPase complex, dihydrostreptomycin caused partial inhibition of proton conductivity. It is concluded that the antibiotic uncouples the hydrolytic activity of the catalytic moiety (F1) from transmembrane proton conduction by the membrane sector (F0) of the ATPase complex. This effect can be followed at higher concentrations of dihydrostreptomycin by inhibition of the catalytic activity of F1 and, when F1 is removed from the membrane, by inhibition of transmembrane proton conduction by F0.

Serosal and mucosal facilitated transport of urea in urinary bladder of of Bufo bufo: evidence for an alleged water uptake.

In Bufo bufo urinary bladder an urea facilitated transport has been localised on the luminal membrane. The transport fulfils the criteria for such a mechanism, i.e. is saturable and is inhibited by phloretin, a specific inhibitor for urea transport. Similarly to that of Bufo marinus and Rana esculenta the luminal membrane of Bufo bufo urinary bladder shows an ADH stimulated facilitated transport. Experiments wtih Amphotericin B, serosal phloretin (with and without ADH), have demonstrated the presence of a facilitated urea transport localised on basolateral membrane. Urea uptake on the isolated epithelial cells of Bufo bufo urinary bladder shows a characteristic feature, different from molecules passively transported such as glycerol yet inhibited by phloretin. Allegedly with urea, water flows in to the cells by a dragging or osmotic effect.

[Evidence of morphological modifications induced by theophylline in the urinary bladder epithelium of Rana esculenta]. / Evidenze di alcune modificazioni morfologiche indotte dalla teofillina sull'epitelio vescicale di Rana esculenta.

This preliminary work concerns the morpho-functional action induced by Theophylline on bladder epithelium. The epithelium treated with Theophylline shows important structural variations. The whole tissue thickness appears reduced with cell disposed in monostratified layer. They show a light grade of swelling with nucleus and cytoplasm less evident and boundary less marked. It is important to note that the permeability of the epithelium does not appear modified.

Seasonal variation in the active transporting ability and in the membrane ATPase activity of the frog intestinal epithelium.

The active sugar and amino acid transport in the small intestine of the American leopard frog (Rana pipiens) and a species of European frog (Rana esculenta) decreases during the winter months. Parallel with this the (Na+, K+)-stimulated ("pump") ATPase activity is markedly depressed. No seasonal changes are observed in the intestine of the tropical bullfrog (Rana catesbeiana). It is assumed that the low pump-ATPase activity is caused by the hibernation of the frogs living in moderate or subtropical areas and is connected to a biological clock. The decreased active transport of non-electrolytes appears to be a consequence of the change of the ATPase activity.